UNITED NATIONS ENVIRONMENT PROGRAMME

INTERNATIONAL LABOUR ORGANISATION

WORLD HEALTH ORGANIZATION

INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY

**ENVIRONMENTAL HEALTH CRITERIA 171**

**DIESEL FUEL AND EXHAUST EMISSIONS**

This report contains the collective views of an international group of experts and does not necessarily represent the decisions or the stated policy of the United Nations Environment Programme, the International Labour Organisation, or the World Health Organization.

Environmental Health Criteria 171

DIESEL FUEL AND EXHAUST EMISSIONS

First draft prepared by the staff members of the Fraunhofer Institute of Toxicology and Aerosol Research, Germany, under the coordination of

Dr. G. Rosner

Published under the joint sponsorship of the United Nations Environment Programme, the International Labour Organisation, and the World Health Organization, and produced within the framework if the Inter-Organization Programme for the Sound Management of Chemicals.

World Health Organization Geneva, 1996

The International Programme on Chemical Safety (IPCS) is a joint venture of the United Nations Environment Programme, the International Labour Organisation, and the World Health Organization. The main objective of the IPCS is to carry out and disseminate evaluations of the effects of chemicals on human health and the quality of the environment. Supporting activities include the development of epidemiological, experimental laboratory, and risk-assessment methods that could produce internationally comparable results, and the development of manpower in the field of toxicology. Other activities carried out by the IPCS include the development of know-how for coping with chemical accidents, coordination of laboratory testing and epidemiological studies, and promotion of research on the mechanisms of the biological action of chemicals.

WHO Library Cataloguing in Publication Data

Diesel Fuel and exhaust emission

(Environmental health criteria ; 171)

1.Air pollutants, Enviromental 2.Air pollution 3.Fueloils I.International Programme on Chemical Safety II.Series

ISBN 92 4 157171 3 (NLM Classification: WA 754)

ISSN 0250-863X

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(c) World Health Organization 1996

Publications of the World Health Organization enjoy copyright protection in accordance with the provisions of Protocol 2 of the Universal Copyright Convention. All rights reserved. The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the Secretariat of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. The 

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CONTENTS

NOTE TO READERS OF THE CRITERIA MONOGRAPHS

PREAMBLE

ENVIRONMENTAL HEALTH CRITERIA FOR DIESEL FUEL AND EXHAUST EMISSIONS

WHO DRAFTING GROUP ON ENVIRONMENTAL HEALTH CRITERIA FOR DIESEL FUEL

AND EXHAUST EMISSIONS

WHO TASK GROUP ON ENVIRONMENTAL HEALTH CRITERIA FOR DIESEL FUEL AND

EXHAUST EMISSIONS

PART A: DIESEL FUEL

A1. SUMMARY A1.1 Identity, physical and chemical properties, and analytical methods A1.2 Sources of human and environmental exposure A1.3 Environmental transport, distribution, and
transformation A1.4 Environmental levels and human exposure A1.5 Effects on laboratory mammals and in vitro test

systems
A1.6 Effects on humans A1.7 Effects on other organisms in the laboratory and the
field
A1.8 Evaluation of human health risks
A1.9 Evaluation of effects on the environment A2. IDENTITY, PHYSICAL AND CHEMICAL PROPERTIES, AND ANALYTICAL

METHODS A2.1 Identity A2.1.1 Fuel components

A2.1.1.1 Alkanes

A2.1.1.2 Alkenes A2.1.1.3 Aromatic compounds

A2.1.1.4 Sulfur
A2.1.2 Fuel additives A2.1.2.1 Cetane number improvers A2.1.2.2 Smoke suppressors A2.1.2.3 Flow improvers A2.1.2.4 Cloud-point depressors A2.1.2.5 Wax anti-settling additives
A2.1.2.6 Other additives

A2.1.3 Quality aspects of diesel fuels A2.1.3.1 Ignition performance and cetane

number A2.1.3.2 Density

A2.1.3.3 Sulfur content A2.1.3.4 Viscosity A2.1.3.5 Cold-flow properties A2.2 Physical and chemical properties A2.3 Analytical methods
A2.4 Conversion factors

A3. SOURCES OF HUMAN AND ENVIRONMENTAL EXPOSURE

A3.1 Natural occurrence A3.2 Anthropogenic sources
A3.2.1 Production and use A3.2.1.1 Production process

A3.2.1.2 Use A3.2.1.3 Production and consumption

levels A3.2.2 Emissions during production and use
A3.2.2.1 Air

A3.2.2.2 Water
A3.2.2.3 Soil
A3.2.3 Accidental releases to the environment A4. ENVIRONMENTAL TRANSPORT, DISTRIBUTION, AND TRANSFORMATION A4.1 Transport and distribution between media A4.1.1 Evaporation from and dissolution in the aqueous phase A4.1.2 Transport in and adsorption onto soil and
sediment
A4.1.2.1 Soil
A4.1.2.2 Sediment
A4.2 Transformation and removal
A4.2.1 Photooxidation A4.2.2 Biodegradation A4.2.2.1 Microbial degradation A4.2.2.2 Phytoplankton and marine algae

A4.2.2.3 Invertebrates and vertebrates
A4.2.3 Bioaccumulation A4.2.4 Tainting A4.2.5 Entry into the food chain
A4.3 Ultimate fate after use
A4.3.1 Use in motor vehicles A4.3.2 Spills A4.3.3 Disposal

A5. ENVIRONMENTAL LEVELS AND HUMAN EXPOSURE
A5.1 Environmental levels A5.2 Exposure of the general population A5.3 Occupational exposure during manufacture, 

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A6. KINETICS AND METABOLISM IN LABORATORY ANIMALS AND HUMANS

A7. EFFECTS ON LABORATORY MAMMALS AND IN VITRO TEST SYSTEMS A7.1 Single exposure A7.2 Short-term exposure A7.2.1 Subacute exposure A7.2.1.1 Dermal exposure
A7.2.1.2 Inhalation A7.2.2 Subchronic exposure A7.2.2.1 Dermal exposure
A7.2.2.2 Inhalation A7.3 Long-term exposure A7.3.1 Dermal exposure
A7.3.2 Inhalation
A7.4 Dermal and ocular irritation; dermal sensitization
A7.4.1 Dermal irritation
A7.4.2 Ocular irritation
A7.4.3 Sensitization A7.5 Reproductive toxicity, embryotoxicity, and teratogenicity A7.6 Mutagenicity and related end-points
A7.6.1 In vitro
A7.6.2 In vivo A7.7 Carcinogenicity A7.7.1 Dermal exposure
A7.7.2 Inhalation

A8. EFFECTS ON HUMANS A8.1 Exposure of the general population A8.2 Occupational exposure

A9. EFFECTS ON OTHER ORGANISMS IN THE LABORATORY AND THE FIELD A9.1 Laboratory experiments A9.1.1 Microorganisms

A9.1.1.1 Water
A9.1.1.2 Soil A9.1.2 Aquatic organisms A9.1.2.1 Plants (phytoplankton)

A9.1.2.2 Invertebrates A9.1.3 Terrestrial organisms

A9.1.3.1 Plants

A9.1.3.2 Invertebrates

A9.1.3.3 Vertebrates
A9.2 Field observations A9.2.1 Microorganisms.

A9.2.1.1 Water
A9.2.1.2 Soil A9.2.2 Aquatic organism A9.2.3 Terrestrial organisms

A9.2.3.1 Plants

A9.2.3.2 Invertebrates

A9.2.3.3 Vertebrates

A10. EVALUATION OF HUMAN HEALTH RISKS AND EFFECTS ON THE

ENVIRONMENT A10.1 Evaluation of human health risks A10.1.1 Exposure of the general population A10.1.2 Occupational exposure A10.1.3 Non-neoplastic effects A10.1.4 Neoplastic effects
A10.2 Evaluation of effects on the environment

A11. RECOMMENDATIONS A11.1 Recommendations for the protection of human health A11.2 Recommendation for the protection of the environment
A11.3 Recommendations for further research

A12. PREVIOUS EVALUATIONS BY INTERNATIONAL BODIES

PART B: DIESEL EXHAUST EMISSIONS
B1. SUMMARY B1.1 Identity, physical and chemical properties, and analytical methods B1.2 Sources of human and environmental exposure B1.3 Environmental transport, distribution, and
transformation B1.4 Environmental levels and human exposure B1.5 Kinetics and metabolism in laboratory animals and

humans B1.5.1 Deposition B1.5.2 Retention and clearance of particles B1.5.3 Retention and clearance of polycyclic aromatic hydrocarbons adsorbed onto diesel

soot B1.5.4 Metabolism B1.6 Effects on laboratory mammals and in vitro test

systems
B1.7 Effects on humans B1.8 Effects on other organisms in the laboratory and the
field
B1.9 Evaluation of human health risks B1.9.1 Non-neoplastic effects B1.9.2 Neoplastic effects
B1.10 Evaluation of effects on the environment

B2. IDENTITY AND ANALYTICAL METHODS B2.1 Identity B2.1.1 Chemical composition of diesel exhaust

gases B2.1.2 Type and composition of emitted particulate

matter B2.2 Analytical methods B2.2.1 Sampling B2.2.1.1 Sampling from undiluted exhaust 

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B2.2.1.2 Sampling from diluted exhaust (dilution tube sampling) B2.2.2 Extraction from particles B2.2.3 Clean-up and fractionation B2.2.4 Chemical analysis
B2.3 Conversion factors

B3. SOURCES OF HUMAN AND ENVIRONMENTAL EXPOSURE B3.1 Anthropogenic sources
B3.1.1 Diesel exhaust emissions
B3.1.1.1 Emission of chemical constituents with the gaseous portion of diesel exhaust B3.1.1.2 Emission of particulate matter and adsorbed components in diesel exhaust gases
B3.1.2 Parameters that influence diesel exhaust
emissions B3.1.2.1 Engine conditions B3.1.2.2 Fuel specification
B3.1.2.3 Malfunction B3.1.3 Total emissions by diesel engines
B3.1.4 Control of emissions B3.1.4.1 Particle traps B3.1.4.2 Catalytic converters B3.2 Regulatory approaches B4. ENVIRONMENTAL TRANSPORT, DISTRIBUTION, AND TRANSFORMATION B4.1 Transport and distribution between media
B4.2 Transformation and removal

B5. ENVIRONMENTAL LEVELS AND HUMAN EXPOSURE B5.1 Exposure of the general population B5.2 Occupational exposure
B5.2.1 Truck drivers and mechanics B5.2.2 Bus garage and other bus workers B5.2.3 Fork-lift truck operators
B5.2.4 Railroad workers
B5.2.5 Mine workers B5.2.6 Fire fighters B5.3 Biomonitoring B5.3.1 Urinary mutagenicity B5.3.2 Other analyses

B6. KINETICS AND METABOLISM IN LABORATORY ANIMALS AND HUMANS B6.1 Deposition B6.2 Retention and clearance of particles B6.3 Retention and clearance of polycyclic aromatic hydrocarbons adsorbed onto diesel soot
B6.4 Metabolism

B7. EFFECTS ON LABORATORY MAMMALS AND IN VITRO TEST SYSTEMS B7.1 Single exposure B7.2 Short-term exposure

B7.3 Long-term exposure and studies of carcinogenicity B7.3.1 Non-neoplastic effects B7.3.2 Carcinogenicity
B7.3.2.1 Inhalation B7.3.2.2 Other routes of exposure
B7.4 Dermal and ocular irritation; dermal sensitization B7.5 Reproductive toxicity, embryotoxicity, and teratogenicity B7.5.1 Reproductive toxicity B7.5.2 Embryotoxicity B7.5.3 Teratogenicity B7.6 Mutagenicity and related end-points
B7.6.1 In vitro
B7.6.2 In vivo
B7.6.3 DNA adduct formation B7.7 Special studies B7.7.1 Immunotoxicity
B7.7.2 Behavioural effects B7.8 Factors that modify toxicity; toxicity of
metabolites B7.9 Mechanisms of toxicity; mode of action B7.9.1 Carcinogenic effects
B7.9.1.1 DNA-reactive mechanisms B7.9.1.2 Cytotoxicity with regenerative cell proliferation B7.9.1.3 Effects of particles B7.9.1.4 Effects of polycyclic aromatic hydrocarbons B7.9.2 Noncarcinogenic effects

B8. EFFECTS ON HUMANS B8.1 General population B8.1.1 Acute exposure: olfactory, nasal, and
ocular irritation B8.1.2 Air pollution B8.2 Occupational exposure B8.2.1 Effects on the respiratory system B8.2.1.1 Symptoms B8.2.1.2 Acute changes in pulmonary
function B8.2.1.3 Pulmonary effects B8.2.2 Epidemiological studies (noncarcinogenic effects) B8.2.2.1 Effects on the respiratory

system B8.2.2.2 Effects on the circulatory
system B8.2.3 Epidemiological studies (carcinogenic effects) 

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B9. EFFECTS ON OTHER ORGANISMS IN THE LABORATORY AND THE FIELD

B10. EVALUATION OF HUMAN HEALTH RISKS AND EFFECTS ON THE

ENVIRONMENT B10.1 Exposure of the general population B10.2 Occupational exposure B10.3 Non-neoplastic effects
B10.3.1 Hazard identification

B10.3.1.1 Humans B10.3.1.1 Experimental animals B10.3.2 Dose-response assessment B10.3.2.1 Epidemiological studies B10.3.2.2 Studies in experimental animals B10.3.3 Exposure assessment
B10.3.4 Risk characterization

B10.3.4.1 Humans B10.3.4.2 Experimental animals B10.4 Neoplastic effects
B10.4.1 Hazard identification B10.4.1.1 Lung cancer: occupational

exposure B10.4.1.2 Urinary bladder cancer: occupational exposure B10.4.2 Dose-response assessment B10.4.2.1 Lung cancer B10.4.2.2 Urinary bladder cancer B10.4.3 Exposure assessment
B10.4.4 Risk characterization B10.4.4.1 Human lung cancer B10.4.4.2 Human urinary bladder cancer
B10.4.4.3 Risk characterization based on studies in experimental animals Appendix B10.1 Construction of a biologically based (alternative) model Appendix B10.2 E-M algorithm Appendix B10.3 A tumour growth model

B11. RECOMMENDATIONS B11.1 Recommendations for the protection of human health B11.2 Recommendation for the protection of the environment
B11.3 Recommendations for further research

B12. PREVIOUS EVALUATIONS BY INTERNATIONAL BODIES

REFERENCES

RESUMÉ

RESUMEN

NOTE TO READERS OF THE CRITERIA MONOGRAPHS

Every effort has been made to present information in the Criteria monographs as accurately as possible without unduly delaying their publication. In the interest of all users of the Environmental Health Criteria monographs, readers are requested to communicate any errors that may have occurred to the Director of the International Programme on Chemical Safety, World Health Organization, Geneva, Switzerland, in order that they may be included in corrigenda.

* * *

A detailed data profile and a legal file can be obtained from the International Register of Potentially Toxic Chemicals, Case postale 356, 1219 Châtelaine, Geneva, Switzerland (Telephone No. 979 9111).

* * *

This publication was made possible by grant number 5 U01
ES02617-15 from the National Institute of Environmental Health Sciences, National Institutes of Health, USA, and by financial support from the European Commission.

Environmental Health Criteria

PREAMBLE

Objectives

The WHO Environmental Health Criteria Programme was initiated in 1973, with the following objectives:

(i) to assess information on the relationship between exposure to environmental pollutants and human health and to provide guidelines for setting exposure limits;

(ii) to identify new or potential pollutants;

(iii) to identify gaps in knowledge concerning the health effects of pollutants;

(iv) to promote the harmonization of toxicological and epidemiological methods in order to have internationally comparable results.

The first Environmental Health Criteria (EHC) monograph, on mercury, was published in 1976; numerous assessments of chemicals and of physical effects have since been produced. Many EHC monographs have 

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Since the time of its inauguration, the EHC Programme has widened its scope, and the importance of environmental effects has been increasingly emphasized in the total evaluation of chemicals, in
addition to their health effects.

The original impetus for the Programme came from resolutions of the World Health Assembly and the recommendations of the 1972 United Nations Conference on the Human Environment. Subsequently, the work became an integral part of the International Programme on Chemical Safety (IPCS), a cooperative programme of UNEP, ILO, and WHO. In this manner, with the strong support of the new partners, the importance of occupational health and environmental effects was fully recognized. The EHC monographs have become widely established, used, and recognized throughout the world.

The recommendations of the 1992 United Nations Conference on Environment and Development and the subsequent establishment of the Intergovernmental Forum on Chemical Safety, with priorities for action in the six programme areas of Chapter 19, Agenda 21, lend further weight to the need for EHC assessments of the risks of chemicals.

Scope

The Criteria monographs are intended to provide critical reviews of the effect on human health and the environment of chemicals, combinations of chemicals, and physical and biological agents. They
include reviews of studies that are of direct relevance for the evaluation and do not describe every study that has been carried out. Data obtained worldwide are used, and results are quoted from original studies, not from abstracts or reviews. Both published and unpublished reports are considered, and the authors are responsible for assessing all of the articles cited; however, preference is always given to published data, and unpublished data are used only when relevant published data are absent or when the unpublished data are pivotal to the risk assessment. A detailed policy statement is available that describes the procedures used for citing unpublished proprietary data,
so that this information can be used in the evaluation without compromising its confidential nature (WHO, 1990).

In the evaluation of human health risks, sound data on humans, whenever available, are preferred to data on experimental animals. Studies of animals and in vitro systems provide support and are used mainly to supply evidence missing from human studies. It is mandatory that research on human subjects be conducted in full accord with ethical principles, including the provisions of the Helsinki
Declaration.

The EHC monographs are intended to assist national and international authorities in making risk assessments and subsequent risk management decisions. They represent a thorough evaluation of risks and are not in any sense recommendations for regulation or setting standards. The latter are the exclusive purview of national and regional governments.

Content

The layout of EHC monographs for chemicals is outlined below.

* Summary: a review of the salient facts and the risk evaluation of
the chemical * Identity: physical and chemical properties, analytical methods * Sources of exposure * Environmental transport, distribution, and transformation * Environmental levels and human exposure * Kinetics and metabolism in laboratory animals and humans * Effects on laboratory mammals and in-vitro test systems

* Effects on humans * Effects on other organisms in the laboratory and the field
* Evaluation of human health risks and effects on the environment * Conclusions and recommendations for protection of human health
and the environment

* Further research * Previous evaluations by international bodies, e.g. the International Agency for Research on Cancer, the Joint FAO/WHO Expert Committee on Food Additives, and the Joint FAO/WHO Meeting
on Pesticide Residues

Selection of chemicals

Since the inception of the EHC Programme, the IPCS has organized meetings of scientists to establish lists of chemicals that are of priority for subsequent evaluation. Such meetings have been held in Ispra, Italy (1980); Oxford, United Kingdom (1984); Berlin, Germany (1987); and North Carolina, United States of America (1995). The selection of chemicals is based on the following criteria: the existence of scientific evidence that the substance presents a hazard to human health and/or the environment; the existence of evidence that the possible use, persistence, accumulation, or degradation of the substance involves significant human or environmental exposure; the existence of evidence that the populations at risk (both human and other species) and the risks for the environment are of a significant size and nature; there is international concern, i.e. the substance is of major interest to several countries; adequate data are available on 

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If it is proposed to write an EHC monograph on a chemical that is not on the list of priorities, the IPCS Secretariat first consults with the cooperating organizations and the participating institutions.

Procedures

The order of procedures that result in the publication of an EHC monograph is shown in the following flow chart. A designated staff member of IPCS, responsible for the scientific quality of the document, serves as Responsible Officer (RO). The IPCS Editor is responsible for the layout and language. The first draft, prepared by consultants or, more usually, staff at an IPCS participating institution is based initially on data provided from the International Register of Potentially Toxic Chemicals and reference data bases such
as Medline and Toxline.

The draft document, when received by the RO, may require an initial review by a small panel of experts to determine its scientific quality and objectivity. Once the RO finds the first draft acceptable, it is distributed in its unedited form to over 150 EHC contact points throughout the world for comment on its completeness and accuracy and, where necessary, to provide additional material. The contact points,

usually designated by governments, may be participating institutions, IPCS focal points, or individual scientists known for their particular expertise. Generally, about four months are allowed before the comments are considered by the RO and author(s). A second draft incorporating the comments received and approved by the Director, IPCS, is then distributed to Task Group members, who carry out a peer review at least six weeks before their meeting.

The Task Group members serve as individual scientists, not as representatives of any organization, government, or industry. Their function is to evaluate the accuracy, significance, and relevance of
the information in the document and to assess the risks to health and the environment from exposure to the chemical. A summary and recommendations for further research and improved safety are also drawn up. The composition of the Task Group is dictated by the range of expertise required for the subject of the meeting and by the need for a balanced geographical distribution.

The three cooperating organizations of the IPCS recognize the important role played by nongovernmental organizations, so that representatives from relevant national and international associations may be invited to join the Task Group as observers. While observers may provide valuable contributions to the process, they can speak only at the invitation of the Chairperson. Observers do not participate in the final evaluation of the chemical, which is the sole responsibility of the Task Group members. The Task Group may meet in camera when it considers that to be appropriate.

All individuals who participate in the preparation of an EHC monograph as authors, consultants, or advisers must, in addition to serving in their personal capacity as scientists, inform the RO if at any time a conflict of interest, whether actual or potential, could be perceived in their work. They are required to sign a statement to that effect. This procedure ensures the transparency and probity of the

process.

When the Task Group has completed its review and the RO is satisfied as to the scientific correctness and completeness of the document, it is edited for language, the references are checked, and camera-ready copy is prepared. After approval by the Director, IPCS, the monograph is submitted to the WHO Office of Publications for printing. At this time, a copy of the final draft is also sent to the Chairperson and Rapporteur of the Task Group to check for any errors.

It is accepted that the following criteria should initiate the updating of an EHC monograph: new data are available that would substantially change the evaluation; there is public concern about health or environmental effects of the agent because of greater exposure; an appreciable time has elapsed since the last evaluation. 

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WHO DRAFTING GROUP ON ENVIRONMENTAL HEALTH CRITERIA FOR DIESEL FUEL

AND EXHAUST EMISSIONS

WHO, Geneva, 6-9 December 1993

Members

Dr J.A. Bond, Chemical Industry Institute of Toxicology, Research Triangle Park, NC, United State

Dr R.P. Bos, University of Nijmegen, Nijmegen, Netherlands

Dr R. Brown, Medical Research Council Toxicology Unit, University of Leicester, Leicester, United Kingdom (Joint Rapporteur)

Dr Chao Chen, Human Health Assessment Group, United States Environmental Protection Agency, Washington DC, United States

Dr I. Farkas, National Institute of Hygiene, Budapest, Hungary

Dr E. Garshick, Pulmonary Section, Brockton/West Roxbury VA Medical Center, West Roxbury, MA, United States

Dr P. Gustavsson, North Western Health Board, Stockholm, Sweden

Dr D. Guth, United States Environmental Protection Agency, Research Triangle Park, NC, United States

Dr U. Heinrich, Department of Experimental Hygiene, Fraunhofer Institute of Toxicology and Aerosol Research, Hanover, Germany

Dr R.F. Hertel, Federal Health Office, Bundesgesundheitsamt, Berlin,
Germany

Professor G. Oberdörster, Department of Environmental Medicine, University of Rochester Medical Center, Rochester, NY, United States

Dr W. Pepelko, United States Environmental Protection Agency, Washington DC, United States

Dr P.J.A. Rombout, Laboratory of Toxicology, National Institute of Public Health and Environmental Protection, Bilthoven, Netherlands (Vice-Chairman)

Dr G. Rosner, Hazardous Substances Documentation Group, Fraunhofer Institute of Toxicology and Aerosol Research, Hanover, Germany

Dr J. Roycroft, National Institute of Environmental Health Sciences, Research Triangle Park, NC, United States 

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United States (Chairman)

Dr B.H. Thomas, Environmental Health Directorate, Ottawa, Canada

Dr L. Turrio, Istituto Superiore di Sanita, Laboratorio Tossicologia Comparata e Ecotossicologia, Rome, Italy

Mr. R. Waller, Department of Health, London, United Kingdom

Secretariat

Dr P. Boffetta, International Agency for Research on Cancer, Lyon, France (6-7 December 1993)

Dr E. Smith, International Programme on Chemical Safety, World Health Organization, Geneva, Switzerland

Representatives/Observers

CONCAWE Dr R.H. McKee, Exxon Biomedical Sciences, East Millstone, NJ,
United States

UNITED KINGDOM DEPARTMENT OF THE ENVIRONMENT Dr P.T.C. Harrison, MRC Institute of Environment & Health, Leicester, United Kingdom

WHO TASK GROUP ON ENVIRONMENTAL HEALTH CRITERIA FOR DIESEL FUEL AND

EXHAUST EMISSIONS

Fraunhofer Institute of Toxicology & Aerosol Research, Hanover 27 June-1 July 1994

Members

Dr J.A. Bond, Chemical Industry Institute of Toxicology, Research Triangle Park, NC, United States

Dr R. Brown, Medical Research Council Toxicology Unit, University of Leicester, Leicester, United Kingdom

Dr Chao Chen, Human Health Assessment Group, United States Environmental Protection Agency, Washington DC, United States

Dr E. Garshick, Pulmonary Section, Brockton/West Roxbury VA Medical Center, West Roxbury, MA, United States

Dr U. Heinrich, Department of Experimental Hygiene, Fraunhofer Institute of Toxicology and Aerosol Research, Hanover, Germany

Dr R.F. Hertel, Federal Health Office, Bundesgesundheitsamt, Berlin,
Germany

Dr Jun Kagawa, Tokyo Women's Medical College, Tokyo, Japan

Professor G. Oberdörster, Department of Environmental Medicine, University of Rochester Medical Center, Rochester, NY, United States

Dr P.J.A. Rombout, Laboratory of Toxicology, National Institute of Public Health and Environmental Protection, Bilthoven, Netherlands

Dr M. Roller, Medizinishes Institut für Umwelthyglene an der Universität Düsseldorf, Düsseldorf, Germany Dr G. Rosner, Hazardous Substances Documentation Group, Fraunhofer Institute of Toxicology and Aerosol Research, Hanover, Germany

Dr A. Sivak, Environmental Health Sciences, Saint Augustine, FL,
United States

Dr L. Turrio, Istituto Superiore di Sanita, Laboratorio Tossicologia Comparata e Ecotossicologia, Rome, Italy

Mr R. Waller, Department of Health, London, United Kingdom

Secretariat

Dr H. Moller, International Agency for Research on Cancer, Lyon,

France

Dr E. Smith, International Programme on Chemical Safety, World Health Organization, Geneva, Switzerland

Dr M. Younes, European Centre for Environment and Health, Bilthoven,

Netherlands

Representatives/Observers

Dr Lutz Von Meyerinck, BP Oil Europe, Brussels, Belgium

GERMAN AUTOMOBILE ASSOCIATION Dr N. Pelz, Mercedes-Benz AG, Stuttgart, Germany

ILSI Dr I.T. Salmeen, Chemistry Department, Ford Motor Company, Dearborn, MI, United States

IUTOX Dr P. Montuschi, Department of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmENVIRONMENTAL HEALTH CRIETERIA FOR DIESEL FUEL AND EXHAUST EMISSIONS

A WHO Task Group on Environmental Health Criteria for Diesel Fuel and Exhaust Emissions met at the Fraunhofer Institute of Toxicology and Aerosol Research, Hanover, Germany from 27 June to 1 July 1994. Dr G. Rosner, Fraunhofer Institute, welcomed the participants on behalf of the Institute and its Director, Professor U. Mohr, and Dr E.M. Smith, IPCS, welcomed the participants on behalf of Dr M. Mercier, Director of the IPCS, and on behalf of the heads of the three IPCS cooperating organizations (UNEP, ILO, and WHO). The Task Group reviewed and revised the draft and evaluated the risks for human health and the environment from exposure to diesel fuel and exhaust
emissions.

The first draft of the monograph was prepared at the Fraunhofer Institute. After international circulation for comment, this draft was extensively revised by a Working/Drafting Group, convened at WHO, Geneva, from 6 to 9 December 1993, and a second draft was prepared for further international circulation for comment. The membership of the drafting group is shown previously. A final draft, incorporating comments received from the IPCS contact points for Environmental Health Criteria monographs and new material, was completed at the Fraunhofer Institute under the coordination of Dr G. Rosner, with important contributions to the text from the following Institute staff

members:

Dr B. Bellman

Dr A. Boehnke Dr O. Creutzenberg
Dr J. Kielhorn

Dr E.M. Smith of the IPCS Central Unit was responsible for the scientific content of the monograph and Mrs E. Heseltine, Lajarthe, France, for the editing.

The efforts of all who helped in the preparation and finalization of the monograph are gratefully acknowledged.

PART A DIESEL FUEL

A1. SUMMARY

A1.1 Identity, physical and chemical properties, and analytical

methods

Diesel fuel is a complex mixture of normal, branched, and cyclic alkanes (60 to > 90% by volume; hydrocarbon chain length, usually

between C 9 and C 30 ); aromatic compounds, especially alkylbenzenes

(5-40% by volume); and small amounts of alkenes (0-10% by volume) obtained from the middle-distillate, gas-oil fraction during petroleum separation. Benzene, toluene, ethylbenzene, and xylenes and polycyclic aromatic hydrocarbons (PAHs), especially naphthalene and its methyl-substituted derivatives, may be present at levels of parts per million in diesel fuel. The sulfur content of diesel fuels depends on the source of crude oil and the refinery process. It is regulated by law in a number of countries and is usually between 0.05 and 0.5 weight percent. Additives are used to influence the flow, storage, and combustion of diesel fuel, to differentiate products, and to meet trademark specifications. At room temperature, diesel fuels are generally moderately volatile, slightly viscous, flammable, brown liquids with a kerosene-like odour. The boiling ranges are usually between 140 and 385°C (> 588°C for marine diesel fuel); at 20°C, the density is 0.87-1.0 g/cm 3 and the water solubility is 0.2-5 mg/litre. The quality and composition of diesel fuel influence the emissions of pollutants from diesel engines considerably. Important variables are ignition behaviour (expressed in terms of cetane number), density, viscosity, and sulfur content. The specifications of commercial diesel fuel differ considerably in
different countries.

Heating fuels and some kerosene jet fuels produced during the refining process may have a composition similar to that of diesel fuel, although with different additives. Biological data on these
mixtures have therefore also been taken into account in the assessments of toxicity and ecotoxicity.

Owing to the complexity of the diesel fuel mixture, there is no specific analytical method, and the analytical techniques used in most environmental assessments are suitable only for measuring the total petroleum hydrocarbon mixture. The methods consist of preliminary solvent extraction, a clean-up procedure to remove naturally occurring hydrocarbons, and subsequent detection by gravimetry, infrared spectroscopy or gas chromatography. Neither the gravimetric nor the infrared technique provides useful qualitative or quantitative information on contaminants and can thus be used only for screening. Gas chromatography combined with detection techniques such as flame ionization and mass spectrometry is the standard procedure for analysing environmental samples. Many other methods are available for the analysis of individual hydrocarbons in diesel fuels.

A1.2 Sources of human and environmental exposure

Diesel fuels are produced by refining crude oils. In order to meet technical specifications for performance, diesel fuels are generally blended; further formulation with additives improves their properties for specific uses. Diesel fuels are widely used as transportation fuels. The more volatile fuels, with low viscosity, are required for high-speed engines and the heavier grades for railroad and ship diesel engines. Much heavy-duty road transport is powered by 

# Pa g e 10 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmdiesel engines. Passenger cars powered by diesel engines are becoming increasingly more common in Europe and Japan (10-25%), whereas in North America the percentage of diesel-fuelled passenger cars is about 1-2%, with a slightly decreasing tendency. Diesel fuel is used in stationary engines and in boilers, e.g. reciprocating engines, gas turbines, pipeline pumps, gas compressors, steam processing units in electric power plants, burner installations, and industrial space and water heating facilities.

Over the last five years, the worldwide demand for diesel fuels has increased steadily. In 1985, the following amounts of diesel fuel were consumed: about 170 000 kt per year in North America; about 160 000 kt per year, including gas oils, in the European Union; and about 46 000 kt per year in Australia, Japan, and New Zealand, equivalent to a total of 1062 kt per day. In 1990, world demand was reported to be about 1110 kt per day.

No information is available on emissions during the production of diesel fuels; however, this source would seem to be of minor importance, because the refining process is carried out in closed systems. Emissions may occur principally during storage and transportation. Diesel fuels are released as a result of spills and at filling stations during the refuelling of vehicles. The atmosphere and the hydrosphere are the most heavily affected environmental compartments. Soil contamination with diesel fuels may occur during accidents and is also a problem in railroad yards. The numerous techniques for cleaning soils contaminated with diesel fuel include excavation, biological methods, and containment.

A1.3 Environmental transport, distribution, and transformation

Very few data are avilable on the environmental fate of diesel fuels, but the mechanisms of their distribution and transformation are considered to be comparable to those of heating fuels, such as No. 2 fuel oil, which have been well studied. Spills of diesel fuel on water spread almost immediately to form a 'slick'. The polar and low-relative-molecular-mass components dissolve and leach out of the slick, and the volatile components evaporate from the surface; microbial degradation also begins. Chemical and biological weathering alter the composition of the spill. These processes are dependent on

temperature; spills that occur in Arctic conditions are more persistent than those that occur in temperate climates. In marine environments, most of the low-relative-molecular-mass aromatic species are dissolved into the water phase, but the primary branched alkanes, cycloalkanes, and remaining aromatic compounds may remain in sediments for more than a year.

Although no information is available on the photooxidation of diesel fuels in water and air, evaporated oil components are degraded photochemically. No. 2 fuel oil has been shown to be photooxidized rapidly in water under environmental conditions.

The individual constituents of diesel fuel are inherently biodegradable, to varying degrees and at different rates. The

n- alkane, n- alkylaromatic, and simple aromatic molecules in the C 10 -C 22 range are the most readily degradable. Smaller molecules are generally rapidly metabolized. Long-chain n- alkanes are more slowly degraded, owing to their hydrophobicity and because they are viscous or solid at ambient temperatures. Branched alkanes and cycloalkanes are relatively resistant to biological breakdown, and PAHs are resistant. The overall rates of degradation of hydrocarbons are limited by temperature, water content, oxygen, pH, inorganic nutrients, and microbial metabolic versatility.

Unicellular algae can take up and metabolize both aliphatic and aromatic hydrocarbons, but the extent to which this actually occurs in nature is poorly understood. Unlike microorganisms that use petroleum carbons as a carbon source, animals generally oxidize and conjugate products, rendering end-products that are more soluble and therefore easier to excrete. All animal species tested can take up petroleum hydrocarbons. PAHs, crude oil, and refined petroleum products are known to induce cytochrome P450 enzymes and to increase the levels of hydrocarbon metabolism in numerous marine and freshwater fish species.

Few data are available on the bioaccumulation of diesel fuel in the laboratory, but there is plentiful evidence from studies of spills and laboratory studies on other oils, particularly No. 2 fuel oil, that aquatic organisms bioconcentrate hydrocarbons. The

n- octanol-water partition coefficient for diesel fuel is 3.3-7.06, which suggests high potential bioaccumulation; however, many of the lower-relative-molecular-mass compounds are readily metabolized, and the actual bioaccumulation of higher-relative-molecular-mass compounds is limited by their low water solubility and large molecular size. Thus, actual bioaccumulation may be low.

Fish have been tainted by diesel fuel after spills. No data are available on the biomagnification of diesel fuel.

No experimental data are available on the movement of diesel fuel through the soil, although a direct correlation between the movement and kinematic viscosity has been proposed. The movement of kerosene through soil depends on the moisture content and nature of the soil.

A1.4 Environmental levels and human exposure

As diesel fuels are complex mixtures, the environmental levels
have not been measured. The individual constituents of diesel fuels can be detected in almost all compartments of the environment, although their source cannot be verified. The general population may 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmbe exposed to diesel fuel at filling stations and as a result of spills.

Occupational exposure to diesel fuel occurs in a large number of activities. Because of their low volatility, diesel fuels should generate only low concentrations of vapours at normal temperatures, but high operating temperatures can result in significant
concentrations.

A1.5 Effects on laboratory mammals and in-vitro test systems

The acute toxicity of diesel fuels is low after oral or dermal exposure or after inhalation. The oral LD 50 value was > 5000 mg/kg
body weight in all species tested (mouse, rabbit, rat, guinea-pig). Dermal application resulted in an LD 50 value of > 5000 mg/kg body
weight in mice and rabbits, although values of > 2000 mg/kg body weight were reported for some kerosenes and middle distillates, with different protocols and lower limit doses. The LC 0 value in rats
exposed by inhalation was about 5 mg/litre, except for one straight-run middle distillate for which a value of 1.8 mg/litre was

seen.

In rabbits treated dermally with up to 8000 μl/kg body weight per day and mice with up to 40 000 mg/kg body weight per day, acanthosis and hyperkeratosis due to severe irritation were seen. Rabbits were
more sensitive than mice. Inhalation of diesel fuel was neuro- depressive in mice at concentrations up to 0.2 mg/litre but not in rats exposed to up to 6 mg/litre. Body and liver weights were
reduced in rats.

Mice, rats, and dogs did not show significant cumulative toxicity after inhalation of up to 1.5 mg/litre subchronically. The specific nephropathy syndrome seen in male rats is linked to an inherent accumulation of hyalin droplets in the renal tubules.

The only effects of long-term exposures were ulceration after dermal application to mice (250 or 500 mg/kg body weight per day) and significant alterations in organ weight after inhalation of 1 or 5 mg/litre by rats. In both studies the mean body weights were

reduced.

Various types of diesel fuels were slightly to severely irritating to the skin of rabbits. Diesel fuels do not irritate the eye, but some kerosenes have been reported to have a slight irritating
effect. Diesel fuels do not cause skin sensitization.

Diesel and jet fuels (kerosene) were neither embryotoxic nor teratogenic in two studies in rats exposed by inhalation to 100 or 400 ppm and in one study in which rats were given up to 2000 mg/kg body weight per day by gavage. In the last study, reduced fetal weight

was observed.

Tests in Salmonella typhimurium did not provide clear evidence of mutagenicity. Some positive findings in S. typhimurium and in mouse lymphoma cells were considered to be equivocal owing to the inconsistency of the results. Tests for genotoxicity in mice in vivo (induction of micronuclei or chromosomal aberrations) also gave equivocal or negative responses.

Diesel fuels induced a low level of dermal carcinogenicity. In the present state of research, it cannot be concluded whether the carcinogenic potency of diesel fuels is mediated by a genotoxic mechanism or by chronic dermal damage.

A1.6 Effects on humans

Non-occupational exposure to diesel fuel can occur during manual filling of fuel tanks. The primary source of dermal exposure is accidental spills, which result in immediate high levels of exposure
but are of short duration.

After accidental dermal contact, anuria, renal failure, gastro- intestinal symptoms, and cutaneous hyperkeratosis have been reported. Toxic lung disease has been observed after accidental ingestion of diesel fuel and subsequent aspiration. Persistent productive cough has been reported after inhalation. In a case-control study of men exposed to diesel fuel, an increased risk for cancer of the lung other than

adenocarcinoma was found; a positive association was also seen with prostatic cancer, although a higher risk was noted for the group with 'nonsubstantial' exposure than for that with 'substantial' exposure. In a cross-sectional study of factory workers exposed to kerosene jet fuels, dizziness, headache, nausea, palpitation, pressure in the chest, and eye irritation were found to be more prevalent than in unexposed controls. The time-weighted average concentration of vapour from the fuel in the breathing zone was estimated to be 128-423 mg/m 3

.

A1.7 Effects on other organisms in the laboratory and the field

Diesel fuel is more toxic than crude oil to aquatic organisms and plants. The ecotoxicity of diesel fuel is generally attributed to soluble aromatic compounds, but insoluble aliphatic hydrocarbons may also be implicated. Of the aromatic compounds, monoaromatics are the least toxic, their acute toxicity increasing with molecular mass up to the four- to five-ring compounds, although these are poorly soluble in seawater. In some animals, e.g. fish and birds, physical coating of the body surface by the fuel can produce toxicity and mortality.

Laboratory experiments have been carried out on diesel fuel, 

# Pa g e 12 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmwater-soluble fractions, oil-water dispersions, and microencapsulated oil. Diesel fuel did not significantly reduce the growth in culture of the green alga Euglena gracilis, whereas a low concentration (0.1%) almost completely inhibited the growth of Scenedesmus quadricauda. Light diesel fuel (0.05%) stimulated the growth, photosynthesis, and chlorophyll asynthesis of Chlorella salina but slightly inhibited respiration; at higher concentrations, the growth rate and photosynthesis were greatly reduced. Long-term exposure inhibited the growth of the benthic algae Ascophyllum nodosum and Laminaria digitata. In blue-green algae, photosynthesis was reduced by the aromatic and asphaltic fractions but not by the aliphatic fraction.

Diesel fuel was acutely toxic to Daphnia spp., chironomid larvae, and the mollusc Viviparus bengalensis (Gastropoda). A concentration of 0.1 ml/litre caused the death of tidepool copepods, Tigriopus californicus, within five days.

Mytilus edulismussels accumulate diesel fuel, have markedly reduced feeding and growth rates, and show reproductive toxicity after chronic exposure to diesel fuel. The EC 50 for spawning in mussels
exposed for 30 days was about 800 μg/litre. The LC 50 of micro-
encapsulated diesel oil after exposure of maturing mussels for 30 days was about 5000 μg/litre. Diesel oil was more toxic to larvae than to juveniles: 10 μg/litre had adverse effects on the growth of

larvae.

Freshwater crabs (Barytelphusa cunicularis) exposed to sublethal concentrations of diesel fuel for up to 96 h generally reduced their oxygen consumption, particularly at lower exposures up to 8 h. With longer exposures, the oxygen consumption was equal to or higher than

that of the controls.

In 96-h tests of acute toxicity in juvenile salmonids under static conditions, diesel fuel was more toxic to pink salmon, Onchorhychus gorbuscha (LC 50 : 32-123 mg/litre), than to cohosalmon,

O. kisutch (LC 50 : 2186-3017 mg/litre), or rainbow trout,
O. mykiss (LC 50 : 3333-33 216 mg/litre), irrespective of water type.

The threshold for detection of behavioural responses of cod
( Gadus morhua L.) exposed to diesel fuel in seawater was within 100-400 ng/litre. The Antarctic fish Pagothenia borchgrevinki
withstood an undiluted water-soluble fraction of diesel fuel oil for up to 72 h but showed signs of stress.

Birds are affected externally and internally by oil contamination. Diesel fuel destroys the waterproof nature of the birds' plumage and is ingested during preening. Diesel and fuel oil fed by gavage at 2 ml/kg body weight to ducks caused lipid pneumonia, extreme inflammation of the lungs, fatty infiltration of the liver, and hepatic degeneration after 24 h. Administration of diesel or fuel oil at 1 ml/kg caused severe irritation of the digestive tract and toxic nephrosis. Higher doses resulted in adrenal enlargement (mainly due to hyperplasia of cortical tissue), depression of plasma cholinesterase levels, ataxia, and tremors. Doses up to 20 ml/kg body weight were not fatal to healthy birds, but the LD 50 for diesel and
fuel oil administered to birds under stress was 3-4 ml/kg body weight.

After spills of diesel fuel, zooplankton appear to be highly vulnerable to dispersed and dissolved petroleum constituents but less so to floating oils. Aquatic organisms may be affected in a number of ways, including direct mortality (fish eggs, copepods, and mixed plankton), external contamination by oil (chorions of fish eggs and cuticles and feeding appendages of crustaceans), tissue contamination by aromatic constituents, abnormal development of fish embryos, and
altered metabolic rates.

A1.8 Evaluation of human health risks

The general population can be exposed to diesel fuel and other middle distillates at filling stations, as a result of accidental spills, during the handling of such fuels, and during use of kerosene for domestic cooking or heating. Workers can be exposed to diesel fuel and other middle distillates while handling and discharging the fuel at terminals, storage tanks, and filling stations; during the manufacture, repair, maintenance, and testing of diesel engines and other equipment; during use of diesel fuel as a cleaning agent or solvent; and in handling and routine sampling of diesel fuel in the laboratory. Owing to the low volatility of diesel fuel, only low concentrations of vapour are likely to occur at room temperature, although in confined spaces at high temperatures significant levels may be found.

Exposure to vapour is minimal during the normal handling of diesel fuel. The most likely effect on human health is dermatitis after skin contact. Diesel fuel is a skin irritant but does not appear to irritate the eye. Acute toxic effects on the kidney can occur after dermal exposure, but the effects of long-term dermal absorption of low
concentrations are unknown.

Diesel fuels are toxic when ingested, sometimes resulting in regurgitation and aspiration, which can cause chemical pneumonia; the same is true for any hydrocarbon in a particular range of viscosity.

In rodents exposed by inhalation to diesel fuel at concentrations up to 0.2 mg/litre, a neurodepressive effect was seen in mice but not in rats at the higher concentrations. Subchronic exposure by inhalation to various distillate fuels induced specific alpha2-microglobulin nephropathy in male rats; this effect is
considered irrelevant for humans. 

# Pa g e 13 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmDiesel fuels were neither embryotoxic nor teratogenic in animals exposed orally or by inhalation.

There is no clear evidence of mutagenic activity in bacteria, and the results of other tests for genotoxicity in vitro and in vivo were equivocal.

A case-control study of workers exposed to diesel fuel suggested an increased risk for cancer of the lung other than adenocarcinoma and for prostatic cancer. In neither case was there an exposure-response relationship. In view of the small number of studies available, the small number of cases, and the correspondingly wide confidence intervals, no conclusion can be drawn about the carcinogenicity to
humans of diesel fuel.

In mice, dermally administered diesel fuels had weak carcinogenic potential. In view of the absence of clear genotoxicity, cancer could be induced by nongenotoxic mechanisms, e.g. by chronic dermal irritation characterized by repeated cycles of skin lesions, causing epidermal hyperplasia.

A1.9 Evaluation of effects on the environment

The environment can be polluted by accidental release of diesel fuel on a large scale, such as during tanker disasters and pipeline leaks, or on a smaller scale from contamination of soil around factories or garages. In water, diesel fuel spreads almost immediately, polar and low-relative-molecular-mass components dissolve and leach out, volatile components evaporate from the water surface, and microbial degradation begins. The extent to which 'weathering' takes place depends on the temperature and on climatic conditions. The chemical composition of spills changes with time: after spillage on water, some fractions evaporate, and the evaporated diesel components are degraded photochemically; in sediment, diesel fuel appears generally to be delivered to bottom sediments by settling particles; in soil, the components of diesel fuel migrate at different rates, depending on the soil type.

The individual constituents of diesel fuel are inherently biodegradable, but the rates of biodegradation depend heavily on physical and climatic conditions and on microbial composition.

Aquatic organisms, in particular molluscs, bioaccumulate hydro-carbons to varying extents, but the hydrocarbons are depurated on transfer to clean water. Diesel fuel may be bioaccumulated; no data are available about biomagnification.

Spills of diesel fuel have an immediate detrimental effect on the environment, causing substantial mortality of biota. Recolonization may occur after about one year, depending on the animal or plant species and the chemical and physical content of the spill residues. Aquatic organisms that survive diesel fuel spills can be affected by
external oil contamination and tissue accumulation: abnormal development and altered metabolic rates are signs of the resulting

stress.

A2. IDENTITY, PHYSICAL AND CHEMICAL PROPERTIES, AND ANALYTICAL

METHODS

A2.1 Identity

Diesel fuels are a gas-oil fraction occurring during petroleum separation and commonly known as middle distillates (International Agency for Research on Cancer, 1989a). Gas oils are generally blended materials formulated to meet technical specifications (CONCAWE, in press). Commercial diesel fuels contain aliphatic, olefinic, cycloparaffinic, and aromatic hydrocarbons (see section A2.1.1), and additives (see section A2.1.2) to improve their fuelling properties (Sandmeyer, 1981).

Four qualities of diesel fuel are available commercially: diesel fuel (general), diesel fuel No. 1, diesel fuel No. 2, and diesel fuel No. 4 (see Table 1). The composition of diesel fuels is comparable to that of heating oils (e.g. No. 1 and No. 2 fuel oils), except for the additives (International Agency for Research on Cancer, 1989a; Agency for Toxic Substances and Disease Registry, 1995).

Diesel fuel No. 2 is used mainly as automobile fuel and corresponds to diesel fuel (general). In Europe (countries of the European Union (EU) and European Free Trade Area (EFTA)), the specifications for diesel fuel for transportation purposes are given in European standard EN 590 (European Committee for Standardization, 1993), which also provides for changes in the specifications to meet the requirements of different climatic conditions. In Sweden, two further qualities of on-road diesel fuel are available: environmental class 1 and class 2 (city diesel), with sulfur contents of 0.05% and 0.001%, respectively (Standardization Board in Sweden, 1991). The sulfur content of city diesel corresponds to that of kerosene. Diesel fuel for ship engines is covered by the International Standards

Organization (ISO) standard 8217 (International Standards Organization, 1987). In the United States of America, three grades of diesel fuel are available: diesel fuel No. 1 (relatively high volatility) for road vehicle engines subject to frequent speed and load changes; diesel fuel No. 2 (lower volatility) for industrial or heavy-duty, high-load engines running at uniform speed; and diesel fuel No. 4 (viscous) for low- and medium-speed engines such as those used in ships (American Society for Testing and Materials, 1988,
1992). 

# Pa g e 14 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmSeveral types of kerosene, also derived from the gas-oil fraction of petroleum separation, are used as aviation turbine fuels (JP fuels, jet fuels). Their hydrocarbon composition is comparable to that of
diesel fuels.

A2.1.1 Fuel components

A2.1.1.1 Alkanes

Normal, branched, and cyclic alkanes (paraffins) are the most abundant components (about 65-85%) of diesel fuels. Pristane (2,6,10,14-tetramethylpentadecane) and phytane (2,6,10,14- tetramethyl-hexadecane) are of particular interest environmentally, as the ratios of pristane to heptadecane and of phytane to octadecane make it possible to identify the source of a fuel spill; furthermore, as these ratios increase during biological degradation, they can be used to estimate the age of an environmental contamination and the degree of elimination. Cycloalkanes and bicycloalkanes constitute a significant portion of the mixture, but individual compounds are present only at low levels and are difficult to analyse. Alkyl derivatives of cyclopentane, cyclohexane, and cycloheptane are common components (Block et al., 1991; Table 2).

A2.1.1.2 Alkenes

Alkenes are not common components of crude oil but may be present in diesel fuel if converted products are added after cracking. These alkenes have predominantly branched and cyclic structures (Block et al., 1991). The total alkene content of diesel fuels is up to 10% (CONCAWE, 1985) (see also Table 2).

A2.1.1.3 Aromatic compounds

Aromatic compounds constitute 5-30% of automotive diesel fuel, 5-40% of marine diesel fuel (CONCAWE, 1985), and 10-30% of diesel fuel No. 2 (Block et al., 1991). Table 2 shows the specifications of commercial diesel fuels in this regard.

Table 1. Synonyms and trade names of commercial diesel fuels

Name CAS CAS Registry Range of Synonyms

name number carbon

numbers

Diesel fuel Diesel oil 68334-30-5 C 9 -C 20 a Auto diesel, automotive

(general) C 10 -C 28 b diesel oil, DERV, diesel,

diesel fuel oil, diesel oil, gas oil

Diesel fuel Not assigned C 9 -C 16 c Diesel fuel oil No. 1,
No. 1 (essentially C 4 -C 16 (for diesel oil No. 1, No. 1

equivalent to wide-cut diesel d
kerosene, aviation) c 8008-20-6)

Diesel fuel No. 2 68476-34-6 Diesel fuel, diesel fuel No. 2 diesel (applicable for oil No. 2, diesel oil fuel specific No. 2, No. 2 diesel e viscosity limits)

Diesel fuel 68476-31-3 C 10 -C 30 f Marine diesel fuel,

No. 4 distillate marine diesel

fuel

Adapted from International Agency for Research on Cancer (1989a) and supplemented

a From CONCAWE (in press)

b From CONCAWE (1985); automotive gas oil (automotive diesel fuel, DERV)

c From CONCAWE (1985, 1995)

d In Europe, fuels similar to US diesel No. 1 are commonly referred to as
'kerosene' or 'Arctic diesel'

e Term uncommon in Europe. In the United Kingdom, distillate fuels are frequently categorized as Class A1 (road diesel) and A2 (off-highway diesel)

f From CONCAWE (1985); distillate marine diesel

Only trace quantities of toxicologically relevant benzene, toluene, ethyl benzene, and xylene compounds (for physicochemical properties, see Table 6) are present in diesel fuel No. 2, but significant levels are found in diesel fuel No. 1 (Arctic diesel), which has lower flash-point specifications (Block et al., 1991); the International Agency for Research on Cancer (1989a) cites a flash-point of 0.25-0.5%. The concentration of benzene in kerosenes is < 0.01% by volume; wide-cut aviation kerosenes may have higher levels, but they are usually < 1% by volume (IPCS, 1986; CONCAWE, 1995; IPCS, 1993).

Table 2. Hydrocarbon specifications of some commercial diesel fuel oils

Specification Diesel Kerosene b Distillate Diesel

fuel a marine fuel d

diesel c 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmParaffins/naphthenes 65-95 e 78-96 60-90 (volume %) (wide-cut aviation, < 1)

n-Hexane (volume %) < 0.01 (wide-cut aviation, < 1)

Saturates (volume %) 59.4-76.6

Olefins (volume %) 0-10 0-5 0-10 0.0-1.0

Aromatics (volume %) 5-30 4-25 5-40 23.4-39.6 (wide-cut aviation, 6-25)

Aromaticity (weight %) 11-15

See also Table 1

a From CONCAWE (1985, in press); fuel oil similar to diesel fuel (general)

b From CONCAWE (1985, 1995); fuel oil similar to diesel fuel No. 1

c From CONCAWE (1985, in press); fuel oil similar to diesel fuel No. 4

d From German Scientific Association for Petroleum, Natural Gas, and Coal (1991); three samples of diesel fuel (general)

e Depending on origin of crude oil

Alkyl benzenes (particularly C 3 and C 4 ) are common components of
diesel fuel. Polycyclic aromatic hydrocarbons (PAHs), e.g. naphthalene, phenanthrene, acenaphthene, acenaphthylene, fluorene, fluoranthene, and pyrene, are also present, as are alkyl- and cycloalkyl-substituted homologues of these substances, the predominant ones being naphthalene and its methyl-substituted derivatives (see Table 6 for physicochemical properties) (Block et al., 1991).

As some PAHs and the benzene, toluene, ethyl benzene, and xylene components have been shown to be toxic and ecotoxic, these classes of compounds are usually included in analytical procedures for environmental contamination by diesel fuel. The PAH content of diesel fuels varies widely, the highest levels being found in low-quality fuel blended for large users (frequently railroad companies). The mid-range aromatic (and PAH) content of diesel blends is limited by the cetane number specification (Block et al., 1991) (see section A2.1.3).

The concentrations of total PAHs in diesel fuel are < 5% by volume, although some marine diesel fuels may contain > 10% by volume (CONCAWE, 1985; International Agency for Research on Cancer, 1989a). In straight-run gas-oil components, which are the major blending material of diesel fuel (see section A3.2.1.1), three-ring PAHs predominate; use of heavier atmospheric vacuum or cracker gas oils in a diesel-fuel mixture leads to an increasing content of four- to six-ring PAHs (CONCAWE, in press).

The concentrations of these constituents in a commercial diesel fuel (unspecified) and, for comparison, in No. 2 fuel oil are shown in
Table 3. It should be noted that the PAH content of No. 2 fuel oil is not limited by cetane number specification, so that it may include a larger proportion of these hydrocarbons (Block et al., 1991). Table 3 also gives the composition of the water-soluble fractions of diesel and No. 2 fuel oils and indicates how the solubility of a compound affects the composition of the fraction and of the whole oil.

A2.1.1.4 Sulfur

The sulfur content of middle distillates depends on the source of crude oil (Booth & Reglitzky, 1991; CONCAWE, 1995). The sulfur content of most diesel fuels is 0.1-0.5% by weight; it is higher than that of gasoline, which is about 0.02% by weight (Scheepers & Bos, 1992a). Only diesel fuel No. 4 (distillate marine diesel) has a sulfur level > 1% by weight (CONCAWE, 1985, in press). ISO standard 8217 (International Standards Organization, 1987) specifies a sulfur content of marine diesel fuel of 1.0-2.0% by weight.

The sulfur content of some diesel fuels ranges from 0.01 to > 3% by weight. In Europe (countries of EU and EFTA), the sulfur content is restricted to a maximum of 0.2-0.3% by weight, and, as of 1996, it will be further reduced to 0.05% by weight (European Commission, 1993). In the United States in 1988, the maximal sulfur content permitted was 0.5% by weight for diesel fuels No. 1 and 2 and 2.0% by weight for diesel fuel No. 4 (American Society for Testing and

Table 3. Concentrations of toxicologically relevant aromatic hydrocarbons in diesel fuel and No. 2 fuel oil and in 10% water- s olubl e fractions prepared from them

Compound Diesel fuel a Diesel fuel b No. 2 fuel c No. 2 fuel d Water-soluble fractions (μg/litre) (% by wt) (% by wt) (% by wt) (% by wt)
No. 2 Diesel

fuel d fuel a

Benzene 0.1 > 0.02 0.006-0.008 NR 550 344

Toluene 0.7 0.25-0.5 0.01-0.08 NR 1040 777 Alkylbenzenese 970 Ethylbenzene 0.2 0.25-0.5 0.01-0.08 NR 139 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmXylene 0.5 0.25-0.5 0.01-0.08 NR 875 Polycyclic aromatic hydrocarbons Naphthalene 0.4 0.273 0.4 840 6.6 1-Methylnaphthalene NR NR 0.82 340 66.2 2-Methylnaphthalene NR 0.67 1.89 480 108 Dimethylnaphalenes e NR NR 3.11 240 NR Trimethylnaphthalenes e NR NR 1.84 30 NR

Fluorenes e NR NR 0.36 20 NR

Phenanthrenes e NR 0.15 0.53 20 NR

NR, not reported

a From Dunlap & Beckmann (1988); the analytical method is not described in detail; the concentration of benzene seems to be unusually high.

b From International Agency for Research on Cancer (1989a); according to CONCAWE (1985), some marine diesel fuels may contain more than 10% polycyclic aromatic hydrocarbons.

c From Stone (1991); summary of several reports

d From Anderson et al. (1974); US National Research Council (1985)

e Total of several isomers Materials, 1988); in October 1993, the maximum was reduced to 0.05% by weight (US Environmental Protection Agency, 1992a; American Society for Testing and Materials, 1992). The permitted sulfur level in Brazil is > 3% by weight (A. Sivak, personal communication, 1993). In Japan, the sulfur content of diesel fuels was reduced to 0.2% by weight in 1994, and a further reduction, to 0.05% by weight, is under discussion (CONCAWE, 1990a). Blended marine diesel fuel may also contain up to about 15% residual components, i.e. material with an initial boiling-point above about 350°C (CONCAWE, in press).

A2.1.2 Fuel additives

Only agents that are added to fuels at a concentration < 1% are described as 'additives'. A more appropriate term for substances present at higher concentrations is 'fuel components'. Fuels are treated with additives for a number of reasons (see Table 4 and below); they also differentiate products and determine the trademark quality of commercial fuels (Fabri et al., 1990).

A2.1.2.1 Cetane number improvers

Cetane number improvers upgrade the ignition characteristics of a base fuel more economically than refinery processes (Fabri et al., 1990) (see section A2.1.3). Primary alkyl nitrates (e.g. isooctyl nitrate) are often used to improve cetane number. Polyethyleneglycol dinitrates, although effective at much lower concentrations, have a number of disadvantages, including their price and the fact that they may not improve the performance of fuel in low-compression engines (Russell, 1989).

A2.1.2.2 Smoke suppressors

Organometallic compounds containing barium, calcium, manganese,
or iron have been used to reduce diesel smoke. With barium-based products, 85-95% of the metal is emitted as particulates in the exhaust (Russell, 1989); however, barium and calcium compounds are no longer used.

A2.1.2.3 Flow improvers

Cold-flow improvers increase the fluidity of the fuel by modifying the growth of wax crystals formed by higher homologues of paraffins at low temperatures. The wax content of diesel fuels is influenced by the origin of the crude oil, the distillation range of the fuel, and the source of blend components (Coley, 1989).

Table 4. Diesel fuel additives

Additive Material Concentration Effect (ppm)

Ignition improvers, Organic nitrates a Enhancement of self-ignition qualities a

cetane enhancers a

Smoke suppressors, Organic compounds of Ca, Ba, Reduction of soot; increase in metal

combustion enhancers a or (sometimes)Mg a sulfate emissions a

Detergents a Amines, imidazolines, Prevention and removal of coke deposits succinimides, etc. b on fuel injector tips, etc. a

Flow improvers Olefin-ester copolymers 50-500 Interaction with wax crystals and modification of their growth; prevention of formation of agglomerates

Cloud-point depressors Olefin-ester copolymers About 1000 Depression of cloud-point; prevention of formation of agglomerates

Wax anti-settlers Modified ethylene-vinyl 100-500 c Reduction of crystal size and rate of acetate copolymers c settling; prevention of formation of agglomerates

Anti-static agents Not reported Not reported Reduction of building up of charges of static electricity

Anti-corrosion chemicals Alkenyl succinic acids and 5-50 Prevention of corrosion or rusting of esters, dimer acids, amine storage tanks, pipelines, and metal fuel salts system components 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTable 4 (contd)

Additive Material Concentration Effect (ppm)

Antioxidants Hindered phenols or amines 25-200 Prevention of aging processes

Anti-foam agents Silicones Up to 20 Reduction of foaming tendency; reduction of risk of ground pollution from oil spi lls

Dehazers Quaternary amine salts 5-50 Reduction of formation of hazes;
acceleration of haze clearance

Biocides Imines, amines, About 200 Prevention of growth of bacteria and imidazolines, etc. fungi in storage tank bottoms

Lubricants Surface-active agents such 50-500 Compensation of lower viscosity of fuels as polyfunctional acids and in low-temperature regions
derivatives

Odour maskers Natural, identical substances, 10-100 Reduction or elimination of smell such as vanillin and terpenes c

From Coley (1989), except as noted

a From Organisation for Economic Co-operation and Development (1993)

b From Russell (1989)

c From Fabri et al. (1990) A2.1.2.4 Cloud-point depressors

Diesel fuels must be easily filterable, as wax crystals formed at low temperatures can clog fuel filters. Cloud-point depressors are therefore added (Fabri et al., 1990) which consist of substances with lower cloud-points, e.g. kerosene. Addition of 10% kerosene lowers the cloud-point of diesel fuel by about 2°C. Olefin-ester copolymers depress the cloud-point by 3-4°C but are not currently in commercial use (Coley, 1989).

A2.1.2.5 Wax anti-settling additives

These additives inhibit the tendency of wax to settle by reducing the crystal size and slowing the settling rate. A five-fold reduction in wax crystal size slows the settling rate by one-twenty-fifth. With increasing temperature, small dispersed crystals redissolve more readily than settled wax (Coley, 1989).

A2.1.2.6 Other additives

Detergents, including amines, amides, imidazolines, and succinates, are used to reduce injector nozzle fouling. Detergents such as polyalkenyl succinimides also improve fuel stability, resistance to corrosion, and combustion efficiency (Russell, 1989). Antistatic agents lower the risk of building up a charge of static electricity during pumping at high rates at bulk terminals or in large-capacity truck fuel tanks.

Other additives used are anti-oxidants (phenols, amines), anti-corrosion chemicals (alkenyl succinic acids, esters, dimer acids, amine salts), anti-foam agents (silicones), dehazers (anti-emulsion agents) (quaternary ammonium salts), biocides, lubricants for cold regions (surface-active polyfunctional acid derivatives), and odour maskers (vanillin, terpenes) (Coley, 1989; Fabri et al., 1990).

A2.1.3 Quality of diesel fuels

A2.1.3.1 Ignition performance and cetane number

The cetane number determines the ignition performance of transport fuels relative to a scale on which methyl naphthalene corresponds to a combustion rate of 0 and cetane to one of 100 (Scheepers & Bos, 1992a). The cetane number is calculated by comparing the ignition quality of a fuel with that of two reference fuel blends of known cetane numbers under standard operating conditions (American Society for Testing and Materials method D 613 CFR). A high cetane number improves cold starting and engine durability and reduces noise, fuel consumption, smoke emissions during warm-up, and exhaust emissions (Russell, 1989). In Europe (countries of the EU and EFTA),

the minimal cetane number must be in the range 45-49, depending on the climatic conditions (European Committee for Standardization, 1993); cetane numbers are usually 49-53 (CONCAWE, 1987). In the United States, the cetane number must be at least 30 for diesel fuel No. 4 and 40 for diesel fuels No. 1 and 2 (American Society for Testing and Materials, 1988, 1992).

A2.1.3.2 Density

The density of diesel fuel influences engine performance: higher density leads to enrichment of the fuel:air mixture, which results in greater engine power output. Enrichment may, however, increase the particulate content of exhaust gas emissions (Fabri et al., 1990) (see section B3.1.2.2). A density range is specified in fuel standards in some countries (American Society for Testing and Materials, 1992; European Committee for Standardization, 1993).

A2.1.3.3 Sulfur content

Gas and particle emissions in diesel engine exhaust are 

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A2.1.3.4 Viscosity

Too low a viscosity can lead to wear in the injection pump; too high a viscosity impairs fuel injection and mixture formation (Fabri et al., 1990). In Europe (countries of the EU and EFTA), the viscosity of commercial diesel fuels at a temperature of 40°C must be

1.5-4.5 mm 2 /s (European Committee for Standardization, 1993). In the United States, the permitted ranges of viscosity at 40°C are

1.3-2.4 mm 2 /s for diesel fuel No. 1, 1.9-4.1 mm 2 /s for diesel fuel
No. 2, and 5.5-45.0 mm 2 /s for diesel fuel No. 4 (American Society for Testing and Materials, 1988, 1992).

A2.1.3.5 Cold-flow properties

Cloud-point and cold filter plugging point characterize the behaviour of diesel fuels at low temperatures (Fabri et al., 1990). These points are lowered by the addition of cloud-point depressors and by special blending techniques, e.g. increasing the kerosene content of the fuel (see section A2.1.2).

Changes in diesel fuel quality have been assessed. Wade & Jones (1984) reported a deterioration in fuel quality with decreasing cetane number and found that a 90% increase in boiling-point led to greater emissions of particulates, nitrogen oxides and PAHs. A decline in diesel fuel quality on the European market was predicted, as the rising demand would lead to greater use of fuels from catalytic or thermal cracking processes (CONCAWE, 1987) (see section A3.2.1.1). A study by the Organisation for Economic Co-operation and Development (1993) indicated an improvement in the quality of diesel fuel in the United States due to a decline in aromaticity.

A2.2 Physical and chemical properties

Diesel fuel is a brown, slightly viscous, flammable liquid at room temperature (Sandmeyer, 1981). It generally has a kerosene-like odour (Agency for Toxic Substances and Disease Registry, 1995). Its physical and chemical properties are listed in Table 5.

The water solubility of diesel fuels varies. The aqueous solubility of crude and fuel oils in the environment is clearly dependent on the salinity of the water and the age of the oil slick (see section A4.1.1) and is of the same order of magnitude as the solubility of fuel oils: at room temperature, 0.37-0.53 mg/litre in sea water (Boehm & Quinn, 1974) and 0.7-11 mg/litre in tap water (Lysyj & Russell, 1974). The solubility of a fuel slick decreases with its age as the concentration of long-chain hydrocarbons increases; the solubility of fresh crude oil is 29.3-32.3 mg/litre, whereas that of weathered crude oil is 0.06-23.2 mg/litre at 25°C (Mackay & Shiu, 1976).

The physicochemical properties of the toxicologically relevant benzene, toluene, ethyl benzene, xylene and PAH components are given
in Table 6.

A2.3 Analytical methods

Exhaustive identification and quantification of the individual constituents of commercial diesel fuel (see section A2.1) is almost impossible owing to their number and complexity. In most environmental assessments, therefore, the mixture is analysed as total petroleum hydrocarbon (Block et al., 1991). All such methods involve preliminary solvent extraction of the matrix, with e.g. trichlorotrifluoroethane. This step may also extract naturally occurring hydrocarbons, which interfere with the analysis; however, some of these compounds are polar and can be removed on silica gel. Three analytical methods are
available:

Table 5. Physicochemical properties of diesel fuels

Property Diesel fuel Diesel fuel Diesel fuel Diesel fuel (general) No. 1 No. 2 No. 4

Melting-point (°C) - 34 a 18 a - 29-9 a

Boiling range (°C) 160-190 b 145-300 (wide-cut 282-338 a 170-420 d

143-384 e aviation, 45-280) c 101- > 588 a

193-293 a

Flash-point (°C) > 56 b > 21 - < 55 52 (closed cup) a > 56 d

58-66 e (wide-cut aviation, > 54 (closed cup) a (Pensky-Martens) < 21) c 38 (closed cup) a

Autoignition temperature (°C) 177-329 a 254-285 a 263 a

Density (g/cm 3 ) 0.81-0.90 (15°C) b 0.805 (wide-cut 0.87-0.95 0.87-0.92 (15°C) d

0.82-0.84 (15°C) e aviation, 0.75-0.801) (20°C) 0.81-0.94 (15°C)

(15°C) c 0.81-0.94 1 (20°C) (15°C)

Kinematic viscosity (mm 2 /s) 2-7.4 (40°C) b 1.5-2.5 (wide-cut 2-7.4 (40°C) d

2.20-3.25 (40°C) e aviation, about 1.1 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm(20°C) c

Vapour pressure (kPa) About 40 (40°C) b About 10 (wide-cut 2.83-35.2 2.83-35.2 (21°C) a

0.04 f aviation, 140-210) (21°C) a (Reid, 37.8°C) c
2.83-35.2 (21°C) a

Table 5 (contd)

Prpoerty Diesel fuel Diesel fuel Diesel fuel Diesel fuel (general) No. 1 No. 2 No. 4

Water solubility (mg/litre) 1 f About 5 (20°C) a About 5 (20°C) a About 5 (20°C) a

0.2 f

Henry's law constant 4.3 × 10 3f 6.03-7.5 × 10 5a 6.03-7.5 × 10 5a 6.03-7.5 × 10 5a

(Pa × m 3 /mol) (20°C)

n-Octanol-water partition 3.3-7.06 a 3.3-7.06 a 3.3-7.06 a coefficient (log K

ow )

Soil sorption coefficient 3.04 f 3.0-6.7 a 3.0-6.7 a 3.0-6.7 a (log K
oc )

Diffusion coefficient in air 4.63 × 10-2 f

(cm 2 /s)

Odour threshold (ppm) 0.7 a 0.5 g

a From Agency for Toxic Substances and Disease Registry (1995)

b From CONCAWE (in press); automotive gas oil

c From CONCAWE (1985, 1995); kerosenes

d From CONCAWE (in press); distillate marine diesel

e From German Scientific Association for Petroleum, Natural Gas, and Coal (1991); average of three samples

f From Custance et al. (1993); water solubility measured, other data from literature; no further details

g From Fraser Williams (Scientific Systems) Ltd (1985)

-- Gravimetric detection: determination of the weight of residue remaining after solvent evaporation. Although this method is useful for measuring gross contamination, it cannot be used in trace analysis.

-- Infrared detection: the absorbance of petroleum hydrocarbons is detected in a solvent matrix at the maximal value, near

2930 cm -1 . This carbon-hydrogen bond stretching absorbance is directly related to the hydrocarbon concentration in the extract. The results are dependent on the hydrocarbon standard used for quantification.

-- Gas chromatographic detection: capillary gas chromatography combined with flame ionization or mass spectrometric detection.

The gravimetric and infrared techniques are fast, relatively simple and widely accepted by regulatory authorities; however, they do not provide sufficient qualitative and quantitative information on composition. Gas chromatography is therefore the standard procedure used to identify and quantify fuel constituents in environmental samples. Different distribution and degradation processes in environmental compartments (see section A4) mean that the composition of petroleum hydrocarbons in air, water, soil, and biota may differ considerably from that of the original fuel. Newton et al. (1991) developed an analytical method for identifying and quantifying traces of diesel contamination in tinned fish products on the basis of the

n- alkane pattern.

Because of the suspected toxicity of aromatic components in diesel fuel, a detailed analysis is often necessary. Two groups of compounds are detected:

-- volatile aromatic compounds: Analysis of these minor components of diesel fuel requires special enrichment techniques, such as purge-trap gas chromatography, and methods of detection including flame ionization and mass spectrometry.

-- PAHs: Gas chromatography with flame ionization or mass spectrometric detection, or liquid chromatography, is used after solvent extraction. As PAHs are poorly resolved from the diesel fuel matrix, gas chromatography-mass spectrometry is necessary in most cases to quantify the components. The target analytes of commonly used analytical methods do not, however, include C 3 and C 4 alkyl-substituted benzenes or most alkyl-substituted

PAHs in diesel fuels.

Table 6. Physicochemical properties of aromatic components of diesel fuel

Aromatic compound Water solubility Henry's law constant Diffusion coefficient (cm 2 /s) Octanol-water partition (mg/litre) (Pa × m 3 /mol) In air In water coefficient (log P

ow )

Benzene 1791 550 0.087 9.8 × 10 -6 1.99

Toluene 534.8 602 0.083 8.6 × 10 -6 2.52 Ethylbenzene 161.0 855 0.076 7.8 × 10 -6 2.94 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmXylene 156.0 778 0.076 1 × 10 -5 2.94 Polycyclic aromatic 0.067 0.007 0.067 2.12 × 10 -6 5.30 hydrocarbons

Adapted from Custance et al. (1993)
A2.4 Conversion factors

As diesel fuel vapour is a complex mixture of gases, it is impossible to give a conversion factor for converting parts per million in the gaseous phase to SI units.

A3. SOURCES OF HUMAN AND ENVIRONMENTAL EXPOSURE

A3.1 Natural occurrence

Diesel fuels are derived from crude oil, which can be considered a 'natural product'. Nevertheless, human and environmental exposure results almost exclusively from anthropogenic activities.

A3.2 Anthropogenic sources

A3.2.1 Production and use

A3.2.1.1 Production process

Diesel fuel is produced during the refining of crude oils but is generally blended to meet the specifications for technical performance. The blending components may be produced by atmospheric distillation of crude oil (straight-run atmospheric gas oil), vacuum distillation of atmospheric residue (vacuum gas oil), thermal cracking (thermally cracked gas oil), or catalytic cracking processes (e.g. light catalytically cracked gas oil; cycle oil). The main components are straight-run gas oils: 80% of European automotive diesel fuel is made up of these components (Booth & Reglitzky, 1991). Secondary processing of heavier fractions is increasingly necessary in order to meet product demand (CONCAWE, in press).

Diesel fuel No. 1 is manufactured by a straight-run distillate process (Agency for Toxic Substances and Disease Registry, 1995); diesel fuel No. 2 is generally made by mixing straight-run and catalytically cracked distillates; and diesel fuel No. 4 is produced by adding blending stocks to distillation residues in order to meet viscosity specifications (International Agency for Research on Cancer, 1989a). Further variations are introduced by formulation with additives to improve fuel properties (see section A2.1.2).

A3.2.1.2 Use

Diesel fuel is widely used as a transport fuel for light- and heavy-duty vehicles; the Organisation for Economic Co-operation and Development (1993) classifies vehicles weighing < 3.5 t as light-duty and those weighing > 3.5 or, occasionally, > 5 t as heavy-duty. Diesel fuel No. 1 is suitable for engines that undergo frequent changes in speed and load (Agency for Toxic Substances and Disease

Registry, 1995). Heavier grades (diesel fuels No. 2 and 4) are used for trucks, railroad and marine diesel engines, and stationary engines in continuous high-load service (Sandmeyer, 1981; International Agency for Research on Cancer, 1989a). Diesel fuels are also used in stationary gas turbines, e.g. to generate electric power during peak-load periods. Residual fuel oils, such as diesel fuel No. 4, are used to generate steam in electric power plants (International Agency for Research on Cancer, 1989a), in commercial and industrial burner installations without preheating facilities, in plants and factories for space and water heating, for pipeline pumping, and in gas compression; they are also sprayed on unmade roads to compact the surface and are used in the manufacture of asphalt cement (Agency for Toxic Substances and Disease Registry, 1995).

A3.2.1.3 Production and consumption levels

The demand for diesel fuel has increased worldwide over time. In 1990, world demand was about 1100 kt/day (Booth & Reglitzky, 1991). The production and consumption of diesel fuel in different regions
over time are shown in Table 7.

Diesel-fuelled passenger cars are relatively common in western Europe. The percentages of passenger cars with diesel engines in various European countries over time are shown in Table 8. In many European countries, taxi-cabs are equipped almost exclusively with diesel engines. In 1985, diesel-fuelled heavy-duty trucks comprised about 86% of the fleet in Norway, 89% in Denmark, 95% in Sweden, and 100% in the former Federal Republic of Germany. In some countries of Europe, diesel fuel consumption has increased steadily over the last decade, and consumption for heavy-duty vehicles is predicted to more than double between 1980 and 2005 (Organisation for Economic Co-operation and Development, 1993). In western Germany, diesel fuel consumption was almost 60% higher in 1990 (about 5100 kt) than in 1984 (about 3200 kt) (Federal Ministry for Transport, 1992).

Diesel-fuelled passenger cars are less common in North America. In the United States in 1986, about 1.6% of passenger cars were diesel-fuelled, and the tendency was predicted to decrease slightly (US Department of Energy, 1988). In contrast, 82% of heavy-duty trucks were diesel-fuelled in 1985 (Organisation for Economic Co-operation and Development, 1993), and 59300 kt of diesel fuel were consumed by highway traffic in 1986, representing 14.8% of all the highway fuel used and about one-half of United States diesel fuel consumption. On-road use of diesel fuel was predicted to increase to 15.5% of all 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmhighway fuel (66600 kt) in 1995. A difference in fuel use patterns is seen between light- and heavy-duty vehicles. In 1986, about 4000 kt were used by passenger cars and light trucks, with an estimate of about 2300 kt for 1995, whereas in 1986 about 55300 kt were used by heavy-duty trucks, with an estimate of 64300 kt for 1995 (US

Department of Energy, 1988). In Canada, only about 1% of passenger cars are equipped with diesel engines. In 1992, 14126 kt of diesel fuel were produced; 13508 kt were sold for domestic purposes (heating) and about 10849 kt for on-road use (Thomas, personal communication,
1994).

The percentage of diesel-fuelled vehicles is increasing in Japan (Table 9).

Table 7. Production and use of diesel fuel (including gas oils in Europe) in different regions, and development over time

Region Production/ 1975 (kt) 1980 (kt) 1985 (kt)

Use

United States Production 134 967 138 323 135 181

Use 133 300 136 161 130 297

Canada Production 19 187

Use 19 130

OECD (all 24 Production 370 240 408 848 372 728 countries) Use 380 181 403 642 386 081

OECD (Europe) Production 173 474

Use 190 960

European Union Production 152 091

Use 163 132

Australia, Japan, Production 44 886

New Zealand Use 45 694

Adapted from International Agency for Research on Cancer (1989a); OECD, Organisation for Economic Co-operation and Development

A3.2.2 Emissions during production and use

A3.2.2.1 Air

No data are available on emissions during the production and use of diesel fuel. Release to the atmosphere during production in the refining process is unlikely, as closed systems are used, but volatilization may occur during storage and transport. Diesel fuel may

be released into the atmosphere as a result of spills during storage and transport and at filling stations during bulk storage and vehicle tank filling. The low-relative-molecular-mass constituents (short-chain alkanes, benzene, toluene, ethyl benzene, and xylene compounds) are the most likely to evaporate under environmental
conditions.

Table 8. Percentage of diesel passenger cars in western Europe, and development over time

Country Diesel passenger cars (%)

1984 1988 1991 1992 a

France 15.8 21.1 25.5 38 Germany (without former 16.9 15.1 15.3 14.9 German Democratic Republic) (1993) b Germany (without former About 8 13.6 13 German Democratic Republic) c Italy 18.7 11.9 7.4 NR Spain (without station wagons) 15.3 10.5 14.6 NR United Kingdom 1.1 4.5 11.3 12.5 United Kingdom d NR 5 9 19 (1993)

Adapted from American Automobile Manufacturers Association (1993), unless otherwise stated; NR, not reported

a From Menne et al. (1994)

b With the former German Democratic Republic

c From German Institute for Scientific Research (1993)

d From Quality of Urban Air Review Group (1993)

Table 9. Percentage of diesel-fuelled vehicles in Japan and development over time

Year Passenger cars (%) Trucks (%) Buses (%)

1984 3.8 23.7 49.5

1988 5.8 28.5 63.2

1991 8.3 40.7 81.7 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmAdapted from American Automobile Manufacturers Association (1993)

A3.2.2.2 Water

No data are available on effluents and emissions during diesel fuel production. Diesel fuel oils may be released to surface waters as a result of leakage from storage tanks or tankers (see section A3.2.3). In the United States, the total volumes of spills of diesel fuel oils in 1991 were (Agency for Toxic Substances and Disease Registry, 1993): diesel fuel No. 1, 10.6 t (20 notations); diesel fuel No. 2, 8.9 t (28 notations); and diesel fuel No. 4, 39.3 t (35 notations).

Groundwater can be contaminated with diesel fuel constituents by leakage from underground bulk storage tanks, but quantitative data are lacking. In the Province of New Brunswick, Canada (Thomas, personal communication, 1993), of a total of 671 gasoline, diesel, and fuel oil tanks examined in 1987, 11% leaked diesel or fuel oil; in 1989, 6% of 1085 tanks had leaks; and in 1991, 13% of 539 tanks leaked fuel. No data were available about the amounts leaked, and diesel contamination was determined on the basis of a cluster of 'middle-range peaks' detected by gas chromatography.

On the basis of chronic petroleum inputs to sediments of Narragansett Bay and Rhode Island Sound, United States, Van Vleet & Quinn (1978) estimated that about 200 kt of petroleum hydrocarbons may be released to surface waters worldwide from municipal treatment plants. The precise source of the inputs cannot be verified in most cases, but they are due, for example, to accidental discharge to sewer systems, disposal of used crankcase and lubricating oils, oil washed from roads, and atmospheric deposition of hydrocarbons.

No data were available from other countries.

A3.2.2.3 Soil

Diesel fuels may be released to soil as a result of accidental spills (see section A3.2.3) and leakage from storage tanks or pipelines. Diesel-contaminated soil is a major problem in railroad yards, where diesel fuel is used in locomotives and as a solvent to clean moving metal parts (the remaining paraffins give a waxy anti-corrosive coat). Spillage during refuelling, engine maintenance, and steam cleaning, leakage from fuel storage tanks, and absorption of diesel fuel on sand used for traction are other possible sources of soil contamination (Dineen, 1991); however, no quantitative data are
available.

A3.2.3 Accidental releases to the environment

Data on the accidental release of diesel fuels are summarized in Table 10. The effects on the environment depend on both the amount and
environmental conditions.

Table 10. Diesel fuel spills and their effects on the environment (see also section A9.2)

Year Place Amount (t); type Effects Referenc e of diesel fuel a

Aquatic environment 1972 North of Puget Sound, > 600; diesel fuel Substantial mortality of some intertidal taxa; Woodin e t al. (1972) Washington State, USA No. 2 larval recruitment within six months

1973 East Lamma Channel 2000-3000; Almost total kill of meiofauna within four days; Wormald (1976); near Hong Kong diesel fuel No. 4 heavy mortality among bivalves and molluscs, Stirling (1977) Nerita albicilla and Monodonta labio, and in
marine fish farms; almost no effect on Clypeomorus humilis or Planaxis sulcatus

1978 Svea, Mijenfjord, Norway 111 Most fuel trapped in ice; transport out of fjord Carstens & during break-up; heavy mortality among shoreline Sendstad (1979) invertebrates in some regions of fjord

1983 Yaquina Bay, Oregon, About 240 (with Decline in population of Rhepoxynius abronius Kemp et al. (1986) USA Bunker C oil) by 75%; influence of spill not clear

1984 Queen Charlotte 130 (plus 30 t Low fuel levels in water during flood tide, high McLaren (1985) Islands, Canada gasoline) levels during ebb as diesel was retained in sediment; high mortality among barnacles

Table 10 (contd)

Year Place Amount (t); type Effects Referenc e of diesel fuel a

1987 Macquarie Island, 230; diesel fuel High mortality among one species of holothuroid, Pople et al. (1990) Australia No. 4 one of isopod, one of limpet, and one of chiton; decreased populations of crab, two species of starfish, and gastropod; small number of dead algae; slow recovery of organisms

1989 Arthur Harbor, 510; diesel fuel High mortality among limpets; partial recovery one Kennicut t et al. Antarctica No. 1 year after spill; small effect on macroalgae (1991a,b ,1992a,b) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTerrestrial environment 1983 San Bernadino County, 3 Hydrocarbon contamination up to 1500 mg/kg; Frankenb erger et al. CA, USA plume migrating towards surface water; in-situ (1989) bioremediation (see also section A3.2.4)

NR Califonia, USA 193; diesel fuel Contamination at 6-34 m below surface; Peters e t al. (1992) No. 2 hydrocarbon levels at 18.3, about 1500 mg/kg; remediation by flooding with surfactants (see also section A3.2.4)

a When available A4. ENVIRONMENTAL TRANSPORT, DISTRIBUTION, AND TRANSFORMATION

Few data are available on the environmental fate of diesel fuels, but there are adequate data on the environmental behaviour of individual hydrocarbon components. The transport, distribution, and transformation of crude oils and some fuel oils (e.g. heating oils No. 1 and 2) have been studied, and the mechanisms of distribution and transformation are considered to be comparable to those of diesel

fuel.

The calculated half-lives for the toxicologically relevant PAH components are listed in Table 11.

A4.1 Transport and distribution between media

Diesel fuels are released into the environment mainly during storage, transport, and use (see section A3.2.3). The hydrosphere and geosphere are the affected compartments.

A4.1.1 Evaporation from and dissolution in the aqueous phase

Oils and fuels spilled on water spread out almost immediately; polar and low-relative-molecular-mass components dissolve and leach out of the slick; and volatile components evaporate from the water surface. With the start of microbial degradation the pattern of substances changes, as was observed, for example, after a major oil spill off the French coast (Atlas et al., 1981). The sum of these processes is known as chemical and biological 'weathering'. At the same time, the oil is emulsified to oil-in-water and water-in-oil ('mousse') emulsions (Atlas & Bartha, 1973). In laboratory experiments with several diesel fuels, however, stable mousses were not formed at any temperature (US National Research Council, 1985). Experiments with South Louisiana crude oil showed that the evaporation rate was more than twice the dissolution rate (Harrison et al., 1975). The loss by evaporation of oil spread on water, which in oil spills amounts to about 25% of the total released mass, is generally dependent on the following factors (US National Research Council, 1985):

-- the exposed area, which increases with time as the film spreads;

-- the oil-phase vapour pressures, which decrease with time as the low-relative-molecular-mass components evaporate. Components with vapour pressures higher than that of n- octane evaporate rapidly from the surface of spilled fuel oil, while those with vapour pressures lower than that of n- octadecane persist and
form a viscous residue that retards the volatilization of other constituents (Regnier & Scott, 1975);

-- the oil-air mass transfer coefficient, which depends on wind speed and hydrocarbon diffusivity (for the diffusion coefficient in air, see section A2.2, Table 5);

-- diffusion barriers, such as emulsions, or formation of a 'skin' on the surface. Many surface waters have a thin layer of organic material of natural origin on the surface, and these films may reduce spread and volatilization, altering the persistence and concentration of the oil in water (Edgerton et al., 1987).

Table 11. Calculated half-lives of polycyclic aromatic hydrocarbon components in environmental compartments

Component Air Water Soil Sediment

Naphthalene 1 day 1 week 2 months 8 months Acenaphthalene 2 days 3 weeks 8 months 2 years Fluorene 2 days 3 weeks 8 months 2 years Phenanthrene 2 days 3 weeks 8 months 2 years Anthracene 2 days 3 weeks 8 months 2 years Pyrene 1 week 2 months 2 years 6 years Fluoranthene 1 week 2 months 2 years 6 years Chrysene 1 week 2 months 2 years 6 years Benz[a]anthracene 1 week 2 months 2 years 6 years Benzo[a]fluoranthene 1 week 2 months 2 years 6 years Benzo[a]pyrene 1 week 2 months 2 years 6 years Perylene 1 week 2 months 2 years 6 years Dibenz[a,h]anthracene 1 week 2 months 2 years 6 years

Adapted from Mackay et al. (1992)

On the basis of the constant of Henry's law (see section A2.2, Table 5) and the classification of Thomas (1990), diesel fuels can be considered to volatilize significantly from the aqueous phase. The constant may change during volatilization, as the composition of the mixture changes with time. Lockhart et al. (1987) observed a clear decrease in the hydrocarbon composition of the 50% water-soluble 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmfraction of diesel fuel in open-aerated test cylinders. Within 48 h, the concentration of hydrocarbons with a very low boiling-point (< 115°C) decreased from 0.73 to 0.04 mg/litre and that of hydrocarbons with a low boiling-point (115-270°C) from 3.2 to 0.17 mg/litre. Under sealed or open non-aerated conditions, there were only minor changes. Regnier & Scott (1975) showed that n- decane volatilizes from diesel fuel with a half-life of 1.65 h at 30°C,
whereas the half-life was 9.67 h at 5°C in a darkened chamber with a constant wind speed of 21 km/h. Edgerton et al. (1987) reported that diesel fuel spilled on ice had lost only 2-4% of its lightest fraction after 10 days.

Dissolution of spilled oils and fuels in water is less important from the viewpoint of mass loss than the ecotoxic effects of the mixture. Lysyj & Russell (1974) conducted long-term experiments (42 days) on the dissolution of different fuel oils and gasoline in water. After a period of stable concentration, during which physical dissolution was probably the main mechanism for oil-to-water transfer, the dissolution of organic compounds accelerated, probably because of chemical modification by oxidation or microbial action of the water-insoluble petroleum fraction.

Under equilibrium mass-transfer conditions, the levels of PAHs (naphthalene and its methyl derivatives, acenaphthene, fluorene, phenanthrene, anthracene, and fluoranthene) in the water phase in
contact with commercial automobile diesel fuel were about four to six orders of magnitude lower than those in the diesel fuel itself, i.e. in the low microgram per litre range. The concentrations of these constituents were likely to be smaller under non-equilibrium conditions but might be higher in the presence of surfactants, emulsifiers, or co-solvents (Lee et al., 1992). When No. 2 fuel oil and kerosene were placed in contact with drinking-water for 17 h, although each fuel contained about 50% by weight of aliphatic hydrocarbons, the water-soluble fractions contained primarily aromatic compounds: total, > 93% by weight; proportion of benzene and substituted derivatives, 19.4% by weight in No. 2 fuel oil and 53.2% by weight in kerosene (Coleman et al., 1984). The chemical composition of the water-soluble fractions of No. 2 fuel oil is compared with that
of the whole oil in Table 3.

During chemical weathering, other changes in composition (e.g.
autoxidation of n- alkanes) may occur that are not necessarily associated with a net mass loss (Davis & Gibbs, 1975).

A4.1.2 Transport in and adsorption onto soil and sediment

A4.1.2.1 Soil

The movement of diesel fuel through soil is directly correlated with its kinematic viscosity (see section A2.2, Table 5). As the viscosity is about one-half that of water and about one-fourth to one-fifth that of gasoline, the rate of percolation of water into soil is about twice and that of gasoline about four to five times that of diesel fuel (Stone, 1991). The retardation coefficient (Rd) is an indicator of the migration of substances into groundwater. In the aqueous phase, the maximal rate of movement of dissolved compounds is that of groundwater, which is represented by an Rd of 1. Stone (1991) calculated the Rd from the adsorption isotherms of individual diesel constituents, and derived the following classification:

-- low mobility (Rd > 100): fluorene, phenanthrene, pyrene, benzanthracene, benzo[ a ]pyrene, fluoranthene;

-- medium mobility (100 > Rd > 10): naphthalene, dimethylbenzenes, ethylbenzene, toluene;

-- high mobility (Rd < 10): benzene, quinoline, cresols, phenol.

In effect, soil contaminated with diesel fuel acts like a chromatographic column, separating individual constituents by their adsorption. Experimental data on the adsorption of diesel fuels on soils and their transport are not available; however, the movement of a synthetic kerosene through a clay-sand soil was dependent on the moisture content. With increasing moisture content, the penetration rate of nonvolatile components through the soil increased and the adsorption of more volatile components decreased. In contrast, upward mobility of the individual constituents (both volatile and nonvolatile) of the fuel decreased with increasing moisture content. In fully water-saturated soil, upward transport of kerosene was completely inhibited (Acher et al., 1989). Desorption of a simulated kerosene applied to three types of soil with the same moisture content but different contents of organic matter was found to be complete after 30 days under environmental conditions. The slowest desorption was from the soil with the highest level of organic matter (Yaron et al., 1989). Transport of kerosene through three types of sand was influenced mainly by volatilization of C 9 -C 13 hydrocarbon
constituents; the increase in the viscosity of the kerosene residue was followed by a decrease in the infiltration rate (Galin et al., 1990).

A4.1.2.2 Sediment

In marine environments, oils and fuels are generally delivered to bottom sediments on settling particles. In areas with low concentrations of suspended material (i.e. terrigenous minerals, plankton, detrital particles, resuspended bottom sediments), the rates of hydrocarbon sedimentation would be low (US National Research Council, 1985).

No experimental data are available on diesel fuels in sediments. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmIn laboratory experiments with fuel oil, binding of the hydrocarbon components to clay minerals of marine sediments was shown to be due to physical adsorption of the van der Waals type. The compounds are associated firmly because, once they are incorporated into sediments, desorption to the overlying water is slow. Saturated hydrocarbon constituents of fuel oils were transported slowly to the sediment (Meyers & Quinn, 1973). In studies of a laboratory marine ecosystem to which No. 2 fuel oil was added for 24 weeks, the concentrations in the sediment remained low until 135 days after the start of the experiment. Then, a steep increase was seen, leading to a level of 9% by weight of the total fuel added (i.e. 12% by weight of the total saturated hydrocarbons). The highest concentrations of saturated

hydrocarbons were found in the surface flocculent layer; 2-3 cm below the sediment surface, the levels were below the detection limit. The smaller particles (< 45 μm) of water-suspended material seemed to adsorb about twice as much fuel oil as larger particles (> 45 μm) (Wade & Quinn, 1980). In a comparable study in a model marine ecosystem with No. 2 fuel oil, the particulate matter contained 40-50% of the total amount of aliphatic hydrocarbons added and only 3-21% of the aromatic fraction, indicating that most of the aromatic hydrocarbons were dissolved in the water phase (see section A4.1.1) (Gearing et al., 1980). Although biodegradation may remove many of the soluble aromatic hydrocarbons (see section A4.2.2), 10-20% by weight of the fuel oil, consisting primarily of branched alkanes, cycloalkanes, and aromatics, may remain in the upper 2 cm of sediment for over a year (Gearing et al., 1980; Oviatt et al., 1982).

A4.2 Transformation and removal

A4.2.1 Photooxidation

No data were available on the photooxidation of diesel fuel in air or water. In the atmosphere, all evaporated oil components are degraded photochemically by hydroxy radicals and other species within hours or days to carbon monoxide, carbon dioxide, and oxygenated compounds (US National Research Council, 1985; IPCS, 1986, 1993). Under laboratory conditions, No. 2 fuel oil was rapidly photo-oxidized in water. Two photooxidant pools were identified: hydrogen peroxide and a mixture of indane and tetralin hydrogen peroxides, the first being the faster acting (Herbes & Whitley, 1983). Peroxides are formed by two mechanisms: the first is the reaction of free radicals with triplet oxygen to produce peroxy radicals and hydroperoxides, which continue the chain reaction; the second is addition of excited singlet oxygen to reactive acceptors such as olefins. The photodegradation products of fuel oils are phenols, naphthols, and carboxylic acids (Larson et al., 1977, 1979).

A4.2.2 Biodegradation

A4.2.2.1 Microbial degradation

The individual constituents of spilled oil are inherently degradable to varying degrees and at different rates. The n- alkane,

n- alkylaromatic, and simple aromatic molecules in the C 10 -C 22 and the most readily degradable. Smaller molecules are generally range are the least toxic rapidly metabolized, although they tend to be more toxic. Long-chain n- alkanes are more slowly degraded because of their hydrophobicity and also because they are viscous or solid at ambient temperatures. Branched alkanes and cycloalkanes are relatively resistant to attack. PAHs are the compounds most recalcitrant to microbial degradation (Morgan & Watkinson, 1988).

A study of the breakdown of heating oils and diesel fuels in a sub-soil demonstrated breakdown rates of 0.014-0.022 g/kg per day, which were slower than the rates of topsoil degradation (Flowers et al., 1984). The mineralization rates for heavy oils, sludges, and crude oils were 0.02-0.6 gram of hydrocarbon per kilogram of soil per day. Under Arctic conditions, the rates were considerably slower.

The relative contribution of bacteria and fungi to hydrocarbon degradation is unclear. At acid pH, fungi are clearly more important, but closer to neutrality hydrocarbon contamination tends to enhance bacterial growth.

The overall rates of hydrocarbon degradation are limited by:

-- temperature (optimal temperature for biodegradation, 30-40°C);

-- water content (in soils, water contents of 50-80% capacity are optimal for microbial activity);

-- oxygen (the metabolism of both aliphatic and aromatic hydro-carbons normally requires oxygen);

-- pH (hydrocarbons are mineralized most rapidly at 6.5-8.0);

-- inorganic nutrients (as petroleum contaminants are extremely deficient in nitrogen and phosphorus, a large quantity of carbon sources tends to result in rapid depletion of the available pools of major nutrients, i.e. enhancement of degradation in fertilized soils);

-- microbial metabolic versatility (topsoils generally contain many hydrogen-degrading bacteria, typically 105 - 106 per gram, but the content of subsoils is unclear) (Morgan & Watkinson, 1988).

A remedial treatment consisting of liming, fertilization, and tilling was evaluated on a laboratory scale for its effectiveness in cleaning up a sand, a loam, and a clay-loam soil contaminated at 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm50-135 mg/g of soil by diesel and other oils. The disappearance of hydrocarbons was maximal at 27°C (Song et al., 1990).

In sandy soils, 50-85% of the hydrocarbons were degraded within a few weeks, depending on the experimental conditions. Degradation efficiency increased with additional soil inoculation (Grundmann & Rehm, 1991).

Frehland et al. (1992) examined the substrate specificity and growth conditions of nine bacterial strains on samples of
diesel-contaminated soil under different conditions. The biodegradative properties of Pseudomonas spp. were superior (59, 60, and 80% in three samples) than those of Micrococcus, Bacillus, Acinetobacter, and other species tested.

A4.2.2.2 Phytoplankton and marine algae

Unicellular algae can take up and metabolize both aliphatic and aromatic hydrocarbons, but the extent to which this occurs in nature is poorly understood. The ability to metabolize naphthalene is widespread (US National Research Council, 1985).

A4.2.2.3 Invertebrates and vertebrates

Unlike microorganisms, which use petroleum as a source of carbon, animals generally oxidize and conjugate products to render them more soluble and therefore easier to excrete. All animal groups tested can take up petroleum hydrocarbons, either through the body surface, across respiratory surfaces, or via the gut, the route varying with the organism and its feeding habits. In copepods (Calanus helgolandicus), dietary uptake was more important than that from water (Corner et al., 1976), but aromatic hydrocarbons accumulated by the polychaete Neanthes arenaceodentata were derived from water and not from sediment (Rossi, 1977). Hydrocarbons are stored in lipid-rich tissues. The principal route of metabolism of hydrocarbons by metazoans is conversion of lipophilic compounds by cytochrome P450-dependent monooxygenases or mixed-function oxidases to derivatives that are more water-soluble. Mixed-function oxygenase activity has been found in all fish species studied. Lee (1981) reported 18 marine invertebrate species belonging to four phyla (Annelida, Arthropoda, Echinodermata, and Mollusca) that are known to contain mixed-function oxidase activity in the hepatopancreas, digestive gland, and other tissues. Sea birds and some other waterfowl also have cytochrome P450 systems, but few estimates have been made of their capacity for hydrocarbon metabolism. Some crude oils, PAHs, and refined petroleum products are known to induce cytochrome P450 enzymes and to increase the levels of hydrocarbon metabolism in marine and freshwater fish, including embryonic and larval forms and polychaetes (US National Research Council, 1985). Induction is usually described in terms of the rate of metabolism of benzo[a]pyrene in vitro.

A4.2.3 Bioaccumulation

Few data are available on the bioaccumulation of diesel fuel itself, although there is adequate evidence from studies on other oils that aquatic organisms bioconcentrate hydrocarbons. The

n- octanol-water partition coefficient (log Pow) for diesel fuel is 3.3-7.1 (see section A2.2, Table 5). Although a higher bioaccumulation potential might be expected from these log P

ow values, the measured
bioconcentration factors are lower than predicted because many compounds of low relative molecular mass, such as substituted benzenes, are readily metabolized, and the actual bioaccumulation of compounds of higher relative molecular mass is limited by their low water solubility and large molecular size.

In a continuous-flow system, diesel fuel was taken up by Mytilus edulis mussels exposed for 41 days to a concentration of 200-400 μg/litre. The hydrocarbon levels in tissue were about 1000 times higher after exposure than before. Depuration was rapid during the first 15-20 days and then decreased rapidly to a minimum (Fossato & Canzonier, 1976).

The accumulation of No. 2 fuel oil has been studied in a number of organisms (Lee, 1977). When No. 2 fuel oil (38% aromatics) was added to a commercial shrimp pond, the concentration of naphthalenes in shrimps, clams, and oysters exceeded the level in ambient water after 38 days. Depuration in clean water was complete after 10 days for shrimps and after 47-96 days for oysters (Cox et al., 1975). In a flow-through system to study accumulation of a No. 2 fuel oil-water dispersion, aromatic petroleum hydrocarbons were accumulated at a rate of 89.6 mg/kg tissue by Rangia cuneata (clams) and 311.7 mg/kg by Crassostrea virginica (oysters) during 8 h of exposure. The bioaccumulation factor for methyl naphthalenes in clams was 2.3-26.7 and increased with increasing alkylation of the naphthalene ring. Depuration in clean water was complete after 28 days (Neff et al., 1976a). The fuel components of a small spill of No. 2 fuel oil had the following biological half-lives in mussels (Mytilus edulis): aliphatic hydrocarbons (C 16 and C 23 ), 0.2 and 0.8 days; alkyl naphthalenes

(C 2 and C 3 ), 0.9 and 1.5 days; phenanthrene, 2.1 days (Farrington
et al., 1982). Lobsters exposed to a simulated small spill of No. 2. fuel oil bioconcentrated PAH constituents in the hepato-pancreas and muscle within three to four days after exposure. The levels remained elevated for up to 10-11 days, and depuration was complete after 20-21 days.

The polychaetous annelid Neanthes arenaceodentata rapidly

accumulated 14 C-naphthalene (to a biomagnification of 40 times) from solution during exposure for 24 h and slowly released the accumulated hydrocarbons within 300 h after return to clean sea water (Rossi, 1977). 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmOral administration of South Louisiana crude oil to mallard ducks (Anas platyrhynchus) at a dose of 5 ml daily led to detectable aromatic petroleum hydrocarbon levels in each of the tissues examined: liver, breast muscle, heart muscle, skin, brain, uropygial gland, and blood. The skin and the underlying adipose tissue had accumulated far more aromatic hydrocarbons than other tissues. The individual components did not accumulate in the same relative concentrations as in the crude oil, suggesting different uptake and/or metabolism; two- and three-membered, condensed ring aromatic compounds were accumulated to a greater extent than alkylbenzenes (Lawler et al., 1978).

A4.2.4 Tainting

Tainting (unpleasant off-flavours) of various aquatic organisms by petroleum hydrocarbons has been reported, and diesel fuel has been suspected of tainting fish in freshwater rivers. Mackie et al. (1972) reported tainted rainbow trout up to at least nine days after a spill of diesel fuel into water (Six Mile Water, Ireland). Ackman & Noble (1973) described tainting of whitefish in the Athabaska River, Canada, which was attributed to diesel fuel from a railroad yard.

A4.2.5 Entry into the food chain

No data are available on the effect of diesel fuel on the food chain. The effects of petroleum compounds on the marine food web have been studied, e.g. on zooplankton Calanus helgolandicus (Corner et al., 1976), on phytoplankton (Mahoney & Haskin, 1980), and on freshwater crayfish, one of the primary foods of waterfowl (Tarshis, 1981). In a study in situ on the distribution, biotransformation, and flux of PAHs in the aquatic food chain (seston --> mussel [ Mytilus
edulis ] --> eider duck [ Somateria molissima ]), the PAH composition changed significantly (Broman et al., 1990). The theoretical inhalation of atmospheric PAHs was insignificant in comparison with exposure from food. PAHs were biotransformed relatively rapidly. Their concentrations did not increase with increasing trophic level.

A4.3 Ultimate fate after use

A4.3.1 Use in motor vehicles

Diesel fuel is combusted in motor vehicles and stationary engines (see section A3.2.1.2). The resulting exhaust is discussed in Part B.

A4.3.2 Spills

The fate of diesel fuel after accidental spills is eventual degradation by abiotic and biotic processes (see section A3.2.3)

A4.3.3 Disposal

In general, incineration is recommended for fuel oils (Fraser Williams (Scientific Systems) Ltd, 1985). This procedure is also useful for disposing of fuel production wastes.

Because diesel fuel is released during use, mainly accidentally into water or onto soil (see sections A3.2.2.2, A3.2.2.3, and A3.2.3), the affected compartments may have to be remediated. In general, oil spills on beaches and into surface waters can be treated by burning

off; absorption onto straw, polyurethane foam, activated carbon, or peat; or addition of sinking agents, gelling agents, or dispersants. Mechanical systems can also be used (Fraser Williams (Scientific Systems) Ltd, 1985). Various operations for treating soil affected by diesel fuel have been reviewed (Dineen, 1991) and are as follows:

-- Disposal: excavation of contaminated soil and disposal at a regulated landfill; the treatment used most commonly during the last decade; advantageous for small volumes.

-- Biodegradation: The natural process of soil microorganisms, which is stopped by a spill as a result of depletion of oxygen or nutrients (mostly nitrogen and phosphorus), is restored by providing sufficient amounts of the depleted substances. The process can be carried out in situ (Frankenberger et al., 1989) (see section A3.2.3, Table 10) or after excavation. Biodegradation is the most widely applicable alternative to landfill disposal, although its duration is uncertain and may be relatively long and a large space is required (see sections A9.1.1.2 and A9.1.3.1).

-- Thermal treatment: incineration of contaminated soil e.g. in cement kilns or in specially designed thermal units, where the soil is baked to remove the fuel, which is subsequently oxidized (thermal desorption). The thermal desorption process is also
feasible in situ. In cement kilns, the desorbed diesel fuel is used for heating the kiln, and the soil is incorporated into the
cement mixture.

-- Road base: The contaminated soil is spread on roads under construction and is compacted and oiled before paving. If the affected soil has a high proportion of expandable clays, it may
not be suitable as a road base.

-- Asphalt batching: The contaminated soil replaces part of the sand and gravel component of asphalt; this is possible only if the content of expandable clays is not too high.

-- Chemical oxidation: A strong oxidant, usually hydrogen peroxide, in combination with a silica polymer stabilizer to 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmcontrol the reaction is used to oxidize diesel hydrocarbons to carbon dioxide and water. The reaction is highly exothermic, and uncontrolled volatilization and leaching of some constituents may

occur.

-- Fixation: addition of a strong base to convert diesel hydrocarbons to organic acids and subsequent treatment with a dissolved silica polymer to effectively encapsulate the acids in amorphous silica; immobilizes diesel fuel in situ and prevents migration into surface or groundwater. The long-term stability of the immobilized chemicals is, however, unknown.

-- Soil washing: Diesel fuel is washed out of soil by a biodegradable, non-toxic surfactant (Peters et al., 1992). The operation is feasible in situ and after excavation and is particularly effective on materials with a low surface:volume ratio (e.g. gravels and coarse sands). The major disadvantage is the need to collect and treat or dispose of the surfactant-diesel mixture after washing.

-- Containment: construction of a low-permeability surface structure ('cap') that prevents infiltration of rain, snow, and other moisture that could lead to leaching of diesel constituents into groundwater. Long-term monitoring of the area of the spill is necessary.

A5. ENVIRONMENTAL LEVELS AND HUMAN EXPOSURE

A5.1 Environmental levels

As diesel fuels are complex mixtures of a great variety of hydrocarbons (see section A2.1), general 'environmental levels' cannot
be described. The individual constituents of diesel fuels should be detectable in almost all compartments of the environment, but the
sources cannot be verified.

A5.2 Exposure of the general population

Nonoccupational exposure to diesel fuel occurs when fuel tanks are filled manually. The primary source of dermal exposure is accidental spills, although these exposures to high levels are of short duration. Drinking-water can be contaminated by components of petroleum products, including diesel fuel, after leakage into groundwater from underground storage tanks. Specific data are not available on the contribution of diesel fuel components to groundwater contamination. Owing to the low volatility of diesel fuels at normal temperatures, there is little exposure to vapours (International Agency for Research on Cancer, 1989a).

A5.3 Occupational exposure during manufacture, formulation, or use

Workers are exposed to diesel fuel in a number of occupations, including manual handling and discharge of fuels; loading and unloading of diesel fuel; retailing at filling stations; manufacture, repair, maintenance, and testing of diesel engines and other equipment; use of diesel fuel as a cleaning agent or solvent; and laboratory handling and routine sampling of diesel fuel (CONCAWE, 1985).

At normal temperatures, low concentrations of vapours are likely to be generated from diesel fuels, owing to their low volatility. Only in confined spaces or at high operating temperatures do significant levels of vapour occur. Hydrogen sulfide gas may evolve from residual fuel oils (CONCAWE, 1985).

A6. KINETICS AND METABOLISM IN LABORATORY ANIMALS AND HUMANS

No quantitative data are available; however, the systemic and local effects observed after exposure to diesel fuels by various routes indicate that they are absorbed dermally, orally, and by
inhalation.

A7. EFFECTS ON LABORATORY MAMMALS AND IN-VITRO TEST SYSTEMS

A7.1 Single exposure

Diesel fuels have a low order of acute toxicity after administration orally, dermally, or by inhalation. Acute LD 50 Table
12. In rats and mice, the values after oral or dermal exposure were values are summarized in generally > 5000 mg/kg body weight, and the values in rabbits after dermal exposure were > 2000 mg/kg body weight. Since the physicochemical properties of kerosenes are similar to those of diesel fuels (see also section A2.1), the LD 50 values
for various types of kerosene are included.

Beck et al. (1984) reported an oral LD 50 value of 7500 mg/kg
body weight (9 ml/kg body weight) for a market-place sample of diesel fuel in male and female rats. Gross necropsy of animals that died during the test showed haemorrhagic gastroenteritis, gastrointestinal tympani, and pneumonia with abscess formation. No deaths were seen among rabbits treated dermally with 5000 mg/kg body weight under an occlusive covering for 24 h.

After oral administration of 12 000 mg/kg body weight kerosene to rats, no change was seen in heart weight or in the relative weights of lung, spleen, and kidney. There were no histopathological changes (Muralidhara et al., 1982). Exposure of rats for 8 h by inhalation to about 0.1 mg/litre deodorized kerosene did not induce adverse effects. Four male cats that inhaled 6.4 mg/litre for 6 h showed no toxic effects on the central nervous system or body weight during the 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmexposure or during a subsequent 14-day observation period (Carpenter et al., 1976).

Male Sprague-Dawley rats given JP-5 jet fuel at a dose of 18 912 mg/kg body weight (24 ml/kg body weight) orally had vacuolization of periportal hepatocytes and serum enzyme alterations, indicating hepatic damage, within two days (Parker et al., 1981). All male and female B6C3F1 mice treated dermally with 5000-40000 mg/kg body weight marine diesel fuel survived a 14-day observation period (US National Toxicology Program, 1986).

Table 12. Acute toxicity (LD 50 and LC 50 values) of diesel and other fuels

Fuel Species Route LD 50 or Dose Reference

LC 50

Diesel fuels Diesel fuel (market-place sample) Rat Oral LD 50 7500 mg/kg bw Beck et al. (1984)
Diesel fuel (market-place sample) Rabbit Dermal LD 50 > 5000 mg/kg bw Beck et al. (1984)
Home heating oil No. 2 a Rat Oral LD 50 14 500-21 100 μg/kg bw American Petroleum

Institute (1980a,b,c) Home heating oil No. 2 a Rabbit Dermal LD 50 > 5000 μg/kg bw American Petroleum

Institute (1980a,b,c) Marine diesel fuel, JP-5, JP-8 Mouse Oral LD 50 > 16 000 mg/kg bw Schultz et al. (1981)
shale, or petroleum oil a Marine diesel fuel, shale, or Mouse Dermal LD 50 > 16 000 mg/kg bw Schultz et al. (1981)
petroleum oil a

Other fuels Straight-run middle distillate b Rat Oral LD 50 > 5000 mg/kg bw American Petroleum

Institute (1985a) Hydrodesulfurized middle distillate b Rat Oral LD 50 > 5000 mg/kg bw American Petroleum

Institute (1982a,b) Straight-run middle distillate b Rabbit Dermal LD 50 > 2000 mg/kg bw American Petroleum

Institute (1985a) Hydrodesulfurized middle distillate b Rabbit Dermal LD 50 > 2000 mg/kg bw American Petroleum

Institute (1982a,b) Straight-run middle distillate b Rat Inhalation LC 50 1.8 mg/litre American Petroleum

Institute (1987a)

Table 12 (contd)

Fuel Species Route LD 50 or Dose Reference

LC 50

Hydrodesulfurized middle distillate b Rat Inhalation LC 50 4.6 mg/litre American Petroleum

Institute (1983a) Light catalytically cracked distillate c Rat Oral LD 50 4660 mg/kg bw (m) American Petroleum
3200 mg/kg bw (f) Institute (1985b) Light catalytically cracked distillate c Rabbit Dermal LD 50 > 2000 mg/kg bw American Petroleum

Institute (1985b) Light catalytically cracked distillate c Rat Inhalation LC 50 4.7-5.4 mg/litre American Petroleum

Institute (1986a,b)

Jet fuel A a Rat Oral LD 50 > 25 000 μl/kg bw American Petroleum

Institute (1980d)

JP-5 shale oil d Mouse Dermal LD 50 8000 mg/kg bw Schultz et al. (1981)

JP-5 petroleum oil d Mouse Dermal LD 50 11 200 mg/kg bw Schultz et al. (1981)

JP-8 shale oil a Mouse Dermal LD 50 6400 mg/kg bw Schultz et al. (1981)

JP-8 petroleum oil a Mouse Dermal LD 50 8000 mg/kg bw Schultz et al. (1981)

Jet fuel A a Rabbit Dermal LD 50 > 5000 μl/kg bw American Petroleum

Institute (1980d)

Kerosene a Rat Oral LD 50 50 000 mg/kg bw Parker et al. (1981)

Kerosene a Guinea-pig Oral LD 50 16 320 mg/kg bw Deichmann et al. (1944)

Kerosene a Rabbit Oral LD 50 22 720 mg/kg bw Deichmann et al. (1944)

Straight-run kerosene a Rat Oral LD 50 > 5000 mg/kg bw American Petroleum

Institute (1985c) Hydrodesulfurized kerosene a Rat Oral LD 50 > 5000 mg/kg bw American Petroleum

Institute (1982b)

Table 12 (contd)

Fuel Species Route LD 50 or Dose Reference

LC 50

Straight-run kerosene a Rabbit Dermal LD 50 > 2000 mg/kg bw American Petroleum

Institute (1985c) Hydrodesulfurized kerosene a Rabbit Dermal LD 50 > 2000 mg/kg bw American Petroleum

Institute (1982c) Petroleum-derived hydrocarbon a Rat Inhalation LC 50 > 5.3 mg/litre American Petroleum

Institute (1987b) Petroleum-derived hydrocarbon a Rat Inhalation LC 50 > 5.2 mg/litre American Petroleum

Institute (1983c) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmDeodorized kerosene e Rat Inhalation LC 50 > 0.1 mg/litre Carpenter et al. (1976)

(saturation)

a Hydrocarbon fraction similar to that of diesel fuel (see section A2.1)

b Hydrocarbon fractions from which also diesel fuels are gained during petroleum separation (see section A3.2.1.1)

c Blending component of diesel fuel

d Narrow boiling range

e Used as a solvent, not a fuel; very low aromaticity Exposure of Sprague-Dawley rats to an aerosol of diesel fuel at a concentration × time product of 8 mg.h/litre caused less than 1% mortality and was estimated to represent the maximal tolerated exposure. On this basis, 12 mg.h/litre was chosen as the highest concentration in a study of repeated exposure (see section A7.2). After exposure to 0.5 mg/litre for 2 h, a transient depression of the startle reflex was observed. Further findings were a dose-related decrease in respiratory frequency and influx of polymorphonuclear cells into the lungs for several days. In a 6-h inhalation test, the no-observed-adverse-effect level (NOAEL) was 2.7 mg/litre (Dalbey et al., 1982, 1987).

A7.2 Short-term exposure

A7.2.1 Subacute exposure

A7.2.1.1 Dermal exposure

Dermal exposure to 0.1 ml/kg body weight per day kerosene for one week was not lethal to male mice. Significant decreases in the relative weights of thymus, spleen, adrenals, and abdominal lymph nodes were observed (Upreti et al., 1989).

Eight rabbits (weighing 2.5-3.5 kg) were exposed dermally (occlusive covering, 24 h/day, five days per week, two weeks) to 4 or 8 ml/kg body weight per day of a market-place sample of diesel fuel. None of the animals given the low dose died, although there was a weight loss of 0.3 kg, while five animals given the high dose died with a weight loss of 0.5 kg; the control group had a weight gain of 0.2 kg. Severe chronic dermal irritation and anorexia were seen, which were followed by cachexia and death (Beck et al., 1984).

Three doses between 1 and 10 ml/kg body weight per day of 10, 30, or 50% fuel oil No. 2 were applied to the skin of New Zealand white rabbits. After a 12-day treatment including a two-day rest, acute
dermal corrosion was seen in all animals treated with the 10% oil and in animals given 10 ml/kg body weight per day of the 30 or 50% oil. There was histopathological evidence of dermal toxicity in all treated groups (American Petroleum Institute, 1980a,b,c).

Application of 8000 μl/kg body weight per day of jet fuel A to four male and four female New Zealand white rabbits for 12 days, including a two-day rest, induced acute dermal corrosion. Histopathological evidence of dermal and hepatic toxicity and hyperplastic changes in the urinary bladder transitional epithelium were seen (American Petroleum Institute, 1980d).

Groups of five male and five female New Zealand white rabbits were treated with 200, 1000, or 2000 mg/kg body weight per day of a straight-run distillate, a light catalytically cracked distillate, or two hydrodesulfurized distillates, three times per week for four weeks. Moderate to severe irritation was seen, the degree of skin irritation being directly related to the number of doses applied. Microscopic examination of the skin at the application sites revealed the presence of acanthosis and hyperkeratosis (American Petroleum Institute, 1983a, 1984, 1985a,b).

In a study of marine diesel fuel containing 12.7% paraffins, 43.7% naphthalenes, and 43.6% aromatic compounds, groups of male and female B6C3F1 mice received dermal applications (with an occlusive covering) of 2000, 8000, 20000, or 40000 mg/kg body weight per day for 14 consecutive days. None of the animals at 20000 or 40000 mg/kg body weight per day survived. Skin lesions were observed in all treated groups, which consisted of moderate acanthosis, parakeratosis, and hyperkeratosis. These findings were accompanied by a mixed cellular inflammatory infiltrate in the upper dermis. The NOAEL was 8000 mg/kg body weight per day. In the same study, JP-5 navy fuel containing 52.8% cycloparaffins, 30.8% paraffins, 15.9% aromatics, and 0.5% olefins (see also Table 2) was applied at doses of 5000-40000 mg/kg body weight per day for 14 days. All animals at 40000 mg/kg body weight per day and females at 30000 mg/kg body weight per day group died before the end of the study. Body weight loss was observed at

doses > 10 000 mg/kg body weight per day. The dermal findings were similar to those in the study of marine diesel fuel. The NOAEL was 5000 mg/kg body weight per day (US National Toxicology Program, 1986).

A7.2.1.2 Inhalation

Rats (strain, age, and sex not specified) were exposed for 6 h/day to a droplet aerosol of deodorized kerosene at a mean concentration of 7.7 mg/litre for four days. After a day with no treatment, the skin of the extremities was dry and flakes had formed. The body weight remained unchanged (Carpenter et al., 1976).

CD-1 mice were exposed for 8 h/day for five days to uncombusted diesel vapour at concentrations of 0.065, 0.135, or 0.204 mg/litre. The dose-response for neurotoxicity (measured by square-box activity) was 50% of the control level at the high dose, 100% at the medium dose, and 150% at the low dose. A drastic deterioration of motor coordination was seen at the high dose in the rotarod test. In the hot plate test, increased sensitivity followed by tolerance was seen at 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmthe high dose, a slight increase at the medium dose, and a substantial increase at the low dose; normalization occurred in all groups within 24 h. No effects were seen in the inclined plane test or the corneal reflex test. Vasodilatation, poor grooming, and tremors were described. In general, the most obvious effects were on the nervous system, diesel fuel acting as a neurodepressant (Kainz & White, 1982,
1984).

Dalbey et al. (1982, 1987) investigated the toxic effects of diesel fuel aerosols to assess the health risks of their military use as smoke screens. Groups of male and female Sprague-Dawley rats were exposed to concentrations of 1.3-6 mg/litre for 2 or 6 h, one or three times per week, for a total of nine exposures; 12 mg.h/litre was chosen as the highest concentration-time product that caused 6.25% mortality. A significant depression of body weight and a slower growth rate were seen initially, and liver weights were significantly decreased. No neurotoxicity was seen. Histologically, there was focal accumulation of free pulmonary cells and increased numbers of leukocytes in bronchial lavage fluid. In pulmonary function tests, a non-dose-dependent change in lung volume was seen, involving a decrease in total and vital capacity and increases in functional residual capacity and residual volume. No other organs were affected.

Groups of 20 male and 20 female Sprague-Dawley rats exposed by inhalation to 0.025 mg/litre of one of three hydrodesulfurized middle distillates for 6 h/day on five days per week for four weeks showed no treatment-related changes. Mild subacute inflammatory changes were observed in the nasal respiratory mucosa of the animals in one group that had been exposed to distillate API 81-09 (American Petroleum Institute, 1986c).

A7.2.2 Subchronic exposure

A7.2.2.1 Dermal exposure

Groups of 10 male and 10 female B6C3F1 mice received dermal applications under an occlusive covering of 250 or 4000 mg/kg body weight per day marine diesel fuel or 500-8000 mg/kg body weight per day JP-5 navy fuel for five days per week for 13 weeks. The marine diesel fuel caused no treatment-related deaths. Males had reduced body weight gain; mild chronic active dermatitis at the site of application was observed at the highest dose. At 1000 mg/kg body weight per day, all animals survived and had weight gains similar to those of the controls. Exposure to JP-5 navy fuel killed five males at the high-dose group and caused dermatosis, splenic extramedullary haematopoiesis, and liver karyomegaly in all groups (US National Toxicology Program, 1986).

A7.2.2.2 Inhalation

The results of studies of subchronic inhalation of various
distillate fuels are summarized in Table 13.

Table 13. Studies of subchronic exposure to distillate fuels by inhalation

Fuel Species Exposure (high dose) Results Reference

Diesel fuels Diesel fuel Rat 4 h/d, 2 d/week, 13 weeks + Reversible loss of body weight; Lock et al. (1984) 8 weeks' recovery (1.5 mg/litre) no other apparent effect

Marine diesel fuel a Mouse, rat 90 d, continuous (0.05 and Nephrotoxic effects specific to Bruner (1984) 0.3 mg/litre) male rats

Other fuels JP-8 jet fuel b Mouse, rat 24 h/d, 7 d/week,90 d + 24 Nephrotoxic effects specific to Mattie et al. (1991) months' recovery (1 mg/litre) male rats

JP-5 jet fuel b Rat, dog 24 h/d, 7 d/week,90 d Nephrotoxic effects specific to Gaworski et al. (1984) (0.75 mg/litre) male rats

JP-4 jet fuel b Mouse, rat 6 h/d, 5 d/week,12 months Nephrotoxic effects specific to Bruner et al. (1993) (5 mg/litre) male rats; increased incidence of hepatocellular adenomas in
female mice

Deodorized Rat, dog 6 h/d, 5 d/week,13 weeks No apparent effects Carpenter et al. (1976)

kerosene c (0.1 mg/litre)

a Hydrocarbon fraction similar to that of diesel fuel

b Narrow boiling range

c Not a fuel; very low aromaticity Groups of 25 male rats and four male dogs were exposed to deodorized kerosene at concentrations up to 0.1 mg/litre, approximately the saturation limit at 25°C, for 6 h/day on five days per week for about 13 weeks. Although dose-related changes were observed in survival, body weight gain, organ weights, and haematological and clinical chemical parameters, none of the findings was statistically significantly different from those in controls. No pathological change was seen in any of the organs examined (Carpenter et al., 1976).

Groups of 12 male and 12 female Sprague-Dawley rats were exposed in whole-body chambers for 4 h/day on two days per week for 13 weeks to 0.25, 0.75, or 1.5 mg/litre aerosolized diesel fuel, followed by an eight-week recovery period. There were no deaths. All treated animals 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmlost weight, indicating slight toxicity, during the first four weeks of exposure, but the effect was reversed during the recovery period in most cases. A slight depression of the startle reflex was seen at the start of the study, and the number of macrophages in bronchial lavage fluid was slightly elevated in all treated animals immediately after cessation of exposure. Further tests of pulmonary function, gas exchange, and dynamic lung function did not reveal any significant, dose-related change. Overall, no significant cumulative toxicity was seen (Lock et al., 1984).

A specific effect of diesel fuel in male rats, the hyaline droplet nephropathy syndrome, was seen in several studies. Gaworski et al. (1984) studied the effects of a 90-day exposure to 0.15 or 0.75 mg/litre JP-5 jet fuel derived from either petroleum or shale oil for 24 h/day, in groups of three male and three female beagle dogs, 75 male and 75 female Fischer 344 rats, and 111-150 female C57Bl/6 mice. The toxicity of the two types of fuel was not significantly different. The only substantial effect at both concentrations was a renal tubular lesion in male rats, probably caused by hyaline droplet accumulation and unique to male rats. Other signs of toxicity were mild liver-cell changes and mild nasal inflammation in rats. Cowan & Jenkins (1980) studied petroleum and shale-oil marine diesel fuel using an identical study design. Reduced body weight gain and hyaline droplet nephropathy were reported in male rats. In another study, groups of 95 male and 75 female Fischer 344 rats and 100 male and 100 female C57Bl/6 mice were exposed for 24 h/day on seven days per week to JP-8 jet fuel at concentrations up to 1 mg/litre for 90 days, and then held for 20-21 months in order to assess the long-term consequences of exposure. The main finding was accumulation of hyaline droplets in the kidneys of male rats (Mattie et al., 1991). In a 90-day study, groups of 75 male and 75 female Fischer 344 rats were exposed continuously to 0.05 or 0.3 mg/litre of marine diesel fuel. The main finding was again hyaline droplet nephropathy in males at both levels (Bruner, 1984). In a study

of the carcinogenicity of JP-4 jet fuel vapour, hyaline droplet nephropathy was observed in groups of 100 male Fischer 344 rats exposed for 6 h/day, five days per week for 12 months to concentrations up to 5 mg/litre (Bruner et al., 1993) (see also section A7.7).

The hyaline droplet nephropathy syndrome is not considered to be relevant for humans. It is linked to an inherent peculiarity of renal protein. The histopathological sequence is: accumulation in the renal proximal tubules of an excessive amount of hyaline droplets containing alpha2-microglobulin, a special protein of male rats which is synthesized in the liver; cytotoxicity and single-cell necrosis of the tubular epithelium; sustained regenerative tubule-cell proliferation; development of intraluminar granular casts from sloughed cell debris; tubular dilatation and papillary mineralization; foci of tubule hyperplasia; and renal tubule tumours (US Environmental Protection Agency, 1991). Mechanistically, a globulin-substance complex that is resistant to hydrolytic degradation is responsible for the protein overload in kidneys.

A7.3 Long-term exposure

A7.3.1 Dermal exposure

Groups of 50 male and 50 female B6C3F1 mice received dermal administrations of 250 or 500 mg/kg body weight per day marine diesel fuel or JP-5 navy fuel on the clipped dorsal interscapular region on five days per week for 103 weeks. (The doses were derived from studies reported in section A7.2.) The high dose of marine diesel fuel caused a dose- and time-dependent reduction in mean body weight in animals of each sex, by up to 23% after week 40 in males and up to 20% after week 72 in females. Survival of treated females was decreased, and exposure to the high dose was stopped at week 84 owing to severe ulceration. Application of 500 mg/kg body weight per day JP-5 navy fuel resulted in a time-dependent reduction in mean body weight in animals of each sex: by up to 22% after week 76 in males and up to 25% after week 60 in females. Treatment of females with the high dose had to be terminated at week 90. Overall, JP-5 navy fuel caused a lesser chronic inflammatory response than marine diesel fuel (US National Toxicology Program, 1986).

A7.3.2 Inhalation

Groups of 100 male and 100 female Fischer 344 rats and C57Bl/6 mice were exposed to JP-4 jet fuel vapour for 6 h/day on five days per week for 12 months at concentrations of 1 or 5 mg/litre (see section A7.7). The sex-specific alpha2-microglobulin nephropathy syndrome was observed in 21% of male controls and in 20% of males at the low dose, and 77% at the high dose (see also section A7.2). No significant, dose-related alteration was seen in clinical findings, mortality,

haematological parameters, or clinical chemistry. All treated male rats had a significant reduction in mean body weight. At the high dose, male rats had significantly increased relative liver and kidney weights and females had a significant decrease in relative spleen weight. These alterations were not significant in comparison with absolute organ weights (Bruner et al., 1993).

A7.4 Dermal and ocular irritation; dermal sensitization

A7.4.1 Dermal irritation

The results of studies of dermal irritation (Table 14) must be interpreted critically, as different protocols were used, e.g. in Europe and the United States. The main difference is in exposure time: 24 h in the American protocols and 4 h in those of the European Union 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmand the Organisation for Economic Co-operation and Development, which may pose a problem when only mild irritation is observed, leading to a classification of 'non-irritant' according to protocols of the latter organizations.

A market-place sample of diesel fuel, tested at a dose of 0.5 ml for 24 h in rabbits, was assessed as 'extremely irritating', corresponding to a Draize score of 6.81 (Beck et al., 1984).

Topical applications three times a week to mice of various types of kerosene and gas oil with boiling ranges similar to that of diesel fuel produced degenerative changes in the skin surface, including necrosis and hyperplasia (CONCAWE, 1991).

Four types of American military fuels were investigated: petroleum JP-5 fuel, shale JP-5 fuel, petroleum marine diesel fuel, and shale marine diesel fuel. None was active in the Draize test, in
which six female New Zealand white rabbits received 0.5 ml of undiluted fuel on intact and abraded skin under an occlusive covering for 24 h (Cowan & Jenkins, 1980). Applications to rabbit skin of 0.5 ml of various shale oil- and petroleum-derived fuels under occlusive dressings for 4 h were not irritating (Schultz et al.,
1981).

Various modifications of fuel oil No. 2 (10, 30, and 50%) were found to be moderately irritating to rabbit skin (American Petroleum Institute, 1980a,b,c), and two hydrodesulfurized middle distillates resulted in primary irritation indices of 4.3 and 5.9 (American Petroleum Institute, 1982a,b). A straight-run middle distillate had a moderately irritating effect, with an irritation index of 3.2 (American Petroleum Institute, 1985d). Jet fuel A was classified as mildly irritating, with an irritation score of 1.96 (American Petroleum Institute, 1980d). A hydrosulfurized kerosene was moderately irritating, with an index of 4 (American Petroleum Institute, 1982c).

A light catalytically cracked distillate (American Petroleum Institute, 1985d) and a straight-run kerosene (American Petroleum Institute, 1985e) had severely irritating effects, with irritation
indexes of 5.5 and 5.6.

Table 14. Studies of dermal irritation in rabbits exposed for 24 h to various fuels

Fuel Classification of Reference

irritation a

Diesel fuel Extremely irritating Beck et al. (1984)

Marine diesel fuel or marine Not irritating Cowan & Jenkins shale fuel (1980)

Hydrodesulfurized middle Moderately irritating American Petroleum distillate Institute (1982a,b)

Straight-run middle distillate Moderately irritating American Petroleum Institute (1985d)

Light catalytically cracked Severely irritating American Petroleum distillate Institute (1985d)

Heating oil No. 2 Moderately irritating American Petroleum Institute (1980a,b,c)

Jet fuel A Mildly irritating American Petroleum Institute (1980d)

Various kerosenes Slightly to severely CONCAWE (1995) irritating

Shale oil- and Not irritating Schultz et al. (1981) petroleum-derived fuels

Hydrodesulfurized kerosene Moderately irritating American Petroleum Institute (1982c)

Straight-run kerosene Severely irritating American Petroleum Institute (1985e)

a Note differences in protocols between regulations of the United States and the Organisation for Economic Co-operation and Development (see section A7.4.1)

A7.4.2 Ocular irritation

All of the available reports indicate no or, at the most, a slight irritating effect of diesel fuels and kerosenes on the rabbit

eye.

Four types of American military fuels (petroleum JP-5 fuel, shale JP-5 fuel, petroleum marine diesel fuel, and shale marine diesel fuel) gave generally negative responses in the Draize test, in which nine female New Zealand white rabbits were given 0.1 ml of undiluted substance (Cowan & Jenkins, 1980). A market-place sample of diesel fuel (0.1 ml, undiluted) was also not irritating in the Draize test (Beck et al., 1984). None of several kerosenes tested was more than slightly irritating to the eye (CONCAWE, 1995), and none of the shale oil- and petroleum-derived fuels tested caused visible irritation in the rabbit eye (0.1 ml, undiluted) (Schultz et al., 1981). 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmThree modifications of fuel oil No. 2 (10, 30, and 50%) were classified as mildly irritating in rabbits (American Petroleum Institute, 1980a,b,c). Two hydrodesulfurized middle distillates did not cause primary irritation in the Draize test (American Petroleum Institute, 1982a,b); a straight-run middle distillate (American Petroleum Institute, 1985d) and a straight-run kerosene also gave negative results (American Petroleum Institute, 1985e). Jet fuel A was classified as mildly irritating, with an average score of 2.67 after one day and 0 after seven days (American Petroleum Institute, 1980d). A hydrodesulfurized kerosene was not irritating (American Petroleum Institute, 1982c).

A7.4.3 Sensitization

Three modifications of fuel oil No. 2 (10, 30, and 50%) were were classified as non-sensitizing for the skin of guinea-pigs (American Petroleum Institute, 1980a,b,c). A straight-run middle distillate (American Petroleum Institute, 1985d), a straight-run kerosene (American Petroleum Institute, 1985e), jet fuel A (American Petroleum Institute, 1980d), a hydrodesulfurized middle distillate (American Petroleum Institute, 1984), and a light catalytically cracked distillate (American Petroleum Institute, 1985c) were also not sensitizing.

Four types of fuels used in the United States Army, petroleum JP-5 fuel, shale JP-5c fuel, petroleum marine diesel fuel, and shale marine diesel fuel, were not sensitizing in a modified Landsteiner test in groups of 24 guinea-pigs (Cowan & Jenkins, 1980). A straight-run kerosene, hydrosulfurized kerosene, jet fuel A (CONCAWE, 1995), and shale oil- and petroleum-derived jet and diesel fuels (Schultz et al., 1981) were also not sensitizing.

A7.5 Reproductive toxicity, embryotoxicity, and teratogenicity

Groups of 20 pregnant CRL:COBS rats were exposed by inhalation on days 6-15 of gestation to 100 or 400 ppm of diesel fuel for 6 h/day (American Petroleum Institute, 1979). A significant reduction in maternal food consumption was seen at the high concentration, and maternal body weight gain and fetal weights were reduced, although not statistically significantly. Overall, no significant, dose-related effects were observed in dams, and the compound had no teratogenic or embryotoxic effect and no effect on fetal growth or development.

No signs of developmental or maternal toxicity were seen when groups of 20 pregnant rats were exposed to jet fuel A or fuel oil for 6 h/day on days 6-15 of gestation (Beliles & Mecler, 1982). Sprague-Dawley rats were fed 500, 1000, 1500, or 2000 mg/kg body weight per day of JP-8 aviation fuel by gavage on days 6-15 of gestation, and fetuses were examined on day 20. Significantly reduced body weights were seen in dams at the two higher doses and in fetuses at 1000-2000 mg/kg body weight per day, but the number of malformations was not altered. The investigators concluded that JP-8 aviation fuel is a developmental toxin at doses > 1000 mg/kg body weight per day but is not teratogenic (Cooper & Mattie, 1993).

A7.6 Mutagenicity and related end-points

A7.6.1 In vitro

Various samples of diesel and related fuels have been tested for mutagenicity in Salmonella typhimurium (Henderson et al., 1981; Conaway et al., 1984; Vandermeulen et al., 1985; US National Toxicology Program, 1986; McKee et al., 1994). Negative results were obtained in direct, unmodified Ames' assays (Conaway et al., 1984; Vandermeulen et al., 1985; US National Toxicology Program, 1986). In one study, refined shale oil, jet fuel, and shale oil- and petroleum-derived marine diesel fuel were not mutagenic (Schultz et al., 1981). Negative results were also found in studies of aliphatic
and aromatic fractions of diesel fuel in S. typhimurium TA98, TA100, and TA1535 in the presence of an exogenous metabolic system (Henderson et al., 1981).

The number of revertants of TA98 increased to 2.3-2.5 times the background levels in a suspension assay (Conaway et al., 1984), but
increases were found at a limited number of doses. In a modified

Salmonella assay with an extraction step to concentrate the mutagenic aromatic compounds, addition of an exogenous metabolic system from Aroclor-induced hamster liver, and use of TA98 as the most responsive strain, increased numbers of revertants were seen (McKee et al., 1994); but the number never exceeded twice the background response, and the results were considered to be equivocal.

A dimethyl sulfoxide extract of an intermediate, catalytically cracked distillate (CAS No. 644741-60-2) was mutagenic in S. typhimurium TA98 with an increased concentration of exogenous metabolic system to optimize detection. Of two dimethyl sulfoxide extracts of light hydrocracked distillate (CAS-No. 64741-77-7) tested in the same system, one induced a weak, non-dose-dependent response, while the other induced a clear dose-dependent positive response (Blackburn et al., 1984). Under the same assay conditions, five samples of cracked middle distillates also gave positive results (German Scientific Association for Petroleum, Natural Gas, and Coal, 1991). These results in a series of assays in Salmonella with a number of variations provide no clear evidence of mutagenic activity. In the unicellular alga Chlamydomonas reinhardtii (streptomycin resistant) tested in a natural aquatic environment, water-soluble fractions of
two crude and two refined oils did not induce forward mutations (Vandermeulen & Lee, 1986). Neither aliphatic nor aromatic dimethyl
sulfoxide fractions of diesel fuel No. 7911 induced mutation in 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmS. typhimurium. After reaction with nitrogen dioxide, which may activate the mutagenicity of diesel exhaust, it was mutagenic in strain TA100, the aromatic fraction being 40-fold more active than the aliphatic fraction (Henderson et al., 1981).

A market-place sample of diesel fuel did not induce mutation in

L5178Y tk +/- mouse lymphoma cells with or without metabolic activation (Conaway et al., 1984). A hydrodesulfurized middle distillate (CAS 64742-80-9) was active in the presence of metabolic activation, but the concentrations were cytotoxic (American Petroleum Institute, 1987c).

A7.6.2 In vivo

Undiluted diesel fuel No. 2 increased the frequency of chromosomal aberrations in the bone marrow of Sprague-Dawley rats given single or repeated (five days; 125, 417, or 1250 mg/kg body weight) oral doses. Diesel fuel given intraperitoneally for one or five days at 0.6, 2, or 6ml/kg body weight significantly increased the number of aberrant cells at the highest dose (Conaway et al., 1984). In order to assess these findings further, McKee et al. (1994) evaluated micronucleus induction by diesel fuel No. 2 and related materials in male and female CD-1 mice treated by gavage at up to 5 g/kg body weight. No effect was seen.

No effect was seen on the frequency of dominant lethal mutations in male CD-1 mice exposed to 100 or 400 ppm of either diesel fuel or jet fuel A for 6 h/day, five days per week for eight weeks (American Petroleum Institute, 1980e, 1981).

A7.7 Carcinogenicity

A7.7.1 Dermal exposure

Diesel fuel has been shown to have at least weak carcinogenic potency after dermal administration in a number of studies. As some of the test samples contained low concentrations of carcinogenic PAHs, it has been suggested (Biles et al., 1988) that the tumours induced may have been a consequence of the dermal damage produced by repeated
treatment with these materials. The available studies are summarized
in Table 15.

Doses of 25 mg per animal of petroleum and shale fuels were applied to groups of 25 male and 25 female C3H/HeN mice three times a week, for 62 weeks for jet fuel A with subsequent observation up to 105 weeks, and for 104 weeks for JP-4 fuel. Controls received mineral oil. The shale and petroleum diesel fuels were characterized for boiling range (jet A, 171-271°C; JP-4, 60-249°C) and sulfur content (jet A: petroleum, 3510 ppm; shale, 89 ppm; JP-4: petroleum, 1830 ppm; shale, 199 ppm). After 60 weeks, increased incidences of squamous-cell carcinomas and fibrosarcomas were reported (jet A: petroleum, 26%; shale, 28%; JP-4: petroleum, 26%; shale, 50%; controls, 0-2%), but no statistical analysis was reported (Clark et al., 1988).

Groups of 50 male and 50 female B6C3F1 mice were given 250 or 500 mg/kg body weight per day marine diesel fuel dermally, on five days per week for 103 or 84 weeks (see section A7.3.). A control group received the vehicle, acetone, only. The survival rates were 26/50 males and 29/50 females at the high dose at 84 weeks, 20/49 males and 12/50 females at the low dose at 104 weeks, and 30/50 male and 40/50 female controls. Males at the high dose had a significantly increased number of hepatocellular adenomas or carcinomas (29%; control, 18%). The total incidences of squamous-cell papillomas and carcinomas at the application site and the adjacent inguinal site were: control, 1/50; low dose, 2/49; high dose, 3/50 in males; and control, 0/50; low dose, 1/45; and high dose, 2/48 in females. No information was available on historical frequencies in acetone-treated mice; the incidence of papillomas and carcinomas in 3500 observations in untreated mice was 0.3-0.4%. In a parallel study with JP-5 navy fuel, 1/50 papillomas were seen at the low dose and 4/49 carcinomas at the high dose in males, and 0/49 carcinomas were seen at the low dose and 1/47 at the high dose in females. The investigators concluded that there was equivocal evidence for the carcinogenicity of marine diesel fuel and no evidence for the carcinogenicity of JP-5 navy fuel (US National Toxicology Program, 1986).

Two middle-distillate fractions of each of two crude oils (boiling range, 170-370°C) were painted at a dose of 50 mg onto the skin of male C3H mice twice a week for 18 months. The yield of benign and malignant tumours (details not specified) was 2-19 % (Lewis, 1983).

Table 15. Studies of chronic dermal exposure of mice (usually 50 mice/group) to distillate fuels

Fuel Exposure Results Referenc e

Marine diesel fuel 0, 250, and 500 mg/kg bw, 1/50, 2/49, and 3/50 (m) and 0/50, 1/45, and 2/48 (f) US Natio nal Toxicolog y 5 d/week, 103 weeks with squamous-cell papillomas and carcinomas after Program (1986)

104 weeks

Straight-run gas oil 25 mg/animal, twice/week, Occasional ulcerations, scarring, epidermal dysplasia; American Petroleum (laboratory sample) a up to 2 years after 2 years, 17/50 with tumours; controls, 0% Institute (1985c)

Straight-run middle 50 μl/animal, twice/week, Mild to moderate irritation (mild desquamation, American Petroleum

distillate a up to 2 years occasional scabbing); after 2 years, 10/50 with Institute (1989a) squamous-cell carcinomas or fibrosarcomas; controls, 0% 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmStraight-run 50 μl/animal, twice/week, 29/50 with malignant and 1/50 with benign tumours; American Petroleum

kerosene b > 2 years latency, 76 weeks; controls, 0% Institute (1989a)

Two cracked 50 μl/animal, twice/week, Mild to moderate irritation; after 24 months, 39/50 American Petroleum

distillates c up to 2 years and 26/50 with tumours Institute (1989a)

Intermediate 50 mg/animal, twice/week, 42/43 with tumours; mean latency, 17 weeks; Blackbur n et al. (198 4 catalytically cracked 80 weeks controls, 0/50

distillate c

Two hydrodesulfurized 50 μl/animal, twice/week, Acanthosis, fibrosis, hyperkeratosis, crusting; at end American Petroleum

middle distillates a lifetime of study, 24/50 and 27/50 with tumours (no controls Institute (1989b) reported)

Table 15 (contd)

Fuel Exposure Results Referenc e

Hydrodesulfurized 50 μl/animal, twice/week, 26 malignant and 1 benign tumours; latency, 77 American Petroleum

kerosene b > 2 years weeks (no controls reported) Institute (1989b)

Jet A and JP-4 fuels 25 mg/animal, three After 60 weeks, increased skin tumour rate; jet A, Clark et al. (1988) from shale or times/ week, 62 weeks 26% petroleum, 28% shale; JP-4, 26% petroleum, petroleum oils b + observation (jet A) 50% shale or 104 weeks (JP-4)

Two middle-distillate 50 mg/animal, twice/week, 2-19% tumour rate (no further information) Lewis (1 983)
fractions 18 months

No. 2 fuel oil b 12, 25, or 50 μl/animal, Overall tumour rate, 10%; control, 0% Witschi et al. (1987) three times/week, up to

100 weeks

Two heating oils a 25 μl/animal, three 7/50 and 9/50 with tumours; control, 0/50 Biles et al. (1988) times/week, lifetime

a Hydrocarbon fractions similar to that of diesel fuel (see section A2.1)

b Hydrocarbon fractions from which also diesel fuels are gained during petroleum separation (see section A3.2.1.1)

c Blending component of diesel fuel Application of American Petroleum Institute No. 2 fuel oil dermally to C3H mice at 12, 25, or 50 μl per animal, three times weekly for up to 100 weeks induced an overall tumour incidence (type not specified) of 10% (15/150 in the three treatment groups; controls, 0/100) (Witschi et al., 1987).

Ten samples of middle-distillate fuels were applied dermally to C3H/HeJ mice at a dose of 25 μl, three times a week for life. A virgin heating oil blending base (142-307°C) and a commercial No. 2 heating oil (156-352°C) had aromatic contents closest to that of diesel fuel. The blending base induced five skin carcinomas and two papillomas in 50 animals, and the commercial heating oil produced eight skin carcinomas and one papilloma in 50 animals. There were no tumours in the respective control groups (Biles et al., 1988).

A7.7.2 Inhalation

Fischer 344 rats and C57Bl/6 mice were exposed for 6 h/day on five days per week for 12 months to 1 or 5 mg/litre of fuel vapours (see section A7.3), followed by a 12-month recovery period. No significant pulmonary neoplastic changes were seen. Female mice at the high dose had a slightly but significantly increased incidence of benign hepatocellular adenomas (10%; controls, 2%); however, the trend was reversed in male mice. No conclusions about carcinogenic potency were drawn from this study (Bruner et al., 1993).

A8. EFFECTS ON HUMANS

A8.1 Exposure of the general population

Like similar petroleum distillates, such as kerosene, diesel fuel can cause dermal irritation and defatting after dermal exposure (Sandmeyer, 1981). Considerable exposure and subsequent absorption may also result in acute renal failure secondary to acute renal tubular necrosis. A 28-year-old man developed progressive oliguria one day after washing his hair with diesel fuel and had acute tubular necrosis on renal biopsy (Barrientos et al., 1977). A 47-year-old man developed acute tubular necrosis after cleaning his arms and hands with diesel oil for several weeks (Crisp et al., 1979). Both recovered renal
function.

Accidental aspiration of diesel fuel resulted in immediate coughing, followed by dyspnoea, cyanosis, and loss of consciousness (Perez Rodriguez et al., 1976). X-Ray examination revealed diffuse alveolar infiltrates. Mildly elevated levels of alkaline phosphatase, serum glutamic-oxaloacetic transaminase, and serum glutamic-pyruvic transaminase were also seen. After 37 days, residual infiltrates remained, but the patient had improved. In another case, abdominal pain and vomiting followed ingestion of a reported 1.5 litres of diesel fuel. Pulmonary infiltrates were noted, presumably from subsequent aspiration of diesel fuel (Boudet et al., 1983).

A8.2 Occupational exposure

The odour threshold values for diesel fuels No. 1 and 4 are 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmreported to be 0.7 and 0.5 ppm, respectively (see section A2.2, Table 5). Six volunteers exposed to 140 mg/m 3 of deodorized kerosene for 15 min did not experience throat irritation. Olfactory fatigue was induced in three subjects, and one reported taste sensation. The investigators suggested that this concentration was the sensory
threshold for kerosene and estimated the odour threshold at 0.6 mg/m 3 (Carpenter et al., 1976).

Acne and folliculitis developed on the arms and thighs of a man who had worked in various automobile workshops for 15 years. He had handled diesel oil with his bare hands and then wiped them on his trousers (Das & Misra, 1988). Hyperkeratosis was noted in 320 drivers working in Russian oil fields with regular dermal contact with diesel fuel (Gusein-Zade, 1974). A 33-year-old man was exposed to diesel fuel vapour (concentration not reported) for 10 days while driving a truck with a fuel injector leak. He developed abdominal cramps, nausea, vomiting, and then acute renal failure with anaemia and thrombocytopenia (Reidenberg et al., 1964). Two aviators who were exposed to JP-5 fuel vapour for 1 h while flying a small airplane experienced eye irritation, difficulties in coordination and concentration, and fatigue (Porter, 1990).

Siemiatycki et al. (1987) conducted a case-control study in Montreal, Canada, and found an increased risk for squamous-cell carcinoma of the lung in men exposed to diesel fuel. When all types of lung cancer except adenocarcinoma were combined, an odds ratio (adjusted for smoking) of 1.6 (90% confidence interval (CI), 1.0-2.4;

n = 39) was found for 'any' exposure. In four subcategories, the odds ratios were 1.9 (short-low), 2.0 (short-high), 1.1 (long-low), and 2.0 (long-high). The possible effects of exposure to combustion products were not taken into consideration. An association with prostatic cancer was also observed, although there was no dose-response relationship, with odds ratios of 2.3 (CI, 1.3-4.0; n = 17) for the subgroup with nonsubstantial exposure and 1.4 (0.7-2.9; n = 8) in the subgroup with substantial exposure.

In another case-control study, no association was found between the occurrence of renal-cell cancer and occupational exposure to fuel oils, including diesel fuel (levels and duration not quantified) (Partanen et al., 1991).

In a cross-sectional epidemiological study, the effects of long-term exposure of factory workers to jet fuels (not specified) was investigated. Dizziness, headache, nausea, palpitation, pressure in the chest, and eye irritation were found to be more prevalent than in unexposed controls. The time-weighted average concentration of jet fuel vapour in the breathing zone was estimated to be 128-423 mg/m 3
(Knave et al., 1978).

A9. EFFECTS ON OTHER ORGANISMS IN THE LABORATORY AND THE FIELD

Comparison and evaluation of studies of the ecotoxicological effects of diesel fuel are complicated by a number of factors:

(1) There are comparatively few ecotoxicological studies on diesel fuel, and most work in the 1970s was concentrated on the effects of No.2 fuel oil, which has a similar but not identical chemical composition to diesel fuel No. 2 (see section A2). The composition of marine diesel fuel differs greatly from both (see Table 1). As described in the same section, diesel fuel itself has various specifications in different countries; in particular, the composition of non-hydrocarbon compounds from e.g. catalytic processes or additives may vary widely. There may also be differences between batches of the same oil (Hedtke & Puglisi, 1982) (see Table 20). The amount of the chemical actually in solution is also pertinent to ecotoxicological studies. Table 3 gives the concentration ranges of toxicologically relevant constituents of diesel and No. 2 fuel oil as
a whole and as 10% water-soluble fractions.

(2) The chemical composition of fuel oils and, as a consequence, their toxicity change with time, as oil can be modified by biodegradation, photooxidation, and volatilization (see section A4). Their toxicity also depends on whether they are mixed with water (sea water or freshwater), sediment, or soil, the temperature (tropical or Arctic), and physical conditions (e.g. storm conditions with continual mixing and breaking up of oil). Oil modified by these processes is usually less toxic than fresh oil (Baker, 1970; Morrow, 1973).

(3) A variety of methods have been used in ecotoxicological
studies on fuel oils. There is an obvious difference between conditions in the laboratory and the field. Most of the field studies that have been reported are related to fuel spills. In laboratory studies, either water-soluble fractions (see below) or oil-water dispersions have been used, and the methods of preparing them vary among laboratories. Oil films and, more recently, microencapsulated diesel oil have also been tested. In none of the laboratory studies reported were test media prepared with 'water accommodated fractions', as described and recommended by CONCAWE (1993) in laboratory protocols for petroleum products. Thus, the amount of product that must be added to a given volume of medium to produce the effects reported cannot be derived from the results of these studies, making comparison of data on ecotoxicity from different studies practically impossible.

Hedtke & Puglisi (1982) tested the lethality of No. 2 fuel oil in two species of freshwater fish and two species of amphibians and found
variable LC 50 values (see Table 20). Oil-water dispersions were very
toxic to fish. It is thought that these tiny oil droplets make contact with the gills and directly damage the respiratory epithelium (Engelhardt et al., 1981; Poirier et al., 1986). With floating oils, 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmthe relationship between toxicity and oil-water contact is a function of the equilibrium time for the soluble components of the oil and water, death occurring more rapidly when the oil is added 96 h before the organisms are exposed. Flow-through tests were more sensitive than static tests, as, in the latter, the chemical nature and toxicity of the oil change with time (Hedtke & Puglisi, 1982). Lockhart et al. (1987) showed that the results of static tests in fish varied considerably with aeration of test containers. Although flow-through techniques give a better estimate of real exposure conditions, many of the results reported in the literature are of static tests.

(4) The susceptibility of organisms to fuel oils and their components depends not only on the species and strain but also on the biological stage and the time of development within stages (Kühnhold, 1977; Winters et al., 1977). As the effects of No. 2 fuel oil and diesel oil have been tested in a wide variety of organisms and life stages under various conditions, direct comparison is difficult.

In spite of these difficulties, it can be concluded that diesel oil is generally more toxic than crude oil, as seen, for example, in barnacle nauplii (Winters et al., 1977), Daphnia (Cladocera), chironomid larvae, and the mollusc Viviparus bengalensis (Gastropoda) (Das & Konar, 1988). Crude oils themselves differ in toxicity, depending on their source (Anderson, 1977a; Winters et al., 1977), as do their corresponding products.

Rice et al. (1977a) concluded that the toxicity of fuel oil is due to the chemical toxicity of soluble aromatic compounds rather than to the physical toxicity of dispersed oil droplets. Physical coating by oil of the gills of fish and the feathers of birds is also a cause of toxicity. Monoaromatic compounds seem to be the least toxic, their acute toxicity increasing with molecular size up to the four- to five-ring compounds (e.g. chrysene and benzo[ a ]pyrene) (Neff et al., 1976b; Rice et al., 1977a), but they are not very soluble in sea water (see Table 6). The toxicity of one-, two-, and three-ring compounds (benzenes, naphthalenes, and phenanthrenes, respectively), seemed to increase with increasing degrees of alkylation of the aromatic nucleus. Neff et al. (1976b) reported 96-h LC 50 values of 0.3-0.6 ppm
for 1-methylphenanthrene, fluoranthrene, and phenanthrene, indicating the high toxicity of these polynuclear aromatic compounds for the polychaete, Neanthes arenaceodentata. Of the pure compounds tested by Winters et al. (1977), the most toxic to barnacle nauplii were indan, naphthalene, xylene, and substituted benzenes and naphthalenes. In four freshwater species, the water-soluble fraction was again associated with the substituted benzenes, and naphthalenes were found to be associated with the observed toxicity (Lockhart et al., 1987). The concentrations of naphthalenes, methylnaphthalenes, and dimethylnaphthalenes are particularly high in fuel oils (see Table 3), and these compounds have been used as analytical markers in several
studies.

Experiments involving microencapsulated oil fractions have shown that insoluble fractions also inhibit growth, e.g. of juvenile mussels (Stromgren & Nielsen, 1991). Anderson (1979) indicated that the larval stage might be the most sensitive to No. 2 fuel oil. Eggs of fish and invertebrates were often tolerant, and adults were sometimes more sensitive than juveniles.

Extensive studies have been carried out on lethal toxicity to marine organisms under the same conditions. The 96-h LC 50 values for
a wide range of invertebrates in static exposure conditions (temperature, 18-22°C) were 1-20 ppm for crude oil in water and 0.4-6 ppm for No. 2 fuel oil (Anderson et al., 1974; Neff et al., 1976b; Rossi et al., 1976). At 4-10°C, the 96-h LC 50 values appeared
to be lower (Rice et al., 1976, 1977a). Table 16 gives the results of lethality studies reported from several laboratories.

Table 16. Lethality (median lethal concentrations) to marine zooplankton of physically dispersed and water-soluble
fractions of No. 2 fuel oil

Zooplankton Oil-water dispersions Water-soluble (mg/litre) fractions (mg/litre)

Ctenophora 0.59 (1 day)

Mollusca

Clams Embryos 0.43 (2 days) Larvae 1.3 (2 days) Larvae 0.53 (10 days) Pteropods < 0.2 (2 days)

Crustacea Barnacles 2.6 (1-h LC 50 )
Copepods 1.0 (3 days) Amphipods 0.3 (2 days) 2.5 (3 days) Decapods Shrimp post-larvae 1.7; 9.4

Lobsters 1.2-6.6

Teleosts 1.5

Adapted from US National Research Council (1985). Values are for total measured (extractable) hydrocarbons, calculated from initial concentrations measured by spectroscopy and chromatography

No. 2 fuel oil (mostly the water-soluble fraction) has been found to have sublethal effects in all phyla examined (Anderson, 1977a,b, 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm1979; US National Research Council, 1985). With increasing concentrations, alterations were seen in the respiratory rate of the
fish Cyprinodon variegatus (Anderson et al., 1974) and Oncorhynchusm gorbuscha (pink salmon) (Rice et al., 1977b), the crustaceans Mysidopsis almyra and Penaeus aztecus, and the glass shrimp Palaemoetes pugio (Anderson, 1979); and changes were found in the pulse rate in embryonic estuarine fish, C. variegatus and Fundulus
heteroclitus (Anderson, 1977a) These effects have been used as a measure of stress caused by exposure. Dysregulation of the ability to regulate internal concentrations of solutes (osmotic and ionic regulation) was demonstrated in the brown shrimp Penaeus aztecus (Anderson et al., 1974; Cox, 1974; Anderson, 1979). The behaviour of
F. similus exposed to a 100% water-soluble fraction of No. 2 fuel oil was correlated with the content of naphthalene in four organs (Dixit & Anderson, 1977). Sublethal studies on No. 2. fuel oil showed reduced growth rates, survival, and reproduction in a range of organisms (Anderson, 1979; Table 17), including the annelid Neanthes arenaceodentata (Rossi & Anderson, 1978) and the estuarine fish F. grandis (Ernst et al., 1977).

A9.1 Laboratory experiments

Diesel fuel is a complex mixture of substances with a range of solubilities in water (see section A2.2). In testing for aquatic toxicity, it is important that the constituents in the medium be characterized (Bennett et al., 1990), but these are seldom reported. The results described in this section refer to diesel fuel, unless
otherwise stated.

As the solubility of diesel fuel in sea and freshwater is low, investigations of the effects of diesel fuel on organisms in the laboratory often involve preparation of a water-soluble fraction or an oil-water dispersion. Water-soluble fractions can be prepared by stirring one part of diesel fuel into nine parts of (sea) water for 20 h (Anderson et al., 1974) or one part of diesel fuel into eight parts of sea water for 24 h (Pulich et al., 1974), and then extracting the water phase. The resulting water phases contain 6-20 μg/ml of total hydrocarbons; the 100% water-soluble fraction of No. 2 fuel oil contains about 7 μg/ml of total hydrocarbons. Results are expressed as the concentration (percent dilution of the water-soluble fraction) and/or as the measured hydrocarbon concentration that causes the specified level of effect. The results cannot, however, be correlated with the original amount of fuel oil that was added to produce the water-soluble fraction or oil-water dispersion (CONCAWE, 1993).

Table 17. Effects of No. 2 fuel oil on the growth and reproduction of marine animals

Species Exposure Concentration (ppm) Growth or reproduction Reference (days)

TH TN TA

Fish Cyprinodon variegatus 7 2.0 0.6 1.7 0% of eggs hatched Anderson et al. (1976)

Decapods Rithropanopeus harrasii 27 1.0 0.3 0.9 Reduced survival and Neff et al. (1976b) extended development to megalopa Palaemonetes pugio 12 0.9 0.3 0.8 Reduced growth rate of Tatem (1977)

larvae 3 1.4 0.6 Reduced viability of eggs from exposed gravid females

Polychaetes Neanthes arenaceodentata 22 1.0 0.3 0.9 Reduced growth of larvae Rossi & Anderson (1976) 28 0.3 0.1 Reduced growth of Anderson (1977a) juveniles by 30% Ctenodrilus serratus 28 2.2 0.5 1.4 Reduced survival and Carr & Reish (1977) reproduction Ophryotrocha sp. 28 1.3 0.3 0.9 Reduced survival and Carr & Reish (1977) reproduction

From Anderson (1977b) TH, total hydrocarbons; TN, total naphthalenes; TA, total aromatic compounds Oil-water dispersions are produced by shaking a measured quantity of oil in water for 5 min (Anderson et al., 1974) or 30 min (Pulich et al., 1974). In some cases, the oil-water dispersion is maintained by continuous mixing, but more usually it is allowed to settle, and the aquatic organisms are exposed to the material in the water and to a
surface film. The effects seen in such tests can be due to both physical action and toxicity, especially in small invertebrates such

as Daphnia.

Insoluble fractions of crude oil and other hydrocarbon formulations strongly inhibited growth when they were microencapsulated and ingested by juvenile mussels (Stromgren et al., 1986; Stromgren & Reiersen, 1988) and larvae (Stromgren & Nielsen, 1991). In microcapsules with a thin acacia-gelatine structure, the total oil is dispersed into particles of 1-10 μm, which are suitable for ingestion by Mytilus larvae. The water-soluble components leak through the wall, providing a low concentration, and are ingested by

the molluscs.

None of the studies involved use of the 'water accommodated fractions' recommended by CONCAWE (1993), which allows the results to be expressed as 'loading rate', defined as the amount of product that 

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A9.1.1 Microorganisms

A9.1.1.1 Water

Marcus & Scott (1989) tested the effects of 0, 10, 20, and 40%
water-soluble fractions of a shale-diesel fuel mixture and a petroleum-diesel fuel mixture on the growth of faecal coliform bacteria. The 20 and 40% fractions of the petroleum-diesel fuel mixture resulted in significantly lower bacterial densities than the shale-diesel fuel mixture. The latter appeared to biostimulate or mediate against toxicity, suggesting a difference in the chemical
characteristics of the two fuels.

The effects of diesel and furnace oils, petrol, kerosene, and Assam crude oil and their paraffinic, aromatic, and asphaltic fractions (higher relative molecular mass, poorly defined) on the photosynthesis and respiration of blue-green algae (Cyanobacteria), in particular Anabaena doliolum were investigated by Singh & Gaur (1990). The chemical composition of the diesel and furnace oils was almost identical, with 37 and 36% paraffins, 51 and 54% aromatic compounds, and 6 and 0.6% asphaltic compounds, respectively. The aromatic fractions of these oils were the most toxic, followed by the asphaltic fractions; photosynthesis was reduced to 40% by the aromatic fractions and to 50% by the asphaltic fractions of both oils at a concentration

of 5.0 mg/litre. The effects were dependent on concentration up to 10 mg/litre. Photosynthesis was stimulated 160% by 5 mg/litre of the paraffinic fractions of crude oil , but only 100% by those of diesel oil and 110% by those of furnace oil. In general, crude oil inhibited photosynthesis and respiration by A. doliolum least and diesel and
furnace oils the most.

A9.1.1.2 Soil

In outdoor lysimeters, the toxicity of spills of 2.3 ml/cm 2 diesel and heating fuels was investigated in soil microbes, using

Microtox (R) measurements (Wang & Bartha, 1990). (For parallel assays of seed germination and plant growth, see section A9.1.3.1.) At time 0, both fuels were moderately toxic (EC 50 = 80-90 mg of contaminated
soil); this was followed by a period of increased toxicity, which started to decrease after 6-12 weeks. The values in soils treated by biodegradation returned to background within 20 weeks, although diesel fuel had significant residual toxicity in untreated soil. Seed germination and plant growth were inhibited in a similar manner, and diesel oil was more toxic and persistent than heating oil (section A9.1.3.1). Biodegradation treatment, in which the conditions for hydrocarbon degradation by microbes were optimized by mixing, aeration, pH control, and addition of mineral nutrients (fertilizers), strongly decreased fuel persistence and toxicity.

A9.1.2 Aquatic organisms

A9.1.2.1 Plants (phytoplankton)

Experiments in both the laboratory and the field have shown that hydrocarbons can inhibit algal growth, although enhancement is occasionally noted at lower concentrations of oil (US National Research Council, 1985).

After 12 days of exposure to 10% diesel oil, the growth of cells

of Euglena gracilis was not significantly reduced, whereas a concentration of 0.1% almost completely inhibited the growth of Scenedesmus quadricauda. Cells of S. quadricauda grown in culture media containing diesel fuel (concentration not given) became chlorotic, suggesting damage to the photosynthesizing system (Dennington et al., 1975).

A concentration of 0.05% light diesel fuel was found to stimulate growth rate, growth yield, photosynthesis, and chlorophyll a synthesis, while at the same time slightly inhibiting the respiration

of Chlorella salina CU-1 (Chan & Chiu, 1985). At higher concentrations (0.5 or 5%), the growth rate, growth yield, and

photosynthesis were greatly reduced (to 50, 50, and 62% and 43, 32, and 41% of the control levels, respectively), but the effect on algal respiration was less severe (81% with 0.5% diesel and 86% with 5%). Diesel fuel containing dispersants caused greater inhibition than did the compounds separately.

The lengthwise growth of the benthic algae Ascophyllum nodosum

and Laminaria digitata and other rocky-shore communities was measured while they were kept in 50-m 3 concrete basins and exposed continuously to diesel fuel for two years (Bokn, 1987). An average hydrocarbon concentration of 130 μg/litre continuously inhibited growth in both species, while a concentration of 30 μg/litre caused periodic inhibition. Under oil-free conditions during the subsequent season, the plants recovered completely.

Boiler fuel was more toxic than diesel fuel to the kelp Macrocystis; both fuels reduced photosynthesis (Baker, 1970).

A9.1.2.2 Invertebrates

(a) Several species

The acute toxicity of diesel fuel to Daphnia (Cladocera), chironomid (insect) larvae, and the mollusc Viviparus bengalensis 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm(Gastropoda) was tested after the organisms were acclimatized to laboratory conditions for four, two, and four days, respectively. The bioassays were conducted at 28 ± 2°C with unchlorinated borehole water (pH 7.0 ± 0.2; oxygen, 8 mg/litre; free carbon dioxide, 1.2 mg/litre; total alkalinity as calcium carbonate, 240 mg/litre; hardness, 260 mg/litre). The results are shown in Table 18. Plankton and insect larvae exhibited erratic, uncoordinated movements before becoming lethargic. In the mollusc, active avoidance and heavy secretion of
mucus were observed. The concentrations at which these behavioural changes occurred were not specified (Das & Konar, 1988).

Fourteen species of five phyla (Echinodermeta, Mollusca, Annelida, Arthropoda, and Urochordata) were exposed to 0.5% No. 2 diesel fuel in sea water. Only the larvae of the echinoderm Crossaster, which were also the largest, survived up to eight days; all of the other larvae died 3-72 h after being placed in the fuel-water mixture (Chia, 1973).

The periwinkles Pachymelania aurita (Muller) and Tympanotonus
fuscatus (Linne) were exposed to refined diesel fuel films for 24 and 48h at concentrations of 2.5, 5.0, 7.0, or 10.0% and to emulsions for 24 h at concentrations of 0.5, 1.5, 2.5, or 3.5%; they were then shaken for 2 min with brackish water. After exposure to the emulsion, the periwinkles were washed and reintroduced into oil-free, brackish water for 24 h and their activity recorded. Both surface films and emulsions were harmful to both gastropods, the emulsions being the
more toxic. T. fuscatus was less susceptible than P. aurita,

possibly because the former can retract further into its shell. The mean percentage survival in the presence of oil film (7.5% diesel fuel) was 100% at 24 h and 80% at 48 h for T. fuscatus and 80% at 24 h and 50% at 48 h for P. aurita. The mean percentage survival in the presence of 3.5% diesel fuel-water emulsion was 90% for T. fuscatus

and 80% for P. aurita (Dambo, 1993).

Table 18. Acute toxicity of diesel fuel to Daphnia (Cladocera), chironomid larvae (insects), and the mollusc Viviparus bengalensis (Gastropoda)

Species LC 5 (mg/litre) LC 50 (mg/litre) LC 95 (mg/litre)

Daphnia magna 1.5 (0.8-2.5) 20 (19.2-21.0) 38.5 (37.5-39.5)

Chironomid 5.0 346.0 (238-455) 865.0 (750-1018)

larvae

Viviparus 2.0 254.0 (185-320) 637.0 (575-700) bengalensis

From Das & Konar (1988)

(b) Molluscs

The feeding rate and growth of mussels ( Mytilus edilus L.) was markedly reduced when they were exposed to 30 or 130 μg/litre diesel fuel for eight months (Widdows et al., 1985). Recovery of physiological responses (clearance rate, respiration, food absorption efficiency, and ammonia excretion) after transfer to unpolluted water was concomitant with the depuration of hydrocarbons from the tissues. Mussels exposed to either dose recovered completely within about 55 days.

The effects on the nutrient storage system and reproductive cell systems of the mussel M. edilus of exposure to a diesel hydrocarbon-sea water emulsion and subsequent depuration were studied in order to assess the capacity of these systems to recover after discontinuation of exposure to the oil. After 144 days of exposure to hydrocarbon concentrations of 27 and 128 units, the volume of storage cell types was significantly reduced and there was a reduction in the volume of ripe gametes. After 53 days of depuration, the volume of the storage cells and of developing gametes increased. Exposure to either concentration also increased the volume of atretic (degenerating)

gametes, but depuration allowed a return to normal (Lowe & Pipe, 1986). The authors suggested that exposure to the hydrocarbons reduced the storage reserves and increased gamete atresia and resorption, so that the storage pool was partially replenished, enabling the organism to tolerate better the hydrocarbon insult.

The sensitivity of M. edulis to pollution with diesel fuel depends on the salinity of the water (Tedengren & Kautsky, 1987). Low salinity and diesel fuel acted synergistically to a greater degree in
Baltic than in North Sea mussels.

Adults and larvae of M. edulis were exposed to microencapsulated diesel fuel at concentrations of 200, 600, 1000, 1300, or 5000 μg/litre and the effects on mortality, larval growth, and spawning frequency were recorded (Stromgren & Nielsen, 1991). The EC 50 for spawning in mussels exposed for 30 days was about 800 μg/litre,

and the LC 50 (30days) for maturing mussels was about 5000 μg/litre.
The longitudinal growth of larvae was significantly reduced at 10 μg/litre; the EC 50 for growth (10 days) corresponded to about
25-30 μg/litre (Stromgren & Nielsen, 1991), which is lower than the

EC 50 of about 1000 μg/litre for growth of juvenile mussels reported
by Stromgren & Reiersen (1988). 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmDiesel fuel is therefore more toxic to mussel larvae than to juveniles.

The EC 50 values (10 days) for growth of larvae of the Quahog

clam, Mercenaria spp., exposed to water-soluble fractions of various crude and refined fuels were 220-4200 μg/litre, indicating that the insoluble fractions of hydrocarbons, ingested as microcapsules, are far more toxic than the water-soluble fractions alone (Byrne & Calder, 1977). Stromgren et al. (1986) drew the same conclusion from their results for the growth of juvenile mussels. Larval mortality during exposure to diesel fuel increased steeply up to 50 μg/litre, at which dose only 20-30% survived 10 days of exposure. At 500 μg/litre, there was 100% mortality. Byrne & Calder (1977) reported LC 50 values (10
days) of 50-2100 μg/litre for larvae of Mercenaria spp.

(c) Crustaceans

Freshwater crabs (Barytelphusa cunicularis) were exposed to sublethal concentrations of diesel fuel (4.5, 3.7, 3.2, and 2.6 ppm) for 24, 48, 72, and 96 h, respectively (Sarojini et al., 1989). The oxygen consumption of the crabs was measured as an index of stress caused by the fuel and compared with that of normal controls. The crabs responded in general by lowering their oxygen consumption up to 8 h (down to 50% at 4 h), particularly at the lower doses; with longer exposures, the oxygen consumption was higher than (approx. 120% at 24 h) or equal to (at 96 h) that of the controls.

Gammarus spp. (Crustacea, Amphipoda) were obtained from marine
and brackish water in the North and Baltic Seas and tested in two laboratories for their sensitivity to diesel-fuel pollution (Tedengren et al., 1988). G. oceanicus was obtained from the Fucus belt at a depth of 1-3 m and G. duebeni from rock pools of various salinities. The respiration, excretion, and atomic O:N and O:P ratios of the two species were compared after a 6-h exposure to 10 mg/litre diesel fuel emulsified in a syringe and injected into experimental aquaria; the resulting hydrocarbons were not measured. Exposure to diesel fuel generally resulted in decreased oxygen consumption and a rise in nutrient excretion, leading to lowered O:N and O:P ratios.
G. oceanicus was the most sensitive, and the response was aggravated by simultaneous lowering or raising of the salinity. No signs of impaired swimming or other physical or mechanical effects were detected in either species. The investigators concluded that
G. duebenihas a greater tolerance to pollutants and changes in salinity, probably because it has broad physiological niches and has evolved in and become adapted to more variable environments.

In a comparison of the effects of an oil-water dispersion of diesel fuel, kerosene, gasoline, and benzene on the tidepool copepod Tigriopus californicus, diesel fuel was the most detrimental: a concentration of 0.10 ml/litre sea water caused 100% mortality within five days (Barnett & Kontogiannis, 1975).

(d) Fish

Freshwater

Table 19 shows the results of a series of 96-h static tests for acute toxicity with diesel fuel in freshwater on several species of juvenile salmonids, Oncorhynchus kisutch (coho), O. gorbuscha (pink), and O. mykiss (rainbow trout), exposed at 14°C in 16 h of light and 8 h of darkness (Wan et al., 1990). The average loading density was 250 mg/litre (range, 100-420 mg/litre) of Canadian diesel fuel with a pour-point of -17.8°C. Three types of water were used for dilution: soft acid city tapwater, hard alkaline lakewater, and intermediate reconstituted deionized city tapwater. The LC 50 values
varied between the species from 32 to 33216 mg/litre. Diesel fuel was more toxic to pink salmon than to coho salmon or rainbow trout, irrespective of the water type.

The LC values for diesel fuel in golden orfe (Leuciscus idus melanotus) in a static test were: LC 0 = 40-120 mg/litre; LC 50 = 120-160 mg/litre; LC 100 = 160-205 mg/litre (Juhnke & Lüdemann,

1978).

Various freshwater fish (Fundulus diaphanus, Roccus saxatilis, Lepomis gibbosus, Roccus americanus, Anguilla rostrata, and Cyrinus carpio) were exposed to No. 2 or No. 4 fuel oil (both dispersed) in a static test at 19°C, pH 7.1, and 60 mg/litre hardness (Rehwoldt et al., 1974). The 96-h LC 50 values for No. 2 fuel oil ranged from
22.2 mg/litre for striped bass (Roccus saxatilis) to 49.1 mg/litre for carp (Cyrinus carpio); those for No.4 fuel oil ranged from 21 mg/litre for banded killifish (Fundulus diaphanus) to 48.1 mg/litre for carp. These concentrations represent total oil added and not the
oil dissolved in the water column.

Table 19. Acute toxicity of diesel fuel to juvenile Pacific salmonids in different types of freshwater

Water type Salmonid LC 50 (mg/litre

24-h 48-h 72-h 96-h

Soft Coho 28 972 28 972 12 345 10 299
Pink 1 972 276 84 74
Rainbow > 32 000 5 525 4 600 3 017 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmIntermediate Coho > 55 560 > 55 560 33 216 33 216 (reconstituted) Pink 1 829 376 133 123
Rainbow 168 363 168 363 28 787 2 186

Hard (lake) Coho > 26 743 26 743 4 845 3 333
Pink 1 404 302 48 32
Rainbow > 23 108 23 108 5 102 2 447

From Wan et al. (1990). Coho, Oncorhynchus kisutch; pink, O. gorbuscha; rainbow trout, O. mykiss

The LC 50 values of water-soluble fractions and emulsions and
the floating layer of No. 2 fuel oil for four freshwater species under static and flow-through conditions are shown in Table 20 (Hedtke & Puglisi, 1982).

Marine

In an investigation of the threshold doses at which cod (Gadus morhua L.) detected samples of Ekofisk diesel fuel, sea water was allowed to gravitate through the diesel fuel. The total content of dissolved materials, estimated by gas chromatography, were 15% benzenes, 2% toluenes, 6% xylenes, 15% naphthalenes, 3% phenols, 42%

n- alkanes (C 10 -C 25 ), and 17% unidentified compounds. There were
no visible surface films or microdroplets in the samples, although the authors suspected the presence of undissolved microdroplets. Behaviour was observed with a sea-water olfactometer, and reaction patterns after injection of samples (snapping, increased activity, coughing, darting, backing) were noted. The most frequent reaction was snapping of jaws followed by a short period of activity. None of the doses produced alarm reactions. The mean activity scores increased with increasing doses of oil compounds. In a set of four experiments, the threshold for detection of diesel oil compounds was 100-400 ng/litre, showing that cod detect fuel compounds at doses lower than those hitherto observed (Hellstrom & Doving, 1983).

The effect of acute exposure to the water-soluble fraction of Arctic diesel fuel (1:9 with sea water) on survival and metabolic rate
was studied in an Antarctic fish (Pagothenia borchgrevinki). The fish proved to be extremely tolerant and withstood an undiluted
water-soluble fraction for at least 72 h. None died as a direct consequence of contact with the water-soluble fraction, but they showed signs of stress. In unpolluted water, little spontaneous swimming activity was seen, but on transfer into water containing diesel fuel they became agitated and increased their swimming activity
for a short time. Their ventilation rates increased but later decreased; the depth of each ventilation increased considerably. Coughing became apparent shortly after transfer and persisted for up to 72 h. The haematocrit increased and remained elevated throughout the experiment. Oxygen consumption was increased to about twice that of the controls (Davison et al., 1992).

In further studies on P. borchgrevinki (Davison et al., 1993), a water-soluble fraction was prepared (1:2 with sea water) and then diluted to 33%; the fish were kept for seven days. On initial placement, behaviour similar to that with the higher concentration in the previous experiment was noted. After seven days, the only noticeable differences between exposed and control fish of similar weight and length were large amounts of mucus streaming from opercula, increased coughing, and slightly deeper ventilation. Plasma chloride and osmolarity were similar in the two groups. Haematocrit and haemoglobin concentration were significantly higher in the treated fish; spleen weights and oxygen consumption were similar. This Antarctic fish can thus survive prolonged periods of exposure to low levels of water-soluble hydrocarbons.

Table 20. 96-h LC 50 values for No. 2 fuel oil on freshwater organisms under various conditions

Species Life stage Test conditions Effect Concentration (μl/litre wat er)

Jordanella floridae (flagfish) Adult Water-soluble fraction of 10% No mortality No dilution
oil-water mixture; static Emulsion; flow-through LC 50 60.5

Pimephales promelas (fathead minnow) Adult Emulsion; flow-through LC 50 38.6
Water-soluble fraction of 10% No mortality No dilution
oil-water mixture; static Floating layer; static LC 50 > 160 000

48 300 (2nd sample)

Rana sylvatica (frog) Larvae Water-soluble fraction of LC 50 413 000

10% oil-water mixture; static
Emulsion; static LC 50 26.4
Emulsion; flow-through LC 50 4.9
Floating layer; static LC 50 < 5000

Ambystoma maculatum (salamander) Larvae Emulsion; flow-through LC 50 86.4

From Hedtke & Puglisi (1982) (e) Amphibians

Larvae of the frog Rana sylvatica were tested in 96-h static and 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmflow-through tests with No. 2 fuel oil as a water-soluble fraction, an emulsion, or a floating layer (Hedtke & Puglisi, 1982) (Table 20). Flow-through exposure to the emulsion was the most toxic (LC 50 = 4.9 μl/litre), and static exposure to the water-soluble fraction was the least toxic (LC 50 = 413 000 μl/litre). The LC 50 of No. 2 fuel
oil emulsion in larvae of the salamander Ambystoma maculatum in a flow-through test was 86.4 μl/litre).

A9.1.3 Terrestrial organisms

A9.1.3.1 Plants

Seed germination and growth of soya beans and ryegrass were inhibited by a diesel fuel spill of 2.3 ml/m 2 (Wang & Bartha, 1990; see sections A3.2.4 and A9.1.1.2). Four weeks after the spill, biodegradation had at least partially restored the ability of the soil to support seed germination and plant growth. Morphological effects, such as reverse geotropism, were noted on some partially emerged seedlings. In untreated diesel-contaminated soil, there was no evidence of seed germination or plant growth. Parallel experiments showed that heating oil was less toxic.

A9.1.3.2 Invertebrates

No data were available.

A9.1.3.3 Vertebrates

Birds are affected externally and internally by oil contamination. The fuel rapidly destroys the waterproofing of the birds' plumage, and as the birds attempt to preen the contaminant off their feathers they ingest fuel. When No. 1 fuel oil was administered by gavage at doses of 1-4 ml/kg body weight to ducks, a dose of 2 ml/kg caused lipid pneumonia, extreme inflammation of the lungs, and fatty infiltration and degeneration of the liver after 24 h (Croxall, 1977). Administration of 1 ml of diesel or No. 1 fuel oil/kg produced severe irritation of the digestive tract (enteritis), with diarrhoea and bile pigments in faeces. Signs of toxic nephrosis were also noted. Adrenal enlargement (mainly hyperplasia of cortical tissue), depression of plasma cholinesterase levels, and incoordination, ataxia, and tremors were reported at higher doses. The treatment was not fatal to healthy birds, even at doses up to 20 ml/kg body weight; however, when No. 1 fuel and diesel fuel were administered to birds under stress, the LD 50 was 3-4 ml/kg body weight (Hartung & Hunt,

1966).

A9.2 Field observations

The following information is derived mostly from observations made after diesel spills (see Table 10).

A9.2.1 Microorganisms

A9.2.1.1 Water

No data were available.

A9.2.1.2 Soil

Six weeks after the ship Bahia Paraiso spilled Arctic diesel fuel into the Antarctic Sea, the impact of the spill on microbial ecology was investigated (Karl, 1992). The acute effects on the metabolic activities of sedimentary microorganisms appeared to be negligible, even in saturated sea water.

A9.2.2 Aquatic organisms

In field studies conducted after fuel spills to investigate the effects on populations and ecosystems, zooplankton appeared to be highly vulnerable to dispersed and dissolved petroleum constituents but less so to floating oils. Field observations on oil spills in general showed that individual organisms were affected in many ways, including direct mortality (fish eggs, copepods, mixed plankton), external contamination by fuel (chorion of fish eggs, cuticles, and feeding appendages of crustacea), tissue contamination by aromatic constituents, abnormal development of fish embryos, and altered metabolic rates (US National Research Council, 1985). A spill of No. 2 diesel fuel was extremely toxic: faunal repopulation of the affected areas did not occur during the subsequent eight months (Blumer et al., 1970). A spill of 750000 litres of No. 2 diesel fuel caused substantial mortality in some taxa of the intertidal population, whereas others showed little or no effect. There was substantial recovery within six months, and there was no observable mortality among the subtidal fauna or flora (Woodin et al., 1972). After a spill of marine diesel fuel (2000-3000 t) off Hong Kong, much of the oil was carried to land by onshore winds. Wormald (1976) described the effects of the oil spill on intertidal meiofauna. Within four days of the spill, there was almost total mortality: nematodes and harpacticoid copepods were observed in an advanced state of decay, and other meiofauna were not detected. The aromatic fractions probably evaporated within four to six weeks but persisted longer in deeper sediment. Harpacticoids and nematodes did not recolonize the beach until nearly eight months after the spill. The density of meiofauna at all stations observed reached a maximum 10 months after the spill and

subsequently showed large fluctuations, which were probably the result of ecological successions during recovery of the beach fauna. One year later, an increase in the number of macrofauna, especially polychaete worms, was noted, which may have been responsible for the decrease in meiofaunal populations. High rainfall 11 months after the spill may 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmhave brought the soluble aromatic compounds nearer to the surface and decreased the harpacticoids and nematode populations. After 15 months, the meiofauna were again well established. Recovery at high shore stations began only when the oil content of the sediment had decreased by at least 50%, primarily through physical dispersion. Recolonization of the lower shore had occurred earlier. Stirling (1977) studied the effects of this spill on rocky shore fauna (see also Table 10), concentrating on large, common species of gastropods (Monodonta labio, Thais clavigera, Nerita albicilla, N. polita, Lunella coronata, Clypeomorus humilis, and Planaxis sulcatus), bivalve molluscs (Donax semignosus, Atactodea striata, Tapes philippinarum, and Circe stutzeri), and crustaceans (shore and hermit crabs). Acute mortality of gastropods was greatest at moderately contaminated sites where oil-dispersing chemicals had been applied to floating oil slicks. Long-term disturbances were most significant at the heavily polluted sites where dispersants had not been used. Recovery of animals taken
from the oiled beaches was studied in clean salt water in the laboratory. Bivalve molluscs and the gastropods Monodonta labioand Thais clavigera were the most sensitive and Clypeomorus humilisand
Planaxis sulcatus the least sensitive. Field observations of acute mortality were consistent with this order of sensitivity. Population studies showed the greatest reductions in Monodonta labio and Nerita albicilla, which were eliminated for at least 13 months from the site of greatest oil pollution.

The release of 600 000 litres of Arctic diesel fuel from the
Bahia Paraiso in 1989 coated intertidal macroalgae, limpets, and birds as well as sediments and shores (Kennicutt et al., 1991b). Macroalgae (Leptosomia simplex and Monostroma spp.) were only moderately affected. Intertidal limpet (Nacella concinna) populations were reduced by 50% within the first few weeks after the spill (Kennicutt et al., 1990), and a year later had still not fully recovered (Kennicutt & Sweet, 1992). Bottom-feeding organisms, such as

clams (Laternula elliptica) and fish (Harpagifer anarcticus and Notothenia coriiceps neglecta) were found to have PAHs in their gut contents and muscle tissues (Kennicutt et al., 1991b). After the spill, all of the chicks of the local population of South Polar skuas (Catharacta maccormicki) died, although few of the adults or the chicks died from direct contact with oil (Eppley, 1992). Within less than two years, and at many locations within a few weeks, the PAH content of sediment and intertidal organisms returned to background levels (Kennicutt & Sweet, 1992), owing partly to the high volatility of the spilled fluid and the high energy environment. Sampling of intertidal limpets showed that the tissue contaminants were primarily naphthalenes, but fluorenes and phenanthrenes were often the most

abundant aromatic contaminants in tissues with higher concentrations of PAHs (Kennicutt & Sweet, 1992; Kennicutt et al., 1992b) After a spill of 8000 litres of diesel fuel into an unspoiled freshwater stream in California, United States, most animals were affected within one to four days. Thousands of aquatic insects perished; some crayfish, aquatic leeches, and freshwater planarians were killed; and over 2500 fish, tadpoles (but no frogs), snakes, a turtle, and common

merganser (Mergus merganser) were found dead (Bury, 1972).

A9.2.3 Terrestrial organisms

A9.2.3.1 Plants

Diesel fuel was applied at 10 litres/m 2 to the soil of five common plant communities in north-east Greenland, comprising wet marsh, grassland, and three types of dwarf-shrub heath. The plant species were observed for up to three years after the application. Within a few days, most species had lost chlorophyll or had abscissing leaves. Species with xeromorphic leaf characteristics reacted more slowly than species with orthophyllous leaves. During three successive growing seasons, shrubs and forbs showed no signs of recovery. Graminoids showed slight resistance and recovered in all mesic and wet plots but were killed in dry plots. Three species of sedge (Carex bigelowii, C. saxatilis, and C. stans) and mosses recovered to some extent in all plots. Recovery of mosses was excellent in the wet plots, good in the mesic plots, and poor in the dry plots. In general, the phytotoxic effects of diesel fuel were much more pronounced than those of crude oil in similar plots (Holt, 1987).

A9.2.3.2 Invertebrates

No data were available.

A9.2.3.3 Vertebrates

No data were available.

A10. EVALUATION OF HUMAN HEALTH RISKS AND EFFECTS ON THE ENVIRONMENT

A10.1 Evaluation of human health risks

Diesel fuel is produced commercially in various qualities with regard to volatility, aromaticity, cetane number, and sulfur content. The composition of diesel fuel, which influences the type and amount of compounds emitted in the exhaust, has not changed greatly during the last few decades, although the cetane number has been slightly increased, resulting in better ignition, and the sulfur content, which

influences the release of particulate matter, has been reduced in some countries. Owing to the lack of data on diesel fuel, some data on heating oils and kerosenes (jet fuels) with compositions similar to that of diesel fuel are included in the evaluations of toxicological
and environmental effects. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmA10.1.1 Exposure of the general population

The general population can be exposed to diesel fuel and other middle distillates at filling stations, as a result of accidental spills, when handling such fuels, or when using kerosene for domestic cooking or heating. No data on exposure were available.

A10.1.2 Occupational exposure

Workers can be exposed to diesel fuel and other middle distillates during manual handling and discharge of the fuel, i.e. during retailing at filling stations; manufacture, repair, maintenance, and testing of diesel engines and other equipment; in jobs where diesel fuel is used as a cleaning agent or solvent; and in the handling and routine sampling of diesel fuel in the laboratory. At room temperature, very low concentrations of vapours are likely to be generated from diesel fuel because of its low volatility; significant levels of vapour are likely to occur only in confined spaces and at high temperatures.

A10.1.3 Non-neoplastic effects

Diesel fuels are toxic when ingested, usually accidentally. Ingestion may result in regurgitation and aspiration, which can cause chemically induced pneumonia. The latter effect is, however, not specific for diesel fuel and can occur with all hydrocarbons of a particular viscosity range.

Exposure to vapours is minimal during normal handling of diesel fuel. The most likely effect on human health is dermatitis as a result of skin contact. Dermal absorption takes place and can result in acute toxic effects on the kidney. The health effects of long-term absorption of low levels are unknown, but the available data on acute human toxicity indicate that practices such as washing hands in diesel fuel should be avoided. Although groundwater contamination and entry into drinking-water are potential sources of adverse health effects, such contamination would be noticeable and affect palatability so that inadvertent ingestion of contaminated drinking-water is unlikely. In experimental animals, diesel fuel has little acute toxicity after exposure orally, dermally, or by inhalation. The oral LD 50 values in
rats were > 5000 mg/kg body weight and those in mice, rabbits, and guinea-pigs even higher. After short-term dermal exposure of mice for 14 consecutive days, the NOAEL for two middle distillates (marine

diesel and JP-5 navy fuel) was 5000-8000 mg/kg body weight per day. Inhalation of up to 0.2 mg/litre had a neurodepressive effect in mice but not in rats. Subchronic exposure to various distillate fuels induced mainly alpha2-microglobulin nephropathy in male rats, which is
not considered relevant for humans.

Female mice that received dermal applications of 250 or 500 mg/kg body weight per day of marine diesel fuel or JP-5 navy fuel on five days per week for 103 weeks showed decreased survival due to severe ulceration. Rats and mice exposed by inhalation to 1 or 5 mg/litre had significant alterations in organ weight.

Diesel fuel irritates the skin but not the eye.

Exposure to diesel or jet fuels orally or by inhalation was neither embryotoxic nor teratogenic in rats; although doses toxic to the dams reduced fetal weight, no effects on viability were seen. No clear evidence of mutagenic activity was seen in a series of tests

in Salmonella typhimurium. A positive response achieved only under special conditions appeared to be equivocal. Other tests for genotoxicity in vitro or in vivo did not show clearly positive

responses.

Owing to lack of data, a quantitative risk assessment is not possible.

A10.1.4 Neoplastic effects

A case-control study of cancers at several sites in relation to exposure to diesel fuel suggested an increased risk for lung cancer other than adenocarcinoma and for prostatic cancer. In neither case was there an exposure-response relationship. As few studies were available, the number of cases was small, and the confidence intervals were correspondingly wide, no conclusion can be drawn at present about the carcinogenicity to humans of diesel fuel.

Twelve studies of dermal carcinogenicity in animals demonstrated that diesel fuels have weak carcinogenic potency in mice. As no clear genotoxicity is seen, cancer may be induced by nongenotoxic mechanisms, such as chronic dermal irritation characterized by repeated cycles of skin lesions and epidermal hyperplasia.

A10.2 Evaluation of effects on the environment

Although there are numerous data on the environmental behaviour of the individual components of diesel fuel, few data are available on diesel fuel as a whole. The environment can be polluted by accidental release of diesel fuel on a large scale, such as during tanker disasters and pipeline leaks, or on a smaller scale, from contamination of soil around factories and garages.

In water, diesel fuel spreads out almost immediately, polar and low-relative-molecular-mass components dissolve and leach out of the slick, volatile components evaporate from the water surface, and microbial degradation begins. The extent to which such 'weathering' takes place depends on the temperature and climatic conditions, and 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmthe chemical composition of spills changes with time. After spillage on water, some fractions evaporate, and the evaporated diesel components are degraded photochemically. In sediment, diesel fuel is generally delivered to bottom sediments by settling particles. In soil, the various components of diesel fuel migrate at different rates, depending on the soil type.

The individual constituents of diesel fuel are inherently biodegradable, but the rates depend greatly on the physical and climatic conditions and microbial composition. PAHs are the most
recalcitrant molecules.

Aquatic organisms, in particular molluscs, bioaccumulate hydrocarbons to various extents, but the hydrocarbons are depurated on transfer to clean water. Bioaccumulation of diesel fuel may occur, but there are few data to indicate biomagnification.

Although there have been a number of spills of diesel fuel, few reports are available on their environmental effects. Generally, diesel fuel is more toxic than crude oil to aquatic and plant species. Spills of diesel fuel have an immediate detrimental effect on the environment, causing substantial mortality of biota; recolonization occurs after about one year, depending on the animal and plant species involved and the chemical and physical content of the spill residues. Aquatic organisms that survive diesel fuel spills can still be affected by external contamination and tissue accumulation; abnormal development and altered metabolic rates are signs of such stress.

There are few data on the effects of diesel fuel on aquatic and terrestrial organisms in experimental situations. In studies of water-soluble fractions and emulsions of diesel fuel, the actual chemical composition has rarely been analysed, although this is crucial to an understanding of the toxicity of diesel fuel. No data are available about flow-through conditions or mesocosms, which better
reflect environmental conditions

A11. RECOMMENDATIONS

A11.1 Recommendations for the protection of human health

Exposure to diesel fuel may cause irritation and dermatitis; therefore, skin contact should be avoided, and diesel fuel should not be used for cleaning purposes.

A11.2 Recommendation for the protection of the environment

As for all petroleum products, accidental releases to the environment should be avoided and should be cleaned up as soon as possible if they occur.

A11.3 Recommendations for further research

A basic data set on the aquatic toxicity of diesel fuels, including complete analysis of test substances, should be generated. The fate and environmental impact of diesel fuel spills and leakages should be investigated in laboratory and controlled field studies. The exposure of humans to diesel fuel in various situations should be quantified.

The mechanism of the carcinogenic action of diesel fuel on the skin of experimental animals should be clarified.

A12. PREVIOUS EVALUATIONS BY INTERNATIONAL BODIES

The carcinogenic risks of diesel fuels for human beings were evaluated by a working group convened by the International Agency for Research on Cancer in 1988 (International Agency for Research on Cancer, 1989a). It concluded that there was inadequate evidence for the carcinogenicity of diesel fuels in humans, and there was limited evidence for the carcinogenicity of marine diesel fuel in experimental
animals.

PART B

B1. SUMMARY

B1.1 Identity, physical and chemical properties, and analytical

methods

Diesel engine exhaust emissions contain hundreds of chemical compounds, which are emitted partly in the gaseous phase and partly in the particulate phase of the exhaust. The major gaseous products of combustion are carbon dioxide, oxygen, nitrogen, and water vapour; carbon monoxide, sulfur dioxide, nitrogen oxides, and hydrocarbons and their derivatives are also present. Benzene and toluene are present in the lower weight percent range in the gaseous part of the hydrocarbon fraction. Other gaseous exhaust components are low-relative-molecular- mass polycyclic aromatic hydrocarbons (PAHs).

A main characteristic of diesel exhaust is the release of particles at a rate about 20 times greater than that from gasoline-fuelled vehicles. The particles are composed of elemental carbon, organic compounds adsorbed from fuel and lubricating oil, sulfates from fuel sulfur, and traces of metallic components. Most of the total particulate matter appears to occur in the submicrometre range, between 0.02 and 0.5 μm. Agglomeration may occur during aging, up to a maximal diameter of 30 μm. The emitted particles have a large surface area. Organic compounds generally contribute 10-30% of the total particulate matter, but poorly designed and maintained engines may result in as much as 90%. Higher-relative-molecular-mass, 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmoxygenated and nitro-PAHs occur at concentrations of parts per million
in this fraction.

Specific transient or steady-state driving cycles are used in measuring vehicle emissions. The exhaust can be sampled from undiluted or diluted exhaust gas. It is difficult to obtain artefact-free samples, as exhaust constituents can undergo chemical reactions, adsorption and desorption processes, and condensation and diffusion. The toxicologically relevant PAHs in diesel particulate matter are usually determined by Soxhlet extraction, clean-up, and fractionation, with subsequent analysis by high-performance liquid chromatography or gas chromatography coupled with mass spectrometry.

B1.2 Sources of human and environmental exposure

Diesel engine exhausts are emitted mainly from motor vehicles; other sources are stationary, railway locomotive, and ship diesel engines. The emissions from diesel motor vehicles have been well described, but the individual results are often not comparable owing to differences in parameters such as driving cycle, engine type, and fuel composition. The individual components are emitted in the

following quantities: carbon dioxide, about 1 kg/km; carbon monoxide, nitrogen oxides, total gaseous hydrocarbons, and particulate matter, 0.1-20 g/km; and aliphatic compounds, alcohols, aldehydes, light aromatics, and PAHs, micrograms per kilometer. The emissions of carbon monoxide, nitrogen oxides, total gaseous hydrocarbons, and particulate matter are regulated by law in a number of countries.

In principle, there is no difference between the quality and quantity of exhaust emissions from light- and heavy-duty engines, although heavy-duty vehicles release larger relative amounts of particulate matter. Exhaust emissions depend on driving cycle (transient or steady state), engine conditions (injection and aspiration techniques, maintenance, total mileage), and fuel composition (sulfur content, aromaticity, volatility); adjustment of the engine plays a major role.

The release of particulate matter increases with decreasing air:fuel ratio, increasing load, and increasing temperature. More particulate is released from older, intensively used engines than from new, low-mileage engines, probably because of a greater consumption of lubricating oil. The emission of particles from light-duty diesel vehicles is also correlated with the sulfur content of the fuel, as the formation of metal sulfates increases the particle mass; particle emissions from heavy-duty vehicles have not yet been established. Increasing fuel aromaticity also increases particle emissions.

PAHs and oxygenated PAHs from diesel and spark-ignition engines are qualitatively similar. Oxygenated and nitrated PAHs are emitted in the low microgram per kilometer range, but the actual concentrations of these compounds are uncertain, as decomposition and formation can occur during sampling. PAH emissions increase with increasing load and temperature and with the age of the engine, probably owing to increased consumption of lubricating oil. PAH emissions also depend on the injection technique of the engine: they increase with increasing air:fuel ratio in engines with direct injection, whereas they decrease in engines with indirect injection. The aromaticity and volatility of the fuel are directly correlated with the emission of PAHs. Malfunction of engine devices, especially the fuel injection system, increases the emission of the main exhaust components. There are few
data on the contribution of diesel motor emissions to the total man-made release of combustion products.

Diesel exhaust emissions can be reduced by improving engine design and by use of particle traps (trap oxidizers) and catalytic converters. While particle traps remove both the soot and soluble organic compounds adsorbed onto the particles, catalytic converters reduce the levels mainly of carbon monoxide and gaseous hydrocarbons. In practice, it is dificult to regenerate particulate traps. Catalytic converters require fuels with a low sulfur content, as sulfur poisons the active centres of the catalyst.

B1.3 Environmental transport, distribution, and transformation

The compartment first affected by diesel exhaust emissions is the atmosphere. The hydrosphere and geosphere are contaminated indirectly by dry and wet deposition. The environmental fate of the individual constituents of diesel exhaust is generally well known: Particles behave like (non-reacting) gas molecules with regard to their mechanical transport in the atmosphere; they may be transported over long distances and even penetrate the stratosphere. The overall removal rate of diesel particles is estimated to be low, resulting in an atmospheric lifetime of several days. During aging, particles may coagulate, with higher fall-out rates, thus reducing the total airborne level. The elemental carbon of diesel particulates may act as a catalyst in the formation of sulfuric acid by oxidation of sulfuric dioxide. The organic components adsorbed on elemental carbon may undergo a number of physical and chemical reactions with other atmospheric compounds and during exposure to sunlight.

B1.4 Environmental levels and human exposure

As diesel exhaust is a complex mixture of a great variety of compounds, general 'environmental levels' cannot be given. The individual components of diesel exhaust should be detectable in all compartments of the environment, although their source cannot usually
be verified. The environmental levels of most of the individual
constituents are known. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmThe general population is most likely to be exposed to diesel exhaust in busy streets or parking areas, particularly underground. The identification of sources is difficult, and the contribution of diesel exhaust to total pollution by traffic combustion products is generally calculated on the basis of emission factors and the percentage of diesel-fuelled vehicles. The levels of exposure of the general population and workers to diesel particulate matter are toxicologically relevant. The daily average ambient concentrations of diesel particulates near roads are 8-42 μg/m 3 . The estimated annual average concentrations were 5-10 μg/m 3 in urban areas and

< 1.5 μg/m 3 in rural areas in Germany and 1-2 μg/m 3 in urban areas and 0.6-1 μg/m 3 in rural areas in the United States of America. The concentrations are directly correlated with traffic density and decrease with increasing distance from roads. Identification of sources may also be difficult in work-places, especially in mines, where the total dust burden is high. Carbon core analysis has been used to determine specific exposure to diesel exhaust in the work-place, and the levels of particulate matter to which workers are exposed have been reported to be 0.04-0.134 mg/m 3 for truck drivers and 0.004-0.192 mg/m 3 for railroad workers. Total respirable particles and total suspended particles have also been used as measures of occupational exposure.

B1.5 Kinetics and metabolism in laboratory animals and humans

B1.5.1 Deposition

Diesel exhaust particles, with a mass median aerodynamic diameter of about 0.2 μm, undergo some filtration in the nose, and the deposition efficiency in the lung is only slightly greater than the minimal value found for particles with a mass median aerodynamic diameter of about 0.5 μm. Thus, 10-15% of inhaled diesel soot particles are deposited in the alveolar region of the lung of rats and guinea-pigs; in humans, about 10% is deposited in the alveolar region.

B1.5.2 Retention and clearance of particles

Mucociliary clearance of particles from ciliated airways is almost complete within 24 h. The long-term clearance of diesel particles from the alveolar region was measured in several studies in rats exposed by inhalation to labelled diesel or test particles. Half-times of 60-100 days were reported for controls with low lung burdens (< 1 mg/lung), whereas the half-times increased to 100-600 days in rats with lung burdens increasing from 1 to 60 mg/lung. In several studies, effects were seen to be caused by particle overload, which has been described in a variety of species and with a number of particulate materials. The phenomenon is generally observed when the deposition rate of particles of low solubility and low acute toxicity
exceeds their clearance rate for a considerable time. The half-time of unimpaired alveolar clearance in humans is several hundred days, which is longer than that in rats.

A dosimetric lung model was developed on the basis of data on the deposition and retention of diesel particles in rats after long-term inhalation and of data on particle deposition and retention in humans. The model can be used to predict retention of diesel particles and adsorbed organic material in the lungs of people of different ages. No data are available on changes in the retention of individual compounds after prolonged exposure to diesel exhaust.

B1.5.2 Retention and clearance of polycyclic aromatic hydrocarbons
adsorbed onto diesel soot

PAHs in diesel soot adhere strongly to the surface of particles. About 50% of the PAHs adsorbed onto diesel particles is cleared from the lung within one day, but the retention half-times for the remaining PAHs were 18-36 days. Studies with 3 H-benzo[ a ]pyrene and

14 C-nitropyrene show that when PAHs are associated with particulate matter, their clearance from the lungs is significantly delayed in comparison with the clearance of inhaled PAHs not associated with particulate matter.

B1.5.4 Metabolism

Benzo[ a ]pyrene coated on diesel exhaust particles was metabolized by oxidation to benzo[ a ]pyrene phenols, diols, and quinones in the lung and in cell cultures of pulmonary macrophages. Nitropyrene adsorbed to diesel particles was metabolized to acetylaminopyrene- phenol after inhalation. More DNA adducts were found in the lungs and type II cells of rats exposed to diesel exhaust than in controls. The oxidative metabolism of some organic compounds to epoxides may be responsible for the formation of these adducts, but adducts were found only after exposure to particles. Certain organic substances in diesel exhaust have been shown to be responsible for the formation of DNA adducts; however, the carbonaceous core itself (without extractable organic material) can also induce adducts, by chronic damage of epithelial cells.

B1.6 Effects on laboratory mammals and in vitro test systems

The few data available suggest that diesel exhaust has little acute toxicity. Mice treated intratracheally with diesel exhaust particles died due to lung oedema; the LD 50 was about 20 mg/kg body
weight. Methanol-extracted diesel exhaust particles were not lethal a concentrations up to about 33 mg/kg body weight. In hamsters, the LD 50 after intraperitoneal administration was 1280 mg/kg body weight. No data are available for exposure by inhalation.

Exposure of rats, guinea-pigs, and cats to diesel exhaust with a 

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accumulations were located near the terminal bronchioles and became larger, due to macrophage attachment (sequestration), during a subsequent recovery period.

Mice, rats, hamsters, cats, and monkeys did not have drastic decreases in body weight or reduced survival times after long-term inhalation of up to 4 mg/m 3 . Dose-related toxic effects seen in all species after long-term inhalation of diesel exhaust were: increases up to 400% in lung weight; pulmonary inflammation measurable by biochemical (cytoplasmic marker enzymes, collagen) and cytological (increase in polymorphonuclear neutrophils) parameters; impairment of lung mechanics; increasing numbers of particle-laden macrophages with focal accumulations (sequestration) under overload conditions; and subsequent proliferative alterations of epithelial cells and onset of
fibrosis.

Limited information on reproductive and developmental toxicity suggests that inhalation of diesel exhaust is not critical. In most experiments, no effects were seen in mice, rats, hamsters, rabbits, or monkeys; however, after intraperitoneal injection, sperm abnormalities were seen in mice given diesel exhaust particles and embryotoxicity occurred in hamsters given diesel exhaust extract.

Most tests for genotoxicity in vitro have been performed with diesel exhaust extracts rather than with total exhaust; positive responses were found in the absence of metabolic activation, i.e. the genotoxic effects appeared to be PAH-independent. About one-half of investigations in vivo gave negative responses; the only positive responses were sister chromatid exchange induction with total diesel exhaust and with organic extracts and micronucleus induction with organic extracts.

Immunotoxic effects were generally not seen after inhalation of diesel exhaust; however, an enhanced anti-ovalbumin immunoglobulin E antibody titre was seen in one study and an increased susceptibility to infections in mice in two experiments. Studies in rats suggested
that inhalation of diesel exhaust affects behavioural and neurophysiological status.

In studies of the carcinogenicity of diesel exhaust administered by inhalation to rats, the gaseous phase (without the particulate fraction) was not carcinogenic. In all valid studies in rats, diesel exhaust was found to be carcinogenic at particle concentrations of > 2 mg/m 3 , corresponding to an equivalent continuous exposure of about 1mg/m 3 . No effect was seen in hamsters or mice. In studies by intratracheal instillation, both diesel exhaust particles and carbon black induced tumours, and the surface area of the carbonaceous particles appeared to be correlated with the tumorigenic potency. Long-term inhalation of carbon black virtually devoid of PAHs at similar concentrations also resulted in lung tumours in rats.

It is not clear whether the carcinogenicity of diesel exhaust involves DNA-reactive or non-DNA-reactive mechanisms (or a combination). Various models have been used to elucidate the carcinogenicity of diesel exhaust.

B1.7 Effects on humans

Diesel exhaust contributes to air pollution in general. Although the role of diesel particles cannot be singled out in acute or chronic studies, they may be partly responsible for the range of health effects found to be associated with air pollution.

Typical diesel exhaust has a characteristic odour, which some people find offensive, particularly at high concentrations. The symptoms seen after acute and chronic exposure to diesel exhaust have been described in studies and anecdotal reports of occupationally exposed subjects. Acute exposure to diesel exhaust has been associated with irritation of the ocular and nasal mucous membranes, and an increased frequency of respiratory symptoms has been observed in occupational cohorts; however, the contribution of diesel particulates is not known. No consistent short-term effect on pulmonary functions has been found, but asthma attacks have been reported.

In a controlled study in which eight healthy non-smoking volunteers were exposed to diluted diesel exhaust in a chamber for 60 min, the phagocytosis rate of alveolar macrophages in broncho- alveolar lavage fluid was reduced.

Some cross-sectional and longitudinal studies on workers with long-term occupational exposure to diesel exhaust show decrements in lung function and an increased prevalence of respiratory symptoms, but these studies are limited by short exposure. Cohort studies in which deaths from cardiovascular and/or cerebrovascular disease due to diesel emissions were investigated did not show a significant excess.

The relationships between cancers of the lung and of the urinary bladder and occupational exposure to diesel exhaust have been evaluated in a number of epidemiological studies. Only those studies that were considered relevant for evaluating the carcinogenic effects of diesel exhaust are included in this monograph. The most relevant studies with regard to lung cancer are those of railroad workers, bus garage workers, and stevedores, who have definite exposure to diesel exhaust. The four most informative studies all reported an increased 

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Several case-control studies have been conducted to examine the relationship between urinary bladder cancer and presumed exposure to diesel exhaust. An increased risk was found, especially for truck drivers; however, all of these studies are limited by poor characterization of exposure. Hence, a causative association between exposure to diesel exhaust and an increased risk for urinary bladder
cancer cannot be established.

B1.8 Effects on other organisms in the laboratory and the field

The effect of diesel emissions as such have been addressed in only one study, of green algae.

B1.9 Evaluation of human health risks

The risk assessment paradigm of the United States National Academy of Sciences (US National Research Council, 1983) was used to assess the risks for both cancer and non-cancer end-points. The four steps in this process comprise: (1) hazard identification, (2) dose-response assessment, (3) exposure assessment, and (4) risk
characterization.

Epidemiological studies of long duration with well-defined exposure and follow-up (> 20 years) are considered to be the most informative. Four studies of lung cancer in occupationally exposed individuals met these criteria. The relative risks for lung cancer associated with exposure to diesel exhaust were generally low and were susceptible to chance, to the effects of unmeasured confounding factours, and to the difficulty in accurately adjusting for known confounding factors. Studies with less precise definitions of exposure support the conclusions of these studies. Overall, it is considered that diesel exhaust is probably carcinogenic to humans; however, no quantitative data are available for estimating human risk.

B1.9.1 Non-neoplastic effects

Two general approaches were used for risk characterization: a no-observed-adverse effect level (NOAEL) divided by an uncertainty factor; and a benchmark concentration. In both approaches, a sophisticated dosimetric model was used which decreases any uncertainty in interspecies extrapolation of dose.

The no-effect level of diesel exhaust particles in humans was calculated to be 0.139 mg/m 3 . The guidance value for the general population calculated from the dosimetric model was 5.6 μg/m 3 , and that calculated without the model was 2.3 μg/m 3

.

The benchmark concentration approach takes into account the entire exposure-response relationship rather than relying on a single data point from studies by inhalation, as in the NOAEL approach. Three sensitive end-points were identified: chronic alveolar inflammation, impaired lung clearance, and hyperplastic lung lesions. The benchmark concentrations, calculated from the same dosimetric model used in the NOAEL approach, were 0.9-2 μg/m 3 for inflammation, 1.2-3 μg/m 3 for impaired lung clearance, and 6.3-14 μg/m 3 for hyperplastic lesions.

B1.9.2 Neoplastic effects

A linearized multistage model was used to estimate unit risks due to exposure to diesel exhaust. Because the results of the available epidemiological studies were considered inadequate for a quantitative estimate of unit risk, data from several studies of long-term inhalation in rats were used in which carcinogenesis occurred at concentrations >2 mg/m 3 . A unit risk of 3.4 × 10-5 μg/m 3 (geometric mean of four risk estimates) diesel exhaust particles was calculated. An alternative biologically based model yielded a similar unit risk, under the assumption that diesel particles affect cell initiation and/or proliferation at low concentrations.

B1.10 Evaluation of effects on the environment

Insufficient information was available to evaluate the specific
effects of diesel exhaust emissions. The effects of combusted diesel
fuel should be similar to those of other fossil fuels and are related to the consumption of diesel fuel.

B2. IDENTITY AND ANALYTICAL METHODS

B2.1 Identity

Diesel engine emissions contain hundreds of chemical compounds, which are emitted partly in the gaseous phase and partly in the particulate phase of the exhaust. These substances form particles or contribute to the gaseous phase of the emissions, depending on vapour pressure, temperature, and the concentration of individual species. The amount and composition of diesel exhaust are related to engine conditions and fuel specifications (see sections B3.1.2.1 and B3.1.2.2). A total of 445 compounds has been identified or tentatively 

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Diesel exhaust has an offensive odour (odour threshold, 320 ppm). Compounds with a relative molecular mass > 80 mainly determine the odour (Scheepers & Bos, 1992b). Partridge et al. (1987) concluded that benzaldehyde and a methylbenzaldehyde isomer are the major substances responsible; unburnt aromatic hydrocarbons may also contribute. Aldehyde levels and odour emissions are directly correlated with the total hydrocarbon content of the exhaust; decreased hydrocarbon levels lead to diesel exhaust with less odour (Organisation for Economic Co-operation and Development, 1993).

B2.1.1 Chemical composition of diesel exhaust gases

The main gaseous products of diesel exhaust are carbon dioxide, oxygen, nitrogen, and water vapour; carbon monoxide, sulfur dioxide, nitrogen oxides, and hydrocarbons and their derivatives are also present. A representative composition of diesel exhaust gas (light-duty engine) is shown in Table 21.

The levels of carbon monoxide, nitrogen oxides, total hydrocarbons, and particulates emitted by diesel engines are regulated by law in a number of countries (see section B3.2). The composition of diesel exhaust gases is similar to that of gasoline engine gases, but because of the relatively higher fuel:air ratio, carbon monoxide and hydrocarbons occur in lower concentrations in diesel exhaust, and the emission of nitrogen oxides, particulate matter, and sulfur compounds is higher (the latter being due mainly to the higher sulfur content of diesel fuels). The relative concentrations in the aldehyde fraction are as follows: about 45% by weight formaldehyde, 17% by weight acetaldehyde, 14% by weight acetone and acrolein, < 10% by weight crotonaldehyde, propionaldehyde, isobutyraldehyde, benzaldehyde, and hexanaldehyde; methylethylketone is also present (Volkswagen AG,
1989).

Table 21. Composition of light-duty diesel engine exhaust

Component Concentration (% by weight)

Carbon dioxide 7.1 Water vapour 2.6 Oxygen 15.0 Nitrogen 75.2
Carbon monoxide 0.03 Hydrocarbons 0.0007 Nitrogen oxides 0.03 Hydrogen 0.002
Sulfur dioxide 0.01

Sulfates 0.00016 Aldehydes 0.0014
Ammonia 0.00005
Particulates 0.006

Adapted from Volkswagen AG (1989)

Diesel exhaust contains toxicologically relevant compounds such as benzene, toluene, ethylbenzene, xylene, and PAHs. In the hydrocarbon fraction, benzene and toluene are present in relative concentrations of 11 and 4% by weight, respectively (Volkswagen AG, 1989).

Possible sources of PAHs in diesel exhaust are unburnt PAHs from the fuel, pyrosynthesis of PAHs during combustion, crankcase oils, and engine and/or exhaust system deposits (Scheepers & Bos, 1992a). These constituents occur partly in the gaseous fraction of total hydrocarbons (benzene, toluene, ethylbenzene, and xylene components, PAHs of lower relative molecular mass) and partly in the particulate fraction due to adsorption (PAHs of higher relative molecular mass). The partitioning of constituents between the particulate and gas phases has been measured by several investigators (Hampton et al., 1983; Schuetzle, 1983; Schuetzle & Frazier, 1986). Most PAHs with five or more rings and aliphatic hydrocarbons with more than 18 carbon atoms are expected to be primarily adsorbed onto particles (Schuetzle, 1983). There is a direct relationship between adsorption behaviour and vapour pressure (Hampton et al., 1983); compounds with vapour pressures < 10-5 mbar (7.5 × 10 -6 mm Hg; 2.09 kPa) at room temperature occur predominantly in the particulate phase. An empirical relationship between boiling-point and particle- to gas-phase partition coefficients has been derived for PAHs and aliphatic hydrocarbons in order to estimate gas-phase emission rates (Schuetzle & Frazier, 1986). According to this equation, the concentration of e.g. anthracene in the gaseous phase is about 20 times higher than that in the particle phase, whereas the levels of e.g. pyrene and fluoranthene are nearly equal in the two phases. Oxygenated PAHs are found in diesel engine exhaust at concentrations as high (Volkswagen AG, 1989) or twice as high (Jensen & Hites, 1983) as those of their parent PAHs, perhaps because these engines have a larger air:fuel ratio than gasoline motors (Scheepers & Bos, 1992a). 9-Fluorenone and 9,10-anthracenequinone are especially abundant in diesel exhaust (Schuetzle & Frazier, 1986). In tests of diesel passenger cars by Volkswagen AG (1989), phenalene-1-one, naphthalene dicarboxylic acid anhydride, cyclopenta[ def ]phenanthrene-4-one, monoaldehydes of phenanthrene and anthracene, mono- and diketones of benzanthracenes and benzo[cd]pyrene, and benzofluorenones were also found (for artefact formation during sampling, see section B2.2.1; for emission 

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Nitrogen-substituted PAHs are formed during combustion as a result of either nitration of PAHs by nitric acid or addition of nitrogen monoxide or dioxide to PAH free radicals generated during combustion (Schuetzle, 1983). In the exhaust of a four-cylinder diesel engine, nitrofluorene, dinitrofluorenone, nitroanthracene, nitrofluoranthene, mono- and dinitropyrenes, and nitrobenzo[ a ]pyrene were detected at concentrations about one order of magnitude lower than those of oxygenated PAHs (Volkswagen AG, 1989).

A number of investigators have found nitrated PAHs in extracts of diesel engine exhaust particles (Handa et al., 1983; Hartung et al. 1984). Many three-, four-, and five-ring structures are involved (Schuetzle et al., 1982). The concentration of 1-nitropyrene, one of the prevalent species, has been reported to be 15-25 mg/kg particulate matter, whereas benzo[ a ]pyrene has been found at concentrations up to 50 mg/kg. The nitrated PAHs are important in determining the health effects of exposure to diesel exhaust, since they are effective mutagens in microbial and human cell systems (Pederson & Siak, 1981; Pitts et al., 1982; Patton et al., 1986). Some nitrated PAHs are carcinogenic to animals (Ohgaki et al., 1985; Imaida et al., 1991).

Combustion products originating from fuel and lubricating oil additives or from corrosion may also occur. For example, nitrate-based cetane improvers (see also section A2.1.2) are assumed to form nitro- PAHs (Organisation for Economic Co-operation and Development, 1993).

B2.1.2 Type and composition of emitted particulate matter

Diesel exhaust is characterized by a higher content (10-20-fold) of particulate matter than that of gasoline-fuelled passenger cars (Egebäck & Bertilsson, 1983; Volkswagen AG, 1989; Williams et al., 1989; CONCAWE, 1990b; Hammerle et al., 1994a). How soot particles are formed is still under discussion, but the precursor is probably acetylene generated during combustion. As a result of the numerous cracking, dehydration, and polymerization processes, quite large amounts of carbon, hydrogen, hydroxyl, and oxygen radicals are available in the combustion chamber, which can produce cyclic and polycyclic aromatic hydrocarbons from acetylene by polymerization and ring closure. With increasing polymerization, the carbon content of the macromolecule increases, finally generating graphite-like soot particles (Klingenberg et al., 1991).

Investigations of two four-cylinder engines with direct and indirect injection (see also section B3.1.2.1) showed that the emitted particles range in shape and size from spherical particles of < 0.01 μm to cluster and chain agglomerates with maximal dimensions of 30 μm (Dolan et al., 1980; Amann & Siegla, 1982; Klingenberg et al., 1991). The formation of agglomerates (coagulation, 'aging') is dependent not only on engine and exhaust conditions but also on environmental factors, such as photochemical processes and humidity. Between 50 and 80% of the total particulate emission occurs in the range of 0.02-0.5 μm (Israel et al., 1982). According to Amann & Siegla (1982), the emitted particles have a concentric, lamellar structure arranged around the centre, similar to the structure of

carbon black.

Emitted soot particles are composed of elemental carbon, adsorbed organic compounds (soluble organic fraction) from fuel and lubricating oil, traces of metallic compounds (iron, calcium, and zinc at < 0.6% by weight; Volkswagen AG, 1989), and sulfates. The organic components of emitted particles are distributed over the surface. Numerous data on the relative proportions of organic compounds are available (Horvath et al., 1987; Williams et al., 1987; Volkswagen AG, 1989; Williams et al., 1989; CONCAWE, 1992). Depending on the engine conditions and testing cycle, the organic compounds comprise 10-90% of the total particulate mass. The ranges of various constituents of particulate matter in nine light-duty and four heavy-duty vehicles (see also section B3.1.2.1) are shown in Table 22. The driving cycles used were combined European Union tests (ECE-15 and 'high speed' extra-urban driving cycle [EUDC]) for the light-duty vehicles and modified ECE-R49 tests for the heavy-duty engines (see also section B2.2.1); four fuels with different sulfur contents and an aromatic content of about 25% by volume were used (CONCAWE, 1992). (For a description of the influence of fuel composition on exhaust emissions, see section B3.1.2.2.)

This study showed considerable variation, and no obvious influence of fuel composition or engine conditions was discernible, although a difference was seen between fuel and lubricating oil hydrocarbons derived from light- and heavy-duty vehicles, respectively. This is probably due to the different driving cycles: transient in the case of light vehicles, steady state in the case of heavy-duty engines. In the individual data, adjustment of the engine appeared to play a major role. In the same study, PAHs were determined in the soluble organic fraction of the exhausts of three light-duty vehicles representing different engine conditions and of the four heavy-duty engines. The data are shown in Table 23. The constituents of the particulate matter of the exhaust are clearly dependent on the driving cycle used. Under steady-state conditions (heavy-duty engines), the PAH content is about one order of magnitude lower than that under transient conditions (light-duty vehicles). In addition,
transient conditions lead to considerable variations in levels. In the study of Volkswagen AG (1989) on a four-cylinder diesel engine in the US-72-test, a hot start was used, and higher levels of fluoranthene, pyrene, benzo[ ghi ]fluoranthene and benzo[ c ]phenanthrene were found, perhaps due to either engine condition or driving cycle. According to Volkswagen AG (1989), some of the PAHs, and especially pyrene, benz[ a ]anthracene and benzo[ a ]pyrene, seemed to decompose 

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Table 22. Particulate matter content of exhausts from light- and heavy-duty vehicles

Constitutent Particulate matter (% by weight) Reference

Light-duty vehicles a Heavy-duty vehicles a

Carbon 32.6-85.1 39.7-81.7 CONCAWE (1992) 82-88 (passenger 68-89 Williams et al.
cars) (1989) b

76-83 (light vans)

60-75 Hammerle et al.
(1994a) c

Fuel-derived 1.1-55.0 9.8-32.5 CONCAWE (1992) hydrocarbons 6.0-18 Hammerle et al.
(1994a) c

Lubricating 4.2-54.4 4.0-25.9 CONCAWE (1992)
oil-derived 5.7-15 Hammerle et al. hydrocarbons (1994a) c

Soluble organic 10.2-64.7 14.0-58.4 CONCAWE (1992) fraction 14-26 (passenger 22-90 Williams et al.

cars) (1989) b

36-48 (light vans) Sulfate (excluding 0.6-12.2 1.4-7.5 CONCAWE (1992) bound water) 3.3 (passenger 0.3-6.9 Williams et al.
cars) (1989) b

2.5 (light vans) 1.5-2.0 Hammerle et al.

(1994a) c

a According to the Organisation for Economic Co-operation and Development (1993), light-duty vehicles are generally classified as vehicles with a weight < 3.5 t, occasionally < 5 t; heavy-duty vehicles are those weighing > 5 t.

b Measurements for four passenger cars and 15 light vans in different Australian driving cycles corresponding roughly to US-FTP-72 and -75 tests for heavy-duty vehicles, and 10 trucks and two buses in one Australian urban traffic driving cycle

c Five passenger cars and light vans in European Motor Vehicle Emissions group cycle

and US-FTP-75 test

Table 23. Polycyclic aromatic hydrocarbon content of the soluble organic fraction of particulate matter in diesel exhaust

Constituent Soluble organic fraction Reference b (mg/kg particulate matter

Light-duty Heavy-duty

vehicles a vehicles

Fluoranthene 8-238 4-52 CONCAWE (1992) 553-560 Volkswagen AG (1989) Pyrene 11-287 7-90 CONCAWE (1992) 579-668 Volkswagen AG (1989) Benzo[ghi]fluoranthene 36-61 3-6 CONCAWE (1992) + benzo[c]phenanthrene 163-179 Volkswagen AG (1989) Benz[a]anthracene 9-285 1-16 CONCAWE (1992) 61-82 Volkswagen AG (1989) Triphenylene and 14-138 4-13 CONCAWE (1992) chrysene 117-138 Volkswagen AG (1989) Benzo[b,j]fluoranthene 4-87 1-10 CONCAWE (1992)

180-193 c Volkswagen AG (1989) Benzo[a]fluoranthen 6-69 CONCAWE (1992) Benzo[e]pyrene 8-40 1-2 CONCAWE (1992) 85-90 Volkswagen AG (1989) Benzo[a]pyrene 2-48 1-3 CONCAWE (1992) 38-54 Volkswagen AG (1989) Perylene 5-32 2 CONCAWE (1992) 4-8 Volkswagen AG (1989) Dibenz[a,j]anthracene 10-78 CONCAWE (1992) Indeno[123-cd]pyrene 6-54 CONCAWE (1992) 55-73 Volkswagen AG (1989) Dinbenz[a,h]- or 3-40 CONCAWE (1992) -[a,c]anthracene Benzo[ghi]perylene 25-65 1-5 CONCAWE (1992) 75-82 Volkswagen AG (1989) Anthracene 3-7 CONCAWE (1992) Dibenz[a,i]pyrene 8-31 CONCAWE (1992) Coronene and 9-52 CONCAWE (1992) dibenzo[a,e]pyrene 33-48 d Volkswagen AG (1989)

a According to the Organisation for Economic Co-operation and Development (1993), light-duty vehicles are generally classified as vehicles with a weight < 3.5 t, occasionally < 5 t; heavy-duty vehicles are those weighing > 5 t.

b From Volkswagen AG (1989): US-72 tests with one four-cylinder engine

c Including benzo[k]fluoranthene

d Excluding dibenz[a,e]pyrene 

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B2.2.1 Sampling

Driving cycles are specified to simulate on-road conditions for measuring vehicle emissions (Grimmer et al., 1988). The test procedures can be divided in two basic types: steady-state and transient cycle tests. In the European Union and Japan, mainly steady-state tests are used, in which emissions from engines are measured at specified combinations of speed and load. The most widely used official tests for light- and heavy-duty vehicles are as follows:

Light-duty vehicles:

* US FTP-75: transient cycle; cold start; driving distance, 17.8 km; 10-min break after 28.7 min (United States) (Volkswagen AG, 1989)

* US SET: transient cycle; 'sulfate emission test'; hot start; simulating rush-hour freeway driving; driving distance, 21.7 km (United States) (Volkswagen AG, 1989)

* ECE 15: transient cycle; cold start; simulating city-centre driving; driving distance, 4.052 km (European Union) (CONCAWE, 1990a)

* US HDC: steady-state cycle; 'highway driving cycle'; hot start; driving distance, 16.5 km; maximal speed, 96.4 km/h (United States) (Volkswagen AG, 1989)

* EUDC: steady-state cycle; 'extra-urban driving cycle'; hot start (after ECE 15); driving distance, 6.755 km; maximal speed, 120km/h (European Union) (CONCAWE, 1990a)

Heavy-duty vehicles:

* HD-FTP: transient cycle; 'heavy-duty federal test procedure'; simulating metropolitan driving (United States) (Organisation for Economic Co-operation and Development, 1993);

* ECE 13: steady-state cycle at 13 different modes; identical to ECE R49 (European Union) (Organisation for Economic Co-operation and Development, 1993). Various bus cycles are also used to test heavy-duty vehicles that are characterized by frequent stop-and-go driving (Westerholm & Egebäck, 1991; Prakash et al.,
1992).

It is difficult to avoid artefacts in sampling. Exhaust gas constituents can undergo chemical reactions, adsorption and desorption processes, and condensation or diffusion. For example, oxygenated and nitrated PAHs are chemically unstable, and artefacts may form on the sampling device, especially if large sampling volumes are needed (Lee & Schuetzle, 1983). Both decreases and increases in the concentrations of PAHs and oxy-PAHs were observed during sampling in a study by Volkswagen AG (1989); in particular, degradation of pyrene and nitropyrene and formation of naphthalene dicarboxylic acid anhydride on the filter occurred. PAHs can be converted into nitroarenes (Pitts et al., 1978; Lee et al., 1980; Gibson et al., 1981; Lee & Schuetzle, 1983; Schuetzle & Perez, 1983; Gaddo et al., 1984; Levsen et al., 1988).

The sampling techniques used for the collection of the gaseous and particulate phases of diesel exhaust comprise dilution tube sampling and raw gas sampling.

B2.2.1.1 Sampling from undiluted exhaust gas (raw gas sampling)

The device for sampling motor vehicle exhaust (total flow method; Kraft & Lies, 1981; Grimmer et al., 1973) consists of a glass cooler, a condensate separator, and a special micron filter of paraffin-impregnated glass-fibre material providing separation of > 99.9% of particles of 0.3-0.5 μm. The cooler is fed with cold tap water, and the filter is placed on top of the cooler to remove any uncondensed liquid or solid exhaust gas constituents that may pass through the cooler coil. The particulate filter retains undissolved particles of the exhaust gas, while the condensate consists of the dissolved components (Volkswagen AG, 1989).

The US Environmental Protection Agency (Lies et al., 1986) defines the 'particulate phase' as all materials (with the exception of condensed water) which, at a temperature of < 51.7°C, are retained on a special filter after dilution with air. Large volumes are collected with this type of sampling, making analysis of trace components difficult. The sample volumes are reduced by diverting the exhaust emission flow and sampling only a part. It is difficult to ensure a representative partial flow, as the amount of exhaust gas produced varies considerably over time (Volkswagen AG, 1989).

B2.2.1.2 Sampling from diluted exhaust (dilution tube sampling)

The exhaust is mixed with ambient air in a dilution tunnel in a constant volume sampler method (Behn et al., 1985). The total flow of air and exhaust is kept constant with a suitable fan in a constant volume sampler unit. This sampling technique more realistically

reflects the immediate dilution of exhaust leaving the tailpipe of vehicles on the road. The concentrations of exhaust components are, however, reduced in accordance with the degree of dilution (magnitude, 1:10), and components that are present only at low concentrations 

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There are various techniques for sampling gaseous constituents. Substances present at sufficiently high concentrations are commonly sampled in a plastic bag; adsorption onto solid materials, consisting of extremely porous, tiny plastic pellets (e.g. Tenax; diameter, 0.1-0.2 mm; specific surface area, 20-50 m 2 /g), or absorption in liquids are also used. Particulate constituents are sampled by filtration (Lee & Schuetzle, 1983; Volkswagen AG, 1989).

B2.2.2 Extraction from particles

PAHs adsorbed onto diesel exhaust particles are of toxicological relevance, and there are numerous methods for extracting these constituents. The most frequently used method is Soxhlet extraction, which permits extraction of 99% of the soluble organic fraction within 8 h; an additional 16 h are necessary to extract the remaining 1%. Extraction is frequently carried out by sequentially changing the solvents, from polar to nonpolar, e.g. toluene, toluene or n- propanol, dichloromethane, and acetonitrile. PAHs may be further extracted by refluxing the exhaust particulates in solvent for 1 h. Sublimation and extraction by ultrasonic agitation are also used (Lee & Schuetzle, 1983; Levsen, 1988), and supercritical fluids such as carbon dioxide are effective (Hawthorne & Miller, 1986). The extraction efficiency of various solvents is still under discussion (Köhler & Eichhoff, 1967; Swarin & Williams, 1980; Schuetzle & Perez, 1981; Grimmer et al., 1982; Lee & Schuetzle, 1983; Schuetzle et al., 1985; Levsen, 1988).
Aromatic solvents and their combinations seem to be best suited for extracting PAHs from diesel particulates (Levsen, 1988).

B2.2.3 Clean-up and fractionation

Complex fractionation schemes for separating the exhaust extract into aliphatic compounds, aromatic compounds, and moderately and highly polar fractions after the removal of acidic and basic fractions have been described by Petersen & Chuang (1982) and Levsen (1992). During fractionation, further artefacts may be produced by the components of a complex sampling mixture like diesel exhaust, especially with acidic and basic extraction and aggressive column separation procedures.

B2.2.4 Chemical analysis

Chemical analysis of separated, cleaned compounds of diesel exhaust involves numerous techniques, including thin-layer chromatography, gas chromatography, gas chromatography with mass spectrometric detection, and high-performance liquid chromatography (US National Institute for Occupational Safety and Health, 1991). Jinno et al. (1986) determined PAHs by supercritical fluid chromatography with a photodiode array detector.

The detection methods vary widely and include ultra-violet, visible, and fluorescence spectrophotometry, mass spectrometry, and more or less specific devices, such as flame ionization, nitrogen flame ionization, and sulfur-specific and electron-capture detectors (US National Institute for Occupational Safety and Health, 1991). A thermal optical method for determining elemental carbon is currently being used as a surrogate for measurement of worker exposure in almost all internal investigations at the US National Institute for Occupational Safety and Health (Niemeier, personal communication, 1994).

Analytical methods are available for determining the organic constituents of diesel particulate matter, with special emphasis on the determination of PAHs and their nitrated and oxygenated derivatives (Levsen, 1988).

B2.3 Conversion factors

Because diesel exhaust emissions are complex mixtures of different gases and particles, it is impossible to give a conversion factor for converting parts per million to SI units.

B3. SOURCES OF HUMAN AND ENVIRONMENTAL EXPOSURE

B3.1 Anthropogenic sources

Diesel exhaust emissions are exclusively man-made and are distributed primarily in the atmosphere. The hydrosphere and geosphere may also be affected owing to dry and wet deposition of the exhaust constituents; however, determination of the source of deposited compounds is difficult. No data are available on the individual components of diesel exhaust and their deposition.

B3.1.1 Diesel exhaust emissions

Diesel fuels are widely used as transport fuels, and exhaust emissions are released mainly by road traffic. Although many investigations of these emissions have been reported, the individual results are frequently not comparable because the experimental parameters, e.g. driving cycles, are different, or different dimensions were chosen. For example, the results of tests on emissions

from light-duty vehicles refer mainly to kilometres driven, whereas those for heavy-duty vehicles are related to the engine power in kilowatt-hours. Furthermore, the statistical power of studies of emission factors is often limited by the small number of vehicles or engines examined. Some investigators change more than one parameter (e.g. sulfur content or aromaticity of the fuel) during their studies, so that the specific cause of changes in exhaust composition is 

# Pa g e 57 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmdifficult to assess. Adjustment of individual engine and fuel parameters plays a major role in determining the quality and quantity of exhaust emissions (see sections B3.1.2.1 and B3.1.2.2).

Further sources of diesel exhaust emissions include off-road and stationary engines (reciprocating or gas turbine), electricity generating plants, pipeline pumping, gas compression, commercial and industrial furnace installations, and space and water heaters in plants and factories (see section A3.2.1.2). Diesel exhaust is also emitted from marine and inland ships' engines; residual fuels and other heavy grades (for example Bunker C) are used in ocean shipping (von Meyerinck, personal communication, 1994).

B3.1.1.1 Emission of chemical constituents with the gaseous portion
of diesel exhaust

The exhaust emissions from diesel-fuelled vehicles are generally given as milligrams of component emitted per kilometre of driving distance (Egebäck & Bertilsson, 1983; Westerholm et al., 1986; Westerholm, 1987; Volkswagen AG, 1989; CONCAWE, 1990b; Westerholm & Egebäck, 1991; Prakash et al., 1992; Scheepers & Bos, 1992a). Data for verifying tests are issued by national regulatory bodies, e.g. the German Federal Office for Motor Vehicles (1993, 1994). Depending on the testing cycle, diesel engine, and fuel (see sections B3.1.1.1 and B3.1.1.2), the data show considerable differences even though the values are of the same order of magnitude. Emission factors used in different regions to estimate the effects of regulated chemical constituents (carbon monoxide, nitrogen oxides, and hydrocarbons) in
air are shown in Table 24.

Like all hydrocarbon fuel-burning engines, diesel engines release significant amounts of carbon dioxide. Westerholm & Egebäck (1991) measured emission factors of about 1 kg/km from a bus and a truck in the US transient cycle and the bus cycle. In measurements of emissions from six buses in four driving cycles (US transient cycle and three bus cycles), Prakash et al. (1992) found emission factors up to about 4 kg/km, especially in the bus cycles. For five European diesel passenger cars and one light van, Hammerle et al. (1994a) determined emission factors of 162-252 g/km carbon dioxide in the European Motor Vehicle Emissions cycle and the US FTP-75 test. Diesel engines are the most efficient of all common types of combustion engines, with peak thermal efficiencies typically > 40%, owing to the relatively large air:fuel ratio during combustion (see section B3.1.2.1) (Organisation for Economic Co-operation and Development, 1993).

Table 24. Emission factors for diesel vehicles in 1985 used as standards for estimating total impact

Region Passenger cars Commercial vehicles

g/km g/kg Average < 5 t > 5 t

g/km g/kg g/km g/kg g/km g/kg

Carbon monoxide

Denmark 1.0 2.0

Germany a 1.5 20 16

Netherlands 2.7 35 3.9 14 2.0 20 4.6 14 European Union 19 19 16
Austria 22.8 22.8 20

Sweden 10.5
Switzerland 4.0

United States b 3.1-6.2

Nitrogen oxides

Germany a 1.0 14 58

Netherlands 0.8 10 18.3 66 0.6 6 22.8 69 European Union 14 18 62

Sweden 39
Switzerland 14.0

United States b 3.1

Hydrocarbons
Germany a 0.5 7 12

Netherlands 0.6 10 4.6 17 0.9 9 6.3 19

Table 24 (contd)

Region Passenger cars Commercial vehicles

g/km g/kg Average < 5 t > 5 t

g/km g/kg g/km g/kg g/km g/kg

Hydrocarbons (contd) European Union 10 13 9

Sweden 3.9
Switzerland 3.0

United States b 1.2

Adapted from Organisation for Economic Co-operation and Development (1993)

a Excluding former German Democratic Republic

b 1975-83, trucks tested in the US transient cycle Cyanides, ammonia, and sulfur dioxide are detectable in the 

# Pa g e 58 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmgaseous portion of diesel exhaust. The average emission factors given by Volkswagen AG (1989) for seven passenger diesel cars with four- and five-cylinder engines in three driving cycles (US 75 test with cold start, US sulfate emission test, US highway driving test) were 0.6 mg/km cyanides, 1.1 mg/km ammonia, and 233.0 mg/km sulfur dioxide.

The emission of individual organic components and classes of compounds, summarized as total hydrocarbons, are shown in Table 25, which provides an overview of the quantities of individual substances emitted in diesel exhausts. Heavy-duty vehicles release more low-boiling hydrocarbons than passenger cars; however, the difference could be due to the driving cycle used, which involves frequent changes of speed and load (see section B2.2.1). The emissions of adsorbed organic components, such as PAHs (see section B3.1.1.2), are correlated with the release of particulate matter.

B3.1.1.2 Emission of particulate matter and adsorbed components in diesel exhaust gases

The emission of total particulate matter from diesel exhaust is shown in Table 26. Analysis of the particulate matter released from heavy-duty vehicles gave the following emission factors (Westerholm & Egebäck, 1991): 170-370 mg/km carbon, 8-71 mg/km soluble organic compounds derived from fuel, 56-167 mg/km soluble organic compounds derived from lubricating oil, 1.4-37 mg/km soluble sulfates, and 0.6-3.7 mg/km nitrate. Comparable values were not available for light-duty vehicles.

As PAHs and oxygenated PAHs from diesel and spark-ignition engines are qualitatively similar, their differentiation in the environment is difficult (Behymer & Hites, 1984); however, benzo[ b ]naphtho-[2,1-d]thiophene is specific for diesel exhaust and can therefore be used as a marker (Grimmer et al., 1979). Emissions of PAH components are shown in Table 27.

In experiments with one bus and one truck, Westerholm & Egebäck (1991) detected 30-220 μg/km of particulate-associated PAHs. The compounds measured included dibenzothiophene and its 4- and 3-methyl derivatives, 2-methylfluorene, phenanthrene and its methylated derivatives, anthracene and its 2-methyl derivative, fluoranthene, pyrene and its 1- and 2-methyl derivatives, retene, benzo[ a ]fluorene, benzo[ ghi ]fluoranthene, cyclopenta[ cd ]pyrene, benzo[ a ]anthracene, chrysene or triphenylene, benzo[ b,k ]fluoranthene, benzo[ e ]pyrene, benzo[ a ]pyrene, perylene, indeno[1,2,3-cd]fluoranthene, indeno[1,2,3-cd]pyrene, picene, benzo[ ghi ]perylene, and coronene. The emission factors for 1-nitropyrene were 0.13-7.3 μg/km.

Table 25. Emission factors for individual organic constituents and classes of compounds in the gaseous portion of diesel

exhaust

Class of compound Emission factor (mg/km)
or constituent

Light-duty vehicles a Heavy-duty vehicles a

**Straight-chain aliphatic hydrocarbons**

Sum of C 1 -C 4 5.3-6.6 (C 1 + C 2 ) b

< 1 - < 143 c

Sum of > C 4 < 1-5 c

Olefins Ethylene 25.6-90.7 d 32-135 e

13.5-16.9 b 19.0-82.9 f

< 1-31 c Propylene 7.3-22 e

8.1-44.0 f
1,3-Butadiene 0.2 g Acetylene 5.7-7.4 b 27.9-106.9 (+ ethane) f

Alcohols Methanol 1.6 d

Ethanol < 1.2-22.2 d
Aldehydes Formaldehyde 8.1 b 60-255 e

7.9-20.7 d 140-150 h

4.2-9.1 g 19.0-207.4 f*
Acetaldehyde 3.1 b 18-137 e

< 22-36 d 40-50 h

2.0-4.4 g Crotonaldehyde 1.2 b

Acrolein < 1-1.8 d 8.5-36 e

4 h
Benzaldehyde 0.6 b 10-14 h

**Light aromatic compounds**

Benzene 2.6-4.3 b 4.5-13.1 e

ND-6 c 6.9-30.5 f

9.5-18.1 d

0.15-3.47 g

Table 25. (cont'd)

Class of compound Emission factor (mg/km)
or constituent 

# Pa g e 59 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmLight-duty vehicles a Heavy-duty vehicles a

Toluene 1.1-1.9 b 2-13.7 e

ND-2 c 7.4-32.5 f

< 1.0 d
Ethylbenzene ND c Xylenes ND c 7.8-41.7 f

Phenols 0.7-1.5 b

ND, not determined

a According to the Organisation for Economic Co-operation and Development (1993), light-duty vehicles are generally classified as vehicles with a weight < 3.5 t, occasionally < 5 t; heavy-duty vehicles are those weighing > 5 t.

b From Volkswagen AG (1989): various numbers of diesel passenger cars in three driving cycles (US-75 test with cold start, US sulfate emission test, US highway driving test); determination of aldehydes only in the US-75 test

c From Scheepers & Bos (1992a): two diesel passenger cars (< 50 000 and > 100 000 km on odometer) in ECE test, 15 min at 90 km/h and 15 min
at 120 km/h

d From Egebäck & Bertilsson (1983): two passenger cars in US-78 test

e From Westerholm & Egebäck (1991): a bus and a truck in bus cycle with
different diesel fuels

f From Prakash et al. (1992): one bus in four driving cycles (US transient cycle, three bus cycles) with the exception (*) of eight buses in the same driving cycles

g From Hammerle et al. (1994a): five passenger cars and one light van in European Motor Vehicle Emissions Group cycle

h From Westerholm et al. (1986): one heavy-duty vehicle on a chassis dynamometer with two diesel fuels

i Mainly phenols, cresols and xylenols

Table 26. Emission factors for total particulate matter from diesel exhaust

Measurement conditions Emission factor (mg/km) Reference

Light-duty vehicles a

Seven passenger cars in three driving cycles 166.4-238.7 Volkswagen AG (1989) (US-FTP-75 test with cold start; US sulfate emission test; US highway driving test)

Four passenger cars, 15 light vans in three 180-250 (passenger cars) Williams et al. (1989) Australian driving cycles (corresponding roughly to 360-440 (light vans) US-72 and US-75 tests)

Seven passenger cars, two light vans in combined 70-365 (passenger cars) CONCAWE (1990b) ECE 15/EUDC test with four diesel fuels 233-609 (light vans)

Various Germany b , 300 Organisation for Economic Netherlands, 600 Co-operation and Development Sweden, 400 (1993)

61 passenger cars and 30 light vans without catalyst 39-188 (mean, 104) Federal Office for Motor Vehicles (optimally tuned) incombined ECE 15/EUDC test (1993)

73 passenger cars and 45 light vans with catalyst 48-127 (mean, 90) Federal Office for Motor Vehicles (optimally tuned) in combined ECE 15/EUDC test (1993)

Table 26 (contd)

Measurement conditions Emission factor (mg/km) Reference

44 passenger cars and 8 light vans without catalyst 52-117 (mean, 95) Federal Office for Motor Vehicles (optimally tuned) inUS FTP-75 test (1993)

118 passenger cars and 34 light vans with catalyst 50-118 (mean, 71) Federal Office for Motor Vehicles (optimally tuned) inUS FTP-75 test (1993)

Five passenger cars and 1 light van in European Motor 52-117 Hammerle et al. (1994a) Vehicle Emissions Group cycle and US FTP-75 test

Heavy-duty vehiclesa

One vehicle on chassis dynamometer (bus cycle); 680-1000 Westerholm et al. (1986)
two diesel fuels with different contents of sulfur and aromatic compounds

10 trucks, 2 buses in one Australian driving cycle 800-7150 Williams et al. (1989) 

# Pa g e 60 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmsimulating urban traffic

One bus, one truck in two driving cycles (bus cycle; 230-600 Westerholm & Egebäck (1991) US transient cycle) with two diesel fuels

106 trucks and buses in ECE-R49 driving cycle 0.056-0.606 g/kWh Federal Office for Motor Vehicles
(1994)

Table 26 (contd)

Measurement conditions Emission factor (mg/km) Reference

Various Netherlands, 800 (< 5 t) Organisation for Economic 3300 (> 5 t) Co-operation and Development Sweden, 400 (< 5 t) (1993) 1700 (> 5 t)

13 trucks and buses (three with particulate trap, 584-804 (without trap) Lowenthal et al. (1994) 10 without) in bus cycle with two diesel fuels 67-206 (with trap)

a According to the Organisation for Economic Co-operation and Development (1993), light-duty vehicles are generally classified as vehicles with a weight < 3.5 t, occasionally < 5 t; heavy-duty vehicles are those weighing > 5 t.

b Without the former German Democratic Republic Table 27. Emission factors for polycyclic aromatic hydrocarbons in
diesel exhaust

Constituent Emission factor (μg/km)

Light-duty vehicles a Heavy-duty vehicles a

Anthracene 17-63 b 0.9-3.2 c ; 9-26 d ;

3-14 d+ ; 1.6 e

Phenanthrene 140 f 4.2-25 c

295-524 b 163-308 d ; 79-186 d+ ;

12.2 e

Fluoranthene 172 f 10-39 c

58-200 g 27-34 d ; 14-17 d+

60.4-79.7 g 13.0 e Pyrene 186 15-102 c

< 0.9-22 28-31 d ; 9-21 d+

66.6-87.1 22.6 e Chrysene 40 9.5-26 c

14-67 8-32 d ; 4-12 d+

10.7-22.3 Not measured e Benzo[b,k]fluoranthene 40 1.6-8.4 c

2.6-47 9-12 d,h ; 2-8 d,h+

10.3-15.2 5.6 e Benzo[ghi]fluoranthene 40 f 1.4-13 c ; 6.9 e Cyclopenta[cd]pyrene 2 f 2.5-11 c

0.81 ag * 1.4 e Benzo[e]pyrene 3-38 b 1.6-23 c

13.8-16.9 5-9 d ; 1-7 d+ ; 2.6 e Benzo[a]pyrene 10 0.8-12 c

< 1-19 2-3 d ; 0.4-2 d+

4.0-5.3 1.3 e
Benzo[a]anthracene 25 3.0-12 c

8-43 5-12 d ; 1-4 d+

2.7-3.9 3.6 e Indeno[123-cd]pyrene 15 f 0.7-15 c

3.5-4.4 g Benzo[ghi]perylene 20 0.6-30 c

< 1-18 2-4 d ; 0.5-1 d+

8.7-11.7 Not detected (detection limit not given)e

Table 27 (contd)

Constituent Emission factor (μg/km)

Light-duty vehicles a Heavy-duty vehicles a

Perylene < 1-2 b Not measured c

0.5-0.6 g 1.0 e

Coronene 10 f < 0.1-12 c

5.43 g * Not detected (detection limit not given) e

a According to the Organisation for Economic Co-operation and Development (1993), light-duty vehicles are generally classified as vehicles with a weight < 3.5 t, occasionally < 5 t; heavy-duty vehicles are those weighing > 5 t.

b From Scheepers & Bos (1992a): two diesel passenger cars (< 50 000 and > 100 000 km on odometer) in ECE test 

# Pa g e 61 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmc From Westerholm et al. (1986): one heavy-duty vehicle on a chassis dynamometer (bus cycle)

d From Lowenthal et al. (1994): 13 trucks and buses (three with particulate trap [d+], 10 without) in bus cycle

e From Rogge et al. (1993): two heavy-duty vehicles in special driving cycle

f From Egebäck & Bertilsson (1983): average of two passenger cars in

US-78 test

g From Volkswagen AG (1989): seven passenger cars in three driving cycles (US-75 test with cold start, US sulfate emission test, US highway driving test) with the exception (*) of one four-cylinder engine, US-72 test with hot start

h Isomers not specified

The emission of oxygenated PAHs by two diesel passenger cars (one four-cylinder and one five-cylinder engine, US 72 test with hot start) was 4-40 μg/km. In a comparable test (one four-cylinder engine, same driving cycle), the emission of nitro-PAHs was 0.1-3.5 μg/km. As a result of possible artefact formation during sampling (see section B2.2.1), these measurements are uncertain (Volkswagen AG, 1989). It is suspected that nitrate-based cetane improvers (see section A2.1.2) could increase nitro-PAH emissions in exhaust (Organisation for Economic Co-operation and Development, 1993).

There are no significant differences in the emissions of adsorbed single substances between light- and heavy-duty vehicles, although heavy-duty vehicles release larger relative amounts of particulate matter (Salmeen et al., 1985).

The chlorine content of diesel fuel is typically < 1 ppm (Oehme et al., 1991). Thus, no dioxins were found in the exhaust of heavy vehicles, at a detection limit of 100 pg/litre of fuel (Marklund et al., 1990). The dioxin emissions from one light-duty (indirect injection) and one heavy-duty (direct injection) diesel engine under different load conditions on a chassis dynamometer were 0.009-0.141 ng of toxicity equivalents per litre of fuel consumed. The highest value was reported in emissions from the light-duty engine at full load. Overall, the emissions were of the same order of magnitude as the dioxin emissions from a petrol-fuelled engine with a catalytic converter (Schwind et al., 1991).

B3.1.2 Parameters that influence diesel exhaust emissions

B3.1.2.1 Engine conditions

In general, diesel engines are operated with excess air; the air:fuel ratio is about 1.5, and at lower air:fuel ratios, much more smoke is emitted (Organisation for Economic Co-operation and Development, 1993). Wide local variations in the equivalence ratio and the temperature of the mixture are caused by the heterogeneous nature of the combustion process. To avoid over-rich mixtures and to improve the air:fuel mixing processes, wall wetting and delay of fuel injection are used (CONCAWE, 1986). In naturally aspirated engines, the amount of air in the cylinder is independent of the power output. For these engines, the maximal power output is limited by the amount of fuel that can be injected without smoke formation. In turbocharged engines, however, increasing the amount of fuel injected increases the energy in the exhaust gas, and the turbocharger pumps more air into the combustion chamber. For this reason, the power output of turbocharged engines is not, in general, limited by smoke. Low air:fuel ratios can occur only during transient acceleration. Most turbocharged engines are further equipped with a cooling system (intercooler) in which compressed air reduces adverse thermal effects. In both types of engines, fuel is injected either directly into the combustion chamber or indirectly into a separate pre-chamber where it is mixed and partly burnt before reaching the main combustion chamber (Organisation for Economic Co-operation and Development, 1993). Scheepers & Bos (1992a) reported that most heavy-duty vehicles have direct injection motors, and indirect injection is used for light-duty
vehicles.

The most important influences of engine conditions on diesel exhaust emissions are summarized in Table 28. Contrasting results are sometimes obtained with regard to the effect of engine conditions on diesel exhaust emissions. Adjustment of the engine plays a major role (CONCAWE, 1990b), which was slightly less in the US FTP-75 test cycle than in the European cycle.

Table 28. Influence of engine conditions on diesel exhaust emissions

Condition Change Influence on exhaust emissions Reference

Air:fuel ratio Increase Increased emissions of hydrocarbons and particulate soluble Organisation for Econ omic fraction; decreased particulate and nitrogen oxide emissions Co-operation and from turbocharged and intercooled engines Development (1993)

Increased PAH emissions, decreased particulate emissions from Barbella et al. (1988 ) direct injection engines; decreased PAH and particulate emissions from indirect injection engines

Increased nitrogen oxide emissions, decreased particulate Iida et al. (1986) emissions from direct injection engines

Injection timing Advance Increased particulate emissions from indirect injection engines Du et al. (1984); Wil liams et al. (1989)

Decreased particulate and hydrocarbon emissions; increased Organisation for Econ omic 

# Pa g e 62 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmnitrogen oxide emissions Co-operation and Deve lopment (1993 )

No influence on concentrations of PAHs and oxy-PAHs adsorbed Jensen & Hites (1983) on particulate matter

Delay Decreased nitrogen oxide emissions, increased volumetric fuel Organisation for Econ omic consumption Co-operation and Deve lopment (1993 )

Table 28. Influence of engine conditions on diesel exhaust emissions

Condition Change Influence on exhaust emissions Reference

Injection timing Delay Decreased particulate emissions from indirect injection engines; Williams et al. (1989 ) (contd) increased particulate emissions from direct injection engines Small effect on PAH emissions from indirect injection engines Schuetzle & Frazier ( 1986)

Small effect on particulate emissions from direct injection engines Campbell et al. (1981 a)

No influence on concentrations of PAHs and oxy-PAHs adsorbed Jensen & Hites (1983) on particulate matter

Load and Increase Increased particulate and PAH emissions at high loads; decreased Pipho et al. (1986) temperature particulate emissions at very high load from indirect injection engines

Decreased concentrations of PAHs and oxy-PAHs adsorbed on Jensen & Hites (1983) particulate matter

Decrease Decreased particulate and nitrogen oxide emissions; extremely Organisation for Econ omic cold charge air may increase hydrocarbon emissions Co-operation and Development (1993)

Lowest PAH emissions at medium load; enrichment of three- Pipho et al. (1986) and four-ring PAHs during idling (indirect injection engines)

PAH, polycyclic aromatic hydrocarbons (a) Engine aging

Increased emissions of hydrocarbons, particulates, and PAHs have been reported from older, more intensively used light- and heavy-duty vehicle engines in comparison with newer ones (Stenberg et al., 1983; Braddock & Perry, 1986; Scheepers & Bos, 1992a). Stenberg et al. (1983) proposed that the combustion of more lubricating oil in older vehicles contributes to the enhanced emissions. Prakash et al. (1992) measured the emission characteristics of several buses and reported that a properly maintained engine would not emit significantly more hydrocarbons and nitrogen oxides during its lifetime, whereas carbon monoxide and particulate emissions could increase with age. These findings are similar to those of Volkswagen AG (1989) on the influence of distance travelled on exhaust emissions from one diesel passenger car over a total distance of about 30 000 km in three driving cycles (US FTP test, US sulfate emission test, US highway driving test). The average deterioration factors, relative to the emissions at about 2500 km, were carbon monoxide, 0.97; nitrogen oxides, 0.96; hydrocarbons, 0.91; particulates, 1.07; fluoranthene, 0.96; pyrene, 1.06; chrysene, 0.62; benzo[ b,k ]fluoranthene, 1.11; benzo[ e ]pyrene, 1.51; benzo[ a ]pyrene, 0.68; perylene, 0.96; indeno[1,2,3-cd]pyrene, 2.91; and benzo[ ghi ]perylene, 3.43.

(b) Lubricating oil

As a result of emission control techniques such as turbocharging, intercooling, and injection timing, the relative importance of emissions caused by lubricating oils has increased (Organisation for Economic Co-operation and Development, 1993). Lubricating oils have been hypothesized to be a sink for incomplete combustion products such as PAHs and their derivatives, which are scrubbed from the exhaust gas. When this 'enriched' lubricating oil leaks into the combustion chamber, the constituents may be emitted in the exhaust (Scheepers & Bos, 1992a; see section B2.1.1). McKee & Plutnick (1989) found, however, that carcinogenic PAHs accumulate in gasoline- but not in diesel-fuelled engine oils.

Oil consumption can be reduced by improving engine manufacturing specifications and engine seals (Organisation for Economic Co-operation and Development, 1993). The particle-bound hydrocarbons derived from lubricating oil contribute up to about 50% by weight in light-duty vehicles and to a maximum of about 26% by weight of the total emitted particulate matter (see section B2.1.2, Table 22). Experiments with 13 C radiolabelled lubricating oil in three light-duty vehicles showed that addition of up to 3% lubricating oil increases the emitted particulate matter by about 50%, and the particulates consist mainly of the soluble organic fraction adsorbed onto the particles (Williams et al., 1989).

B3.1.2.2 Fuel specification

Fuel parameters influence the particulate emissions of diesel engines but seem to be less important than other parameters like engine adjustment, load, and maintenance (Weidmann et al., 1988). The actual trend in legislation of the quality of diesel fuel, i.e. to a lower sulfur and aromatic content, will improve emissions, especially of particles. In addition, the more stringent emission standards in many countries will decrease diesel-related emissions.

(a) Sulfur content 

# Pa g e 63 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmThere have been numerous investigations on the influence of sulfur content in diesel fuel on the release of particulate matter (CONCAWE, 1990b; Van Beckhoven, 1991; Westerholm & Egebäck, 1991; Scheepers & Bos, 1992a). The sulfur content is presumed to be directly correlated with the release of particulate matter. About 3-5% of the fuel sulfur in light-duty vehicles and 2-3% of that in heavy-duty
vehicles reacts with metal sulfates which are adsorbed onto the particulate matter, increasing its mass and its hygroscopic properties, as metal sulfates tend to absorb significant amounts of water from the atmosphere (Organisation for Economic Co-operation and Development, 1993).

A reduced sulfur content leads to decreased levels of particulates in the exhaust of light-duty vehicles. For example, investigations on seven passenger cars and two light vans in a combined ECE 15/EUDC testing cycle with four diesel fuels of different sulfur contents (0.055, 0.12, 0.22, and 0.31% by weight) showed a statistically significant reduction in the emission of particulates up to a maximum of 7.4%. The investigators noted variations in particulate levels between the different vehicles, however, that were about two orders of magnitude higher than could be attributed directly to the influence of the sulfur content (CONCAWE, 1990b).

Reduction of the emission rates of heavy-duty vehicles by lowering the fuel sulfur content is still under discussion. In a study of four heavy-duty engines in the ECE R49-13 mode driving cycle with the same four fuels as described above, no statistically significant reduction in emitted particulate matte was seen (CONCAWE, 1990b). In experiments with one heavy-duty vehicle on a chassis dynamometer bus cycle with two different fuels, however, a considerable reduction in the release of particles was found (Westerholm et al., 1986). This result was confirmed by Westerholm & Egebäck (1991) who investigated one bus and one truck in three driving cycles (ECE 13 test, bus cycle, US transient test) with eight diesel fuels of different sulfur and aromatic contents. Comparable results were found by Gairing et al. (1994) in tests with one heavy-duty engine (direct injection) in ECE R49 with 12 different fuels. Adjustment of the engines again appeared
to be a decisive factor.

(b) Aromaticity (density)

Aromaticity can be directly associated with density, as a higher density is frequently achieved by a larger content of aromatic compounds. Obviously, increasing the aromaticity leads to increased emissions of particulate matter, although volumetric fuel consumption is decreased (CONCAWE, 1986). This finding was confirmed by Hare (1986) for light- and heavy-duty vehicles on the basis of data from the automobile industry. The presumed increase in the release of particulates is 0.5-1.9% for a 1% by volume change in aromatic content. In a study of four light-duty vehicles in three Australian driving cycles (resembling roughly the US 72 and 75 tests), Williams et al. (1989) found reductions in particulate emissions of 22-27% at a 10% difference in fuel aromaticity. Unfortunately, the two fuels also differed in sulfur content and volatility. Van Beckhoven (1991) achieved comparable results with passenger cars with indirect injection but found no significant influence for heavy-duty direct injection engines. Experiments by Westerholm & Egebäck (1991) on one bus and one truck showed a reduction in the emission of particulates
when fuel with low levels of sulfur and aromatics was used. The increased aromatic content is probably also the source of higher levels of PAHs and nitro-PAHs in the exhaust (Schuetzle & Frazier, 1986; Westerholm, 1987; Westerholm & Egebäck, 1991; Organisation for Economic Co-operation and Development, 1993).

(c) Volatility

The volatility of diesel fuels is described by the boiling range, and especially by the 10% boiling-point (low boiling end fraction) and the 90% boiling-point (high boiling end fraction) of the mixture. The influence of the boiling range, particularly the 90% boiling-point, on the release of particulate matter is still under discussion. While some investigators (Hare, 1986; Williams et al., 1989) assume that emissions of particulate matter increase with an increased 90% boiling-point, others have not been able to confirm this effect, suggesting that other parameters, such as sulfur content and aromaticity, are responsible for the increase (Westerholm & Egebäck, 1991; Organisation for Economic Co-operation and Development, 1993).

(d) Cetane number

An increase in cetane number lowers the emission of diesel particulate matter. A slight decrease in cetane number, combined with a small increase in the density of diesel fuels (see section A2.1.3), was predicted in 1987 to result in an increase in emissions from 3 to 12% for carbon monoxide, and to maxima of 2% for nitrogen oxides, 7% for hydrocarbons, and 6% for particles (CONCAWE, 1987).

B3.1.2.3 Malfunction

Malfunctions or defective components of the engine or exhaust gas system may result in considerably higher emissions than those from a properly maintained engine. Up to twofold increases in emissions of carbon monoxide, hydrocarbons, and particulate matter have been measured even in the absence of a significant effect on power or fuel consumption (Ullman et al., 1984). Malfunction of the fuel injection system may contributed markedly to higher emissions (Organisation for Economic Co-operation and Development, 1993). Volkswagen AG (1989) tested a diesel passenger car with defective injection nozzles in three driving cycles (US FTP test, US sulfate emission test, US 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmhighway driving cycle) and found statistically significant increases in the emissions of carbon monoxide (up to about 25%), hydrocarbons (up to about 45%), and particulate matter (up to about 15%), whereas the release of nitrogen oxides was almost unaffected. The amounts of benzene and PAHs emitted were slightly increased. Comparable results were obtained by Williams et al. (1989) for the release of particles by several light-duty vehicles equipped with defective injectors. Malfunction of US medium- to heavy-duty vehicle diesel engines increased the amount of nitrogen oxide emitted to about 4%, that of hydrocarbons to about 75%, and that of particulates to about 140% (Organisation for Economic Co-operation and Development, 1993).

B3.1.3 Total emissions by diesel engines

Few data are available on the emissions of different constituents of diesel exhaust into the atmosphere and their contribution to the total release by traffic and industry. The contribution of heavy-duty vehicles seems to be much greater than that of passenger cars, despite lower percent kilometres. For example, in westen Germany, only 8% of the total annual urban mileage was attributable to heavy-duty vehicles (Fromme, 1990). In contrast, in 1987, power plants contributed 15.3% (85 000 t), industry 63.5% (360 000 t), domestic and minor consumers 8.1% (44 000 t), and total traffic 13% (72 000 t) to the particles released (Metz, 1989). The contribution of diesel particulate matter to the total particle burden of the atmosphere is highly dependent on local conditions (Table 29).

Diesel vehicles, particularly in the light-duty category, represent a smaller proportion of motor traffic in the United States than in most European countries. Measurements from two heavy-duty vehicles in 1982 indicated that light- and heavy-duty diesel vehicles contributed about 6% to the total fine aerosol, organic carbon emissions in the Los Angeles basin, about 1200 kg/day being emitted by heavy-duty vehicles and 600-870 kg/day by light-duty vehicles (Hildemann et al., 1991; Rogge et al., 1993).

B3.1.4 Control of emissions

The automotive industry has made efforts to reduce diesel exhaust emissions (Obländer, 1991; Fortnagel, 1992; Hammerle et al., 1994b), and technology is advancing.

B3.1.4.1 Particle traps

Particle traps are placed in the exhaust system to collect particulate matter and to clean the filter by burning the trapped particles (trap oxidizers). Most traps are based on cellular ceramic monolith material or ceramic silica fibre coils. Many regeneration techniques have been developed for trap oxidizers (Organisation for Economic Co-operation and Development, 1993), including thermal regeneration, which is the burning of collected particulate matter, and catalytic regeneration, in which fuel catalysts, e.g. iron and copper, containing organic compounds form finely dispersed metal particles during combustion to reduce the burning temperature of the diesel particles so that there is more efficient regeneration (Volkswagen AG, 1989; Hammerle et al., 1994b). Traps remove the soluble organic fraction adsorbed onto particles and the dry carbonaceous soot, while catalysts can remove substances only from the soluble organic fraction. In catalytic regeneration, over 95% of the metal used as fuel catalyst is retained in the trap (Hammerle et al., 1994b).

Studies of the emissions of one light-duty diesel passenger car by Volkswagen AG (1989) showed that the release of particulates was reduced by a thermal trap by at least 80% during the collection phase and by about 60% during the regeneration phase; the reduction was 80% for the whole cycle. Sulfate emissions decreased to about 50%; however, emission of carbon monoxide increased by about 10%, probably due to additional burning and/or incomplete oxidation of particulate matter on the trap. Comparable experiments with a catalytically regenerated trap, with addition of a manganese compound, also showed a trap efficiency of about 80% for particulate matter. Hydrocarbon and carbon monoxide emissions were slightly increased, probably due to incomplete oxidation on the trap. Sulfate emissions were reduced to less than half, emissions of PAHs were reduced to about 25%, and those of phenol and aldehydes to about 25-33%. About 10% of the manganese catalyst was detected in the exhaust.

Measurements with a heavy-duty vehicle on a chassis dynamometer to test two particle traps showed decreases of 79 and 86% in the release of particles, depending on the trap and fuel characteristics. The emissions of hydrocarbon, carbon monoxide, aldehydes, and PAHs were slightly reduced (Westerholm et al., 1986).

Table 29. Diesel exhaust emissions and their contribution to total anthropogenic emissions

Region Year Annual emissions (kt/year) Reference

NO

x CO HC PM SO 2

**Light-duty vehicles**

Germany a 1986 52 (3.3) b 76 (1.2) b 23 (2.2) b 15 (26.3) b NR Fromme (1990) 1987 NR NR NR 12 (2.2)c NR Metz (1989)

United Kingdom 1991 (0.7) c (0.4) c (1.1) c (2) c,d (0.2) c Quality of Urban Air Review Group (1993) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm**Light- and heavy-duty vehicles**
Europe NR NR NR NR NR (1.6) c OECD (1993)

United States 1984 NR NR NR NR 620 (3) c OECD (1993)
1986 NR NR NR 54.8 (20) b NR Carey (1987)
(4) c

Germany a 1989 640 240 150 55 NR Federal Environment Agency(1991)

**Heavy-duty vehicles**
OECD (Europe) 1980 1548 (13) c NR 393 (4) c NR 361 (2) c OECD (1993)

Germanya 1986 502 (32.2) b 130 (2.1) b 104 (10.0) b 37 (64.7) b 35 (2) c OECD (1993); Fromme (1990)
(17) c (2) c (4) c (7) c
1987 NR NR NR 32.5 (5.9) c NR Metz (1989)

Table 29 (cont'd).

Region Year Annual emissions (kt/year) Reference

NO
x CO HC PM SO 2

**Heavy-duty vehicles** (contd) Germany 1988 614 351 1.31 NR 38 Federal Environme nt Agency (1992) United Kingdom 1991 (19.9) c (1.9) c (5.4) c (14) c,d (1.0) c Quality of Urban Air Review Group (199 3)

Canada e 1985 265 104 37 20 21 Environment Canada
(1990)

NO

x , nitrogen oxides; CO, carbon monoxide; HC, hydrocarbons; PM, particulate matter; SO 2 , sulfur dioxide; NR, not reported; OECD,
Organsiation for Economic Co-operation and Development

a Excluding the former German Democratic Republic

b Percentage of total traffic emissions

c Percentage of total man-made emissions

d PM 10

e Sum of light- and heavy-duty trucks The effect of a ceramic wall-flow monolith particulate trap was studied on the emissions of a 1979 heavy-duty diesel engine with direct injection in two steady-state modes. Reductions of 70% (50% at full load) and 86% (75% at full load) in particle release were found. There was also a decrease in the soluble organic fraction, whereas the emitted portion of volatile organic carbons was only slightly reduced. The trap had almost no effect on the release of nitrogen oxides (Dorie et al., 1987).

The measurements of Lowenthal et al. (1994) on 13 trucks and buses (three with particulate traps, 10 without) in a bus cycle showed a similar reduction in the release of particles (Table 26). There was a significant decrease in the emission of particle-bound PAHs.

B3.1.4.2 Catalytic converters

Diesel catalytic converters reduce gaseous and particle-bound hydrocarbon constituents of the exhaust, e.g. aldehydes, PAHs, nitro- PAHs, and carbon monoxide. Regeneration systems are not needed, as particulate matter is not collected by common flow-through catalytic converters. The efficiency in controlling particulates is, however, much lower than that of traps. Disadvantages of catalytic converters are further conversion of nitrogen monoxide to the more toxic nitrogen dioxide and of sulfur dioxide to particulate sulfates (Organisation for Economic Co-operation and Development, 1993). Catalysts that selectively oxidize organic compounds and oxidize almost no sulfur dioxide are currently in use. Removal of sulfur from diesel fuels would allow the use of more active catalysts; use of catalysts in diesel vehicles run on sulfur-containing fuel is limited because the strong adsorbing tendency of sulfur partly inactivates the catalyst (Hammerle et al., 1994b; see section A2.1.1).

B3.2 Regulatory approaches

The levels of carbon monoxide, nitrogen oxides, total hydrocarbons, and particulates emitted by diesel engines are regulated by law in a number of countries (Table 30). Most other industrialized countries have adapted United States or European regulations (for details, see CONCAWE, 1990a and Mercedes-Benz AG, 1994a).

B4. ENVIRONMENTAL TRANSPORT, DISTRIBUTION, AND TRANSFORMATION

Few data are available on the fate of diesel exhaust in the atmosphere, which is the first environmental compartment affected. In general, the effects on the overall environment of diesel fuel combustion are typical of those of fossil fuel burning. The contribution of diesel fuel emissions is proportional to diesel fuel consumption both locally and globally.

Table 30. Limit values for components of diesel exhaust

Region Carbon Nitrogen Hydrocarbons Particulates Comments 

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**Light-duty vehicles (g/km)**

Austria 2.1 0.62 0.25 0.124 < 3.5 t; since 1991; from 1995, adoption of European Union standards planned
Canada 2.1 0.62 0.25 0.12 Since 1987 European Union 2.72 0.97 (with 0.14 Since 1992 hydrocarbons)

1.0 0.7 0.08 From 1996
Finland Since 1993 Japan 2.1 0.7 0.62 None Since 1986
2.1 0.5 0.4 0.2 Since 1994 Sweden, Norway 2.1 0.62 (city) 0.25 0.124 < 3.5 t; from motor year 1992 0.76 (highway) Switzerland 2.1 0.62 (city) 0.25 0.124 < 3.5 t; since 1988; from 1995, 0.76 (highway) adoption of European Union standards planned USA (California) 2.1-5.2 0.2-0.6 0.2-0.3 (except 0.05 (up to Depending on mileage methane) 31 000 km) US Environmental 2.1-2.6 0.6-0.8 0.2 0.05-0.12 Depending on mileage Protection Agency

Table 30 (contd)

Region Carbon Nitrogen Hydrocarbons Particulates Comments
monoxide oxides

**Heavy-duty vehicles (g/kWh)**

Austria 4.9 9.0 1.23 0.4 Canada 15.5 5.0 1.3 0.25 g/bhp-h 15.5 5.0 1.3 0.1 g/bhp-h; from 1995-97 European Union 4.5 8.0 1.1 0.36 Since 1992

4.0 7.0 1.1 0.15 From 1995-96 Japan 7.4 5.0 2.9 0.7 Indirect injection engines 7.4 6.0 2.9 0.7 Direct injection engines

Sweden 4.9 9.0 1.23 0.4 USA 15.5 5.0 1.3 0.07 g/bhp-h; bus 15.5 4.0 1.3 0.1 g/bhp-h; truck 15.5 5.0 1.3 0.05 g/bhp-h; bus; from 1998 15.5 4.0 1.3 0.1 g/bhp-h; truck; from 1998

Adapted from Mercedes-Benz AG (1994b); reference year, 1994, unless otherwise stated; bhp-h, brake horsepower-hour Thus, the major components of diesel exhaust contribute to acid deposition (nitrogen and sulfur oxides), tropospheric ozone formation (carbon monoxide, nitrogen oxides, and hydrocarbons), and global warming (carbon dioxide and particulates; carbon monoxide, nitrogen oxides and hydrocarbons by ozone formation).

The environmental fate of released particulate matter and of organic compounds, particularly the PAHs, is important environmentally.
The mechanisms of the distribution and transformation of individual hydrocarbons in diesel exhaust have been well described (IPCS, 1986, 1993).

B4.1 Transport and distribution between media

The major fraction (50-80%) of the particulate emissions of diesel engines is in the submicron size, ranging from 0.02 to 0.5 μm (see section B2.1.2). Once particles have been emitted, their mechanical transport in the atmosphere is like that of gas molecules (nonreactive). Together with carbon particles from other combustion processes, they may be transported over long distances and even penetrate the stratosphere (Muhlbaier Dasch & Cadle, 1989).

Only one study was available on the agglomeration behaviour of the particles in the atmosphere. Albrechcinski et al. (1985) measured the aging behaviour of particulate matter from a 1978 diesel passenger

car in a 600-m 3 model chamber with a nominal exhaust gas dilution of 150:1 and found peak mean aerosol diameters, based on volume size distributions, of 0.1-0.18 μm for 'fresh' aerosol. The concentrations of total particulate decreased during aging of the aerosol due to coagulation, fall-out, and perhaps losses to the walls of the chamber. The average decrease was 20% after 8 h and 60% after 24 h. Excess nitrogen dioxide seemed to inhibit particle growth. Particles with an average diameter of 1-10 μm constituted approximately 10% of the fresh aerosol, but this percentage decreased during aging as a result of apparently rapid fall-out of larger particles. It is not clear whether these results are also representative of the particle emission pattern of modern diesel passenger cars.

Model experiments of the gas-phase chemistry of nitrogen oxides, total hydrocarbons, and sulfur dioxide from diesel exhaust were performed by varying parameters such as irradiation, nitrogen dioxide concentration, and ozone concentration (Albrechcinski et al., 1985). Irradiation alone had only a minimal effect, but excess nitrogen dioxide led to higher rates of decay of nitrogen monoxide in the dark than under irradiation, probably because of photodissociation of nitrogen dioxide to nitrogen monoxide. As nitrogen monoxide is rapidly converted to nitrogen dioxide in the presence of excess ozone, the

nitrogen oxide concentration decreased as the ozone concentration reached a maximum, presumably partly because of the formation of 

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The hydrosphere and geosphere may be affected indirectly by diesel exhaust emissions after dry or wet deposition of particulate matter or individual constituents (see section B4.2). In experiments with dysprosium-traced diesel fuel, Horvath et al. (1988) (see section B5.1) found 0.012-0.07 g diesel particulate matter per gram of road

dust.

B4.2 Transformation and removal

Atmospheric removal of airborne carbon particles consists mainly of dry deposition and scavenging by precipitation (wet deposition). The rate of wet removal is directly correlated to the ratio of organic to elemental carbon and is low for small ratios (Muhlbaier Dasch & Cadle, 1989). As the overall removal rate of diesel particulates is estimated to be low, the atmospheric life-time is several days (Jaenicke, 1986).

The elemental carbon of diesel particulates may act as a catalyst in the formation of sulfuric acid by oxidation of sulfur dioxide. The
oxidation reaction is assumed to occur on the surface of solid soot particles or in liquid drops containing soot particles (Novakov, 1984; Mészáros & Mészáros, 1989). Furthermore, diesel particles themselves contain sulfate, adsorbed in the form of metal sulfates, at a concentration of about 1-12% by weight, depending on the sulfur content of the fuel (see section B2.1.2), which may be washed off if the particles come into contact with precipitation. The organic compounds adsorbed on the emitted particulate matter may undergo a number of physical and chemical reactions (volatilization of low-relative-molecular-mass compounds, formation of secondary aerosols, oxidation, nitration) with other atmospheric compounds or during exposure to sunlight (Pitts et al., 1985). The estimated half-lives for unbound PAHs in the atmosphere range from one day (naphthalene) to one week (e.g. pyrene, chrysene, benzofluoranthenes, benzo[a]pyrene) (Mackay et al., 1992; see Table 11); however, PAHs adsorbed onto particles may be more stable to nitrogen dioxide and ozone than airborne PAHs (Gibson et al., 1981).

B5. ENVIRONMENTAL LEVELS AND HUMAN EXPOSURE

Because diesel exhaust is a complex mixture of a great variety of compounds (see also section B2.1), general 'environmental levels'
cannot be described. The individual constituents of diesel exhaust should be detectable in all compartments of the environment. Since diesel exhaust is only one of numerous other sources, the levels of individual compounds cannot be correlated directly to diesel exhaust.

Particulate matter appears to be the most relevant fraction of diesel exhaust for the general population and workers exposed to diesel exhaust emissions from a toxicological point of view. The concentration of particles is about 10- to 20-fold higher in diesel exhaust than in the exhaust from gasoline-fuelled vehicles, and the particle diameters are in the respirable range.

B5.1 Exposure of the general population

The general population is exposed to high levels of diesel exhaust in certain microenvironments, such as busy streets and parking areas. Exposure to the main gaseous components of diesel exhaust -- carbon dioxide, carbon monoxide, nitrogen oxides, and total hydrocarbons -- can be estimated from the emission factors given in section B3, the kilometrage of diesel vehicles in relation to total traffic, and atmospheric dispersion models (Cuddihy et al., 1984; Volkswagen AG, 1989). In view of the fact that measured emission factors vary widely (see section B3.1.1) and are mainly dependent on adjustment of individual engines, the validity of these estimations is
limited.

On a global basis, there are wide variations in the contribution of diesel emissions to total airborne particulate matter, which depends on the following factors: percentage of diesel vehicles in the total volume of traffic; type, age, and maintenance of individual engines (see section B3.1.2.1); fuel quality (e.g. sulfur content and aromaticity; see section B3.1.2.2); and emission control techniques. Particle concentrations in environmental air samples are usually given as total particle concentrations, with no differentiation by source. The contribution of diesel particulate matter is then estimated on the basis of the percentage of diesel vehicle emissions in the total
emissions from traffic or total man-made emissions. Emissions from running stationary diesel engines have up to now not been taken into account. Data on emissions from light- and heavy-duty vehicles in some industrialized countries are shown in Table 30 (section B3.1.3). The proportion of the concentrations to which people are exposed that is attributable to diesel vehicles varies widely, depending on traffic flow and composition in the vicinity and the magnitude of background pollution from other sources.

In a study in the United Kingdom (Waller et al., 1965), measurements were made at a sampling site at the centre of a busy street canyon, where traffic flows were typically 1200 gasoline and 600 diesel vehicles per hour, in order to assess the maximal concentrations of traffic pollutants in urban areas. The contributions of airborne particulate matter, expressed as seasonal means for daytime working hours, were 200-421 μg/m 3 , after allowance for the urban background measured at nearby control sites. The additional particulate matter was attributed primarily to the diesel vehicles, while the corresponding contribution to carbon monoxide, averaging 

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In France, Joumard & Perrin (1988) measured atmospheric particulate matter (diameter, 0.01-25 μm) in a 30-40-m wide street open to pedestrians, with two central lanes reserved for use by bus services involving weekday traffic of more than 1000 vehicles per day in each direction. The measurement point was in the immediate vicinity of two bus stops, 3 m from the edge of the pavement and at a height of 2.5 m. The particle concentrations, determined by measuring ß radiation, were 34 μg/m 3 during the period 23.00-05.00 h and 67 μg/m 3 during 07.00-19.30 h.

Airborne particle concentrations were calculated for an eight-lane American freeway and for four- and six-lane European autoroutes. The one-dimensional Gaussian model used neglects chemical reactions during transport and possible deposition on the ground and assumes turbulent mixing in the air. The calculated maximal concentrations were 7-13 μg/m 3 close to the road, and these decreased with distance from the measuring point. About 100 m away, the estimated particle concentration at a height of 1.5 m was three to four times lower than that next to the motorway (Volkswagen AG, 1989).

Particle emissions, especially from diesel vehicles, were estimated roughly for various locations by the lead surrogate method,
in which historical ambient lead concentrations in urban areas are used as indices of mobile source pollutant levels. Particle emission factors for light- and heavy-duty vehicles and the distribution characteristics of lead and diesel particulate are also taken into consideration (Rykowski & Brochu, 1986). The results are shown in

Table 31.

Concentrations of particles from diesel engines in the environment were determined in Vienna, Austria, at several measuring points during a four-week period. As there was only one distributor of diesel fuel for the whole network of service stations in Vienna, the rare earth element dysprosium was added as an indicator to the fuel at this central point, which was distributed to diesel vehicles at service stations and then emitted with the exhaust. Dust samples collected from the air were subjected to neutron activation analysis and the level of dysprosium was determined. The proportion of dysprosium in diesel soot was calculated on the basis of emission factors for diesel engines in the literature and from laboratory experiments, assuming that the marker is totally transferred from fuel to exhaust during combustion. At seven of the 11 measuring points for different traffic densities, the concentrations were almost linearly correlated to traffic density. Ambient particulate levels increased by 5.5 μg/m 3 per 500 diesel vehicles per hour, from 10.3 in a residential area to 23.6 μg/m 3 near an urban freeway; the mean value was 14.4 μg/m 3 . The diesel particulate concentration in the centre of Vienna was calculated to be 11 μg/m 3 by correlating the measured values and traffic density. At four measuring stations, significant deviations from linearity were found which were due to local

conditions. The concentration of the marker was below the detection limit before the testing period and one year later (Horvath et al., 1988). A deficiency of the study is that the dysprosium concentrations of diesel fuel before the addition were not taken into account, so that the emission factors used may not reflect the true situation (Holländer, personal communication, 1994). Nevertheless, the measured ambient particle concentrations fit well with estimated data for diesel exhaust emissions by other investigators.

Table 31. Estimated particle concentrations emitted from diesel engine exhaust

Location and year Concentration (μg/m 3 )

United States a , cities with > 1 million 1.3-3 residents (1984)

United States a , cities with 100 000-250 000 residents 0.7-1.7 Naples, Italy, urban area (1982) 7.2-35.1 Birmingham, United Kingdom, urban area (1982) 3.2-20.3 Birmingham, United Kingdom, university area (1982) 0.8-5.5 Stockholm, Sweden, inner city (1982) 2.5-16.4

From Rykowski & Brochu (1986)

a According to Cuddihy et al. (1984), the proportion of diesel-fuelled light-duty vehicles in the United States was

about 2%.

In a programme to measure airborne diesel soot levels in Düsseldorf, Germany, the total carbon content of fine dust (particle diameter, < 7μm) at a busy crossing was 8-42 μm 3 as a daily average, 18 μg/m 3 as an annual average, 21 μg/m 3 on workdays, and 14 μg/m 3 on weekends. The carbon content after solvent extraction (elemental carbon) was 6-32 μg/m 3 as a daily average, 16 μg/m 3 on workdays, and 10 μg/m 3 on weekends. In a residential area in Duisburg, Germany, the concentration of elemental carbon was 3 μg/m 3

. The method of determination was as follows: a defined volume of air was sucked through a fibreglass filter, and the respirable particle fraction was separated. This fine dust was further separated into an incinerable portion (according to German standard method TRGS 900), total carbon, and the carbon content after liquid extraction (elemental carbon). Total and elemental carbon were determined by coulometry. By correlation with diurnal changes in carbon and nitrogen 

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1991 and 1992 confirmed the results (Elbers & Richter, 1994). Validation of the analytical method gave detection limits of 0.39 for elemental carbon (relative standard deviation, 14% or 4.4 μg/m 3 ) and 0.65 μg/m 3 for total carbon (relative standard deviation, 13% or 7.8 μg/m 3 ).

The weekly average ambient concentrations of diesel particulate in Germany were estimated to be 5-10 μg/m 3 in urban areas, 15-25 μg/m 3
in urban areas close to busy streets, and < 1.5 μg/m 3 in rural areas; however, the method used for the estimations was not reported (State Committee for Immission Protection, 1992).

Estimates of the US Environmental Protection Agency for 1986 gave annual mean ambient concentrations of diesel particle in the United States of 2.6 μg/m 3 in urban areas and 2.4 μg/m 3 in rural areas. A prediction for 1995 gave values of 1.5 μg/m 3 for urban and 1.2 μg/m 3 for rural areas (Mauderly, 1992). Higher concentrations were estimated by the Motor Vehicles Manufacturing Association (Sienicki & Mago, 1992), which predicted a mean value of 2.7 μg/m 3 for the city of Los Angeles in 1995. Lower levels were estimated in a study by the US Environmental Protection Agency (1993) in which national average emission factors for diesel vehicles for 1988 were multiplied by urban and rural mileage. For 1990 and 1995, the annual exposure levels were estimated to be 2.0 and 1.2 μg/m 3 in urban air and 1.1 and 0.6 μg/m 3 in rural air, respectively. These estimates were confirmed by ambient monitoring. Using data from the early 1980s, when the proportion of diesel vehicles, the number of kilometres travelled, and emission standards differed from present characteristics, McClellan (1986) derived ambient diesel particulate concentrations of 10-30 μg/m 3 on American urban freeways and in urban street canyons.

B5.2 Occupational exposure

The levels of particulate matter to which workers are exposed when their predominant exposure to exhaust is that from diesel engines
are described in this section. It is difficult to determine the contribution of diesel exhaust to respirable particles, as they include not only those from diesel engines but also others generated in the worker's environment, such as sand, dirt, coal dust, and cigarette smoke particles. Thus, the levels of total and respirable particulates reported in jobs with potential exposure to diesel exhaust may be significant overestimates of the actual exposure. Only two occupational groups have been studied in this regard: railroad workers (Woskie et al., 1988a,b) and mechanics and drivers in the trucking industry (Zaebst et al., 1991). The specific exposures of these groups to diesel particles range from 0.026 to 0.192 mg/m 3 . In the railroad workers, cigarette smoke was found to account for a substantial proportion of total respirable particulate matter, and the

geometric mean exposure to respirable particulates of workers not exposed to diesel exhaust was reduced by over 80% when the contribution of cigarette particulate was considered (Woskie et al.,
1988a).

The principal marker chosen to measure exposure of workers in the trucking industry (including road drivers, local drivers, stevedores, and mechanics) to diesel exhaust was elemental carbon, which is derived from the core of the diesel exhaust particle and represents about 20% of respirable particulate; cigarette smoke does not contribute appreciable amounts of elemental carbon. Mechanics and stevedores had the highest exposure to diesel exhaust, and truck drivers had lower levels (Zaebst et al., 1991). The degree to which a marker of diesel exhaust exposure actually represents true exposure depends on the nature of all sources of respirable particles in the work-place. Occupational exposure to particulates is summarized in

Table 32.

Table 32. Reported occupational exposure to particulates among workers exposed to diesel exhaust

Occupational group Particles (mg/m 3 )

Total Respirable Diesel

Railroad workers 0.01-1.99 0.06-0.41 0.051-0.192 a
Mine workers 0-23.0 0-16.1

Truck drivers 0.1-1.0 0.13-1.2 0.025 b
Bus garage and bus 0.15-1.2 0.01-0.61

workers

Stevedores 0.157 b

Truck mechanics 0.133 b

a Estimated using nicotine to account for exposure to cigarette smoke

b Based on carbon core analysis, with an estimated 20% of respirable particulate due to carbon (Steenland et al., 1992)

Because 1-nitropyrene is associated with the particle fraction of diesel exhaust, this compound was used as an indicator of the particulate fraction of diesel exhaust derived from various sources 

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extracted with acetone by sonication and detected by gas chromatography-mass spectrometry. The following concentrations of respirable diesel exhaust were determined: 0.044-0.33 mg/m 3 in various work-places with fork-lift trucks, 0.10-0.37 mg/m 3 in railroad engine repair shops, 0.080mg/m 3 in a river vessel, and 0.67 or 3.33 mg/m 3 in the breathing zone of railroad workers.

B5.2.1 Truck drivers and mechanics

In a study of 15 truck drivers in Geneva, Switzerland, exposure to particulate matter was measured during one working day with a direct-reading instrument and by sampling beside the drivers in the truck cabin (Guillemin et al., 1992). Local drivers had greater exposure to particulate matter (0.3 mg/m 3 ) than long-distance drivers (0.1 mg/m 3 ) and spent more time in polluted areas, such as streets with heavy traffic and construction sites. Smoking did not influence the exposure of professional truck drivers to particulate matter, probably because the ventilation rate of the truck cabins was relatively high, even on cold days.

Industrial hygiene surveys were conducted during warm and cold weather at eight terminals and truck repair shops to determine exposure to submicrometre particles of elemental carbon among American workers in one of four jobs -- road drivers, local drivers, stevedores, and mechanics -- all of whom were exposed mainly to diesel aerosol (Zaebst et al., 1991). The overall arithmetic mean exposure was 5.1 μg/m 3 for 72 long-distance road drivers, 5.4 μg/m 3 for 66 local drivers, 26.6 μg/m 3 for 80 mechanics, and 31.3 μg/m 3 for 54 stevedores exposed to diesel fork-lift trucks. The background concentrations in the same cities were 3.4 μg/m 3 on 21 major highways and 1.4 μg/m 3 in 23 residential areas. The exposures of the truck drivers were significantly higher than the background concentrations in residential areas but could not be distinguished from the background highway concentrations.

Area air sampling in an American truck repair shop in 1987 gave the following results, depending on the sampling location: 0.225 and 0.32 μg/m 3 submicrometre particles, 0.26 and 0.51 μg/m 3 respirable particles, and 0.63 and 0.66 μg/m 3 total particulate matter. The values for elemental carbon were 86-118 μg/m 3 submicrometre particles and 159-214 μg/m 3 total carbon. In personal samples from four mechanics, 79-193 μg/m 3 submicrometre elemental carbon were found (US National Institute for Occupational Safety and Health, 1990a).

The concentrations of particles collected on fibreglass filters at 25 work-places in Germany where mainly fork-lift trucks and trucks were used were 0.02-0.8 mg/m 3 in area samples and 0.13-1.2 mg/m 3 in personal samples (Lehmann et al., 1990). Area measurements in two German truck repair garages (five samples from each) showed concentrations of incinerable fine dust (according to German standard method TRGS 900) of 0.11-0.40 mg/m 3 (Blome et al., 1990).

Personal samples from truck drivers who loaded diesel-engined trucks onto car ferries in Sweden showed particle concentrations of 0.1-1.0 mg/m 3 (Ulfvarson et al., 1987). The drivers may also have been exposed to diesel exhaust from the ferries.

B5.2.2 Bus garage and other bus workers

In area samples taken in an American bus garage in 1989, the concentrations of particles were 0-0.1 mg/m 3 submicrometre particles, 0-0.3 mg/m 3 respirable particles, 0-0.22 mg/m 3 total particulate matter, and 0.014-0.326 mg/m 3 elemental carbon (US National Institute for Occupational Safety and Health, 1990b).

The concentrations of smoke were measured in two diesel bus garages in London, United Kingdom, from high-volume samplers for successive periods through the day and night, which were linked to bus movements and the exposure of the maintenance staff. The contribution from the buses was assessed by subtracting the background values at control sites outside the garages. In the most polluted garage, 0.3-1.2mg/m 3 of smoke were measured during working periods (Waller et al., 1985).

In Sweden, a particle concentration of 0.46 mg/m 3 was found during personal sampling in periods of high activity (arrival and departure of buses) (Ulfvarson et al., 1987).

Area air sampling of the concentrations of incinerable fine dust (according to German standard method TRGS 900) in two bus depots (six samples in each) and one bus garage (three samples) in Germany showed concentrations of 0.22-0.37 mg/m 3 (Blome et al., 1990).

The respirable dust levels in bus garages in Italy and the United States ranged from <0.01 to 0.73 mg/m 3 (Gamble et al., 1987a). The total dust levels in bus garages in Denver, Colorado, United States, were 0.01-0.81 mg/m 3 (Apol, 1983; Pryor, 1983).

B5.2.3 Fork-lift truck operators

Area samples taken in one American trucking terminal where diesel-powered fork-lift trucks were used in 1987 contained 48-62 μg/m 3 submicrometre particles, 34-57 μg/m 3 respirable 

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elemental carbon in area samples from another American trucking terminal in 1988 were 40-44 μg/m 3 , and those in personal samples from fork-lift operators were 33-77 μg/m 3 (US National Institute for Occupational Safety and Health, 1989b). Area samples taken in an American freight dock contained concentrations of <12 and 294 μg/m 3 respirable dust. The levels were significantly higher in areas where unfiltered rather than filtered diesel motors were used (US National Institute for Occupational Safety and Health, 1990c).

A mean concentration of 31.3 μg/m 3 of submicrometre elemental carbon was found in personal samples from stevedores operating diesel-powered fork-lift trucks. About 10 times less was found for operators of gasoline- or propane-powered trucks (Zaebst et al.,
1991).

The concentrations of particles in the breathing zone during use of diesel-powered fork-lift trucks for loading, unloading, and warehousing operations in American ammunition storage magazines were < 0.01 to1.3 mg/m 3 (Ungers, 1984) and 0.5-5.0 mg/m 3 (Ungers, 1985).

B5.2.4 Railroad workers

The mean levels of total particulate matter were 0.38 mg/m 3 (range, 0.1-0.8) in Finnish locomotive cabs and 1.99 mg/m 3 (range, 0.07-8.7) in roundhouses (Heino et al., 1978). In three German engine sheds, the ambient concentrations of incinerable fine dust were 0.1-0.41mg/m 3 (Blome et al., 1990).

A summary of American literature showed time-weighted average levels of 0.05-0.07 mg/m 3 (8-h) in tunnels, 0.16 mg/m 3 (7.5-h) during freighting, 0.01 mg/m 3 (5-h) in switchyards, and 0.27 mg/m 3 (8-h) in cabooses (Hobbs et al., 1977). The arithmetic mean concentrations of total respirable particles in personal samples from shop workers, engineers, brakemen, conductors, and farmers in the American railroad industry (after correction for the estimated contribution of cigarette smoke particulates) were 51-192 μg/m 3 (Woskie et al., 1988a).

B5.2.5 Mine workers

Diesel vehicles and machines have been used in nearly all American mines since the 1960s (Wichmann & Brüske-Hohlfeld, 1991). The mean concentration of diesel particulate matter in personal and area samples in mines where diesel equipment was used was 4.6 mg/m 3 (range, 0.2-14) (Holland, 1978). Total respirable dust levels of 0.6-1.7 mg/m 3 were found by personal sampling (Wheeler et al., 1981); average concentrations of 0.9-2.7 mg/m 3 were found in full-shift personal samples, and the range for full-shift area samples was 0-16.1 mg/m 3 (Reger et al., 1982). The 8-h time-weighted average level of respirable particulate matter in personal samples taken in an

American molybdenum mine was 0.2-1.9 mg/m 3 (Cornwell, 1982). Total respirable dust concentrations of 1.1-4.4 mg/m 3 were reported in an American gold mine (Daniel, 1984). The US Mine Safety and Health Administration reported that the concentrations of diesel particulate matter in American mines other than coal mines were 0.3-1.5 mg/m 3 (Mauderly, 1992).

Data from a salt mine in Canada showed a diesel particulate level of about 0.5 mg/m 3 (Johnson & Carlson, 1986).

In area samples from different work-places in eight German coal mines, the levels of airborne diesel particulate matter (incinerable fine dust) were 0.19-0.70 mg/m 3 (Blome et al., 1990). In German potash-salt mines, depending on the district, 0.2-0.4 mg/m 3 of diesel particles were measured (Bauer et al., 1990).

B5.2.6 Fire fighters

The average concentration of total particulate matter in personal samples from fire-fighters in Boston, New York City, and Los Angeles, United States, in 1985 was between < 0.1 and 0.48 mg/m 3 . On a typical day in Boston or New York City, exposure to diesel exhaust particles was estimated to be about 0.225 mg/m 3 after correction for the contributions of background and smoking. Simulation of 'worst-case' exposures in Los Angeles fire stations gave a maximal concentration of 0.748 mg/m 3 total particles (Froines et al., 1987).

B5.3 Biomonitoring

B5.3.1 Urinary mutagenicity

The occurrence of mutagens, measured by the Ames test, in the urine and faeces of eight car mechanics occupationally exposed to diesel exhaust was compared with that of a reference group of nine office workers. The samples were collected over 48 h; information was also gathered on dietary intake, smoking habits, and medication. No enhancement of the incidence or degree of faecal or urinary mutagenicity was seen (Willems et al., 1989).

Post-shift urinary mutagenicity was measured by the Ames test in 306 samples from 87 railroad workers with a range of exposures to diesel exhaust (Schenker et al., 1992). Exposure was measured throughout the work shift by constant-flow personal sampling pumps. In separate analyses of smokers and nonsmokers, no independent 

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In studies of the effect of diesel exhaust on stevedores on 'roll-on-roll-off' ships (Ulfvarson et al., 1987), no difference in mutagenicity to Salmonella typhimurium TA98 or Escherichia coli WP

2 uvr A was seen in urine collected during periods of exposure and of no exposure. Similarly, no increase in urinary mutagenicity was found in samples from six volunteers before and after exposure to diesel exhaust collected from an experimental automobile run for 3.7 h at 60 km/h and 2580 rpm.

B5.3.2 Other analyses

Samples of total suspended particulate matter were collected in a repair shop for train engines, and the diesel exhaust concentrations were determined using 1-nitropyrene as a biomarker (see section B5.2). Urinary metabolites and the corresponding nitro-substituted derivatives were detected by immunoassay in the urine of three diesel-engine mechanics. They had a significantly increased excretion of metabolites in comparison with two office clerks (Scheepers et al., 1994b).

B6. KINETICS AND METABOLISM IN LABORATORY ANIMALS AND HUMANS

B6.1 Deposition

The mass median aerodynamic diameter of diesel exhaust particles in long-term studies in animals exposed by inhalation was 0.17-0.25 μm (Lewis et al., 1986; Lee et al., 1987; Wolff et al., 1987; Creutzenberg et al., 1990). The deposition efficiency of particles of this size range is lower than that of smaller particles, for which the deposition by diffusion increases with decreasing diameter. The deposition efficiency of particles with a diameter > 0.5 μm increases with the aerodynamic diameter, owing to sedimentation and impaction. The nose has hardly any filter effect for diesel particles because the fraction of particles < 0.4 μm deposited in the nose is very low (Heyder et al., 1986).

After short-term inhalation by rats of 0.29-μm fused aluminosilicate particles, 8% was deposited in the bronchi and 13% in the lungs (Raabe et al., 1988). A total of 10-20% diesel exhaust particles was deposited in rats and guinea-pigs (Chan et al., 1981; Lee et al., 1983; Dutcher et al., 1984). In humans, the alveolar deposition efficiency of 0.2-μm particles was 10-20%, and no bronchial deposition was detected (Heyder et al., 1986).

These experimental results agree well with those of a predictive model for deposition of inhaled diesel exhaust particles in laboratory animals and humans (Yu & Xu, 1987). In humans, the deposition efficiency was about 7% in the alveolar region and 4% in the tracheobronchial region. The model can also predict the fractions that will be deposited in different airway generations; maximal deposition occurs in the alveolar region (twentieth generation) of adults and of children over two years of age.

The length and concentration of exposure had no significant effect on deposition in rats exposed to 0, 0.4, 3.5, or 7.1 mg/m 3 diesel exhaust particles for 6, 12, 18, or 24 months, when deposition of 0.1-μm gallium oxide particles was used as the indicator (Wolff et al., 1987).

B6.2 Retention and clearance of particles

The kinetics of inhaled substances are conveniently described by their pulmonary clearance and retention. These terms are often used interchangeably, but it should be kept in mind that the fraction cleared from the lung plus the fraction retained in the lung account for the total deposited amount (retention = deposition - clearance). Thus, in the following paragraphs, the term 'clearance' refers to a clearance rate ( r, per unit of time) and the term 'retention' to a biological half-time ( t  ́), provided there is a constant rate of exponential clearance (ln2/ t  ́).

Clearance of diesel particles from the alveolar region is important in the long-term effects of particles on cells, as it is more than two orders of magnitude slower than mucociliary clearance,
measured in several studies in rats administered labelled diesel or surrogate particles by inhalation. The pulmonary retention half-times in studies in which the lung burden of diesel soot was also reported are summarized in Table 33. The equivalent continuous exposure concentration is calculated by multiplying the actual concentration by the number of hours of weekly exposure divided by 168 h. This figure allows a comparison of exposure levels in studies with different exposure protocols. The assumption is that Habers' law ( c × t = constant) is applicable. This is probably valid until a certain burden of particulate is reached in the lung; however, when that level is exceeded (particle overload), the calculation is increasingly invalid.

The retention half-time in these studies was 60-100 days in control rats with low lung burdens and up to 600 days in animals with high lung burdens. In a double logarithmic plot of retention half-times and lung burden, a linear relationship can be seen for lung burdens of diesel particles above 1mg per lung (Figure 1).

The gaseous phase of diesel exhaust appears to have no effect on alveolar clearance in rats or hamsters (Heinrich et al., 1986a).

The retardation of lung clearance is not specific to diesel 

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deposition kinetics at low concentrations, (iii) with associated pathophysiological changes including altered macrophage function, inflammation, and pulmonary fibrosis, and (iv) an uncertain association with an increased incidence of lung tumours in rats (McClellan, 1990).

The effect of overloading lung clearance mechanisms was investigated in NMRI and C57Bl mice, Wistar and Fischer 344 rats, and Syrian golden hamsters exposed to particulates, including test toner (polymer pigmented with carbon black), polyvinyl chloride, carbon black, diesel exhaust, and two crystalline forms of titanium dioxide (anatase and rutile), at concentrations of 0.8-64 mg/m 3 for up to two years. In rats, lung overloading generally occurred after exposure to 0.5-1.5 mg per lung or about 1 mg of material per gram of lung
tissue. A two- to 10-fold decrease in the clearance rate was seen after heavy particulate loading in all of these rodent species (Muhle et al., 1990).

In comparisons of the effects of particles of different densities, the retained volume of particles appears to be a more useful parameter than the retained mass, as alveolar macrophages have an upper volumetric uptake limit. The particulate overload effect appears to be initiated in rats when the average particle volume per alveolar macrophage exceeds about 60m 3 , and alveolar macrophage-mediated particle clearance virtually ceases when the phagacytosed particulate volume exceeds an average of 600m 3 (Morrow, 1988).

At a lung burden above threshold, signs of lung overloading in Fischer 344 rats persisted for 15 months after cessation of exposure (Bellmann et al., 1992). Retardation of alveolar clearance was also observed in hamsters and mice at a lung burden above about 1 mg per lung. For mice, no data were available with regard to lung burdens below 1 mg per lung, but retardation of alveolar clearance is probably already present at lower lung burdens. Lung overloading is thus seen in a variety of species with various materials. It is generally observed after the threshold lung burden of particles of low solubility and low acute toxicity is exceeded for a considerable time (Muhle et al., 1990). The effects of overloading should be considered in extrapolating the health effects seen at high concentrations in experiental animals to the low concentrations occurring in the environment or the work-place.

The alveolar clearance rates of highly insoluble particles differ among species, with retention half-times ranging from 50 to 100 days in rodents to several hundred days in dogs and man (Figure 2). In human lung, the retention half-time of insoluble, nontoxic particles may be >500 days (Bailey et al., 1985a,b).

Table 33. Pulmonary retention of insoluble particles in rats after exposure to diesel exhaust, and lung burden of dies el soot

Strain and sex No./group Duration of Exposure Pulmonary retention Reference

exposure (h/week) Concentration (mg/m 3 ) Duration Lung burden Material Half-time (weeks) (mg/lung) studied (days) Actual Equivalent
continuous

Fischer 344, 4 140 0 0 0 0 14 C-Diesel 77 Chan et al.

male 0.25 0.21 7 0.2 exhaust 90 (1984)

0.25 0.21 16 0.5 92

6.00 5.00 1 0.7 166

6.00 5.00 9 6.5 562 6.00 5.00 16 11.8 (> 1000)

Wistar, female 5-7 90 0 0 13 0 59 Fe 2 O 3 61 Creutzenberg 0.84 0.45 0.63 (0.3 μ m ) 94 et al. (199 0

2.5 1.34 2.5 119 Heinrich et

6.98 3.74 8.2 330 al. (1995)

0 0 52 0 72

0.84 0.45 2.8 121

2.5 1.34 10.7 254

6.98 3.74 35.7 541

0 0 78 0 96

0.84 0.45 5.5 a 221

2.5 1.34 12.2 272

6.98 3.74 58.8 687

Table 33 (contd)

Strain and sex No./group Duration of Exposure Pulmonary retention Reference

exposure (h/week) Concentration (mg/m 3 ) Duration Lung burden Material Half-time (weeks) (mg/lung) studied (days) Actual Equivalent
continuous

Fischer 344, 8 35 0.15 0.03 18 0.035 Diesel 87 Griffis et a 

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female 4.10 0.85 1.89 165

Fischer 344, 4 140 6.00 5.00 1 1.7 14 C-Diesel 61 Lee et al.

male 6.00 5.00 3 4.4 exhaust 124 (1987)

6.00 5.00 6 10.4 192

Fischer 344, 15-16 (lung 35 0 0 104 0 134 Cs- F used 79 Wolff et al. male and burden) 0.35 0.07 0.6 aluminosilicate 81 (1987) female 10 3.50 0.73 11.5 particles 264 (retention) 7.00 1.46 20.5 (2 μm) 240

a Measured; value derived by interpolation from data at 52 and 104 weeks

Retention of diesel particles in the lungs and lymph nodes has been reported in Fischer 344 and Wistar rats after long-term inhalation (Brightwell et al., 1986; Ishinishi et al., 1986; Mauderly et al., 1987; Strom et al., 1988; Creutzenberg et al., 1990; Strom et al., 1990). Some results with regard to lung burden are included in Table 33. Because clearance of lung burdens above about 1 mg per lung was impaired after long-term, continuous inhalation of a concentration equivalent to about 0.5mg/m 3 , no steady-state lung burden was found at serial sacrifice at up to two years. About 20% of the lung burden of diesel particles was found in lung-associated lymph nodes at that
time.

Mathematical models have been developed to calculate the retention of diesel particles in rats. Yu & Yoon (1990) and Yu et al. (1991) developed a dosimetry model to predict the retention of diesel particles and associated organic compounds in rats and humans of different ages. The model indicates that lung clearance would not be impaired in humans exposed to a concentration < 0.05 mg/m 3 for 24 h/day. The highest lung burden per unit of lung weight was calculated for five-year-old children. This model was used to assess the risk of inhaling diesel particles (Pepelko & Chen, 1993).

B6.3 Retention and clearance of polycyclic aromatic hydrocarbons
adsorbed onto diesel soot

The clearance of particles from ciliated airways by the mucociliary escalator after bronchial deposition is virtually complete within 24 h. For soluble organic material like PAHs on the surface of carbon particles, diffusion-limited clearance through the airway epithelium to capillary blood was found to be slower in the bronchial region than in the alveoli (Gerde et al., 1991a, 1993a,b).

PAHs in diesel soot adhere strongly to the surface of the particles. Several studies have shown that the association of PAHs with particles significantly slows the clearance of the PAHs from lungs in comparison with the clearance of inhaled PAHs unassociated 

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After inhalation by Fischer 344 rats of 3 H-benzo[ a ]pyrene adsorbed onto diesel soot (0.1% by mass) for 30 min, biphasic lung clearance was noted. In a first, fast phase, about 50% of the inhaled compound was cleared with a half-time of about 1 h, predominantly by mucociliary clearance. At the end of exposure, about 15% of the 3 H label was found in blood, liver, and kidney, indicating rapid, direct absorption of this fraction of 3 H-benzo[ a ]pyrene and its metabolites into blood. The remaining 50% was retained in the lung, with a half-time of 18 days; this is one-third of the retention half-time of diesel particles. Thus, removal of benzo[ a ]pyrene and its metabolites from the particle carrier may be the rate-limiting step of this clearance process (Sun et al., 1984). In contrast, more than 95% of

pure 3 H-benzo[ a ]pyrene was cleared from the lung within 12 h after inhalation (Sun et al., 1982).

These results were confirmed and extended to another PAH, 1-nitropyrene (Bond et al., 1986a,b), when the clearance of

14 C-labelled compound adsorbed to diesel soot was determined by the same method in Fischer 344 rats. After exposure to 490 ng/litre of pure labelled compound, about 90% was cleared in the first, fast clearance phase, with a retention half-time of 1 h; after exposure to 650 ng/litre of 14 C-1-nitropyrene adsorbed onto diesel particles, 45% of the labelled compound was cleared from the lungs in the first phase, with a half-time of about 2 h. The remaining 55% was retained with a half-time of 36 days, which was twice the corresponding value

for 3 H-benzo[ a ]pyrene in the study of Sun et al. (1984).

On the basis of their data on the clearance of 3 H-benzo[ a ]pyrene

and 14 C-1-nitropyrene, Bond et al. (1986b) predicted equilibrium lung burdens in humans after long-term inhalation. Assuming inhalation of 3.5 g/m 3 of particles containing 0.1% benzo[ a ]pyrene and 1-nitropyrene over an 8-h working day, they predicted equilibria of 160 ng benzo[ a ]pyrene and 31 ng 1-nitropyrene per lung. No data are available on changes in the retention of individual compounds after prolonged exposure to diesel exhaust.

The clearance of a very low concentration (2.6 g/g particles) of

3 H-benzo[ a ]pyrene adsorbed onto diesel particles was measured in Sprague-Dawley rats after intratracheal instillation (Bevan & Ruggio, 1991). Only 25% was cleared in the early, fast phase; a long-term retention half-time of eight days was calculated for the remainder during a three-day follow-up, which was similar to the clearance in that period in the study of Sun et al. (1984).

B6.4 Metabolism

In the studies of Sun et al. (1984) and Bond et al. (1986a,b),

the levels of 3 H-benzo[ a ]pyrene and 14 C-1-nitropyrene and their metabolites in extracts of lung homogenates after inhalation of diesel exhaust particles were measured by high-performance liquid chromatography. Sun et al. detected oxidized metabolites 30 min after

inhalation: 35% of the 3 H-benzo[ a ]pyrene had been metabolized in equal amounts to phenols (3-hydroxy- and 9-hydroxybenzo[ a ]pyrene) and quinones (1,6 and 3,6 isomers). Twenty days later, only 13% phenols and 5% quinones were detected. In the study of Bond et al.
(1986b), about 30% of the 3 H-1-nitro-pyrene had been metabolized to acetylaminopyrenephenol (6 and 8 isomers) and about 40% remained
unmetabolized 1 h after inhalation. Oxidative metabolism of

14 C-benzo[ a ]pyrene, in solution or adsorbed onto diesel particles at a concentration of 500 pmol/106 cells, was also detected in cell cultures of pulmonary macrophages obtained from beagle dogs. The quantity of metabolites increased with incubation time up to 45 and

125 pmol/10 6 cells in extracts and in the media, respectively, after 48 h of incubation. After incubation for 6 h, the major metabolites
found in extracts of cells were benzo[ a ]pyrene-7,8-diol (0.5pmol/10 6

cells) and benzo[ a ]pyrene-4,5-diol (0.2 pmol/10 6 cells); those in
extracts of culture medium were benzo[ a ]pyrene-7,8-diol (1.7 pmol/10 6

cells) and benzo[ a ]pyrene-9,10-diol (1.4 pmol/10 6 cells), in comparison with a total concentration of metabolites of 2.5 pmol/10 6 cells in the extracts and 6 pmol/10 6 cells in the medium. Minor metabolites were benzo[ a ]pyrene phenols (3-hydroxy- and 9-hydroxy-benzo[ a ]pyrene) and benzo[ a ]pyrene quinones (6,12, 1,5, and 3,5 isomers). The quantities of metabolite were similar when macrophages were incubated with benzo[ a ]pyrene in solution or adsorbed onto diesel particles (Bond et al., 1984).

Using the 32 P-postlabelling technique, Wong et al. (1986) found increased DNA adduct formation in the lungs of Fischer 344 rats after exposure for 31 months to diesel exhaust containing 7.1 mg/m 3 particles. After exposure of Fischer 344 rats and Syrian golden hamsters to dilutions of diesel engine exhaust for six months to two years, the level of haemoglobin adducts (2-hydroxyethylvaline and 2-hydroxypropylvaline), corresponding to the metabolic conversion of 5-10% of inhaled ethylene and propylene to its oxides, increased dose-dependently (Törnqvist et al., 1988). An adduct of benzo[ a ]pyrenediol epoxide with deoxyguanosine was identified after inhalation of benzo[a]pyrene adsorbed onto carbon black (20 mg/g) (Wolff et al., 1989), demonstrating the production of epoxides of PAHs by oxidative metabolism.

Methods for extracting diesel soot before testing for genotoxicity in vitro have been discussed. As dichloromethane and dimethyl sulfoxide do not correspond to leaching conditions in the lung, it can be assumed that the bioavailability of genotoxic materials is lower in lungs than in vitro. An investigation of dipalmitoyl phosphatidylcholine, the primary component of 

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B7. EFFECTS ON LABORATORY MAMMALS AND IN VITRO SYSTEMS

B7.1 Single exposure

Data on the acute toxicity of diesel exhaust are very limited; the two available studies suggest that diesel exhaust particles and their extracts have little acute toxicity.

Male ICR mice were administered diesel exhaust particles by intratracheal instillation, and mortality due to lung oedema was assessed for up to 24 h. At a dose of 0.9 mg per animal, all animals
died within 24 h. The LD 50 value was 0.6 mg per animal, equivalent
to about 20 mg/kg body weight. Diesel exhaust particles washed three times with methanol did not cause death at a dose of 1 mg per mouse, suggesting that the primary source of toxicity is the extractable organic compounds rather than the particles themselves. These compounds may produce superoxide radicals that damage endothelial cells (Sagai et al., 1993).

The LD 50 value for diesel exhaust extract in adult Syrian
golden hamsters after intraperitoneal injection was 1280 mg/kg body weight (Pereira et al., 1982).

B7.2 Short-term exposure

Tracer-monitored DNA synthesis, indicating a proliferative response, was observed in whole lung tissue of groups of 3-10 Fischer 344 rats exposed to diesel exhaust by inhalation at 6 mg/m 3 for 20h/day on seven days per week for periods of 1-14 days. The response was maximal after two days of exposure, showing a fourfold increase. The type II cell labelling index was significantly higher (fourfold) than that in controls after two and three days of exposure to diesel

exhaust. Both values returned to control levels after a one-week recovery period. Experiments involving incorporation of 14 C-palmitic acid revealed transient changes in lipid metabolism after one day of exposure, with a threefold increase in the amount of phosphatidyl- choline in lung tissue (Wright, 1986).

Exposure of various species by inhalation to 6 mg/m 3 resulted in slight alterations in lung function and histopathological changes.

Groups of 10 male Chinese hamsters were exposed for 8 h/day on seven days per week for six months to particle concentrations of either 6 or 12 mg/m 3 diesel exhaust.

Measurements of pulmonary function indicated dose-related increases in lung weight and dose-related decreases in vital and diffusion capacity. Pathological evaluation revealed oedema, thickening of the alveolar lining, a possible increase in collagen, and the presence of numerous particle-laden macrophages, which almost filled the alveolar spaces (Vinegar et al., 1981).

Groups of 41-51 male and female Hartley guinea-pigs were exposed for 20 h/day on seven days per week to exhaust from a light-duty diesel engine diluted to attain a concentration of 6 mg/m 3 particulate matter. Separate groups were exposed to raw exhaust and exhaust that had been irradiated with ultra-violet light for an average of about 1 h. Pulmonary function was measured, and electrocardiography was conducted on animals exposed for four weeks;

those examined for histological changes were exposed for eight weeks. The only changes noted after four weeks were a 35% increase in pulmonary flow resistance and a small, but significant decrease in heart rate in animals exposed to irradiated exhaust. No changes in the electrocardiogram were noted. The pathological changes were limited to hyperplasia of alveolar lining cells and slight focal thickening of
the interstitium in occasional alveoli with accumulation of particle-laden macrophages. Growth rates were unaffected (Wiester et al., 1980).

As part of the same study, cats (Pepelko et al., 1981) and male rats (15 per group) (Pepelko, 1982) were exposed for four weeks to unirradiated exhaust under identical conditions. The pulmonary function of rats was unchanged; the only changes were significant increases in vital and total lung capacity, but these differences were quite small and of questionable significance. The only functional change noted in the cats was a decrease in maximal expiratory flow rate at 10% of vital capacity. The pathological changes were again limited to focal thickening of the alveolar walls near accumulations of particle-laden macrophages. In general, the functional and pathological changes were limited under these experimental conditions.

Kaplan et al. (1982) exposed male Fischer 344 rats (168 per group), A/J mice (672 per group), and male Syrian golden hamsters (236 per group) to 1.5 mg/m 3 of diesel exhaust particles for 20 h/day on seven days per week for three months or to clean air. Animals were killed at the end of exposure and after an additional six-month recovery period. Mortality was not increased during the observation period, and body weight was not significantly altered in any species. At the end of exposure, the relative weight of the lung was significantly increased in rats. In mice, the pulmonary adenoma response was increased but not significantly. In all three species, 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmgrey to black discolouration of the lungs and lung-associated lymph nodes were observed. Microscopically, most of the particulate was found in macrophages, but some was free. After six months' exposure to clean air, the lungs of all species had considerably less pigment
accumulated in foci.

Sixteen male Fischer 344 rats were exposed to 6 mg/m 3 diesel exhaust for periods ranging from 6 h to 63 days, with subsequent histological examination of the lungs. The numbers of diesel particulate-containing alveolar macrophages, type II alveolar cells, and inflammatory cells were found to be increased over those in eight controls. After a nine-week exposure, accumulation of macrophages was observed near the terminal bronchioles (White & Garg, 1981). In rats that inhaled 6 mg/m 3 diesel exhaust for two weeks and were either

killed immediately or allowed to recover for six weeks before sacrifice, there was no statistical difference in the number of macrophage aggregates (240 000 and 220 000 per lung); however, the aggregates in the recovery group had significantly higher average diameter and volume, demonstrating macrophage attachment after the end of exposure (Garg, 1983).

The observations of Wiester et al. (1980) and Garg (1983) demonstrate the importance of alveolar macrophages in the clearance of particles of low solubility. These are engulfed by alveolar macrophages and eliminated via the mucociliary escalator and gastrointestinal tract, or, in the case of lung particle overloading, increasing fractions are sequestered in focal areas of the lung tissue and transferred to the lymph nodes (Muhle et al., 1990).

B7.3 Long-term exposure and studies of carcinogenicity

B7.3.1 Non-neoplastic effects

No obvious signs of toxicity were observed in groups of 183 male and 183 female Fischer 344 rats exposed by inhalation to diesel exhaust at 0.35, 3.5, or 7.1 mg/m 3 for 7 h/day on five days per week for up to 30 months (Mauderly et al., 1987). The body weights did not differ significantly from those of controls, and the exposures did not affect the life span of animals of either sex. Focal fibrotic and proliferative lung disease was observed in parallel with progressive accumulation of diesel soot in the lung. Henderson et al. (1988) reported the results of biochemical and cytological analyses of lung lavage fluid and lung tissue from rats and mice exposed in the same study, including determinations of lactic dehydrogenase, glutathione reductase, ß-glucuronidase, glutathione, and hydroxyproline, and the numbers of macrophages and neutrophils. No changes were seen in rats or mice exposed to 0.35 mg/m 3 , but at higher concentrations dose- and duration-dependent increases in biochemical parameters and cell numbers in lavage fluid were observed, indicating a chronic inflammatory response. Lung weights were also increased in male and female mice exposed to the two higher levels for any length of time, in rats exposed to 7.0 mg/m 3 for 12 months or longer, in female rats exposed to 3.5 mg/m 3 for 18 months, and in males exposed to 3.5 mg/m 3 for 24 months. Tissue enzyme levels, measured in lung lavage fluid as an indicator of damage, were consistently raised in rats and mice at the two higher exposure levels and in lung tissue of rats and mice at various times (most pronounced at 7 mg/m 3 ). These changes probably indicate an increase in lung tissue due to increased inflammatory cells and epithelial cell proliferation.

In the same study, Mauderly et al. (1988) described focal proliferative, fibrotic, and emphysematous changes in the lungs of animals at the higher doses. Measurements in vivo revealed a reduction in lung volume and compliance.

In a follow-up study, Mauderly et al. (1990) exposed male Fischer 344 rats that were either normal or had pre-existing pulmonary emphysema (elastase-treated) to diesel soot at 3.5 mg/m 3 for 7 h/day on five days per week for 24 months, to determine whether emphysematous rats were more susceptible to diesel exhaust. No increase in susceptibility was seen. The emphysematous lungs had a lower accumulation of particles than the normal lungs, which prevented an overproportional response of the compromised animals.

Male and female Fischer 344 rats exposed to diesel exhaust particles at 0.7, 2.2, or 6.6 mg/m 3 for 16 h/day on five days per week for two years or to filtered diesel exhaust showed an exposure-related reduction in the rate of weight gain throughout the study at the highest dose and an increase in absolute lung weight at the two higher doses (Brightwell et al., 1986, 1989).

Groups of 72 male and 72 female Fischer 344 rats exposed to diesel exhaust particles at 2 mg/m 3 for 7 h/day on five days per week for 24 months showed alveolar type II cell hyperplasia, inhibition of long-term clearance of particles and pulmonary lipidosis. No immunological abnormalities, no induction of pulmonary or hepatic xenobiotic metabolizing enzymes, and no effects on mortality, body weight gain, organ:body weight ratios or clinical chemical parameters were seen (Lewis et al., 1986, 1989). In the same study, a dose-related depression in phagocytic activity was seen in alveolar macrophages (Castranova et al., 1985), and a trend to increased pulmonary arterial wall thickness was observed in comparison with controls (Vallyathan et al., 1986).

In groups of 15 male monkeys, exposure to diesel exhaust (as described above) resulted in mild obstructive airway disease, but there was no evidence of restrictive lung disease. The effects were observed throughout the exposure period, after 6, 12, 18, and 24 

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Reduced body weight, significantly increased lung:body weight ratios, and type II cell proliferation were seen in female Fischer 344 rats exposed to whole-fraction diesel exhaust at 5 mg/m 3 for 8 h/day on seven days per week for 24 months (Iwai et al., 1986).

Groups of 96 male and 96 female Syrian golden hamsters, female NMRI mice, and female Wistar rats were exposed to diesel engine emissions at 4.4 mg/m 3 for 19 h/day on five days per week for 120-140 weeks (Heinrich et al., 1986a; Stöber, 1986). The body weights of mice were significantly reduced, by about 12%. Lung weights were increased after two years of exposure, by two- to threefold in rats and mice and by 50-70% in hamsters. Different levels of histological response were seen in the three species. Hamsters had thickened alveolar septa and bronchioalveolar hyperplasia. Of the mice, 64% had

bronchioalveolar hyperplasia (controls, 5%), 71% had multifocal alveolar lipoproteinosis, and 43% had multifocal interstitial fibrosis (controls, 4%). The nose, larynx, and trachea were not affected. Treated rats showed severe inflammatory changes in the lungs, thickened septa, foci of macrophages, and hyperplastic and metaplastic lesions, but no significant changes in respiratory rate, minute volume, compliance, or resistance were seen in 14 exposed rats. Significantly increased airway resistance and a significant decrease in dynamic lung compliance were observed in rats after two years of exposure. The hamsters had a significant increase in airway resistance and a nonsignificant reduction in lung compliance at termination of a one-year exposure; these changes persisted during the second year of exposure. In the same experiments, filtered diesel exhaust did not have the same effects as total exhaust in any of the three species (Heinrich et al., 1986a).

Groups of 27 female Fischer 344 rats and about 50 male and 50 female C57Bl/6N and ICR mice were exposed to particle concentrations of 2-4 mg/m 3 for 4 h/day on four days per week for up to 24 months. After 12 months, the incidence of alveolar hyperplasia was increased in all animals. Exposed rats had more severe inflammation than controls, with inflammatory exudate into the nose, and bronchitis and pneumonia, but this effect was ascribed to the housing, which was not specific pathogen-free (Takemoto et al., 1986).

Groups of nine male Hartley guinea-pigs were exposed at concentrations of 0.25, 0.75, or 1.5 mg/m 3 for 20 h/day on 5.5 days per week for up to two years. An analysis of the alveolar capillary membrane by quantitative morphometry showed that it was thickened as a
result of an increase in the absolute tissue volume of interstitium and type II cells. Exposure to 0.75 mg/m 3 for six months caused fibrosis in regions of macrophage clusters and focal type II cell proliferation, as observed under the light microscope. No appreciable change in morphometric parameters was seen after exposure to the lowest dose, but the higher doses increased the thickness of alveolar septa and the numbers of various types of alveolar cells (Barnhart et al., 1981, 1982).

As part of this investigation, Fischer 344 rats were exposed by inhalation to 0.25 or 1.5 mg/m 3 diesel exhaust particles, and various biochemical and cytological studies were conducted up to 36 weeks (Misiorowski et al., 1980). At the high dose, normalized lung weight, the rate of collagen synthesis, and the cellular and lipid content of lung tissue were significantly increased after six months. It was concluded that the reactivity of fibrogenic cells had been enhanced. In a long-term study of respiratory end-points in male guinea-pigs, including light and electron microscopy, lavage cytology, and lung tissue biochemistry, the lowest-observable-adverse-effect level (LOAEL) was 0.796 and the no-observable-adverse-effect level (NOAEL) was 0.258 mg/m 3 (US Environmental Protection Agency, 1992b).

Groups of 120 male Fischer 344 rats, 450 male A/J mice, and 120 male Syrian hamsters were exposed to 0.25, 0.75, or 1.5 mg/m 3 diesel exhaust particles for 20 h/day on seven days per week for 15 months. Mortality was not increased in any species. A dose-related deposition of pigment was observed histopathologically in alveolar macrophages and regional lymph nodes. Accumulation of alveolar macrophages was associated with an increase in connective tissue in alveolar walls, with mild fibrosis and proliferation of type II pneumocytes; these effects were termed pneumoconiosis. No treatment-related responses were found in organs other than the respiratory tract. The LOAEL was 0.735 and the NOAEL, 0.242 mg/m 3 (Kaplan et al., 1983).

Groups of 120 male and 95 female Fischer 344/Jcl rats were exposed to light-duty diesel engine exhaust particulates at 0, 0.1, 0.4, 1.1, or 2.3 mg/m 3 or heavy-duty diesel engine exhaust particulates at 0, 0.5, 1.0, 1.8, or 3.7 mg/m 3 for 16 h/day on six days per week for up to 30 months (Ishinishi et al., 1986, 1988; Suzuki et al., 1990). The body weights of females exposed to the highest dose of heavy-duty engine exhaust were decreased throughout the study by 15-20% in comparison with controls. No changes were observed histopathologically in the lungs at any concentration. Accumulation of particle-laden macrophages was observed at levels > 0.4 mg/m 3 . In areas of macrophage accumulation, bronchiolar epithelium replaced alveolar epithelium in the ducts. Proliferation of bronchiolar epithelium and type II cells and oedematous thickening and fibrosis of the alveolar septum were observed, which developed into small fibrotic lesions classified as hyperplastic. The incidences of hyperplastic lesions were 4, 4, 6, 12, and 87 per 124 rats exposed to light-duty engine exhaust and 1, 3, 7, 14, and 25 per 124 rats in those exposed to heavy-duty engine exhaust. The LOAELs were 1.2 mg/m 3 for light-duty and 1.0 mg/m 3 for heavy-duty exhaust, and 

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In the same experimental series, Kato et al. (1992) studied non-neoplastic lesions in the respiratory organs of the rats every six months. The marked changes consisted of intake of diesel particles by type I epithelium, hypertrophy, and glandular proliferation of type II epithelium, with many extended microvilli and increased numbers and size of lamellar inclusion bodies, focal increase of collagen fibres in the interstitium, and infiltration of particle-laden macrophages, neutrophils, mast cells, and plasma cells into the interstitium of alveolar septa. The most marked changes in the airway were focal shortening of cilia and protrusions of nonciliated cells. These morphological changes appeared in all groups exposed to particle concentrations > 1 mg/m 3 after six months of exposure and were more marked with increasing particle concentration and with time.

Exposure of Syrian golden hamsters to 4 or 11 mg/m 3 diesel exhaust for 7 h/day on five days per week for five months resulted in enlargement of liver sinusoids and loss of cristae in mitochondria (Meiss et al., 1981).

The induction of pulmonary adenomas and carcinomas in rats after long-term inhalation of carbon black is proposed to be due partly to inflammatory cells and their mediators (Driscoll et al., 1995; Oberdörster et al., 1995; Driscoll et al., in press). After exposure of rats to carbon black at 1, 7, or 50 mg/m 3 for 6 h/day on five days per week for 13 weeks, 353, 1923, and 7112 μg of carbon, respectively, were deposited per lung. After exposure to air or to 1 mg/m 3 of carbon black, no macrophage inflammatory protein-2 (MIP-2) mRNA expression and no increase in inflammatory cells was observed; with 7 or 50mg/m 3 , both MIP-2 mRNA and the number of neutrophils in bronchoalveolar lavage fluid were increased, and the hprt frequencies were 3.2 and 4.2 times greater than in controls.

Groups of 25 cats were exposed to either clean air or diesel exhaust for 8 h/day on seven days per week for two years at a concentration of 6 mg/m 3 through week 62 and to 12 mg/m 3 for weeks
62-124. Some of the cats were then held in clean air for an additional six months. No response was seen after 61 weeks, but after 123 weeks signs of restrictive lung disease were observed. Total lung capacity, forced vital capacity, functional residual capacity, peak expiratory flow rates, and diffusion capacity were significantly decreased (Moorman et al., 1985). Morphological evaluation revealed exposure-related lesions in the centriacinar region of the lungs, bronchiolar epithelial metaplasia associated with the presence of ciliated and basal cells, and alveolar macrophages containing diesel particle-like inclusions. After six months' recovery in clean air, the metaplasia had disappeared but the fibrosis had progressed (Hyde et al., 1985). In exposed cats, the epithelium of the terminal and respiratory bronchioles consisted of ciliated, basal, and Clara cells, whereas controls had only Clara cells (Plopper et al., 1983). These
observations indicate that the foci of the lesions were the epithelium, the interstitial compartments, and the centriacinar region of the lung. The metaplastic changes suggest that cancer may be induced by continuous exposure, while the other major effect is pulmonary fibrosis resulting in restrictive disease which is not ameliorated and in fact progresses after removal from exposure. Progression of effects is probably due to continued lung overloading with particulate matter, despite cessation of exposure.

The dose-dependent effects on lung morphology, histopathology, biochemistry, and cytology after long-term exposure to diesel exhaust described above are reflected in the following findings seen in rats at a lung burden of about 0.5 mg/lung (Henderson et al., 1988; Muhle et al., 1990):

-- The wet and dry weights of lungs increase by up to fourfold in comparison with those of concurrent controls. This is due to severe functional overloading in the lung, resulting in enhanced leukocyte influx and tissue proliferation.

-- Pulmonary inflammation is manifested by cytological (increased numbers of polymorphonuclear neutrophils) and biochemical (lactic dehydrogenase, collagen in lung lavage fluid) parameters.

-- The retention of material is characterized by an increasing number of particle-laden macrophages and by black discolouration of lungs due to sequestration of accumulated particles. Additionally, increased transfer of material to the lung-associated lymph nodes is observed.

-- Later findings during inhalation are proliferative alterations of the epithelial cells and onset of fibrosis.

B7.3.2 Carcinogenicity

B7.3.2.1 Inhalation

Rats were exposed in several studies to diesel emissions containing > 2 mg/m 3 of particles for more than two years, resulting in the induction of pulmonary tumours (Table 34). Many organic and inorganic gaseous components were also present in the test atmospheres, and the relatively high concentration of some pulmonary irritants like nitrogen dioxide, aldehydes, and sulfur dioxide may have contributed to the inflammatory response and to epithelial cell damage and subsequent cell proliferation. Exposure to gaseous PAHs, like naphthalene which is toxic to Clara cells, may influence the metabolic state of pulmonary cells. It is not clear however, whether the combined exposure to particles and gases in these experiments 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmfavoured tumour induction. It should be noted that Brightwell et al. (1986), Heinrich et al. (1986a), Iwai et al. (1986), Ishinishi et al. (1986), and Takaki et al. (1989) have reported that the gaseous phase, i.e. the proportion of exhaust without particles, does not induce pulmonary tumours.

Table 34. Carcinogenicity of diesel engine exhaust in rats

Strain No. and Exposure concentration Duration Lung Pulmonary tumours Probability Reference sex (mg/m 3 ) a (observation burden of error (%) b period) (mg/lung) No. % Mean Actual Continuous (f+m) (%)

Wistar 40 m 8.3 1.5 6 h/d, 5 d/week, up NR 1 17c NR Karagianes
40 m Clean air to 20 months 0 et al. (20 months) (1981)

Fischer 72 f 6.6 3.1 16 h/d, 5 d/week, 24 NR 39 54 38.5 < 0.1 Brightwell 344 71 m months (30 months) 16 23 et al.

72 f 2.2 1.1 11 15 9.7 < 0.1 (1986, 198 9

72 m 3 4

71 f 0.7 0.4 0 0.7

72 m 1 1
Filtered exhaust 0

0
142 f Clean air 1 1 1.4

140 m 3 2

Wistar 95 f 4.4 2.5 19 h/d, 5 d/week, 32 NR 17 18 < 0.1 Heinrich e t 92 f Filtered exhaust months (up to death) 0 al. (1986a )
96 f Clean air 0

Table 34 (contd)

Strain No. and Exposure concentration Duration Lung Pulmonary tumours Probability Reference sex (mg/m 3 ) a (observation burden of error (%) b period) (mg/lung) No. % Mean Actual Continuous (f+m) (%)

Fischer
344/Jcl Light-duty 60 f 2 1.1 16 h/d, 6 d/week, 30 NR 1 2 2.4 NR Ishinishi engine 64 m months (30 months) 2 3 et al.

59 f 1 0.6 2 3 4.1 (1986,1988 ; 64 m 3 5 Takaki et a 61 f 0.4 0.2 0 0.8 (1989)

64 m 1 2

59 f 0.1 0.06 2 3 1.6

64 m 0
59 f Clean air 2 3 3.3

64 m 2 3 Heavy-duty 60 f 4 2.3 3 5 6.5 1.8 engine 64 m 5 8

59 f 2 1.1 1 2 3.3

64 m 3 5

61 f 1 0.6 0 0

64 m 0

59 f 0.4 0.2 1 2 0.8

64 m 0
59 f Clean air 1 2 0.8

64 m 0
16 m Filtered exhaust 0

Table 34 (contd)

Strain No. and Exposure concentration Duration Lung Pulmonary tumours Probability Reference sex (mg/m 3 ) a (observation burden of error (%) b period) (mg/lung) No. % Mean Actual Continuous (f+m) (%)

Fischer 19 f 4.9 1.6 8 h/d, 7 d/week, 24 NR 8 42 < 1 d Iwai et al

. 344 16 f Filtered exhaust months (24 or 30 0 (1986) 22 f Clean air months) 1 5

Fischer 15 f 2.0-4.0 0.2-0.4 4 h/d, 4 d/week, 24 NR 0 NR Takemoto 344 15 f Clean air months (24 months) 0 et al. (1986)

Fischer 69 f 7 1.5 7 h/d, 5 d/week, 30 NR 13 19 16.1 < 0.1 (f) Mauderly e t 344/Crl 74 m months (up to death) 10 14 1.7 (m) al. (1986)

68 f 3.5 0.7 2 3 4.6

63 m 4 6

68 f 0.35 0.07 0 0.7

70 m 1 1
68 f Clean air 0 1.4

73 m 2 3

Fischer 227 f+m 7 1.5 7 h/d, 5 d/week, 30 20.8 12.8 < 0.1 Mauderly e t 344/N Crl 221 f+m 3.5 0.7 months (up to death) 11.5 3.6 4.8 al. (1987)

223 f+m 0.35 0.07 0.6 1.3 230 f+m Clean air (24 months) 0.9 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTable 34 (contd)

Strain No. and Exposure concentration Duration Lung Pulmonary tumours Probability Reference sex (mg/m 3 ) a (observation burden of error (%) b period) (mg/lung) No. % Mean Actual Continuous (f+m) (%)

Wistar 100 f 7.0 3.8 18 h/d, 5 d/week, 24 63.9 22 22.0 < 0.1 Heinrich e t 200 f 2.5 1.3 months (30 months) 23.7 11 5.5 < 1 al. (1992,

198 f 0.8 0.5 6.3 0 1995); 217 f Clean air (24 months) NR 1 0.5 Heinrich (1994)

Fischer 212 f+m 6.3 3.0 16 h/d, 5 d/week, 24 85.4 38 17.9 17.9 NR Nikula et a 344/N 210 f+m 2.4 1.1 months (24 months) 40.7 13 6.2 6.2 (1994) 214 f+m Clean air (23 months) 3 1.4 1.4

Fischer 49 f 4.7 1.3 15 h/d, 3 d/week, 24, NR 6 12.2 NR Kawabata 344/N 42 f 4.7 1.3 12, or 6 months 8 19.0 et al.

45 f 4.7 1.3 1 2.2 (1993)
48 f Clean air 5 10.4

Fischer 183 f+m 2.0 0.4 7 h/d, 5 d/week, 24 NR 8 4.4 4.4 NR Lewis et a l 344/N 180 f+m Clean air months 6 3.3 3.3 (1989)

From Pott & Heinrich (1987) and supplemented; NR, not reported

a Mass median aerodynamic diameter, 0.17-0.25 μm; 'Continuous', equivalent continuous value

b Unilateral Fisher's exact test

c Only six animals investigated

d With Yates' correction No difference in lung tumour incidence was seen in groups of male and female rats exposed for 7 h/day on five days per week for 24 months either to diesel exhaust particulates at 2 mg/m 3 or to filtered air (Lewis et al., 1989).

Exposure of six male Wistar rats to diesel exhaust soot at 8.3 mg/m 3 and coal dust for up to 20 months gave inconclusive results (Karagianes et al., 1981). The study had little statistical power owing to the small group size, and the insufficient duration of exposure may have prevented detection of tumours, although one mammary fibroadenoma and one lung bronchiolar adenoma were detected. One subcutal fibrosarcoma and one renal lymphoma were seen in controls.

Eight bronchioalveolar adenomas and nine squamous-cell tumours were seen in 95 rats (18%) that had inhaled 4.4 mg/m 3 diesel exhaust for 19 h/day on five days per week for 32 months. No tumours were observed in controls (Heinrich et al., 1986a). The dose-dependent carcinogenic potency of diesel exhaust was confirmed in a study in which rats were exposed to diesel exhaust at concentrations of 0.8, 2.5, or 7.0 mg/m 3 for 18 h/day on five days per week for 24 months (Heinrich et al., 1992; Heinrich, 1994; Heinrich et al., 1995).

In male and female rats exposed to diesel exhaust at 6.6 mg/m 3

, a high incidence of lung tumours was observed in animals dying or sacrificed after 24 months (24/25 females, 12/27 males). The incidence of lung tumours (Table 34) was derived by pooling pulmonary adenomas; squamous-cell carcinomas; adenocarcinomas; mixed ademomas, adenocarcinomas, and squamous-cell carcinomas; and mesotheliomas. Non-neoplastic pulmonary lesions were not described (Brightwell et al., 1986, 1989).

Groups of 24 female Fischer 344 rats were exposed to clean air, to whole-fraction (4.9 mg/m 3 particulates) fuel, or to filtered diesel exhaust for 8 h/day on seven days per week for 24 months. After six months of exposure, the rats exposed to whole exhaust had lower body weights than those in the other two groups; after 18 months, both exposed groups showed body weight reduction. The relative lung weight was increased after 12 months' exposure to whole exhaust, and epithelial proliferation and significantly elevated numbers of lung tumours were seen in animals exposed to the whole fraction after six months. Increased incidences of leukaemia and mammary gland tumours were also reported in animals exposed to whole or filtered exhaust; however, because of the small numbers of animals, the significance of the results cannot be determined (Iwai et al., 1986).

A total of 140 female and 140 male Fischer 344/N rats (in groups of three males and three females, five males and five females, or eight males and eight females, depending on the end-point) were exposed for 16 h/day on five days per week for up to 24 months, beginning at eight weeks of age, to particles of diesel exhaust or carbon black at 2.5 or 6.5 mg/m 3 of air, or to clean air (Mauderly et al., 1994). Rats were killed after 3, 6, 12, 18, or 23 months of exposure for histopathological assessment and measurement of lung and lung-associated lymph node burdens of particles, lung weight, bronchoalveolar lavage indicators of inflammation, DNA adducts in whole lung and alveolar type II cells, and chromosomal anomalies in circulating lymphocytes. Diesel exhaust and carbon black had nearly identical effects, with a similar relation to exposure. There was a dose-related slowing of particle clearance after three months, with progressive accumulation of particles in lungs and lymph nodes. Inflammation, epithelial proliferation, and fibrosis were progressive and related to dose. Diesel exhaust slightly increased the number of lung DNA adducts, and the numbers of DNA adducts in type II cells were increased by both diesel exhaust and carbon black. No exposure-related chromosomal damage was found in circulating lymphocytes. The incidence 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmof primary lung neoplasms was increased significantly and was related to the doses of both diesel exhaust and carbon black. The types of neoplasms were identical and included benign adenomas, malignant adenocarcinomas, squamous-cell carcinomas, and adenosquamous carcinomas; squamous cysts were also seen. The neoplasms had similar growth characteristics when transplanted; 50-67% of the transplanted squamous-cell carcinomas and 25-40% of the transplanted adenocarcinomas grew after injection into athymic mice (Table 35), whereas the squamous cysts did not.

Fischer 344/Jcl rats were exposed to a range of concentrations of exhaust emission particulates from light-duty (up to 2 mg/m 3 ) or heavy-duty diesel engines (up to 4 mg/m 3 ) for 16 h/day on six days per week for up to 30 months. After 18 months, marked hyperplasia of type II cells was noted in the groups at higher exposures. After 30 months, treatment-related pulmonary hyperplastic lesions were detected, and a dose-response relationship was seen for tumours in the groups exposed to heavy-duty but not light-duty diesel exhaust. The lung tumours were diagnosed as adenocarcinomas, squamous-cell carcinomas, or adenosquamous carcinomas. A significant correlation was
observed between the incidence of anthracosis and areas with hyperplastic lesions; furthermore, anthracosis and hyperplastic lesions did not appear in lungs exposed to particle-free exhaust. The authors concluded that the occurrence of hyperplastic lesions was dependent on the presence of carbon particles (Ishinishi et al., 1986, 1988; Takaki et al., 1989).

Table 35. Growth in athymic mice of lung tumours from rats exposed to
diesel exhaust and carbon black

Type of tumour Carbon black Diesel exhaust

Number Growing Number Growing implanted No. % implanted No. %

Adenocarcinom 8 2 25 10 4 40 Squamous-cell 2 1 50 3 2 67
carcinoma Squamous cyst 19 0 0 7 0 0

From Mauderly et al. (1994)

In a large-scale study of unfiltered diesel exhaust, 1097 Fischer 344 rats were exposed for up to 30 months to 0.35, 3.5, or 7.0 mg/m 3 of diesel soot. A group of 365 rats exposed to clean air served as controls. Exposure to exhaust did not cause overt signs of toxicity, and no significant, exposure-related alterations in body weight were observed. Rats were killed at six-month intervals, examined for their lung burdens of diesel soot, and subjected to complete necropsy. Diesel soot accumulated at all doses but minimally in the low-dose group. An increased incidence of tumours was observed in the groups at the medium and high doses; most (81%) of the tumours were observed after two years of exposure. The inhaled soot concentration and the lung burden of diesel soot were highly significantly related to tumour incidence ( P < 0.001) (Mauderly et al., 1987). In a follow-up study with longer daily exposure (16 h/day), the dose-dependency of the tumour incidence was reproduced (Nikula et al., 1994). In this study, the carcinogenicity of diesel exhaust soot (8% extractable organic material; 2.5 mg/m 3 ) was compared with that of carbon black (0.12% extractable organic material; 6.5 mg/m 3 ) in 1152 male and female Fischer 344 rats exposed for 16 h/day on five days per week for up to 24 months and examined histopathologically. The lung burden of particles was greater after exposure to diesel exhaust than to carbon black, but the neoplastic response in the lung was similar and usually occurred late in the study. When the lung burden of particles was used as the dose parameter, the carcinogenicity of carbon black was greater than that of diesel exhaust soot.

Groups of 50 four-week-old female Fischer 344 rats were exposed to diesel soot at a mean concentration of 4.73 mg/m 3 for 15 h/day three times per week for 6, 12, or 24 months, and animals that died after 18-30 months were evaluated for tumours. After 30 months, all animals, including controls, were sacrificed and examined for lung

tumours. The incidences of lung tumours were 10.4% in controls and 2.2% after six months' exposure to diesel exhaust, 19.0% after 12 months, and 12.2% after 24 months. No statistical analysis was presented, but it seems unlikely that there was a significant effect (Kawabata et al., 1993).

Concurrent administration of known carcinogens with diesel exhaust has also been studied. Exposure of rats to diesel exhaust given an intraperitoneal injection of N- nitrosodiisopropylamine or

N- nitrosodiethylamine did not influence the rates of tumours induced by the nitrosamines (Takemoto et al., 1986). The effects of treatment
of hamsters with N- nitrosodiethylamine or benzo[ a ]pyrene were not influenced by subsequent administration of diesel exhaust, whereas pretreatment of mice with benzo[ a ]pyrene or dibenzanthracene followed by diesel exhaust gave inconsistent results. Exposure to diesel exhaust did not affect the total tumour rates in rats pretreated with 6.25 or 12.5 g/kg body weight of N- nitrosodipentylamine; however, the incidence of squamous-cell tumours was increased in both pretreated groups (Heinrich et al., 1986a).

Hamsters were exposed for two years to unfiltered diesel exhaust at 0, 0.7, 2.2, or 6.6 mg/m 3 or to filtered diesel exhaust, and 

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N- nitrosodiethylamine subcutaneously before the beginning of exposure. The rate of pulmonary tumours was not increased by diesel exhaust, with or without pretreatment with the nitrosamine. Several hamsters that developed 'wet tail' infection after six months on test, resulting in significant mortality (46%), were treated with oxytetracycline, chloramphenicol, and dimetridazole (Brightwell et al., 1986, 1989).

Overall, the only clear effect in these studies of co-carcinogenicity in rats was an increase in the percentage of malignant tumours in animals treated with diesel exhaust plus carcinogens in comparison with the carcinogen alone (Heinrich et al., 1986a).

The results of studies in other species (mice, hamsters, and monkeys) treated by inhalation are given in Table 36. Heinrich et al. (1982) found significant increases in lung tumour incidence in mice, but this result could not be repeated in later studies (Heinrich et al., 1986a, 1992, 1995). Small but statistically significant increases in lung tumour incidence were reported in ICR and C57Bl mice (Takemoto et al., 1986) and in female Sencar mice (Pepelko & Peirano, 1983); however, because only small increases in non-malignant tumours were seen only in females in the study of Pepelko & Peirano (1983), the results must be considered to be weakly positive or equivocal. Hamsters showed no response to diesel exhaust.

Table 36. Long-term studies of diesel exhaust by inhalation in species other than rats

Strain, species No. and sex Exposure Results Reference

Material Particles Duration (mg/m 3 )

NMRI mouse 2 × 96 f (a) Exhaust 4 19 h/d, 5 d/week, Lung tumours (adenomas and Heinrich et (b) Filtered 27-28 months adenocarcinomas) (a) 24/76 (32%), (1986a) (c) Clean air including 13 (17%) adenocarcinomas; (b) 29/93 (31%), including 18 (19%) adenocarcinomas; (c) 11/84 (13%), including 2 (2.4%) adenocarcinomas Incidence in historical controls,

< 32%

ICR and 315 ICR, Exhaust or 2.0- 4 .0 4 h/d, 4 d/week, Lung tumours (adenomas and Takemoto et C57Bl/6N 297 C57Bl/6N clean air 13-28 months adenocarcinomas). ICR: 14/56; (1986) mice, newborn (sex 7/60 in controls; C57Bl/6N: unspecified) 17/150; 1/51 in controls

Sencar mouse 2 × 130 f (a) Exhaust 12 8 h/d, 7 d/week, Adenomas and carcinomas Pepelko & 2 × 130 m (b) Clean air to 15 months of (a) m: 5.9%; f: 16.3% Peirano (19 8 age (from (b) m: 3.8%; f: 7.2% conception to sacrifice)

NMRI mouse 2 × 80 f (a) Exhaust 7.5 18 h/d, 5 d/week, Lung tumours Heinrich et (b) Clean air 13.5 months, 9.5 (a) 32.1% (1992, 1995 ) months' recovery (b) 30%

Table 36 (contd)

Strain, species No. and sex Exposure Results Reference

Material Particles Duration (mg/m 3 )

NMRI and 3 × 120 f (a) Exhaust 4.5 18 h/d, 5 d/week; Lung tumours Heinrich et C57Bl/6N mice 3 × 120 f (b) Filtered NMRI, 23 months; NMRI: (a) 23.3% (1992, 1995 ) (c) Clean air C57Bl, 24 months (b) 46.7% + 6 months' (c) 30% recovery C57Bl: (a) 8.5% (b) 3.6% (c) 5.1%

Syrian golden 3 × 48 f (a) Exhaust 3.9 ± 0.5 7- 8 h/d, 5 d/week, No effect on survival, no lung Heinrich et hamster (b) Filtered up to 30 months tumours (1982) (c) Clean air

Syrian golden 3 × 48 f (a) Exhaust 4 19 h/d, 5 d/week, Normal life span, no lung tumours Heinrich et hamster 3 × 48 m (b) Filtered 27-28 months (1986a) (c) Clean air

Syrian golden 52 f + 52 m (a) Exhaust 0.7, 2.2, 16 h/d, 5 d/week, No significant increase in Brightwell hamster 104 f + 104 m (b) Filtered 6.6 24 months frequency of tumours in et al. (controls) (c) Clean air respiratory tract (1986, 1989 )

Cat 2 × 25 m (a) Exhaust 6 (1st 8 h/d, 7 d/week, Lung function: decrease in closing Pepelko & yr), 12 24 months volume after 1 year; reduction Peraino (19 8 (2nd yr) in inspiratory, vital, and total (b) Clean air lung capacity after 2 years; diagnosis: pulmonary fibrosis of
the interstitial or intra-alveolar type 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTable 36 (contd)

Strain, species No. and sex Exposure Results Reference

Material Particles Duration (mg/m 3 )

Cynomolgus 4 × 15 m (a) Coal dust 2.0 7 h/d, 5 d/week, No difference in tumour incidence Lewis et al

. monkey (b) Exhaust 2.0 24 months among groups (1986) (c) Coal dust 1.0

+ exhaust 1.0 (d) Clean air

It is clear that repeated inhalation of diesel exhaust at concentrations of more than about 2 mg/m 3 (actual value), corresponding to a calculated equivalent continuous concentration of about 1 mg/m 3 , increases the incidence of pulmonary tumours. These tumours occur late in life after exposures of 24 months.

B7.3.2.2 Other routes of exposure

Studies have also been conducted by intratracheal instillation in rats and hamsters and by painting on mouse skin (Table 37). While the former are especially useful for hazard identification and for evaluating specific mechanistic aspects, they are less useful for the purposes of risk characterization and no attempt was made to identify critical studies. The results of the studies by intratracheal instillation, however, confirm that both diesel exhaust particles and carbon black induce lung tumours and demonstrate that the specific surface area of carbonaceous particles is correlated with the tumorigenic potency. Both of these findings support the suggestion that a nonspecific particle effect is of crucial importance for the induction of lung tumours by diesel exhaust. The dermal experiments, conducted by Nesnow et al. (1982a,b, 1983), have been used to estimate risk quantitatively, by the comparative potency method.

B7.4 Dermal and ocular irritation; dermal sensitization

No data were available.

B7.5 Reproductive toxicity, embryotoxicity, and teratogenicity

B7.5.1 Reproductive toxicity

In a two-generation study of reproduction, 100 male and 100 female CD-1 mice were exposed by inhalation to exhaust from a light-duty diesel engine for 8 h/day on seven days per week, at a concentration of 12 mg/m 3 . Most treatment-related effects were minimal. Overall fertility and survival rates were not significantly altered (Pepelko & Peirano, 1983).

(C57Bl/6 × C3H)F mice (number not given) showed sperm abnormalities (reduced sperm count and weight of testis; teratospermia) after daily intraperitoneal injections of 50, 100, or 200 mg/kg body weight of diesel exhaust particles for five days. The highest dose caused an eightfold increase in abnormalities over that in controls and a significant decrease in sperm number (Quinto & De Marinis, 1984).

Table 37. Carcinogenicity of diesel exhaust after exposure other than by inhalation

Strain, No. and Exposure Schedule, Results Refer e species sex duration
Route Material Dose (mg)

Fischer 344 (a) 31 f Intratracheal (a) Activated carbon 1/animal 10/week, Survival rate, 71-83% Kawab a rat (b) 59 f instillation (b) Diesel particles 1/animal 30 months' (lowest ingroup b). Malign ant et al

. (c) 53 f (c) None observation lung tumours: (a) 7/23; (1986 ) (d) 27 f (d) Vehicle (b) 20/42; (c) 0/44; (d) 1 /23; P < 0.01. Benign and malignant tumours combined : (a) 11/23; (b) 31/42

Osborne- Groups Lung Organic material from Observation (a) 1 bronchioalveolar ade noma; Grimm e Mendel rat of 35 f implant diesel exhaust: until (b) 5 squamous-cell carcin omas; et al

. (a) Hydrophilic fraction 6.7 spontaneous (c) 1 bronchioalveolar ade noma; (1987 ) (b) Hydrophobic fraction 20.0 death (d) 6 carcinomas; (c) Nonaromatic compounds 19.2 (e) No tumours + PAHs (2 or (f) 1 carcinoma 3 rings) (g) 7 carcinomas, 1 adenom a (d) PAHs ( > 4 rings) 0.2 (h) No tumours (e) Polar PAHs 0.3 (i) 1 adenoma (f) Nitro-PAHs 0.2 (g) Reconstituted hydrophobic fraction 19.9 (h) None (i) Vehicle

Table 37 (contd)

Strain, No. and Exposure Schedule, Results Refer e species sex duration 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmRoute Material Dose (mg)

Wistar rat (a) 40 f Intratracheal (a) Diesel soot (34 m 2 / g) (a) 3 × 15 Primary lung tumours (%) Pott & (b) 58 f instillation (b) Diesel soot (70 m 2 / g) (b) 3 × 10 (a) 65 Rolle r (c) 38 f (c) Diesel soot (70 m 2 / g) (c) 3 × 20 (b) 60 (1994 ) (d) 37 f (d) Carbon black (270 (d) 3 × 15 (c) 66 Pott e

m 2 /g) (d) 65 al.(1 9 (e) 37 f (e) Activated charcoal (e) 3 × 10
(f) 39 f (860 m 2 /g) 3 × 20 (e) 27 (g) 40 f (f) NaCl solution (f) 3 × 10 (f) 0 (g) NaCl solution (g) 0.4 × 20 until (g) 0 ml spontaneous

death or 131

weeks

Wistar rat Groups Intratracheal (a) Diesel soot (native) (a) 1 × 15 Primary lung tumours (%): Heinr i of 48 f instillation (b) Diesel soot (toluene (b) 2 × 15 (a) 17 (1994 )

extract; 130 m 2 /g) 1 × 15 (c) Carbon black (c) 1 × 15 (b) 23 (toluene extract; (d) 1 × 15 4

270 m 2 /g; primary (e) Vehicle control (c) 21 particle, 15 nm) (d) Carbon black (toluene (d) 8

extract; 270 m 2 /g; (e) 0 primary particle,
15 nm) (e) NaCl solution

Table 37 (contd)

Strain, No. and Exposure Schedule, Results Refer e species sex duration
Route Material Dose (mg)

Syrian Groups Intratracheal (a) Diesel particles 1.25, 1/week, 15 1 lung adenoma at high dos e Shefn e golden of 50 m; instillation (b) Diesel particles + 2.5, in groups (a) and (c) afte r 61 et al

. hamster various same amounts of weeks weeks; no lung tumours in (1982
controls ferric oxide controls (c) Diesel particle

extract + ferric oxide

Syrian (a) 3 × Intratracheal (a) Exhaust extract 0.1, 0.5, 1/week, 15 Survival rates: (a) 95, 92 , Kunit a golden 62 m instillation (b) Vehicle 1 weeks 71%; (b) 98%. No differenc e in et al

. hamster (b) 59 m (c) 0.5 mg tumour incidence between ( a) (1986 ) (c) 62 m benzo[a]pyrene and (b); 88% respiratory tumours in positive contro ls

C57Bl 12 m, Dermal Acetone extract of 0.5 ml 3 times/week 16 mice dead by 10 weeks, 33 Kotin mouse 40 f, particles for life or skin tumours by 13 months, 2 et al

. 69 22- 2 3 months skin papillomas, no tumours in (1955 )

controls controls

A mouse 50 m, Dermal Acetone extract of 0.5 ml 3 times/week Males: 8 skin tumours by 1 6 25 f, particles for life or 1 papilloma, 3 squamous-ce ll
34 22-23 months carcinomas. Females: 20 sk in controls tumours by 13 months,17 tu mours at 13-17 months, no tumour s in

controls

Table 37 (contd)

Strain, No. and Exposure Schedule, Results Refer e species sex duration
Route Material Dose (mg)

Sencar Groups Dermal Dichloromethane 0.1, 0.5, 1/week, 50- 5 2 Skin carcinomas: Nesno w mouse of 40 m extracts of particles 1.0, 2.0, weeks Engine A: 3% (m), 5% (f) a t et al

. + 40 f from engines (A, B, E) or 4.0 4 mg Engine B: 3% (m) at 0 .5 mg (1982 a Benzo[a]pyrene 12.6- Engine E: 3% (f) at 0.1 mg 1983) 202 μg Positive control: 10-90%

C57Bl/6N Groups Subcutaneous Particles in olive oil 10, 25, 1/week, 5 First tumours palpated in: Kunit a mouse of injection containing 5% dimethyl 50, 100, weeks; 18 week 47 at 25 mg/kg bw et al

. 15-30 f sulfoxide 200, or months' week 30 at 50 mg/kg bw (1986 ) 500 mg/kg observvation week 27 at 100 mg/kg bw bw week 39 at 200 or 500 mg/k g bw Malignant fibrous histiocy tomas in 5/22 mice at 500 mg/kg bw; 0/38 in controls

PAH, polycyclic aromatic hydrocarbon; TPA, 12-O-tetradecanoylphorbol 13-acetate Groups of 15 male cynomolgus monkeys were exposed by inhalation to 2 mg/m 3 diesel particulates for 7 h/day on five days per week for two years. Sperm motility and velocity were similar to those of controls (Lewis et al., 1989).

In a test for dominant lethal mutation, male Fischer 344 rats 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htminhaled 2 mg/m 3 diesel exhaust for 7 h/day on five days per week for six months and were subsequently mated with untreated females. Live and dead implants and preimplantation losses were analysed on days 19-20 of gestation: no significant effects were observed (Lewis et al., 1989). In a similar test, 100 male and 54 female T stock mice were exposed to 6 mg/m 3 diesel exhaust for 8 h/day on seven days per week for seven weeks. There were no significant dominant lethal effects in males or females. A reduced number of corpora lutea (reproductive function) was the only significant result (Pepelko & Peirano, 1983).

B7.5.2 Embryotoxicity

No increase in the frequency of sister chromatid exchange was seen in the livers of fetuses of Syrian golden hamsters that inhaled diesel exhaust particles at 12 mg/m 3 for 8 h/day on days 5-13 of gestation. An intraperitoneal injection of 300 mg/kg body weight of diesel particulates on day 12 of gestation also had no significant effect on this end-point; however, intraperitoneal injection of 23% particulate mass extracted with methylene chloride on day 12 resulted in a dose-dependent increase in sister chromatid exchange frequency in fetal liver on day 13, and a doubling was seen at 320 mg/kg body weight. It was concluded that chemicals must be eluted from diesel particles in order for the genotoxic material to cross the placenta (Pereira et al., 1982).

B7.5.3 Teratogenicity

Sprague-Dawley rats and New Zealand white rabbits were exposed by inhalation to exhaust from a light-duty diesel engine, at a particulate matter concentration of 6 mg/m 3 , for 8 h/day on seven days per week.

Twenty rats were exposed on days 5-16 of gestation and 20 rabbits on days 6-18. The numbers of viable fetuses per litter, dead fetuses per litter, resorptions per litter, implantation sites per litter, corpora lutea per litter, and average fetal weight did not differ significantly from those of controls (Pepelko & Peirano, 1983).

Reproductive and developmental toxicity are considered unlikely to be critical end-points for diesel exhaust.

B7.6 Mutagenicity and related end-points

B7.6.1 In vitro

The genetic effects of particles and particle extracts have been described (Lewtas & Williams, 1986; Morimoto et al., 1986; Henschler, 1987). Diesel exhaust extracts rather than particles were used in most
of these studies. Point mutations were observed in vitro in bacteria and mammalian cells. Extracts caused chromosomal aberrations, DNA damage, sister chromatid exchange, and cell transformation (Table 38). The mutagenic potency of diesel engine exhaust depended in several studies on the characteristics of the diesel fuel and the type, age, and operating conditions of the engine (Henschler, 1987). As a rule, organic extracts of particles were mutagenic in the absence of an exogenous metabolic activating system (S9) (Ball et al., 1990); addition of S9 decreased (Lewtas, 1983) or suppressed (Morimoto et al., 1986) the mutagenic activity, perhaps by increasing protein binding of mutagenic material or by metabolic detoxification of directly acting mutagens, such as the nitroarenes, in the Ames test. It can therefore be concluded that PAHs and thioarenes, which must be metabolically activated, do not account for the mutagenic potency of diesel particles. Salmeen et al. (1985) and Beland et al. (1985) showed that nitroarenes are the main genotoxic agents in Salmonella typhimurium T98 in the absence of S9. A number of other investigators have reported the presence of nitrated PAHs in extracts of diesel engine exhaust particles (Handa et al., 1983), including many three-, four-, and five-ring structures (Schuetzle et al., 1982). The concentration of 1-nitropyrene, one of the prevalent species, has been reported to be 15-25 mg/kg particulate matter, whereas benzo[ a ]pyrene has been found at concentrations up to 50 mg/kg. The nitrated PAHs are important in the health effects of diesel exhaust since they are effective mutagens in microbial and human cell systems (Patton et al., 1986). Some nitrated PAHs are also carcinogenic in animals (Imaida et al., 1991). Ball & Young (1992) found that strain T102, which does not respond to the mutagenic action of nitroarenes, responds to a class of oxidizing compounds that can interact with DNA; these compounds are not nitroarenes or typical PAHs.

The effects of diesel soot particles with and without adsorbed organic substances, diesel soot extract without particles, an isolated fraction of PAHs, and titanium dioxide and carbon black (Printex 90) as reference particles were investigated in vitro on hamster lung epithelial cells (Mohr & Riebe-Imre, 1992). The substances were added to the cells at concentrations of 100-300 mg/litre. Diesel soot extract, but not the PAH moiety isolated from the extract, stimulated mixed-function oxygenases. The diesel soot extract was more cytotoxic than the PAH fraction for the epithelium of the hamster respiratory tract, but both mixtures induced the development of micronuclei. Under the study conditions, only the PAH fraction led to transformation of cells. The respiratory epithelium of hamsters was more sensitive than

human lung epithelial cells investigated in parallel, perhaps because hamster cells are generally more easily transformed than human cells. Diesel soot particles, titanium dioxide, and Printex 90 had hardly any cytotoxic effect but triggered transformation. The authors concluded that a combination of effects of particles and PAHs are responsible for the degeneration of cells. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmSchiffmann & Henschler (1992) studied the effects of diesel soot extract and isolated fractions of PAHs, oxy-PAHs, and nitro-PAHs on hamster embryo fibroblasts in vitro. The end-points investigated were induction of micronuclei, unscheduled DNA synthesis, and cell
transformation. Diesel soot extract induced micronuclei at concentrations of 80-400 mg/litre, whereas the fractions of various PAHs were strongly cytotoxic. Diesel soot extract and the PAH fraction transformed the fibroblasts; the nitro- and oxy-PAH fractions had a weaker transforming effect, but their cytotoxic potency was clearly stronger. Kinetochore analysis showed a high percentage (37%) of chromosomes in micronuclei. No unscheduled DNA synthesis was induced. Micronuclei were also induced in human embryonic lung fibroblasts treated with diesel soot extract under comparable experimental
conditions.

B7.6.2 In vivo

The genotoxicity of total diesel exhaust, diesel particles, and
diesel soot extracts in vivo has been investigated in somatic cells (Table 38). All of the substances induced genotoxic responses. Total exhaust induced only sister chromatid exchange in Syrian golden hamsters. The frequency of micronuclei was not increased significantly in mice, even at high doses (up to 640 mg/kg body weight). Organic extracts had clear genotoxic effects in various assays (Henschler, 1987).

No mutagenic activity was found in urine taken from rats exposed to diesel exhaust emission by inhalation at 2 mg/m 3 for 7 h/day on five days per week for up to two years. A slight but nonsignificant
increase in micronuclei was observed in the bone marrow of mice that had been exposed for six months; no increase in micronuclei was detected in rats over a 24-month period (Ong et al., 1985).

In studies of heritable mutations in T stock mice, males were examined for point mutations, induction of dominant lethal mutations, translocations, and spermatogonial survival; females were examined for oocyte death and dominant lethal mutations. No significant effect was seen on germ cells. A reduced number of corpora lutea (reproductive function) was the only significant result (Pepelko & Peirano, 1983).

B7.6.3 DNA adduct formation

Extensive research has been done to determine whether the DNA adducts induced in lungs by diesel exhaust are related to later tumorigenesis.

In rats that inhaled benzo[ a ]pyrene absorbed to carbon black (20 mg/g), an adduct of the diol epoxide with deoxyguanosine was identified (Wolff et al., 1989), demonstrating that epoxides of PAHs are produced by oxidative metabolism. Haemoglobin and albumin adducts were also investigated.

Methods have been developed to detect DNA adducts of 1-nitropyrene or 1,6-dinitropyrene, which are markers of diesel engine exhaust. The adducts are analysed in rat tissue or peripheral blood lymphocytes by the 32 P-postlabelling method and may be useful for the dosimetry of 1-nitropyrene or diesel exhaust particulates in occupational settings (El-Bayoumy et al., 1994). Studies on adduct formation in lung DNA induced by inhaled diesel exhaust have been conducted by Wong et al. (1986), Wolff et al. (1990), Bond et al. (1988, 1989, 1990a,b), and Gallagher et al. (1993, 1994), all involving the 32 P-postlabelling technique. In general, these studies suggest that DNA adducts are good measures of the 'effective dose' of carcinogenic compounds.

Wong et al. (1986) found increased DNA adduct formation in the lungs of Fischer 344 rats exposed to diesel exhaust particles at 7.1 mg/m 3 for 31 months. After Fischer 344 rats and Syrian golden hamsters were exposed to dilutions of diesel engine exhaust for six months to two years, the level of haemoglobin adducts (2-hydroxy- ethylvaline and 2-hydroxypropylvaline) increased dose-dependently, corresponding to metabolic conversion of 5-10% of inhaled ethylene and propylene to their oxides (Törnqvist et al., 1988).

Bond et al. (1988) designed experiments to determine the location of DNA adducts in the respiratory tract of rats exposed to exhaust. Fischer 344 rats were exposed for 12 weeks to diesel exhaust soot at 10 mg/m 3 ; they were then sacrificed, and various regions of the respiratory tract were removed and analysed for DNA adducts. Adducts were found only in peripheral lung tissue and nasal tissue; the total levels were highest in the peripheral tissue (about 18 adducts per 10 9 bases). Thus, the levels of total DNA adducts and exhaust-induced adducts are highest in the region of the rat respiratory tract where tumours are formed after exposure to a carcinogenic concentration of diesel exhaust (Mauderly et al., 1987).

Bond et al. (1990c) exposed groups of Fischer 344 rats to soot at 0.35, 3.5, 7.0, or 10 mg/m 3 for 12 weeks and found that the levels of DNA adducts were similar, at about 14 adducts per 10 9 bases. This level is nearly twice as high as that found in sham-exposed rats.

Thus, DNA adduct formation in lungs is independent of the concentration of exhaust at the levels tested. One explanation for these results (Bond et al., 1990a) is that the lung enzymes responsible for the metabolism of soot-associated chemicals to
metabolites that bind to DNA were saturated at the concentrations tested. These data also suggest that other factors are important in the carcinogenicity of diesel exhaust, since the number of adducts was elevated at an exposure level (0.35 mg/m 3 ) that did not increase lung tumour incidence. The induction of DNA adducts at low 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmconcentrations is likely to result in tumour initiation by the organic compounds present; at higher concentrations, the particles will induce cell death, and subsequent proliferation may well act as a promotional event. The combination may then result in detectable increases in
tumour incidence.

Bond et al. (1990a) also investigated the time course for DNA adduct formation and persistence. Fischer 344 rats were exposed to soot at 7 mg/m 3 for up to 12 weeks and were sacrificed at 2, 4, 8, 12, 14, and 16 weeks after the start of exposure. DNA adducts accumulated slowly in the lung, and the number was highest at the end of exposure, representing about 160% of the level seen in controls exposed to air only. The levels declined rapidly after termination of exposure and were not significantly different from those of controls four weeks later. Thus, steady-state levels of DNA adducts would be reached during long-term exposure to diesel exhaust.

Bond et al. (1989, 1990c) and Wolff et al. (1990) investigated whether exposure to carbon particles without associated mutagenic organic chemicals also increases DNA adduct levels. Rats were exposed either to diesel exhaust or carbon black at 0, 3.5, or 10 mg/m 3 for 12 weeks. Solvent-extractable organic compounds made up about 30% of the diesel soot but only about 0.04% of the carbon black particles. Exposure to the highest levels of both carbon black and diesel exhaust increased the DNA adduct levels in lungs, although the levels in rats exposed to diesel exhaust were about 30% higher than those in rats exposed to carbon black. Heavy exposure to carbon particles can therefore increase DNA adduct formation, although the concentrations

are lower than those of exhaust-related adducts.

Bond et al. (1990b) investigated whether DNA adducts could be formed in specific cells of the rat lung, i.e. alveolar type II cells. After exposure of rats to 6.2 mg/m 3 diesel exhaust or carbon black particles for 12 weeks (16 h/day on five days per week), type II cells were isolated and their DNA analysed for adducts. Both diesel exhaust
and carbon black resulted in about a fourfold increase in total DNA adducts in type II cells. DNA adduct levels in peripheral lung tissue were increased by 60-80% in male and female Fischer 344 rats and female cynomolgus monkeys but not in female B6C3F1 mice or female Syrian hamsters after inhalation of diesel soot at 8.1 mg/m 3 for 12 weeks (Bond et al., 1989, 1990c).

Gallagher et al. (1993, 1994) exposed female Wistar rats to diesel soot at 7.5 mg/m 3 and to carbon black at 11.3 mg/m 3 for 24 months and measured DNA adducts in lungs. The mean adduct levels were similar in the two groups but were not significantly greater than
those in controls. Time-course studies indicated that the levels in the lungs of rats exposed to diesel exhaust were lower at 24 months than after two or six months of exposure, presumably as a result of dilution due to increased cell proliferation. The level of a single DNA adduct, thought to be derived from a nitro-PAH in diesel exhaust and not observed in rats exposed to carbon black or titanium oxide, was elevated over that in controls after two, six, and 24 months. The
DNA adduct levels in control rats increased over the duration of the 24-month study as an effect of age.

Gallagher et al. (1993) compared DNA adduct formation after exposure to diesel emissions in vitro and in vivo. After exposure of human lymphocytes to diesel extract, five major DNA adducts were detected, one of which was characterized as an adduct with
benzo[ a ]pyrene. One was also detectable in reaction products of calf thymus DNA and diesel particle extract in vitro and in skin and lung DNA from mice treated dermally with 50 mg diesel extract in vivo. The differences in DNA patterns in vitro and in vivo (skin and lung) may reflect differences in metabolic pathways.

Taken together, the studies of DNA adducts suggest that some organic chemicals in diesel exhaust can form DNA adducts in lung tissue and may play a role in the carcinogenic effects. As pointed out by Bond et al. (1989), however, DNA adducts alone cannot explain the carcinogenicity of diesel exhaust, and other factors, such as chronic inflammation and cell proliferation, are also important.

B7.7 Special studies

B7.7.1 Immunotoxicity

During the clearance process, one possible pathway is translocation of diesel particulates into the lymphatic channels. After male guinea-pigs were exposed to diesel exhaust at a particle concentration of 1.5mg/m 3 for four or eight weeks, the B- and T-cell counts in lymph nodes were not altered, and there were no significant changes in blood or spleen (Dziedzic, 1981).

Fischer 344 rats were exposed to 2 mg/m 3 diesel exhaust for 7h/day on five days per week for 12 or 24 months and the immunological function of splenic B and T cells was measured by enumerating antibody-producing cells in the spleen or monitoring the proliferative response. No changes were observed (Mentnech et al., 1984).

Table 38. Genotoxicity of diesel exhaust, particles, and extracts

Test organism End-point Exhaust Particles Extract Reference

**Mutagenicity**
**Bacteria** S. typhimurium Point mutation (his) + + Huisingh et al. (1978) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmS. typhimurium Point mutation (his) + + Loprieno et al. (1980) S. typhimurium Point mutation (his) + Clark & Vigil (1980) S. typhimurium Point mutation (his) + + Li & Royer (1982) S. typhimurium Point mutation (his) + Salmeen et al. (1984) S. typhimurium Point mutation (his) + Ong et al. (1985) S. typhimurium Point mutation (his) + Bechtold et al. (1986) S. typhimurium Point mutation (his; + Whong et al (1986) SOS umu test) S. typhimurium Point mutation (his) + Wallace et al. (1987) S. typhimurium Point mutation (his) + a Wallace et al. (1990) S. typhimurium Point mutation (his) + Lewis et al. (1989) S. typhimurium Point mutation (his) + Rasmussen (1990) S. typhimurium Point mutation (his) + + Keane et al. (1991) S. typhimurium, E. coli Point mutation (trp) + Crebelli et al. (1991) E. coli Point mutation (trp) + Lewtas (1983)

Mammalian cells L5178Y mouse Point mutation (tk) + Lewtas (1983) lymphoma cells L5178Y mouse Point mutation (tk) + Mitchell et al. (1981) lymphoma cells

Table 38 (contd)

Test organism End-point Exhaust Particles Extract Reference

**Mutagenicity** (contd) Chinese hamster Point mutation (hprt) + Chescheir et al. (1981) ovary cells Chinese hamster Point mutation (hprt) (+) Chescheir et al. (1981) ovary cells Chinese hamster Point mutation (hprt) (+) Casto et al. (1981) ovary cells Chinese hamster Point mutation (hprt, + Morimoto et al. (1986) lung V79 cells ATPase) Human xeroderma Point mutation (hprt) + + McCormick et al. (1980) pigmentosum fibroblasts Human lymphoblast Point mutation (tk) + Barfknecht et al. (1981)

TK6 cells Balb/c3T3 mouse Point mutation (ATPase) + (+) Curren et al. (1981)
fibroblasts

**DNA damage** Syrian hamster DNA chain breaks - Casto et al. (1981) embryo cells (alkaline elution) Human xeroderma DNA damage + + McCormick et al. (1980) pigmentosum fibroblasts Rat primary Unscheduled DNA + Lewtas (1983) hepatocytes synthesis Chinese hamster lung Unscheduled DNA + a + Gu et al. (1994) V79 cells synthesis

Table 38 (contd)

Test organism End-point Exhaust Particles Extract Reference

**Chromosomal effects** Chinese hamster Sister chromatid exchange + Lewtas (1983) ovary cells Chinese hamster Chromosomal aberrations + Lewtas (1983) ovary cells

Chinese hamster Chromosomal aberrations + b Hasegawa et al. (1988) lung V79 cells - c Chinese hamster Sister chromatid exchange + Hasegawa et al. (1988) lung V79 cells Chinese hamster Sister chromatid exchange + + Keane et al. (1991) lung V79 cells Human lymphocytes Chromosomal aberrations + Lewtas (1983) Chinese hamster Sister chromatid exchange + Morimoto et al. (1986) lung V79 cells Human lymphocytes Sister chromatid exchange (+) Tucker et al. (1986) Human lymhoblastoid Sister chromatid exchange + Morimoto et al. (1986)

cells Hamster embryo Micronuclei + Schiffmann & Henschler fibroblasts (1992) Chinese hamster lung Micronuclei + a + Gu et al. (1992) V79 and ovary cells

Table 38 (contd)

Test organism End-point Exhaust Particles Extract Reference

**Cell transformation** Balb/c3T3 mouse fibroblasts Cell transformation (+) Curren et al. (1981)
Balb/c3T3 mouse fibroblasts Cell transformation + b Hasegawa et al. (1988)
(+) c Hamster lung epithelial cells Cell transformation + + Mohr & Riebre-Imre (199 2) Hamster embryo fibroblasts Cell transformation + Schiffmann & Henschler (1992)

**In vivo a**  Mouse Micronuclei, bone marrow - - + Pereira (1982) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmMouse Micronuclei, bone marrow - Lewis et al. (1989) Mouse Micronuclei, bone marrow - Morimoto et al. (1986) Mouse Micronuclei, bone marrow - - - Pepelko & Peraino (1983 ) Mouse Micronuclei, bone marrow (+) Ong et al. (1985) Rat Micronuclei, bone marrow - Ishinishi et al. (1988) Rat Micronuclei, bone marrow - Ong et al. (1985) Chinese hamster Micronuclei, bone marrow (+) - - Pepelko & Peraino (1983 ) Mouse Sister chromatid exchange, - + + Pereira (1982)

bone marrow Rat Sister chromatid exchange, - Ishinishi et al. (1988)

bone marrow Rat Sister chromatid exchange, - Morimoto et al. (1986)

bone marrow Rat Sister chromatid exchange, - Ong et al. (1985) peripheral blood leukocytes

Table 38 (contd)

Test organism End-point Exhaust Particles Extract Reference

Rat Sister chromatid exchange, - Lewis et al. (1989) lymphocytes Syrian golden Sister chromatid exchange, + Pereira (1982) hamster lung cells (inhalation) Syrian golden Sister chromatid exchange, + Guerrero et al. (1981) hamster lung cells (instillation) Syrian golden Sister cromatid exchange, + + Pereira (1982) hamster lung cells (instillation) Syrian golden Sister chromatid exchange + Morimoto et al. (1986) hamster liver cells (transplacental) Syrian golden hprt mutation + Morimoto et al. (1986) hamster (transplacental) Mouse Mutation, S. typhimurium - Morimoto et al. (1986) (host-mediated assay) Mouse Heritable effects on germ - Pepelko & Peirano (1983) cells (various assays; m/f) Drosophila Heritable effects (sex-linked Schuler & Niemeier (1981 ) melanogaster recessive lethal mutation) -

his, histidine independence; trp, tryptophane independence; hprt, hypoxanthine-guanine-phosphoryltransferase (8-aza- or 6-thioguanine resistance); tk, thymidine kinase (bromodeoxyuridine or trifluorothymidine resistance); ATPase, Na+/K+-ATPase (ouabain resistance); +, positive; (+), weakly positive

a Dispersed in artificial surfactant

b Light-duty engine

c Heavy-duty engine

d Modified from Henschler (1987) and supplemented Fischer 344 rats and CD-1 mice were exposed to diesel exhaust particles at 0.35, 3.5, or 7 mg/m 3 for 6, 12, 18, or 24 months to investigate whether the accumulation of diesel particulates in lymph nodes (indicated by black discolouration) influences the subsequent response to immunization by sheep red blood cells, evaluated by the presence of immunoglobulin (Ig) M antibody-forming cells. In rats, the total number of lymphoid and antibody-forming cells was significantly increased at the medium and high aerosol concentrations. In mice, the number of lymphoid cells was increased only at the high concentration. The authors concluded that diesel exhaust particles have only a minimal effect on the immune and antigen filtration functions in lung-associated lymph nodes because the relative numbers of antibody-forming cells and specific IgM, IgG, and IgA antibodies in rat sera were not significantly changed (Bice et al., 1985).

Five intranasal inoculations of various doses of a suspension of diesel engine exhaust particles in ovalbumin were administered to BDF1 mice at intervals of three weeks. Ovalbumin IgE antibody titres, assayed by passive cutaneous anaphylaxis, were enhanced by doses as low as 1 μg (Takafuji et al., 1987, 1989). Similarly, primary IgE responses were increased after intraperitoneal administration to mice of ovalbumin or cedar pollen allergen mixed with diesel exhaust particulates (Muranaka et al., 1986).

Female CR/CD-1 mice were exposed by inhalation to 6-7 mg/m 3 diesel exhaust for 2-6 h (acute), 8 h/day for 2-16 days (subacute), or 8h/day for about 300 days (chronic). Immediately after the end of exposure, animals were exposed to the infectious aerosols S. typhimurium, Streptococcus pyogenes, or A/PR8-34 influenza virus. Increased susceptibility to post-infection mortality was seen with the bacterial but not the viral pathogens. Nitrogen dioxide and acrolein vapour had specific effects (Campbell et al., 1981b).

CD-1 mice were exposed to 2 mg/m 3 diesel exhaust for one to six months. After three months of exposure and subsequent infection with influenza virus, an enhanced severity of response was seen, involving significant lung consolidation and focal macular collections of particle-laden macrophages (Hahon et al., 1985).

B7.7.2 Behavioural effects

Sprague-Dawley rats were exposed for 8 h/day on seven days per week for 16 weeks to diesel exhaust diluted to a particulate matter concentration of 6 mg/m 3 . Spontaneous locomotor activity, measured weekly on Wahman LC-34 running wheels, was significantly decreased during 8, 9, 11, and 12 weeks of exposure. In rats exposed for 20 h/day on seven days per week for six weeks, forced activity on a motorized treadmill was measured during the last week. Exhaustion
occurred in less than half the time that it occurred in control animals. Groups of 10 rats were exposed to diesel exhaust at 6 mg/m 3 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmfor 20 h/day on days 1-7 post partum and were then held in clean air until 15 months of age. In bar pressing acquisition training carried out at five-day intervals for the next 42 days, the rate of learning was much slower than in control rats. The difference was highly statistically significant (Pepelko & Peirano, 1983).

As these three studies were conducted at a high concentration, 6 mg/m 3 , the practical consequences of the effects observed are not known. The results, while limited in scope, indicate, however, that behavioural and neurophysiological effects may be important toxic end-points which should be investigated further.

B7.8 Factors that modify toxicity; toxicity of metabolites

Little information is available, but chemicals and other factors may modify the toxicity of diesel emissions. For instance, chlorophyllin, a derivative of the green pigment chlorophyll, has been reported to inhibit or reduce the mutagenic activity of diesel particulate extracts in the Ames test (Ong et al., 1986).

B7.9 Mechanisms of toxicity; mode of action

It is not clear whether DNA-reactive or non-DNA-reactive mechanisms, or a combination of the two, are responsible for the carcinogenic action of inhaled diesel exhaust in laboratory animals. The results of several studies indicated that many PAHs (e.g. benzo[ a ]pyrene) cause a carcinogenic response in rodents, but it is
not known whether the same PAHs when associated with diesel exhaust also induce a carcinogenic response. One hypothesis for the mechanism of the tumorigenic response is that organic chemicals (e.g. PAHs, nitro-PAHs) desorb from soot particles, are metabolized to reactive metabolites, and interact with lung DNA to initiate carcinogenesis. This would be a predominantly DNA-reactive mechanism. The observation that organic chemicals associated with diesel soot can be metabolized by lung cells to metabolites that can form DNA adducts after long-term exposure to diesel exhaust supports this hypothesis. Studies with carbon black essentially devoid of PAHs but which also form DNA adducts do not, however, corroborate the PAH-DNA-reactive concept.

Another hypothesis is that the lung tumours that arise in rats exposed to high levels of diesel exhaust are due to overloading of the normal lung particle clearance mechanisms, accumulation of soot particles, and cell damage followed by regenerative cell proliferation. Enhanced cell proliferation may increase the mutation frequency of key target genes. In this hypothesis, genotoxic chemicals may not be causative factors in tumorigenicity. This view is supported by the observations that lung cancer can be induced in rodents by inhalation of highly insoluble particles of low toxicity that are virtually devoid of organic chemicals (e.g. talc, carbon black, coal dust, titanium dioxide), although high concentrations of these particles (overload) are typically necessary.

A third hypothesis, which draws upon the above two mechanisms, is that carcinogenesis is initiated by exposure to organic compounds associated with diesel soot and promoted by the chronic inflammation, cytotoxicity, and cell proliferation arising from the high concentrations of particles deposited and retained in the lungs. Again, however, the tumorigenic response observed with particles alone (carbon black, titanium dioxide) does not support this hypothesis.

A key consideration is the relative contribution of different mechanisms at different levels of exposure. As discussed later in this section, high concentrations of particles, including diesel exhaust, leading to high lung burdens, compromise normal clearance mechanisms. Therefore, certain mechanisms may be invoked at high lung burdens that may not occur at lower lung burdens. Genotoxic aromatic compounds may take on increasing importance at lower concentrations.

Figure 3 is a diagram outlining the possible mechanism of action of diesel exhaust, including the effects of overload conditions.

B7.9.1 Carcinogenic effects

B7.9.1.1 DNA-reactive mechanisms

Diesel exhaust contains hundreds of chemicals, including PAHs, nitro-PAHs, alkyl-PAHs, oxy-PAHs, oxy-nitro-PAHs, and aldehydes (Scheepers & Bos, 1992b), many of which are known mutagens and carcinogens in laboratory animals. For example, inhaled benzo[ a ]pyrene (in tar and pitch) is carcinogenic in rat lung (Heinrich et al., 1994). Nitro-PAHs are potent mutagens and, in some instances, carcinogenic. In pharmacokinetic studies cited in this monograph, clearance of PAHs and nitro-PAHs associated with the diesel particulate fraction is retarded or delayed. Other studies have shown clearly that the organic compounds associated with diesel soot are bioavailable to lung cells. Mechanisms for the delayed clearance of the PAHs have been discussed (Bond et al., 1986b; Gerde et al., 1991b). PAHs require metabolic activation to metabolites (e.g. epoxides) that can potentially bind to DNA. In the case of the PAHs associated with diesel soot, it is likely that alveolar lung cells (e.g. type II cells) are responsible for their metabolic activation, so that they can bind to lung cell DNA (Bond et al., 1983).

Additional evidence for a DNA-reactive mechanism in the carcinogenicity of diesel exhaust is that exposure of laboratory animals, including rats and monkeys, to diesel exhaust results in the formation of DNA adducts in lung cells (see section B7.6). DNA adducts and subsequent DNA replication can result in mutations that play a key 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmrole in initiating the carcinogenic response. These mutations could involve activation of oncogenes or inactivation of tumour suppressor genes. There may be some as yet unidentified chemicals in diesel exhaust with high mutagenic potential; furthermore, chemical interactions (additive or inhibitory) may occur among the components of diesel exhaust (Ball & Young, 1992).

B7.9.1.2 Cytotoxicity with regenerative cell proliferation

Another mechanism by which diesel exhaust may induce carcinogenesis involves cell killing, or cytotoxicity, by the particles or the reactive gases in exhaust, followed by regenerative proliferation of lung cells. Enhanced cell replication may result in an increased frequency of spontaneous mutations. Mutations could also arise from PAH-DNA adducts or from oxidative DNA damage caused by the reactive oxygen and nitrogen species in diesel soot, resulting in the inflammatory response. Driscoll et al. (1994) showed that the hprt gene mutation frequency in rat lung epithelial cells was significantly
increased when the cells were incubated in vitro with inflammatory cells obtained by bronchoalveolar lavage from rats exposed to quartz particles. This appears to be an important mechanism in lung tumour induction by particles that elicit a chronic inflammatory response in the lung. Sagai et al. (1993) showed that diesel exhaust particles could produce superoxide anions and hydroxyl radicals in the absence of biological activation (detection by cytochrome c and electron spin resonance). Reactive oxygen species may induce specific DNA adducts, such as 8-hydroxydeoxyguanosine. As mentioned above, mutations could involve activation of oncogenes or inactivation of tumour suppressor

genes.

Several data sets cited in this monograph support this hypothesis. For example, carbon black particles, which are virtually devoid of PAHs but are morphologically similar to diesel soot particles, induce lung cancer in rats. High particle burdens in rodent lungs result in an inflammatory response (Henderson et al., 1988). Release of cytokines and reactive oxygen and nitrogen species during the inflammatory response to particles may result in cell death. Some evidence exists (Wright, 1986) that diesel exhaust enhances lung cell replication.

B7.9.1.3 Effects of particles

The importance of pulmonary particle burden on lung tumour induction has been demonstrated clearly in long-term studies by inhalation in rats. As reviewed above, rats have similarly increased lung tumour incidences when exposed to diesel exhaust or carbon black particles by inhalation for 24 months. Similarly, carbon black particles practically devoid of PAHs induce pulmonary tumours after

intratracheal instillation. The very large surface area of carbon black and of diesel exhaust particles after desorption of adsorbed organic compounds in vivo may be involved mechanistically in a tumorigenic effect. Heinrich (1994) showed that the tumour response to different types of carbon black particles instilled intratracheally correlated well with their respective surface areas. Pott (1991) and Pott et al. (1993) suggested that particles with a large surface area have greater effects in humans than in rats because insoluble particles with no specific toxicity remain significantly longer in the terminal airways of humans than of rats (retention half-time, about 70 days in rats and about 500 days in humans).

The correlation between particle surface area and lung tumour incidence was examined (Oberdörster & Yu, 1990) by evaluating published studies of inhalation of diesel and other particles. Tumour
induction in rats was best correlated with the surface area of the particles retained in the lung rather than with the particle mass, particle volume, or number of particles, regardless of the PAH content. It was suggested that particle surface area and surface properties play a decisive role ('critical surface area') and that absorbed PAHs are not responsible for the tumour response in rats exposed to diesel exhaust. In the human situation, however, it could not be excluded that organic compounds and gas-phase components are also involved, since the human particulate lung burden is much lower than those achieved in rats after long-term inhalation. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmInhalation of diesel engine exhaust can result not only in pulmonary tumours but also in inflammation and fibrosis and in a delay in alveolar (not bronchial) pulmonary clearance. The mechanism by which tumours develop due to particle overload and its associated pathological and anatomical changes may be restricted to rats and may not occur under environmental conditions in humans, since the lung burdens of humans do not reach the levels that induce lung tumours in rats. This is of importance for quantitative risk assessment. Only occupational exposure to diesel exhaust may result in lung burdens near or at overload conditions, particularly if the lung is already compromised by exposure to other dusts. Bohning et al. (1982) reported retarded particle clearance in smokers; in these people, additional exposure to diesel exhaust may induce overload and associated toxic effects. It is not known, however, whether the mechanisms of particle-induced lung tumours are the same in rats and humans, and this information is necessary for extrapolating data from rats to

humans.

Not only diesel soot (0.8, 2.5, or 7.0 mg/m 3 ) but also carbon black nearly completely devoid of organic compounds (Printex 90, 7.5-12 mg/m 3 ; particle size, 10 nm) and ultrafine titanium dioxide particles (7.5-15 mg/m 3 ; particle size, 20 nm) caused lung tumours in female Wistar rats exposed by inhalation for 18 h/day on five days per week for 24 months. The tumour rate increased with increasing

particle concentrations, independently of the type of particles inhaled. No lung tumours were observed in the rats exposed to the lowest concentration of diesel particles. The authors concluded that the carcinogenic component of diesel exhaust is in the inner part of the diesel soot particle, the carbon core, and is not the relatively small amount of carcinogenic PAHs (3.9 g benzo[ a ]pyrene per gram of diesel soot) (Heinrich et al., 1992; Heinrich, 1994; Heinrich et al., 1995). These results were confirmed in another two-year study, in which male and female rats were exposed by inhalation to various concentrations of carbon black and diesel exhaust. Lung tumours were observed with both particle types (Nikula et al., 1994).

Diesel soot does not appear to have a specific carcinogenic effect in rats; rather, there is a nonspecific effect of particles.

B7.9.1.4 Effects of polycyclic aromatic hydrocarbons

Exposure of rats by inhalation to 2.6 mg/m 3 of an aerosol of tar-pitch condensate with no carbon core but containing 50 μg/m 3
benzo[ a ]pyrene and other PAHs for 10 months caused lung tumours at a rate of 39%. The same amount of tar-pitch vapour condensed onto the surface of carbon black particles at 2 and 6 mg/m 3 resulted in tumour rates that were roughly two times higher (89 and 72%). Since exposure to 6 mg/m 3 carbon black almost devoid of extractable organic material caused a lung tumour rate of 18%, the tumour rate of 72% seen after combined exposure to tar-pitch vapour and carbon black particles indicates a syncarcinogenic effect of PAHs and carbon black. A possible mechanism is an effect of deposition of PAHs (Heinrich et al., 1994). As the level of benzo[ a ]pyrene in the coal-tar pitch was about three orders of magnitude greater than those in diesel soot, PAHs may play a negligible role in the tumorigenicity of diesel soot in rats. The PAH profile in diesel soot is, however, quite different from that in coal-tar pitch, as diesel soot contains highly mutagenic, carcinogenic nitro-PAHs and other poorly characterized mutagens that are not present in coal-tar pitch or on some of the carbon black particles used in experimental studies (Heinrich et al., 1994; Nikula et al., 1994).

In a study of various extracts of diesel exhaust particles, 30-40% of the total mutagenicity could be attributed to a group of six nitroarenes (Salmeen et al., 1984).

A diesel exhaust particle extract was separated into a water- and a lipid-soluble fraction, and the latter was further separated into a PAH-free, a PAH-containing, and a polar fraction by column chromatography. These fractions were then tested in Osborne-Mendel rats by pulmonary implantation at doses corresponding to the composition of the original diesel exhaust. The water-soluble fraction did not induce tumours; the incidences induced by the lipid-soluble

fractions were 0% with the PAH-free fraction, 25% with the PAH-containing fraction, and 0% with the polar fraction. The PAH-containing fraction, comprising only 1% by weight of the total extract, was shown to be responsible for the carcinogenic activity (Grimmer et al., 1991).

Various dichloromethane extracts, each representing a complex mixture, were obtained from particulate emissions of four diesel-fuelled and one gasoline-fuelled automobiles (combustion), a coke oven battery (pyrolysis), and a roofing tar pot (evaporation), and their tumorigenic potency was compared in Sencar mice. It was concluded that the benzo[ a ]pyrene content alone could not explain the tumorigenic activity of the mixtures (Nesnow et al., 1982a, 1983).

The lung tumour rates in rats exposed to atmospheres containing PAHs depend not only on the PAH concentrations of the exhaust gas but also on parameters such as the composition of the carrier particle (mass ratio of carbon core to adsorbed layer of organics), the dissolution rate of particle-attached organic compounds, the retention half-time, and the cytotoxic effect of the carrier particle in the lung (Heinrich et al., 1991).

Extracted diesel soot given intratracheally to rats induced lung tumours, but native, unchanged diesel soot resulted in higher tumour

rates than extracted soot. Carbon black also caused tumours after 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmintratracheal administration, and the rate increased with decreasing size of the primary particles (Heinrich, 1994).

Carbon black particles almost completely devoid of organic compounds (< 0.046% extractable organic compounds and 0.6 ng/g benzo[ a ]pyrene) caused tumours in the lungs of 17% of rats after exposure to a concentration of 6 mg/m 3 for 18 h/day on five days per week for 10 months. No lung tumours occurred in controls exposed to clean air. Thus, the particle effect may be responsible for induction of lung tumours in rats exposed to diesel engine exhaust, and the effect may be related to the surface area of the carbon particle. A PAH depot effect could lead, however, to retarded dissolution of PAHs from the carrier, resulting in very efficient use of the extremely small amount of carcinogenic PAHs retained in the lung after exposure to diluted diesel engine exhaust, which contained benzo[ a ]pyrene at about 10 ng/m 3 (Heinrich et al., 1991). Since diesel particles induce DNA adducts at lower concentrations than carbon black (Bond et al., 1989, 1990c; Wolff et al., 1990), PAHs may have tumour initiating properties that are too weak to be detected at low doses; at higher doses, promotional effects of the particles, perhaps with additional initiation, may synergize to produce detectable carcinogenic effects.

B7.9.2 Non-carcinogenic effects

Diesel exhaust contains various respiratory irritants in the gas phase and in particulate matter. Both can induce inflammatory responses in the airways and alveolar regions of the lung. Airway inflammation involves damage to epithelial cells, including lipid peroxidation of cell membranes by oxidizing gaseous pollutants such as nitrogen dioxide. Indirect effects of particles, resulting from phagocytosis, can include the formation and release of various mediators, including oxidants, such as superoxide anions and hydroxyl radicals, and cytokines. These mediators may play a role in focal loss or shortening of cilia, type II cell hypertrophy, and hyperplasia. The latter changes can lead to the hyperplastic lesions seen in animals exposed to diesel exahaust (Ishinishi et al., 1986, 1988; Suzuki et al., 1990; Kato et al., 1992). Phagocytosis and subsequent clearance of particles by alveolar macrophages can be compromised by high particle burdens (Wolff et al., 1989; Creutzenberg et al., 1990), which may also increase the access of particles to the interstitium, leading to focal fibrosis (Henderson et al., 1988).

Inflammation, altered lung clearance, and hyperplastic lesions can be considered early markers of exposure to diesel exhaust and are the basis of the non-neoplastic effects used to determine both the
NOAEL and the 'benchmark concentration' described in section B10.1.3.

B8. EFFECTS ON HUMANS

B8.1 General population

B8.1.1 Acute exposure: olfactory, nasal, and ocular irritation

Acute exposure to diesel exhaust has been associated with irritation of the eyes and nose. Signs and symptoms reported after acute exposure to diesel exhaust are described in section B8.2.1.

The exhaust from e.g. poorly maintained engines or engines under load may be visible as a black, smoky cloud. WHO (1987) considered that sensory effects were parameters that could be used in setting occupational exposure limits. The characteristic odour of diesel exhaust provides a warning of its presence. At higher concentrations and under certain operating conditions, the odour can be offensive. The odour-causing agents are not known; however, compounds with a relative molecular mass > 80 are probably those mainly responsible, and aliphatic olefins (C 1 -C 6 , including acetylene), hydrocarbons
with more than six carbons, and aliphatic aldehydes are probably not involved. Substances with low odour thresholds, such as nitrogen dioxide (0.3 ppm) and acrolein (0.5 ppm), appear to contribute to the odour of diesel exhaust to minor extents (5.3 and 0.25%, respectively) (Oelert & Florian, 1972).

The ability to detect the odour and the irritating effects of diesel exhaust vary. For example, when six subjects smelled diesel exhaust diluted with air, the dilution factors needed to achieve the
odour detection threshold varied from 140 to 475. The concentrations
of different constituents of the exhaust at the odour threshold also varied: formaldehyde, 0.012-0.088 ppm; acrolein, 0.011-0.046 ppm; and nitrogen dioxide, 0.11-2.28 ppm (Linnell & Scott, 1962). When six subjects were exposed to three concentrations of diesel exhaust for 10 min, the mean concentrations at the odour threshold were 1.3-4.2 ppm nitrogen dioxide, 0.2-1 ppm sulfur dioxide, < 0.1 ppm formaldehyde, and < 0.05 ppm acrolein. Three of the subjects exposed to the highest level discontinued exposure before 10 min, while none at the lowest level discontinued exposure although some experienced eye irritation (Battigelli, 1965).

The health effects of inorganic gases present in diesel exhaust are not specific and are therefore not discussed in detail; however, it must be noted that these gases contribute to the environmental

burden.

B8.1.2 Air pollution

The concentrations of some air pollutants, such as particulates and sulfur dioxide from the burning of coal for domestic heating and industrial purposes, are in many parts of the world much lower than they were several decades ago, and the associated adverse health

effects have diminished accordingly. A number of epidemiological 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmstudies have demonstrated, however, that the relatively low remaining levels of air pollutants are associated with a range of health indices, including day-to-day changes in mortality (Schwartz, 1993), visits to hospital emergency departments (Schwartz et al., 1993), and changes in measures of lung function (Pope & Dockery, 1992). In most studies, the associations have been strongest with the fine particulate component of pollution, some of which may be derived from
diesel exhausts.

Vehicle traffic, both gasoline and diesel, also contributes to the nitrogen dioxide content of urban air, and there is evidence, primarily from experimental studies (Bauer et al., 1986; Koenig et al., 1988; Grant et al., 1993) that it can induce short-term decrements in lung function in asthmatic and non-asthmatic patients. Nitrogen dioxide and volatile hydrocarbons are involved in the formation of ozone and associated photochemical pollutants.

Associations have also been demonstrated between long-term exposure to urban air pollutants and death rates from certain chronic conditions. Confounding factors often make interpretation difficult, but in an investigation conducted in the United States (Dockery et al., 1993), air pollution attributable to fine particles was positively associated with lung cancer and cardiopulmonary disease, after adjustment for smoking and other relevant risk factors, but not with other causes considered collectively.

In neither short- nor long-term studies can the possible role of diesel particulates be specified, but diesel emissions contribute to urban particulates, especially in Europe and developing countries. The acidic and sulfate components of particulates, which are derived primarily from stationary fuel-burning sources, appeared to be implicated in a number of studies.

B8.2 Occupational exposure

B8.2.1 Effects on the respiratory system

B8.2.1.1 Symptoms

Most of the information about symptoms after acute exposure to diesel exhaust comes from anecdotal reports of occupationally exposed individuals. It was noted in a bus garage in London, United Kingdom, that the buses produce a 'lachrymatory mist' when they were started up from cold (Commins et al., 1956). In a review of 13 cases of acute overexposure to diesel exhaust in five underground coal mines in Utah and Colorado, United States, between 1974 and 1985, interviews in 1986 with the miners revealed that 12 had experienced symptoms of mucous membrane irritation, headache, and light-headedness; eight had reported nausea, and four reported a sensation of unreality and 'heartburn' (Kahn et al., 1988).

In a study designed to investigate the acute effects of diesel exhaust on respiratory symptoms, a questionnaire was administered to 232 male workers in four diesel bus garages, and personal samples of nitrogen dioxide and respirable particulate were obtained. After adjustment for age and smoking (current, ex-, never), workers with the highest exposure to respirable particulate (0.31 mg/m 3 ) reported significantly more cough, itchy or burning eyes, headache, difficult or laboured breathing, a feeling of chest constriction, and wheeze than workers with lower exposures. Those exposed to nitrogen dioxide at >0.4 mg/m 3 (> 0.3 ppm) also more frequently reported itchy or burning eyes, difficult or laboured breathing, a feeling of chest constriction, and wheezing. In comparison with a group of lead acid battery workers, the reporting of the following symptoms as 'sometimes' or 'often' was significantly more prevalent among the workers in the diesel bus garage: eyes itch, burn, or water (49.5 versus 23.5%), headache (24.2 versus 12.1%), difficult or laboured breathing (13.5 versus 3.6%), nausea (13.5 versus 4.5%), and wheeze (13.7 versus 4.5%) (Gamble et al., 1987a).

In comparison with 11 office workers, 17 ferry stevedores had a greater prevalance of wheezing (24 versus 9%), chest tightness (29 versus 9%), nasal complaints (47 versus 0%), chest pain (24 versus 0%), and eye irritation (59 versus 27%); however, more of the stevedores smoked. The concentrations of nitrogen dioxide (0.3 mg/m 3 ) and sulfur dioxide (0.7mg/m 3 ) were both < 0.25 ppm; respirable particulate was not measured (Purdham et al., 1987). In a study in an iron ore mine where diesel engines were used underground, more workers who were smokers reported pressure over the chest or difficulty in getting air than the nonsmokers. No such difference was observed among smoking and nonsmoking surface workers. The underground workers who were smokers also had more episodes of a productive cough lasting three weeks during several winters (Jorgensen & Svensson, 1970).

B8.2.1.2 Acute changes in pulmonary function

The pulmonary function of 60 coal miners who worked in mines equipped with diesel engines was compared with that of 90 coal miners not exposed to diesel exhaust. There were similar proportions of current smokers (45% in the exposed group and 43% in the unexposed group). Measurements made with personal dust samplers and passive nitrogen dioxide dosimeters showed average concentrations of 0.04 mg/m 3 (0.2 ppm) nitrogen dioxide, 0.4mg/m 3 (0.3 ppm) formaldehyde, and 2.0 mg/m 3 respirable dust in the exposed group; the unexposed group was exposed to 1.4 mg/m 3 respirable dust. Forced vital capacity (FVC) and forced expiratory volume in 1 sec (FEV) were reduced during the work shift for both the diesel-exposed and unexposed groups. The reduction was slightly but not significantly greater in smokers than in ex- and nonsmokers. An additional analysis 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmwith adjustment for age, exposure to respirable dust, and years of underground mining still revealed no difference in shift-related pulmonary function between miners exposed and unexposed to diesel exhaust (Ames et al., 1982).

No significant shift-related change in FVC or FEV was seen among workers on ferries transporting diesel trucks and other vehicles in comparison with controls (Purdham et al., 1987), and no significant change in spirometry measured over a shift was seen in 232 workers in four diesel bus garages (Gamble et al., 1987a). Ulfvarson et al. (1987), however, found significant decrements in FVC and FEV 1 across a shift in 23 ferry workers who had had no exposure to diesel for 10 days, with no difference between smokers and nonsmokers. The concentration range of total particles was 0.13-1 mg/m 3 , that of formaldehyde 0.04-0.6mg/m 3 (< 0.03-0.5 ppm), and that of nitrogen dioxide 0.12-4.6mg/m 3 (0.06-2.3 ppm). In a repetition of the study, a significant shift-related decrement was noted in FVC but not in

FEV 1 . In a later study in which filters were put on the diesel
trucks, a shift-related decrement in FVC was seen, but no subsequent shift-related changes in FEV were noted, with or without filters (Ulfvarson & Alexandersson, 19901).

Three male railroad workers developed asthma after heavy exposure to locomotive emissions (Wade & Newman, 1993). The workers, none of whom were current smokers, had no previous history of asthma or of any respiratory disease, except one who had seasonal rhinitis. They were riding immediately behind the lead engines of the trains, where diesel exhaust was blown almost continually into the cab. In two, the acute onset of asthma occurred within the first hours of exposure.

B8.2.1.3 Pulmonary effects

In a study to investigate the effects of diesel exhaust on the cells found in bronchoalveolar lavage (BAL) fluid (Rudell et al., 1990), eight healthy, nonsmoking volunteers were exposed to diesel exhaust (for 60 min according to Rudell et al., 1989) at least three weeks after an initial BAL. The median steady-state concentrations measured in the exhaust were 4.6mg/m 3 (3.7 ppm) nitric oxide, 3.1 mg/m 3 (1.6 ppm) nitrogen dioxide, 22.5mg/m 3 (27 ppm) carbon monoxide, 0.5 mg/m 3 formaldehyde, and 4.3 × 10 5 particles/cm 3 ; according to Rudell et al. (1989), this particle concentration corresponds to a mass concentration of approximately 100 μg/m 3 . BAL was performed again 18 h after exposure. A significant reduction (P < 0.02) in the total number of mast cells was observed; the number of neutrophils was slightly but significantly (P < 0.05) increased in comparison with the values before exposure. The ratio of T-helper:suppressor cytotoxic cells was elevated ( P < 0.02), and the rate of phagocytosis of opsonized yeast cells by alveolar macrophages
in vitro was reduced (P < 0.02). The number of lymphocytes remained unchanged.

B8.2.2 Epidemiological studies (noncarcinogenic effects)

Some but not all of the results from cross-sectional and longitudinal studies on workers with occupational exposure to low levels of diesel engine exhaust show decrements in lung function and an increased prevalence of respiratory symptoms.

B8.2.2.1 Effects on the respiratory system

A group of 550 underground workers and 273 surface workers in six coal mines where diesel-powered equipment was used were matched for smoking status, age, height, and years of underground mining to workers at other underground coal mines where diesel units were not used. The workers at the mines with diesel engines reported significantly more persistent cough (23.6%) than controls (16.5%) and more frequently had exacerbations of cough and phlegm (21.7 and 16.2%, respectively). Pulmonary function (FVC and FEV ) was decreased in both surface and underground workers in mines where diesel engines were used in comparison with surface and underground miners in other mines. There was no consistent relationship between years of underground mining and respiratory symptoms and pulmonary function; however, the mean time spent in underground mining was only 4.7 years. Full-shift area samples showed concentrations of respirable dust ranging from 0.4 to 16.1 mg/m 3 for various jobs; the mean values from personal samples were 0.93-2.73 mg/m 3 , and the nitrogen dioxide levels in personal samples were 0.16-0.26 mg/m 3 (0.13-0.22 ppm) (Reger et al., 1982).

Pulmonary function and respiratory symptoms were studied in 1976 in 630 miners in six potash mines where diesel-powered vehicles had
been introduced between 1950 and 1964. There was no consistent relationship with exposure to diesel exhaust, considered in several ways, including years of exposure, cumulative exposure to dust, nitrogen dioxide concentration, and prevalence of diesel use in the mine. The mean length of exposure to diesel engines was, however, relatively short: 5-14 years across mines. In addition, cumulative exposure was measured only to total dust rather than respirable dust. The mean values for total dust in the mines ranged from 9 to 23 mg/m 3 (personal sampling); the ratio of total dust to respirable dust ranged from 2 to 11 (area sampling), and the nitrogen dioxide concentration was 0.2-6.3mg/m 3 (0.1-3.3 ppm) (personal sampling) (Attfield et al., 1982).

A total of 259 miners in five salt mines where diesel equipment was used in some of the mines were studied with regard to the number of years worked underground and cumulative exposure to respirable particulate and nitrogen dioxide. These parameters were associated with phlegm after adjustment for age and smoking but not cough. The 

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0.2-0.7 mg/m 3 , and the average exposure to nitrogen dioxide was 0.6-4.8mg/m 3 (0.3-2.5 ppm). Pulmonary function (FEV 1, FVC, peak flow) was not related to the three parameters but was slightly reduced in comparison with that of other blue-collar workers. In another analysis in the same population, phlegm but not pulmonary function was associated with years of work underground in mines where diesel engines were used, after adjustment for age and smoking. There was a nonsignificant trend for an association between cough and shortness of breath with years of exposure (Gamble & Jones, 1983).

Longitudinal changes in pulmonary function and chronic respiratory symptoms were studied between 1977 and 1982 in 280 miners exposed to diesel exhaust and 838 miners not exposed to diesel exhaust in American underground coal mines. The mean number of years of underground work ranged from 6.6 to 17.4 years across mines. Area samples taken in 1977 revealed 'low' levels of dust and diesel exhaust constituents, which were reported not to exceed 25% of current standards, but the actual values were not given. The exposed workers had smaller average decrements in FVC and FEV 1 than workers not
exposed to diesel exhaust after adjustment for age, smoking, and years of underground work. Additional analysis revealed no relationship with cumulative years of underground work, but the level of pulmonary function was not considered as a predictor of longitudinal change. The exposed miners also had a lower five-year incidence of cough, phlegm, and breathlessness than the miners not exposed to exhaust (Ames et al., 1984).

Pulmonary function and respiratory symptoms were studied in 283 male diesel bus workers in four garages in two cities. The number of years worked was a significant predictor of lower FVC and FEV 1 adjustment for age, height, race, and smoking status. In comparison after with another blue-collar population, the bus garage workers had a higher prevalence of cough and phlegm after adjustment for age and smoking. The prevalence of these symptoms was not related to the number of years worked (Gamble et al., 1987b).

B8.2.2.2 Effects on the circulatory system

No consistent excess of deaths due to cardiovascular disease has been identified in cohort studies of workers with potential exposure to diesel emissions. Motor vehicle examiners (Stern et al., 1981), who are probably exposed to exhaust from a variety of vehicles, had a slight, nonsignificant increase in deaths due to cardiovascular disease (standardized mortality ratio [SMR] = 10 5 ). No difference in mortality was seen among men working in potash mines with diesel-powered equipment in comparison with men in potash mines with no diesel engines (Waxweiler et al., 1973). In a pilot study of 129 men employed in a bus company in 1951-59 for whom mortality was ascertained until 1978 (Edling & Axelson, 1984), a fourfold increase in cardiovascular deaths was seen in garage workers. The increase was

calculated on the basis of at least 15 years since first exposure and 10 or more years of exposure, but was based on only four deaths. This finding was not confirmed in a follow-up of 694 men through 1983 (Edling et al., 1987). In a study of 8490 garage maintenance workers with at least one continuous year of service between 1967 and 1975, a deficit of deaths due to cardiovascular disease was seen (Rushton et al., 1983).

B8.2.3 Epidemiological studies (carcinogenic effects)

The relationship between cancers of the lung and bladder and occupational exposure to diesel exhaust has been evaluated in a number of epidemiological studies. The occupations examined included diesel truck drivers, bus garage workers, railroad workers, heavy equipment operators, and stevedores. The risk for lung cancer in relation to exposure to diesel exhaust, either self-reported or assumed from occupational categories, has also been evaluated in case-control studies. Only those studies that were considered relevant for an evaluation of the carcinogenic effects of diesel exposure are included
in this assessment.

B8.2.3.1 Lung cancer

The epidemiological studies are summarized in Tables 39-41. Table 39 is a summary of nine case-control and cohort studies of workers exposed to diesel exhaust, Table 40 summarizes nine population- and hospital-based studies, and Table 41 presents two studies based on information on exposure derived from registries (surveillance studies).

Until recently, the epidemiological study of lung cancer and exposure to diesel exhaust was limited by failure to consider the latency and duration of exposure necessary for the development of lung cancer, both generally considered to be about 20 years (Schenker & Speizer, 1979). Studies of exposed workers have also been limited by a lack of worker- and industry-specific data on exposure. Such data were available only for the American railroad and trucking industries. For other occupational groups, such as bus garage workers and stevedores, indices of exposure were derived from vehicle use, fuel consumption, or years of exposure, or simply job title unsubstantiated by exposure assessment. In studies based in hospitals, the general population, or registries, information on usual work in a job with exposure to vehicle exhaust was derived from self- or surrogate reporting; self-reporting of exposure to diesel fumes has also been used. Studies with short durations of exposure and follow-up are not useful for determining whether diesel exhaust is a human carcinogen. Studies in which there is an imprecise definition of exposure to diesel exhaust 

# Pa g e 98 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm(such as self-reported job title) do not allow detection of an effect, since the extent of exposure is unknown. In studies in which an elevated risk for lung cancer is noted, such imprecision can result in
wide confidence intervals.

The major potential confounding factor in occupational studies of lung cancer is tobacco smoking. Such a factor must be related not only to the outcome but also to the exposure. In order to assess whether tobacco use is a true confounder in a study of occupation, the smoking rates among individuals exposed and unexposed to diesel exhaust must be known; however, owing to the retrospective nature of many occupational studies, smoking histories are often not available. It is unlikely that the smoking habits of workers exposed and unexposed to diesel exhaust within the same occupational cohort are differentially related to the exposure, but when the effect of exposure is small, in the order of a relative risk of 1.5 (such has been found for diesel exhaust and lung cancer in most studies), differences in smoking that
are unaccounted for could reduce the relative risk attributable to exposure to diesel exhaust. Interpretation of the results of studies lacking specific information about smoking is more uncertain when small relative risks are seen. Despite imprecise exposure histories, one strength of hospital- and registry-based studies of lung cancer is that smoking histories are often available.

(a) Occupationally based cohort and case-control studies

Three studies have been conducted of railroad workers exposed to diesel exhaust. The transition from steam- to diesel-powered locomotives occurred in the United States during the 1950s. By 1959, 95% of the locomotives in service were diesel powered. In a retrospective cohort study of American railroad workers, the mortality of 55 407 white male workers aged 40-64 in 1959 who had worked on the railroads for 10-20 years was ascertained through 1980, providing a 22-year period of follow-up (Garshick et al., 1988). A yearly three-digit job code up to the time of death or retirement was
available from the Railroad Retirement Board. The workers had held jobs in 39 selected categories, and an industrial hygiene survey was conducted to categorize the job codes into those associated with regular exposure to diesel exhaust (engineers, conductors, firemen, brakemen, and diesel locomotive repair shop workers) and those not exposed to diesel exhaust (clerks and signal maintainers). The concentration of respirable particulate, adjusted for cigarette smoke, was used as an indicator of exposure to diesel exhaust (Woskie et al., 1988a,b). A potentially confounding exposure in the railroad industry is asbestos, which was used to insulate the steam locomotives run previously. Thus, workers in steam locomotive repair shops would have been exposed to asbestos. Workers who had the longest potential duration of exposure, i.e. those aged 40-44 in jobs with exposure to diesel exhaust in 1959, had a relative risk of dying of lung cancer of 1.45 (95% confidence interval [CI] = 1.11-1.89). Workers aged 45-49 in 1959, who would have had slightly less exposure, had a relative risk of 1.33 (95% CI = 1.03-1.73). Exclusion of the workers with potential past exposure to asbestos did not appreciably change these relative
risks.

The US Environmental Protection Agency has supported an effort to obtain more detailed information on exposure in this study in order to estimate unit risk (Pepelko & Chen, 1993). Adequate exposure-response curves could not be drawn for years of exposure or for measurements of current exposure. Possible reasons include an imprecise assessment of past exposure, since only current measurements of exposure were available; changes in exposure over time, since improved ventilation and new diesel locomotives have been introduced, resulting in reduced exposure; and the lack of exposure histories for the workers who would have been exposed to diesel exhaust before 1959. In addition, the industrial hygiene survey was performed in four smaller railroads, where exposure may not reflect that throughout the industry; the presence of non-diesel particulate in the respirable samples collected could have led to exposure misclassification. Nevertheless, the industrial hygiene survey validated the classification of jobs into those associated and not associated with exposure.

In a case-control study of American railroad workers by the same investigators (Garshick et al., 1987), deaths occurring between 1 March 1981 and 28 February 1982 were recorded, and 1256 from lung cancer were matched by age to deaths from causes other than cancer or accidents. Smoking histories were obtained from next-of-kin. After adjustment for smoking and past exposure to asbestos, workers aged 64 at the time of death and with 20 years of work in a job with exposure to diesel exhaust had an odds ratio of 1.41 (95% CI = 1.06-1.88).

A cohort study addressed mortality between 1965 and 1977 among
all 43 826 male railroad workers who had retired from the Canadian National Railway Company before 1965 but who were still alive and those who retired between 1965 and 1977. Occupation at the time of retirement was used to classify workers as unexposed, possibly exposed, or probably exposed to diesel fume. The relative risk of dying of lung cancer was 1.20 for workers with possible exposure (P = 0.013) and 1.35 for those probably exposed (P < 0.001). Similar
relative risks were obtained after exclusion of workers involved in locomotive repair, who were most likely to have had past exposure to
asbestos. These results are consistent with those of the studies of American railroad workers, although the degree to which the workers were actually exposed to diesel fumes is not known. Only retired workers were studied, excluding younger, active workers who would have had the most exposure. No information was given about the duration of exposure of the retired workers to diesel exhaust since many would have worked primarily before the transition to diesel locomotives (Howe et al., 1983). 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTable 39. Occupation-based case-control and cohort studies of lung cancer in groups exposed to diesel engine exhaust

Population Exposure assessment Results Limitations Reference

43 826 male pensioners of the Exposure classified by RR = 1.20 (P = 0.013) Incomplete exposure Howe et al. Canadian National Railway experts on occupation at and 1.35 (P = 0.001) for assessment owing to (1983) time of retirement: possible and probable lack of lifetime occupational unexposed, possibly exposure, respectively. history. Difficult to exposed, probably Significant trend in RR separate combustion products exposed with increasing likelihood from diesel exhaust, of exposure since the cohort would
have worked in both the
steam and diesel eras; many would have had relatively short exposure
to diesel. No data on smoking; method of categorizing exposure
not validated or described
in detail

Incident deaths between Personal exposure Adjusted for smoking and Possible misclassification Garshick et a 1 March 1981 and 28 assessed by industrial to asbestos, OR = 1.41 of exposure to diesel (1987) February 1982 in US railroad hygiene sampling in 39 (1.06-1.88) (< 64 years of exhaust since job title might workers. 1256 cases of job categories; yearly age) for 20 years of work not reflect exposure in each lung cancer and 2385 matched job title used to in a diesel-exposed job. case; however, this should controls (up to two each, on dichotomize exposure For > 20 years of work in a reduce the OR. age and date of death). Cases into exposed and diesel-exposed job relative and controls had worked for unexposed between 1959 to 0-4 years, OR = 1.64
the railroad for > 10 years. and retirement (1.18-2.29)

Table 39 (contd)

Population Exposure assessment Results Limitations Reference

55 407 white male railroad Industrial hygiene data RR = 1.45 (1.11- 1 .89) in No data on smoking Garshick et a workers, aged 40-64 and used to categorize jobs workers aged 40-44 in (1988) with 10-20 years of work in as exposed or unexposed 1959; RR = 1.33 (1.03- 1959. Mortality due to lung to diesel exhaust; jobs 1.73) in workers aged 45- cancer (1694 deaths) well categorized for 49. Unexplained by exposure determined retrospectively most of the cohort to asbestos. Workers with through 1980 highest potential cumulative exposure to diesel exhaust had highest
risk for cancer

About 20 000 male London Five job categories used SMRs for all five job Duration of work not Waller (1981 ) Transport workers aged 45-64. to define exposure, categories < 100 for lung considered; impossible to 667 cases of lung cancer validated by cancer (healthy worker follow individuals after ascertained among active environmental effect), but highest SMR in retirement; no adjustment for workers > 25 years (1950-74) concentrations of highest exposure group, not smoking benzo[a]pyrene and significantly greater than particles measured least exposed (underground in 1957 and 1979 train crew)

8490 male London Transport 100 job titles grouped SMR = 101 overall, but job No validation of exposure Rushton et a l maintenance workers (cohort into 20 broad categories category 'general hand' by air sampling; short (1983) mortality study). Mortality of showed increased SMR for (average, 6 years) follow-up; workers employed for > 1 year lung cancer with no no adjustment for smoking between 1 January 1967 apparent link to diesel

and 12 March 1975 exhaust

Table 39 (contd)

Population Exposure assessment Results Limitations Reference

695 workers in bus garages Intensity of exposure to SMR = 122 (71- 1 96); for No adjustment for smoking Gustavsson e t
who had worked for > 6 months exhaust and asbestos increasing exposure index, al. (1990) in 1945-70. 20 deaths from according to industrial RR = 1.34 (1.09-1.64), 1.81 lung cancer identified (nested hygiene estimate; (1.20-2.71), 2.43 case-control study) duration of exposure (1.32-4.47) based on company records

Cohort study of 107 563 US Usual occupation (truck SMR = 153 (23 cases) No cases among non-smokers, Walrath et a l veterans alive in 1954 and driver) among people who had but SMR calculated from (1985) followed-up to 1 January 1970 ever smoked; no statistics smoking-specific rates given

994 male lung cancer deaths Longest job held: diesel OR = 1.55 (0.97-2.47) for Extent of actual exposure Steenland et in 1982-83 and 1085 controls truck driver, gasoline long-haul drivers with > 18 to diesel exhaust unknown al. (1990, 1 9 (excluding lung cancer, truck driver, both years of employment; OR = bladder cancer, accidents). types, truck mechanic, 1.89 (1.04-3.42) for diesel Cases and controls from stevedore. Exposure truck drivers with > 35 Teamsters Union who had filed validated by industrial years of employment claims (requires 20-year hygiene survey (adjusted for smoking) tenure)

Retrospective cohort study of Cohort assumed to be Lung cancer: SMR = 168 No adjustment for smoking; Gustafsson e t stevedores in Sweden first exposed on basis of job (136-207) for incidence; exposure assumptions not al. (1986) employed before 1974 for a duties, but no formal SMR = 132 (105-166) for fully justified; no continuous period of 6 months. assessment made mortality calculations done to explore 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm6071 workers employed in 1961 employment time as a surrogate with mortality determined for exposure through 1980. 89 cases of lung cancer noted in Swedish Cancer Registry (71 deaths)

Table 39 (contd)

Population Exposure assessment Results Limitations Reference

Stevedores in Swedish Estimated from use of Adjusted for smoking Crude adjustment for Emmelin et a l population defined by fuel in each port and (yes/no) depending on smoking; no industrial (1993) Gustafsson et al. but limited years of work since use index of exposure used; hygiene data to validate to workers in ports for which of diesel equipment exposure-response estimates of exposure to data data on diesel fuel relationship noted, diesel exhaust consumption were available. relative odds for medium Case-control study with 50 exposure ranging from 1.5 cases of lung cancer detected to 2.7 and that for high in 1960-82 matched to 154 exposure ranging from 2.9 controls on port and date of to 6.8. Lower limit of 90% birth confidence interval < 1, except for one high-exposure category

RR, relative risk; OR, crude odds ratio; SMR, standardized mortality ratio; in parentheses, 95% confidence interval (CI), u nless
otherwise stated Three studies reported findings among bus garage workers. Waller (1981) studied lung cancer occurrence among London Transport staff on the basis of job category. Diesel buses were introduced in the 1930s, gradually replacing gasoline-powered vehicles and, in the 1950s, electric trolley-buses. Mortality from lung cancer was ascertained for active workers aged 45-64 between 1950 and 1974. A lower risk was seen for people in all job categories in comparison with the general population of the London area, indicating a 'healthy worker effect'. The highest SMR was seen for workers who had the most exposure to diesel exhaust (bus garage engineers), although the SMR was not significantly different in workers with no exposure. Although this study showed no relationship between job category and mortality from lung cancer, the study had significant limitations. No retired workers were followed up, so that workers who would have had significant exposure were excluded. Since the results were not presented on the basis of years of work (years of exposure to diesel fumes), workers with little exposure were grouped with those with long exposure, making it more difficult to detect an effect of diesel exhaust.

Rushton et al. (1983) ascertained the mortality of 8490 London Transport bus garage workers who had been employed for at least one year between 1967 and 1975. One hundred job titles were grouped into 20 broad categories. The overall SMR for lung cancer was not elevated, and there was no relationship with any job with exposure to diesel exhaust; however, the SMRs were not presented on the basis of years of work in a job with exposure, and the duration of follow-up was short, with a mean of 5.9 years.

Using a nested case-control design, Gustavsson et al. (1990) studied a cohort of 695 men who had worked in five bus garages in Stockholm, Sweden, in 1945-70. Diesel-powered buses were first introduced in Stockholm in the 1930s, and after 1945 all of the internal combustion engines in buses were diesel-powered. No measurements of exposure were available, so exposure to diesel exhaust was estimated by industrial hygienists on the basis of garage ventilation, work practices, and the number of buses in the garages and was graded on a five-point scale, each increase corresponding to a 50% increase in intensity. An index of cumulative exposure to diesel exhaust was calculated for each worker by multiplying the exposure level for each work period by the duration in years. Past exposure to
asbestos was estimated on the basis of historical measurements of asbestos fibres obtained during brake-repair operations. Twenty cases of lung cancer were identified and matched by age to six controls. The relative risk for lung cancer increased with increasing index of exposure to diesel exhaust: That at the lowest exposure level was 1.34 (95% CI = 1.09-1.74), that at an intermediate level was 1.81 (95% CI = 1.20-2.71), and that at the highest level was 2.43 (95% CI = 1.32-4.47). No such increase was observed with a similar index of exposure to asbestos. No information was available on smoking.

Diesel-powered trucks were introduced in the United States in the 1950s and 1960s. Trucking companies had completed the transition to diesel trucks by 1960, while independent drivers and non-trucking companies would have completed the transition later. Steenland et al. (1990, 1992) studied 994 deaths from lung cancer occurring in 1982 and 1983 among male members of the Teamsters Union who had filed claims for pension benefits (requiring at least 20 years of membership). Work histories and smoking histories were obtained from next-of-kin; job categories were also available from the Union records. The odds ratio for lung cancer among long-haul drivers, who drove mostly diesel trucks, with 18 or more years of employment after 1959 was 1.55 (95% CI = 0.97-2.47) after adjustment for age, smoking, and exposure to asbestos. Teamsters with 35 years or more of employment whose main job was a diesel truck driver had a relative risk for lung cancer of 1.89 (95% CI = 1.04-3.42). For drivers of gasoline trucks, the relative risk was only 1.34 and did not achieve conventional levels of significance (95% CI = 0.81-2.22).

In an industrial hygiene survey (Zaebst et al., 1991; Steenland et al., 1992) that accompanied this study, measurements of elemental 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmcarbon were used to estimate exposure to diesel exhaust. The truck drivers appeared to have received much of their current exposure to diesel exhaust from background levels on the road rather than directly from their engines. No historical measurements of exposure were
available.

Gustafsson et al. (1986) studied the mortality of 6071 stevedores in Sweden who had been employed for at least six months between 1961 and 1974 through 1980. The stevedores had been exposed to both gasoline and diesel exhausts; diesel-powered trucks were first used in Swedish ports in the late 1950s, with a rapid increase in use in the 1960s. The SMR for lung cancer was 132 (95% CI = 105-166). The year-specific lung cancer rates in the stevedores between 1961 and 1980 were greater than those in the Swedish male population.

In order to refine the assessment of exposure among the stevedores and to adjust for smoking, a case-control study was conducted in this cohort (Emmelin et al., 1993). Cases and controls were selected from among male stevedores who had been employed for at least six months in 1950-74; the cases were those occurring in 1960-82. On the basis of 50 cases and 154 controls, with adjustment for smoking (yes/no), the odds ratio for lung cancer was seen to increase with increases in three indices of exposure: years since diesel equipment was used in the port, estimates of cumulative fuel consumption, and years that fuel use was above a minimal level in the port. The increases were consistent with an exposure-response relationship. Only one of the point estimates of the odds ratios for lung cancer reached statistical significance.

Table 40. Population- and hospital-based studies of lung cancer in groups exposed to diesel engine exhaust

Population Exposure assessment Results Limitations Referen c

7518 (3539 men, 3979 Occupation determined OR = 1.52 for truck drivers Exposure estimate based on William s women) incident invasive at interview (P > 0.05) self-reporting; not validate d et al. cancers from the Third 47% non-response; controls (1977) National Cancer Survey. Lung consisted of other cancers, cancer cases: 432 in men, probably diluting risk 128 in women. Combined other estimate; few cause-specific
cancer sites used as cancers and individual controls occupations

Cohort study of 107 563 US Usual occupation (truck SMR = 153 (23 cases) No cases in nonsmokers, Walrath veterans alive in 1954 and driver) among people who had but SMR calculated using et al. followed-up to 1 January 1970 ever smoked; no statistics smoking-specific rates (1985)

589 cases of lung cancer Job title from next-of- k in Adjustment for smoking; No validation of exposure Damber & reported to the Swedish for occupations held for (0.6-2.2) for professional groupings, so actual exposur e Larsson Cancer registry 1972-77 with > 1 year drivers (> 20 years of to diesel exhaust uncertain (1987) death before 1979; 562 employment) with dead matched dead controls (sex, controls; OR = 1.1 (0.6-2.2) age, year of death, with living controls municipality) drawn from National Registry of Causes of Death; matched living controls (sex, year of birth, municipality) drawn from National Population Registry

Table 40 (contd)

Population Exposure assessment Results Limitations Referen c

Montreal, Canada, Interview with subject; Adjustment for smoking; Job title used to indicate Siemiat y hospital-based cases and job title translated into OR = 1.2 for squamous-cell exposure to diesel exhaust; et al. controls; 857 cases of lung likely exposure to lung carcinoma (90% CI, not validated; exposure (1988) cancer and 1523 controls (other combustion products by 1.0-1.5) for any exposure ill-defined cancers than lung) industrial hygienist but not elevated for other cell types. No relationship with years of exposure or intensity

461 981 male volunteers Self-reported occupation Adjustment for smoking; Exposure information Boffett a enrolled in the American and exposure to diesel > 16 years' exposure to based on self-reporting; et al. Cancer Society prospective exhaust diesel exhaust, OR = 1.21 not validated (1988) mortality study in 1982; aged (0.94-1.56); for 1-15 years, 40-79 at enrolment; 2-year OR = 1.05 (0.80-1.39). follow-up; 378 622 men with Lung cancer mortality also known exposure to diesel elevated in miners and heavy exhaust equipment operators

1260 cases of lung cancer Job title obtained from For motor vehicle drivers Crude classification of Benhamo u and 2084 controls matched occupational history (adjusted for smoking), exposure et al. for age, sex, hospital obtained at interview; OR = 1.42 (1.07-1.89); for (1988) admission in France in 1976-80 exposure based on any all transport equipment work in a job operators, OR = 1.35 (1.05-1.75). Increase in risk with duration of employment

Table 40 (contd)

Population Exposure assessment Results Limitations Referen c 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmPooled data from three Job title from interviews Smoking-adjusted OR = No validation of exposure Hayes case-control studies with with subjects or 1.5 (1.1-1.9) for truck classification; unclear if et al. 2291 male lung cancer cases next-of-kin; years of work drivers with > 10 years of most exposure was to (1989) and 2570 controls. Analysis in motor exhaust-related work and job history obtained diesel exhaust or if job limited to 1444 cases and occupation calculated by direct interview. For all titles selected as 'exposed' 1893 controls who provided motor exhaust-related were actually exposed to information at interview occupations and > 10 years diesel exhaust of work, OR = 1.5 (1.2-1.9)

Hospital-based study of 2584 Occupational titles and Smoking- a djusted OR for Exposure histories not Boffett a cases in 18 hospitals in six self-reporting of exposure occupations with probable validated et al. US cities; 5099 controls exposure to diesel exhaust (1990) = 0.95 (0.78-1.16). For self-reported exposure, OR = 1.21 (0.78-2.02)

Detroit, USA, area; 3792 Interview to obtain full For drivers of heavy trucks Exposure histories not Swanson cases and 1966 controls occupational history; with > 20 years of work, validated et al. identified through cancer analysis by job title OR = 2.5 (1.4-4.4), with (1993) surveillance system adjustment for smoking;

exposure-response relationship with years of work. For drivers of light
trucks with > 20 years of work, OR = 2.1 (0.9-4.6)

OR, odds ratio; SMR, standardized mortality ratio; in parentheses, 95% confidence interval (CI), unless otherwise stated (b) Population- and hospital-based studies

In a study of 107 563 American veterans who were alive in 1954 and were followed to the end of 1969, the SMR among smokers who reported their occupation as truck or tractor driver was 153, based on 23 cases. Most of these individuals would have had little exposure to diesel exhaust, however, since diesel trucks were introduced in the United States only in the 1950s and 1960s (Walrath et al., 1985).

In the Third National Cancer Survey in the United States, self-reporting of work as a truck driver and adjustment for smoking resulted in an odds ratio of 1.52, which was not statistically significant (Williams et al., 1977). In a study of 589 cases of lung cancer reported to the Swedish Cancer Registry between 1972 and 1977 among people who had died before 1979, the odds ratio for > 20 years of work as a professional driver was 1.2 (95% CI = 0.6-2.2) after adjustment for smoking (Damber & Larsson, 1987). In a hospital-based study of 857 cases of lung cancer and controls, occupational histories obtained by interview were interpreted by an industrial hygienist. After adjustment for smoking, the odds ratio for squamous-cell carcinoma of the lung was 1.2 (90% CI = 1.0-1.5) in association with any exposure to diesel exhaust (Siemiatycki et al., 1988). The mortality of 378 622 men between 1982 and 1984 was analysed in the American Cancer Society prospective study of cancer on the basis of self-reporting of exposure to diesel exhaust. After adjustment for age, smoking, and other occupational exposures, including asbestos, coal-tar and pitch, and gasoline exhaust, the relative risk for lung cancer for men with 16 years of exposure was 1.21 (95% CI = 0.94-1.56). For truck drivers who reported exposure to diesel exhaust, the relative risk was 1.22 (95% CI = 0.77-1.95) (Boffetta et al., 1988).

In a study in France, 1260 male cases of lung cancer and 2084 controls matched on age and hospital were collected between 1976 and 1980; job titles and smoking histories were obtained by interview, and odds ratios were calculated on the basis of any work in a particular occupation. After adjustment for smoking, the odds ratios were 1.35 (95% CI = 1.05-1.75) for transport equipment operators and 1.42 (95% CI = 1.07-1.89) for motor vehicle drivers. There was no increase in risk with increasing years of work in a job, but details of the analysis were not presented (Benhamou et al., 1988).

The data from three case-control studies carried out by the American National Cancer Institute between 1976 and 1983 were pooled in order to study lung cancer in people with occupations associated with motor vehicles. The analysis was limited to 1444 male patients

with lung cancer and 1893 controls who provided information on smoking and occupation at an interview. The odds ratio for lung cancer for 10 or more years of employment in any occupation related to exhaust from either diesel or non-diesel engines was 1.5 (95% CI = 1.2-1.9), after adjustment for age and smoking. In truck drivers, the odds ratio was 1.5 (95% CI = 1.1-1.9) (Hayes et al., 1989).

In a case-control study of patients with lung cancer in 18 hospitals in six American cities, 2584 cases seen between 1969 and the late 1980s were matched to at least one control on the basis of age, sex, hospital, and year of interview. A smoking history and usual occupation were obtained by interview. After 1985, reports of self-reported exposure to diesel exhaust were obtained for 477 lung cancer patients and 946 controls. Exposure was graded as 'probable' if the individual had usually worked on railroads (the specific job was not used) or in a variety of jobs related to motor vehicles. For these individuals, the odds ratio for lung cancer was 1.31 (95% CI = 1.09-1.57); for those reporting usual occupation as a truck driver, the odds ratio was 1.31 (95% CI = 1.03-1.67). After adjustment for cigarette use, age, race, and date of interview, however, the odds ratios for lung cancer were reduced to 0.95 (95% CI = 0.78-1.16) for individuals with probable exposure and 0.88 (95% CI = 0.67-1.15) for truck drivers. For self-reported exposure to diesel exhaust, the odds 

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In a study of 3792 cases of lung cancer in men in Detroit, United States, newly diagnosed in 1984-87, an occupational history and information on smoking were collected by interview with the subject or a surrogate. Controls were men with colon or rectal cancer. After adjustment for smoking, the odds ratio for lung cancer was 2.5 (95% CI = 1.4-4.4) among white men who had driven heavy trucks for 20 years and 2.1 (95% CI = 0.9-4.6) for drivers of light trucks for 20 years. There was a significant increase in the odds ratio for lung cancer with increasing years of work for drivers of both types of truck (Swanson et al., 1993).

(c) Registry-based, surveillance studies

Table 41 summarizes two studies in which information on exposure was based on occupational titles obtained from censuses. No information was available on smoking or on length of employment. Both studies reported an elevated risk for lung cancer. Ahlberg et al. (1981) found a relative risk of 1.33 (95% CI = 1.13-1.56) for professional drivers, and Hansen (1993) found an SMR for all respiratory cancer in truck drivers of 160 (95% CI = 128-198).

Table 41. Surveillance studies of lung cancer in groups exposed to diesel engine exhaust

Population Exposure assessment Results Limitations R eference

Swedish census-based cancer Listing of occupation in RR = 1.33 (1.13-1.56) No validation of exposure; A hlberg et al. incidence registry; 154 cases census no adjustment for smoking ( 1981) among professional drivers

Retrospective cohort study of Job title in census 84 cancers in truck drivers; No adjustment for smoking; H ansen (1993) 14 225 truck drivers in 1970; SMR =160 (128-198); information on specific mortality determined until expected number based exposure or length of 1980. Referent group of persons on rates in referent employment with no exposure to combustion population products on basis of census job title

RR, relative risk; SMR, standardized mortality ratio; in parentheses, 95% confidence interval

Table 42. Case-control studies of urinary bladder cancer in individuals with possible exposure to diesel exhaust

Population Exposure assessment Results Reference

Incident cases in 480 men Any exposure to diesel and Based on 15 cases and controls with discordant Howe et al. and 152 women in three traffic fumes obtained at exposure histories, OR for exposure to diesel and (1980) Canadian provinces, April interview traffic fumes = 2.8 (0.8-11.8); not adjusted for 1974-June 1976; each case smoking matched by age and sex to a neighbourhood control

Incident cases in 303 white Occupation and industry For any employment in trucking, RR = 2.2 (1.1-4.4) Silverman et a men in metropolitan Detroit, obtained at interview based on 28 cases and 13 controls. For any work in (1983) USA, December 1977-November railroad and railway express services, RR = 1.9 1978; 2986 controls (0.9-3.8) based on 22 cases and 12 controls, not selected by random-digit adjusted for smoking. For any employment as a truck dialling but of same age driver (42 cases and 18 controls), adjusted for distribution as cases smoking, RR = 2.1 (1.4-4.4). For > 10 years as truck driver, smoking-adjusted RR = 5.5 (1.8-17.3), and for
truck drivers who ever drove vehicles with diesel engines, smoking-adjusted RR = 11.9 (2.3-61.1). Elevated risk for truck drivers with employment after rather than before 1950, when diesel truck use became more prevalent

Table 42 (contd)

Population Exposure assessment Results Reference

White residents of New Occupation and exposure For any employment as a truck driver (35 cases, 53 Hoar & Hoov e Hampshire and Vermont, to diesel fuel or engines controls), OR = 1.5 (0.9-2.6), not adjusted for smoking. (1985) USA, who died of bladder by next-of-kin Those employed 1930-49 had the highest OR (2.6; cancer 1975-79 (on death 1.3-5.1) after adjustment for coffee drinking and certificate) matched to one smoking. For 26 cases and 39 controls who reported control with cause of death exposure to diesel fuel or engines, OR = 1.5 (0.8-2.8), other than suicide, matched not adjusted for smoking. Significant trend for on state, sex, race, and age, increased relative odds up to 39 years of exposure
and a second control with to diesel exhaust
same criteria but also matched on county. Information obtained

for 325 cases and 673 controls

Cases in 512 men in Turin, Occupation obtained at In truck drivers, based on 16 cases and 16 controls, Vineis & Italy, matched by age to 596 interview OR = 1.2 (0.6- 2 .5), not adjusted for smoking Magnani (19 8 hospital controls without cancer

Cases in 194 men in 18 hospitals Usual occupation obtained; Only 16 cases in occupations with potential exposure Wynder et a l 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmin six US cities, January likelihood of exposure to to diesel exhaust. ORs all < 1 for warehousemen, bus (1985) 1981-May 1983. Controls diesel exhaust determined and truck drivers, and heavy equipment operators. In were persons hospitalized at on basis of estimates of railroad workers, based on two cases and one control, the same time as the case but percentage of workers with OR = 2. For any exposure, with adjustment for smoking, for diseases not related to potential exposure in that OR = 0.87 (0.47-1.58). For high exposure, OR = 1.68 cigarette smoking. Each case occupation: high exposure, (0.49-5.73), not adjusted for smoking

matched to three controls > 20% of workers; moderate (total, 582) by age, race, year exposure, 10-19% of workers of interview, and hospital of
admission

Table 42 (contd)

Population Exposure assessment Results Reference

1909 new cases in white men Occupation obtained at After adjustment for smoking, RR for men usually Silverman from 10 metropolitan areas in interview working as a truck driver = 1.5 (1.1-2.0), based on 99 et al. (198 6 the USA, 1977-78. Up to two cases and 123 controls. For those ever employed as controls (total, 3569) selected truck driver, increased risk with increasing years of by random-digit dialling and employment; however, truck drivers first employed matched for age and < 40 years since diagnosis had no increased risk. OR geographic area with case for bus driver as usual occupation = 1.5 (0.6-3.9), based on 9 cases and 13 controls, adjusted for smoking

Cases in 99 men in La Plata, Occupational history Based on 20 cases ever employed as a truck or railway Iscovich et Argentina, matched on age to obtained at interview driver, RR = 4.31 (P < 0.05); reduced whenr (1987) 99 hospital controls without adjusted for smoking, but no details provided cancer and 99 neighbourhood

controls

Cases in 371 men and women Occupational history After adjustment for age and sex, RR = 1.55 (1.06-2.28) Jensen et a l in the Danish Cancer Registry. at interview any employment as a land transport worker; based (1987) Controls (771) selected randomly on 51 cases and 73 controls. After adjustment for from general population smoking, age, and sex, years of work as a land transport worker, and years of work as a bus, taxi, or truck driver significantly associated with increased risk

Deaths in 731 men in Ohio, Yearly listing of Work as a truck driver > 20 years, OR = 12 (P < 0.01), Steenland USA, 1960-82, identified on occupation obtained from six cases and one control; for > 20 years as a railroad et al. (198 7 death certificates and city commercial directories worker, OR = 2.21 (P < 0.01), based on 22 cases, not matched to six controls without adjusted for smoking tumours of the urinary tract or pneumonia on age, year of death, and race

Table 42 (contd)

Population Exposure assessment Results Reference

Cases in 826 men and women Occupational history and For any exposure of men to exhausts, OR adjusted for Risch et al

. diagnosed 1979-82 in exposure to 'exhausts' cigarette smoking = 1.16 (0.91-1.48). For men ever (1988) Edmonton, Calgary, Toronto, obtained at interview exposed 8-28 years before diagnosis, OR = 1.21 and Kingston, Canada, (0.93-1.58). For men in jobs with any contact with matched by age, sex, and diesel or traffic fumes 8-28 years before diagnosis, area of residence to 792 adjusted for smoking, OR = 1.69 (1.24-2.31); randomly selected population significant trend with increasing years of exposure

controls

Cases in 486 men and women Interview with subject; After adjustment for smoking, OR = 1.0 (90% CI, Siemiatycki in Montreal, Canada, hospitals job title translated into 0.8- 1 .2) for any exposure to diesel exhaust, based et al. (198 8 and 2196 controls with likely exposure to on 82 cases with exposure diseases other than lung combustion products by or kidney cancer industrial hygienists

Cases in 136 men in 18 Usual occupation and For any exposure, OR = 1.24 (0.77-2.00), based on Iyer et al. hospitals in six US cities. self-reporting of exposure 41 cases, with adjustment for smoking (1990)
Two controls without to diesel exhaust obtained tobacco-related diseases matched at interview; grouped into to each case on age (< 2 years), probable and possible race, hospital, and year of exposure
interview

Table 42 (contd)

Population Exposure assessment Results Reference

256 cases and 287 controls in Job titles for all jobs held For any exposure to diesel exhaust, RR = 1.7 Steineck et a l population-based study of all obtained by postal (0.9-3.3), based on 25 cases and 19 controls, adjusted (1990) urothelial cancer in Stockholm, questionnaire; industrial for year of birth and smoking. For subjects considered Sweden, 1985-87, among hygeienists reviewed titles to have moderate or high exposure, RR = 1.1 (0.3-4.3), men born 1911-45 and categorized subjects as but based on only four cases and five controls exposed to various

substances

Cases in 658 men in seven Interview with subject; job After adjustment for smoking, OR for work in road Cordier et al

. French hospitals, 1984-87, title and duties translated transport = 1.02 (0.62-1.69), based on 36 cases and (1993) matched for age, race, and into likely exposure to 35 controls. For stevedores, OR = 1.31 (0.87-1.98); place of residence to 658 diesel exhaust by industrial for material-handling equipment operator, OR = 7.67 controls selected randomly hygienists (0.96-61.4); and for transport equipment operators, among patients admitted OR = 0.88 (0.62-1.26). For diesel fumes, OR = 0.99 for diagnoses other than (0.32-3.03), based on seven cases and two controls 

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Cases in 153 men in one Occupational history For any work as a road transport worker, Notani et al. hospital in Bombay, India, obtained at interview smoking-adjusted OR = 1.36 (0.5-3.7), based on eight (1993) 1986-90, matched by age to cases and nine controls
212 controls with oral or pharyngeal cancer or benign
oral disease

OR, odds ratio; RR, relative risk; in parentheses, 95% confidence interval (CI), unless otherwise stated B8.2.3.2 Urinary bladder cancer

Fifteen case-control studies of urinary bladder cancer in relation to presumed exposure to diesel exhaust are summarized in Table 42. The limitation of all of these studies is that exposure to diesel exhaust was not clearly characterized. Howe et al. (1980) used any self-reported exposure to fumes that included diesel fumes, while Hoar & Hoover (1985) used reports by next-of-kin of work with any diesel fuel or engines or work as a truck driver. Risch et al. (1988) obtained a history of exposure to exhaust by interview. Wynder et al. (1985) estimated the likelihood of exposure to diesel exhaust during a worker's lifetime on the basis of usual occupation, obtained at interview, and attempted to divide exposure into high and moderate. In a later study, self-reported exposure to diesel exhaust was recorded (Iyer et al., 1990). Silverman et al. (1983), Vineis & Magnani (1985), Silverman et al. (1986), Jensen et al. (1987), Iscovich et al. (1987), and Notani et al. (1993) based their analyses of the risk for bladder cancer on employment in an occupational category with a high likelihood of exposure to diesel exhaust, such as truck driver, railroad worker, transport worker, bus driver, and other related occupations. In the studies of Siemiatycki et al. (1988) and Steineck et al. (1990), occupational histories were reviewed by industrial hygienists to link occupational titles to possible exposure. Cordier et al. (1993) also used expert review of occupational histories to estimate exposure to diesel exhaust on the basis of job title, but additionally examined specific job titles. Furthermore, exposure was usually assumed to have occurred if the person had ever been employed in an occupation or had ever been exposed to diesel exhaust or other fumes. Silverman et al. (1983, 1986) and Jensen et al. (1987) used self-reported years of work as a truck driver. Self-reporting of exposure poses a potential problem of differential recall among cases and controls, as cases may recall exposure to diesel fumes or work in a diesel-exposed job more readily than controls. Steenland et al. (1987) used city directories listing occupation to obtain independent occupational histories.

As the available occupational histories were crude, it was also difficult to assess latency or duration of exposure. Silverman et al. (1983) found that the relative risk (adjusted for smoking) was 5.5 (95% CI = 1.8-17.3) for truck drivers with 10 or more years of work experience and 11.9 (95% CI = 2.3-61.1) for those with a history of driving vehicles with diesel engines. Truck drivers employed after 1950, when diesel truck use became prevalent, had the highest risk. In a later, larger study (Silverman et al., 1986), men who usually worked as a truck driver or deliveryman had a relative risk of 1.5 (95% CI = 1.1-2.0), adjusted for smoking. The risk increased with increasing years of work; however, truck drivers who had started work fewer than 40 years before diagnosis, who would have driven primarily diesel engines, did not have an increased risk for bladder cancer. Hoar & Hoover (1985) also noted increased relative odds with increasing years of employment in jobs with reported exposure to diesel exhaust.

An additional limitation of the 15 case-control studies is that results indicating an effect of presumed exposure to diesel exhaust or a specific occupation are based on relatively few cases and controls, even when conventional levels of statistical significance were achieved. The study of Silverman et al. (1986), in which exposure was defined as working as a truck driver or deliveryman, had the largest number of exposed individuals (99 cases and 123 controls). Four additional cohort studies not listed in the table (Howe et al., 1983; Rushton et al., 1983; Schenker et al., 1984; Boffetta et al., 1988), which were designed mainly to examine lung cancer and exposure to diesel exhaust, also had too few cases of bladder cancer on which to base meaningful conclusions. One cohort study of Danish bus drivers (Netterstrom, 1988) showed an elevated SMR of 153, which was of borderline statistical significance (95% CI = 91-217), but this result

was based on 13 cases.

Misclassification of exposure to diesel exhaust would hide any increase in risk for bladder cancer, since many unexposed individuals would be included with those classified as exposed. Diesel exhaust is responsible for most of the respirable particles in mixed vehicle exhausts, and exposure to respirable particles with adsorbed PAHs is presumed to result ultimately in an increased risk for bladder cancer due to metabolic transformation and urinary excretion of carcinogens. The risk for bladder cancer is also increased in cigarette smokers and in workers exposed to dyes containing aromatic amines (Ruder et al., 1990). Coffee has been suggested to be a risk factor for bladder cancer but has not been clearly implicated (Howe et al., 1980). The classification of exposure to diesel exhaust is sufficiently crude in the papers summarized in Table 42, however, that even when smoking was considered, the effect of other, unmeasured confounding exposures
cannot be excluded. The actual contribution of diesel exhaust to an increased risk of bladder cancer is uncertain, even when an effect of presumed exposure was noted (Silverman et al., 1983; Hoar & Hoover, 1985; Silverman et al., 1986; Iscovich et al., 1987; Steenland et al., 1987). Although the literature suggests that truck drivers have an increased risk for bladder cancer, the specific exposure responsible
has not been determined. 

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Virtually no data are available relating specifically to the
effects of diesel fuel exhaust emissions.

The growth and photosynthesis of liquid cultures of the green alga Chlamydomonas reinhardtii were examined after incubation with iso-octane extracts of diesel particulate exhaust containing 51 compounds (mainly polynuclear aromatic ketones and pure PAHs) identified by gas chromatographic-mass spectrometric analysis. At concentrations up to 0.125% by volume, dose-dependent growth retardation of the algae was observed. Higher concentrations (no

details given) caused death. The algae adapted to sublethal concentrations of the extract over a period of several days; thereafter, toxic extracts at concentrations up to 2.5% by volume affected neither growth nor photosynthesis. It was noted that the amounts of certain components decreased during incubation, suggesting uptake into the cells (Liebe & Fock, 1992).

B10. EVALUATION OF HUMAN HEALTH RISKS AND EFFECTS ON THE ENVIRONMENT

Hundreds of chemical compounds are released during the combustion of diesel fuel. The characteristics and amount of exhaust depend on the type of diesel engine, its operating conditions and adjustment, and the composition of the fuel. The following evaluation focuses mainly on the risks to human health associated with exposure to diesel particulate matter, since the large amount of soot particles emitted
is the main characteristic of diesel exhaust.

In general, the risk assessment paradigm proposed by the National Academy of Sciences of the United States (US National Reserach Council, 1983) will be followed. The four steps in this process are: hazard identification, dose-response assessment, exposure assessment, and risk characterization. The outcome of these individual steps is critical for taking risk management decisions, including setting exposure standards that have consequences for public health, and for social, economic, and political issues.

The epidemiological studies in humans and the studies in experimental animals considered useful for human health risk assessment are discussed in this section. Both carcinogenic and other effects are considered, but a threshold is assumed to exist only for non-cancer effects. An IPCS method for deriving health-based guidance exposure limits provides further details (IPCS, 1993).

B10.1 Exposure of the general population

The exposure of the general population to diesel exhaust varies with proximity to diesel-powered vehicles. For example, higher levels of diesel exhaust constituents are present in busy streets and parking areas than in rural areas. The concentrations of the main gaseous components resulting from diesel exhaust can be estimated from specific emission factors, the contribution of diesel vehicles to total traffic, and atmospheric dispersion models. As assumptions are necessary to attribute the emissions of the gaseous components to diesel sources, the validity of such estimates is considered to be
limited.

On a global basis, the contribution of diesel exhaust to the total emissions of particulate matter varies widely, depending on the percentage of diesel vehicles in the total volume of traffic, maintenance of individual engines, fuel quality, and emission control techniques. Such factors also lead to wide local variations in the concentrations to which humans are exposed.

Measurements and calculations of ambient concentrations of diesel particulates near roads provide a daily average range of 8-42 μg/m 3

. The estimated annual average concentrations in Germany are 5-10 μg/m 3
in urban areas and 1.5 μg/m 3 in rural areas, and those in the United States are 1-2 μg/m 3 in urban areas and 0.6-1 μg/m 3 in rural areas. As the proportion of diesel vehicles is smaller in the United States than in Europe, the concentrations in 1990, estimated from national averages, were 2.0 μg/m 3 in an urban area and 1.1 μg/m 3
in a rural area, with predictions of 1.2 g/m 3 for urban areas and 0.6 μg/m 3 for rural areas in 1995.

Diesel particulates form part of the fine particle range (< 2.5 μm diameter). The average concentration of this fraction of suspended particulate matter as a whole, measured in six cities in the United States during 1979-85, was 11-30 μg/m 3 or, in terms of inhalable particles (<10-15 μm diameter; equivalent to particulate

matter with a diameter < 10 μm [PM 10 ]), 18-46 μg/m 3 (Dockery et
al., 1993). In these locations, diesel particulates probably represented less than 10% of the suspended particulates measured as

PM 10 ; considerably higher proportions were found in London and other
large cities in the United Kingdom on the basis of data on emissions (Quality of Urban Air Review Group, 1993), but few data are available that are related directly to ambient concentrations of the diesel component. The one experimental study that has been reported (Horvath et al., 1985), carried out in Vienna, Austria, using chemically labelled fuel, yielded a value for the background concentration of diesel particulates within the city of 11 μg/m 3

.

B10.2 Occupational exposure

Occupational exposure to airborne diesel particulate matter has been well determined for only two occupational groups, railroad 

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the railroad industry, exposure to diesel exhaust was greater in the years after the introduction of diesel equipment. Reports of working conditions in the 1950s and 1960s in diesel repair shops cited 'smoky' atmospheres that were probably related to use of diesel equipment and poor ventilation (Woskie et al., 1988b). No direct information is available that allows reconstruction of accurate levels of past

exposure.

Reports of concentrations of diesel exhaust in other industries are not accurate because of the presence of other dusts, although attempts have been made to measure the contribution of diesel exhaust (Daniel, 1984; Lehmann et al., 1990). Even for American railroad workers (Woskie et al., 1988a), the contribution of other dusts could not be determined, although cigarette smoke particulate was taken into consideration. Consequently, the concentrations of diesel exhaust reported in some jobs on railroads where other dusts (such as sand) were present might have been lower.

B10.3 Non-neoplastic effects

B10.3.1 Hazard identification

B10.3.1.1 Humans

Diesel exhaust contains gases that irritate the nose, throat, and eyes. It also contains particles which, together with the gases, can cause airway irritation. Diesel exhaust may be recognized by its characteristic odour. After acute exposure, it may induce mucous membrane irritation, headache, and light-headedness. When volunteers were exposed to diesel exhaust with an estimated particle concentration of 100 μg/m 3 for 1 h, several indicators of an inflammatory response were observed in bronchioalveolar lavage fluid (see section B8.2.1). In a well-conducted study on diesel-bus garage workers, those with the highest exposures to respirable particulate (> 0.31 mg/m 3 ) reported significantly more cough, itchy or burning eyes, headache, and wheeze, after adjustment for age and smoking, than people with lower exposure. The contribution of diesel exhaust to the respirable particulate is not known. Exposure to nitrogen dioxide at > 0.3 ppm also resulted in more eye irritation and wheeze. Studies of exposure indicate, however, variability in the ability of individuals to detect the odour and the irritating effects of diesel exhaust. Several cases of persistent asthma and asthma attacks have been reported after acute exposure.

No consistent change in pulmonary function has been reported over a work shift, although in one study of ferry workers, a decline in FVC

and FEV 1 was noted. Specific measurements of diesel exhaust are
lacking in these studies, although total dust and respirable dust were sometimes measured. Cases of persistent asthma have been reported in railroad workers acutely exposed to apparently high levels of whole exhaust, but the mechanism is unknown.

Studies of the possible chronic effects of exposure on respiratory parameters are limited to occupational cohorts who are still at work and thus have relatively short durations of exposure. Such cohorts are likely to be healthier than older cohorts, which include retirees who had longer exposure. Although excess respiratory symptoms and reduced pulmonary function have been reported in some studies, it is not clear whether these are long-term effects of

exposure.

B10.3.1.2 Experimental animals

Long-term studies in laboratory animals are described in section B7.3. Many end-points were measured in the respiratory system of several species of rodent after two or more years of exposure and at
several intermediate times. Adverse effects were seen after a sufficient particle load had accumulated in the lung; and the appearance of the effects is determined by both the concentration and the duration of exposure. The results of these studies illustrate the response of the respiratory tract in terms of biochemistry, histopathology, cytology, pulmonary function, inflammation, and clearance of particles. Early events in the pathogenesis of this response include phagocytosis of inhaled particles. As the lung burden increases, particle-filled macrophages are observed in the alveoli. At lung burdens associated with particle overload, migration of these macrophages out of the lung appears to be inhibited, and an early event in the development of lung damage is the accumulation and aggregation of clusters of particle-filled macrophages. Degeneration of macrophages is observed, and the numbers of macrophages and neutrophils in the alveolar and interstitial spaces increase.

Damage to the alveolar epithelium in areas surrounding these macrophage clusters includes cell damage and proliferation. Biochemical changes are seen in lavage fluid and lung tissue. The alterations to particle-filled macrophages result in decreased function and impaired particle clearance, the latter leading to a more rapid increase in the lung burden of particles. Late effects include inflammatory and proliferative responses, leading to fibrotic responses. The sequence of these events has not been distinguished 

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The possible mechanism of action of diesel exhaust (discussed in section B7.9) includes a role of the gas phase, as opposed to the particle phase, in causing lung damage. The non-carcinogenic toxicity of diesel exhaust is considered to be due to the particle content, since no effects are seen in rodents exposed to diesel exhaust that has been filtered to remove particulate matter. Another issue in the

dosimetry of diesel exhaust is the role of the insoluble carbon core in relation to that of the adsorbed organic material. As discussed in section B7.9, studies that show similarities in non-carcinogenic responses to exposure to diesel exhaust and to carbon particles without an organic component indicate a role of long-term retention of
the insoluble carbon core.

B10.3.2 Dose-response assessment

B10.3.2.1 Epidemiological studies

Variability in the reporting of odour and irritation indicates that a subsegment of the population is sensitive to diesel exhaust; however, the proportion of the population with this characteristic
cannot be estimated. Chronic effects have not been seen in occupationally exposed people, but excess respiratory symptoms have been reported after acute exposure. It is not possible to determine the dose of diesel exhaust (on the basis of exposure to gas or particles) that produces these symptoms.

B10.3.2.2 Studies in experimental animals

In order to select the pivotal study for establishing the NOAEL or uncertainty factor, the most sensitive, relevant studies must be identified. All of the long-term studies focus on effects on the respiratory tract, which are clearly those most relevant. Studies of relatively low concentrations are listed in Table 43, which gives the target and actual exposure concentrations, the equivalent continuous exposure of the animal, the equivalent continuous human exposure, and the effect level. The equivalent continuous exposure of animals is calculated by multiplying the actual exposure concentration by the (number of hours of daily exposure/24) and by the (number of days per week of exposure/7). This value is necessary for comparing exposure concentrations among studies with different protocols, i.e. different numbers of hours of exposure per day and/or days per week.

The equivalent continuous human exposure is calculated from the particle deposition-clearance model of Yu & Yoon (1990), discussed in section B6.2. This model is based on studies of rats since only those studies provide data on the exposure-response relationship for inhibition of particle clearance.

The model of Yu & Yoon (1990) is a sophisticated approach to defining the relationship between the inhaled concentration and the dose in lung tissue. It combines detailed models of rat and human lung structure, aerodynamic models of the air flow in the airways, particle deposition dynamics, and information on clearance of particles from rat and human lung. The model is specific to diesel particles because the characteristics of the particles used in the model are derived

from studies of diesel particles and because the information on lung
clearance is based on studies of diesel exhaust in rats. The model has been adapted for diesel exhaust particles by allowing evaluation of the insoluble carbon core and of the tightly and loosely bound organic compounds separately. Values for deposition and clearance are combined with input from experimental studies to calculate the burden of particles in the lungs of the animals at the end of the study. The continuous concentration that would result in the same lung burden in humans is then calculated, as the mass of particles per unit of alveolar surface area or unit of lung weight. This lung burden is
assumed to result in a similar effect in humans and in rats. With these considerations, the equivalent human concentrations are predicted on the basis of the assumption that the retained particle mass per unit alveolar surface area is the appropriate dose measure for extrapolation between species. The effect level used in the risk assessment is based on an evaluation of the adversity of the effect.

For the purposes of risk characterization, the NOAEL or, if that is not available, the LOAEL for the critical effect is related to exposure. The critical effect is the first adverse effect that appears when the critical concentration is reached at the target site. A decision about whether an effect is critical is a matter of expert judgement. An adverse effect is defined as a change in morphology, physiology, growth, development, or the life span of an organism which results in impairment of functional capacity or of capacity to compensate for additional stress or an increase in susceptibility to the harmful effects of other environmental influences (International Union for Pure and Applied Chemistry, 1993).

The NOAEL is defined as the greatest concentration or amount of a substance, found by experiment or observation, that causes no alteration in morphology, functional capacity, growth, development, or the life span of target organisms that are distinguishable from those observed in normal (control) organisms of the same species and strain under the same defined conditions of exposure (International Union for Pure and Applied Chemistry, 1993).

The LOAEL is defined as the lowest concentration or amount of a substance, found by experiment or observation, that causes an adverse alteration in morphology, functional capacity, growth, development, or 

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Table 43. Exposure of experimental animals, equivalent continuous human exposure, and effect levels of long-term inhal ation in stud i
of rats described in section B7.3

Length of End-points Target Actual Equivalent Equivalent Effect Stud y exposure evaluated a exposure exposure continuous continuous level b
(mg/m 3 ) (mg/m 3 ) animal exposure human exposure (mg/m 3 ) (mg/m3)

7 h/d, HP, LL, 0.35 0.353 0.0735 0.042 N Maud erly 5 d/week, LTB, TC, 3.5 3.47 0.723 0.360 A et a l. (1987)

30 months LB 7 7.08 1.47 0.582 A

Light-duty, HP 0.1 0.11 0.063 0.038 N Ishi nishi 16 h/d, 0.4 0.41 0.23 0.139 N et a l. (1986,
6 d/week, 1 1.18 c 0.67 0.359 A 1988)

30 months 2 2.32 1.3 0.571 A

Heavy-duty, HP 0.4 0.46 0.26 0.155 N Ishi nishi 16 h/d, 1 0.96 0.55 0.303 A et a l. (1986, 6 d/week, 2 1.84 1.05 0.493 A 1988 )

30 months 4 3.72 2.13 0.911 A

110 h/week, HP 0.25 0.258 0.17 NA N Barn hart

6 months 0.75 0.796 0.52 NA A et a l. (1981)

1.5 1.53 1.0 NA A

7 h/d, HP, SB 2 1.95 0.57 0.336 A Lewi s et al. 5 d/week, (198 9)

24 months

Table 43 (contd)

Length of End-points Target Actual Equivalent Equivalent Effect Stud y exposure evaluated a exposure exposure continuous continuous level b
(mg/m 3 ) (mg/m 3 ) animal exposure human exposure (mg/m 3 ) (mg/m3)

19 h/d, HP, LL, 4 4.24 2.40 1.02 A Hein rich et 5 d/week, TC, LB al. (1986a)

32 months

18 h/d, HP, LL, 0.8 0.84 0.45 0.23 A Hein rich et 5 d/week, TC, LB 2.5 2.50 1.34 0.59 A al. (1995)

30 months 7.5 6.98 3.74 1.56 A

NA, not available (guinea-pigs were used)

a HP, histopathological examination; LL, lung lavage fluid; LTB, lung tissue biochemistry; TC, tracer clearance; SB, serum bioc hemistry ; LB, lung burden

b N, no effect; A, adverse effect

c According to Suzuki et al. (1990), 1.08 mg/m 3 The selected NOAEL is divided by uncertainty factors to account for data gaps. For these data, uncertainty exists in two areas: extrapolation from animals to humans and accounting for sensitive subpopulations. The factor for accounting for sensitive subpopulations is a default value of 10, as it is considered that there are no data suggesting a different factor. An additional default value of 10 is usually applied for interspecies extrapolation, which is nominally considered to consist of 100.4 (2.5) for toxicodynamics and 100.6 (4.0) for toxicokinetics (IPCS, 1994).

The LOAELs and NOAELs are selected on the basis of the following reasoning. The LOAEL is the lowest level at which adverse effects are seen in studies of adequate quality. The NOAEL, the highest level at which no effect is seen in the available studies, must be lower than the selected LOAEL. The LOAELs and NOAELs are identified by a comparison with the equivalent continuous human exposure level. The LOAELs in these studies are strikingly similar, with values of 0.360, 0.359, 0.303, 0.336, and 0.23 mg/m 3 in studies from the Inhalation Toxicology Research Institute (United States; Mauderly et al., 1987), the Health Effects Research Program on light- and heavy-duty diesel engine exhausts (Japan; Ishinishi et al., 1986, 1988), the National Institute for Occupational Safety and Health (United States; Lewis et al., 1989), and the Fraunhofer Institute for Toxicology and Aerosol Research (Germany; Heinrich et al., 1986a, 1995), respectively (see Table 43). The consistency of these levels of effect, in view of the diversity of end-points represented, gives a high level of confidence in the end-points.

The study at the Inhalation Toxicology Research Institute was selected as the most representative because it covers the greatest variety of end-points, including histopathology, bronchiolar lavage biochemistry and cytology, lung tissue biochemistry, and particle clearance; LOAELs were observed for all of these end-points. The studies of the Health Effects Research Program showed a similar LOAEL but only for histological changes in the lung. The value for equivalent continuous human exposure is not available from the study carried out by the General Motors Corporation (Barnhart et al., 1981) 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmbecause the main results were for guinea-pigs; although rats were included in the study, detailed results were not presented. The appropriate NOAEL in these studies is not selected on the basis of the LOAEL. As close dosing intervals were used in the studies of of the Health Effects Research Program, the NOAEL was selected on the basis of studies of light-duty diesel engines, as 0.139 mg/m 3

.

B10.3.3 Exposure assessment

Exposure to diesel exhaust is discussed in sections B10.1 and

B10.2.

B10.3.4 Risk characterization

B10.3.4.1 Humans

A quantitative assessment of the risk for humans of non-carcinogenic effects of exposure to diesel exhaust cannot be made
on the basis of studies in humans. A substantial volume of literature shows an association between acute exposure to fine particulates and morbidity and mortality in the general population. It is somewhat uncertain whether there is a direct causal link and, if so, whether it is related to a specific component of the suspended particulates. Attention has been focused on fractions of suspended particulate matter, currently measured as PM 10 (diameter < 10 μm) or PM 2.5 (diameter < 2.5 μm), comprised mainly of diesel particulates in the submicrometre range, which represent an increasing proportion in some countries. There is also evidence that exposure to particulates is involved in respiratory symptoms and in mortality from certain chronic respiratory conditions.

B10.3.4.2 Experimental animals

Three approaches were used in assessing the risk for non-carcinogenic effects of exposure to diesel exhaust, on the basis of exposure (dose)-response assessment in experimental studies (see section B10.3.2.2), and for deriving guidance values for exposure. Approaches 1 and 2 are based on the NOAEL from studies in rats. The difference between the two is that approach 1 involves a dosimetric extrapolation model (Yu & Yoon, 1990) for converting the actual concentration to which animals were exposed to an equivalent continuous human exposure, thereby reducing the uncertainty in interspecies extrapolation.

In approach 2, no dosimetric conversion is performed but the usual uncertainty factors for interspecies and intraspecies extrapolation are applied. In contrast to these NOAEL-defined approaches, approach 3 is based on the principle of the 'benchmark dose', in which a concentration of exposure to diesel exhaust is derived from dose-response relationships observed in rats.

The model of Yu & Yoon (1990) takes into consideration the specific characteristics of particle deposition and retention in rat and human lung. Application of the model to studies of inhalation in experimental animals is shown schematically in Figure 4. The deposited and retained doses in the lung are calculated from the concentrations in rats and expressed per unit lung weight or per unit alveolar surface area. Under the assumption that the retained dose per lung will lead to the same effect in rodent and human lung, a human equivalent concentration can be calculated (as shown in Table 43), to obtain an equivalent continuous human exposure. The equivalent

continuous exposure of animals shown in Table 43 was derived either by calculating the continuous exposure at which the lung burden attained in a study by inhalation (if measured) would have been reached, or by calculating the lung burden in rats at the end of exposure on the basis of the actual exposure parameters (also shown in Table 43) and then calculating as above. The correlation between continuous exposure in animals and humans in the model of Yu & Yoon (1990), assuming that overload induces prolonged lung clearance, is shown in Figure 5. Use of this approach, rather than simple adjustment of discontinuous to continuous exposure by the c x t constant used in other assessments, should reduce the uncertainty associated with the transformation.

Approach 1. The NOAEL of 0.41 mg/m 3 from the study in rats exposed by inhalation to light-duty engine exhaust (Ishinishi et al., 1986, 1988; Table 43) is converted to an equivalent continuous exposure of 0.23 mg/m 3 in rats and then to an equivalent continuous exposure of 0.139 mg/m 3 , assumed to be the NOAEL in humans. Application of a sophisticated dosimetric model decreases the uncertainty in interspecies extrapolation from 10 to 10 0.4 (IPCS, 1994). Application of the usual uncertainty factor of 10 for intraspecies differences results in a total uncertainty factor of

10 × 10 0.4 = 25.

The guidance value derived from this approach is

0.139 mg/m 3 = 5.6 μg/m 3

.

--------------- 25

No additional correction factor is required, resulting in a guidance value (for the general population) of 5.6 μg/m 3

.

The inherent uncertainties in the dosimetric model are due to the assumptions that must be made. A principal assumption is necessary to estimate inhibition of clearance in humans, since data on this aspect
do not exist. It is assumed that clearance in humans is inhibited at the same lung burden (mass per alveolar surface area) as in rats. The 

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Approach 2. In this approach, the equivalent continuous animal exposure based on the NOAEL of 0.41 in rats (light-duty diesel exhaust; Ishinishi et al., 1986, 1988; Table 47) is used to derive the guideline value. Because the dosimetric model is not used, the default value of 10 is applied for interspecies uncertainty and an uncertainty factor of 10 is added for intraspecies differences. The guidance value obtained with this approach is

0.23 mg/m 3
------------- = 2.3 μg/m 3

100

Approach 3. An alternative to using the NOAEL is to derive a 'benchmark dose', as described by Crump (1984). In studies by inhalation, the concentration rather than the dose is considered to be more precise, and the 'benchmark concentration' is used. Like the benchmark dose, this term covers the entire exposure-response relationship in a given study rather than relying on only one data point representing the NOAEL or LOAEL. This approach reduces the uncertainty inherent in defining an NOAEL or LOAEL by considering the upper 95% confidence interval of the full exposure-response curve from a study in experimental animals, in which the lower 95% confidence limit of a concentration corresponds to a 1, 5, or 10% increase in response, defined as the percentage of animals responding in a specific group. This exposure concentration is then the benchmark concentration of the study. It is converted to the human benchmark concentration on the basis of differences in pulmonary dosimetry between the two species, which additionally reduces the use of uncertainty factors. Because the full range of experimental results from a specific study is used, the benchmark concentration approach reduces statistical uncertainty.

In principle, this approach involves three steps. The first is selection of the appropriate experimental study and end-point for establishing an exposure-response curve. The second is calculation of
the benchmark concentration for the animals from a mathematical description of the exposure-response curve and determination of the 95% confidence interval. Thirdly, the human benchmark concentration is calculated; for exposure by inhalation, the exposure of the animals is extrapolated dosimetrically to the human situation, with application of uncertainty factors the size of which is determined as discussed above. These factors generally consist of one for interspecies extrapolation and one for sensitive subpopulations. The first two steps of the benchmark concentration approach are shown schematically in Figure 6. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTwo sets of data were used to derive the benchmark concentration for diesel exhaust. The first was that of Ishinishi et al. (1988) from a study in which rats were exposed to exhaust from a light-duty diesel engine for two years at four concentrations. Hyperplastic lesions, shown histopathologically to be a sensitive indicator of lung damage, were used to establish a well-described exposure-response relationship. The second data set was that of Creutzenberg et al. (1990) for female Wistar rats exposed for 96 h per week for a total of
78 weeks to three concentrations of diesel exhaust. The most sensitive end-points for exposure-related non-neoplastic changes were lung clearance of particles and the occurrence of polymorphonuclear neutrophils (PMN) in lung lavage fluid as indicators of inflammation. Since these measurements are laborious, however, they were performed only in a subset of the exposed animals, so that only six to eight rats were studied per group.

The responses in the study of Ishinishi et al. (1988) were expressed in terms of individual animals affected per total number of exposed animals in each group, whereas the data of Creutzenberg et al. (1990) were reported as group mean values plus or minus the standard deviation, which is less useful for the benchmark concentration approach. Individual responses in the latter study were, however, provided by Bellmann (personal communication), so that the data from this study could also be expressed in terms of percentage of animals with impaired lung clearance. Impairment of lung function was considered to be significant when the calculated pulmonary retention half-time of the administered particles was at least 3.5 times greater than their normal average half-time in the lungs of control animals.

Evaluation of the end-point chronic inflammation (PMN in lung lavage fluid) proved to be more difficult, since all of the rats at the lowest exposure level (0.8 mg/m 3 ) in the study of Creutzenberg et al. (1990) had increased PMN levels (100% response). Lung lavage was performed at 22 and 24 months of exposure, and since the responses at these two times were similar, the data were combined to increase the numbers of animals per group to 11-14. The individual responses (percentage of PMN among the total number of cells in the lavage fluid) were used to derive the curve and the 95% confidence interval. An excess of PMN of up to 3% of the total number of cells over the background level was used to define the benchmark concentration. The highest value observed in the control group was 2.75% PMN after 22 months of exposure.

The three data sets and the results of the probability function, using a Weibull model to calculate the benchmark concentration, are given in Tables 44, 45, and 46 and are illustrated in Figures 7 and 8.

Table 44. Benchmark concentration for rats after long-term exposure to diesel exhaust by inhalation in study by Ishinishi et al. (1988); end-point: hyperplastic lesions of the lung

Concentration (mg/m 3 ) Rats that responded (equivalent continuous exposure of rats) Actal/observed Predicted Percent

numbers number

0 4/125 4.6 3.2 0.063 4/125 4.6 3.25 0.23 6/125 4.7 4.8
0.67 12/123 12.1 9.76 1.3 87/124 87.0 70.2

Probability function [p(conc)] - A 0 ]/(1 - A 0 ) = [1 - exp(-A 1 *

Conc) K ]

A 0 = 3.7196 × 10 -2

A 1 = 3.740

K = 4.3504

Lower confidence limit of the benchmark concentration (BC) for a given
risk: 

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0.1 0.634

0.05 0.511

0.01 0.313

The respective benchmark concentrations from the data sets for rats are 0.634, 0.119, and 0.090 mg/m 3 , corresponding to a 10% response (the lower 95% confidence limit on the exposure concentration for hyperplastic lung injury and impaired lung clearance) or a 3% excess of PMN in lavage fluid (the lower 95% confidence limit on the exposure concentration for chronic alveolar inflammation).

The resulting human benchmark concentrations for the three end-points were calculated from the model of Yu & Yoon (1990), applying uncertainty factors of 10 to account for sensitive subpopulations (human intraspecies differences) and 10 0.4 (2.5) for potentially different toxicodynamics (see above), to be: 14 μg/m 3
for hyperplastic lung lesions, 3 μg/m 3 for impaired lung clearance, and 2 μg/m 3 for chronic alveolar inflammation. Although the mathematical description of the exposure-response relationships in rats is very good for all three sets of data, they should be viewed as hypothetical since they are based on dosimetric conversions from rats to humans and on the application of uncertainty factors.

Table 45. Benchmark concentration for rats after long-term exposure to diesel exhaust by inhalation in study by Creutzenberg et al. (1990); end-point: impaired lung clearance

Concentration (mg/m 3 ) Rats that responded (equivalent continuous exposure of rats) Actal/observed Predicted Percent

numbers number

0 0/6 0 0
0.45 2/8 1.7 25 1.34 3/6 3.0 50 3.74 5/6 5.2 83.3

Probability function [p(conc)] - A 0 ]/(1 - A 0 ) = [1 - exp(-A 1 * Conc - D 0 )A 2 ]

A 0 = 0.0

A 1 = 0.522

A 2 = 1.00

D 0 = 0.0 (threshold)

Lower confidence limit of the benchmark concentration (BC) for a given
risk:

Risk Rat BC (mg/m 3 )

0.1 0.119

0.05 0.058

0.01 0.011

The non-cancer guidance values and the benchmark concentrations derived from these approaches are summarized in Table 47.

More sensitive end-points with respect to the adverse effects of diesel exhaust on the lower respiratory tract are impaired clearance in the deep lung and chronic alveolar inflammation, rather than hyperplastic lung lesions. Chronic alveolar inflammation was also a significant finding in the long-term study of rats exposed to diesel exhaust by Henderson et al. (1988), who found a significantly increased percentage of PMN in lavage fluid from the group with the lowest exposure (0.35 mg/m 3 ) after two years. For all three types of end-points, however, there may be a threshold below which no change is to be expected. The threshold concept is not included in the mathematical model, and it would be difficult to do so at this point in the absence of specific experimental data and data on variation between human subjects.

These estimates are for the effects of long-term exposures, and the concentrations are expressed as annual means over a lifetime. They could apply to long-term effects on health, such as the association between mortality from certain chronic conditions and exposure to particulates in air pollution seen in a study in six cities in the United States (Dockery et al., 1993). Their relevance to human experience must also be considered in the context of particulate air pollutants in general, however, and they have no direct bearing on the acute effects of exposure to particulates, for which guidance values are required in terms of 24-h mean concentrations.

B10.4 Neoplastic effects

B10.4.1 Hazard identification 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmB10.4.1.1 Lung cancer: occupational exposure

In the 1970s and 1980s, the recognition that diesel exhaust contains respirable particles and that known carcinogenic substances are adsorbed on the surface of these particles led to the hypothesis that inhalation of diesel exhaust could result in lung cancer in humans. As lung cancer develops slowly, over many years, studies of individuals with long, well-defined exposure and follow-up (> 20 years) were considered to be the most informative. Four studies of occupationally exposed individuals meet these criteria (Garshick et al., 1987, 1988; Gustavsson et al., 1990; Emmelin et al., 1993; see Table 43).

Table 46. Benchmark concentration for rats after long-term exposure to diesel exhaust by inhalation in study by Creutzenberg et al. (1990); end-point: chronic alveolar inflammation

Concentration (mg/m 3 ) PMN in lung lavage fluid (%) No. rats (equivalent continuous responding exposure of rats Actual/observed Predicted per no. exposed

0 0.7 0 0/11 0.45 11.1 11.7 13/13
1.34 29.9 27.6 14/14 3.74 50.8 51.9 14/14

Dose-response function: F(dose) = Q(0) × (Dose - (Dose - D 0 ) Q(2)

Dose = log(1 + d) (d = rat exposure concentration)

Q(0) = 0.6.53 Q(1) = 77.010 Q(2) = 1.056

D 0 = 0.0 (threshold)

Lower confidence limit of the benchmark concentration (BC) for a given excess polymorphonuclear neutrophil (PMN) response (% of total lavaged cells):

PMN in excess Rat BC of control (%) (mg/m 3 )

3 0.090

2 0.059

1 0.029

Transition from steam to diesel locomotives occurred in the American railroad industry during the 1950s. In a case-control study of American railroad workers (Garshick et al., 1987), exposure was estimated on the basis of yearly job (exposed or unexposed to diesel exhaust) from 1959 to death or retirement; deaths were identified for 12 months in 1981-82. In an additional study of these workers (Garshick et al., 1988), a retrospective cohort was defined on the basis of work in a job with exposure to diesel exhaust in 1959, and 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmdeaths were recorded for 1959-80. Although the previous exposure levels of the railroad workers were not available, the classification of workers into groups on the basis of exposure was validated by an industrial hygienist and a review of work practices. Air sampling was conducted to establish current exposure levels in various jobs (Hammond et al., 1988; Woskie et al., 1988a,b).

Gustavsson et al. (1990) studied lung cancer among bus garage workers in Stockholm, Sweden, where all buses with internal combustion engines have been diesel-fuelled since 1945. A cohort was established of people who had worked in bus garages for at least six months between 1945 and 1970, and mortality was determined for 1952-86. A nested case-control study was performed within this cohort. Although actual exposure was not measured, relative exposure to diesel exhaust was graded on the basis of a scale established by an industrial hygienist in a review of work practices, bus engine operation, and shop ventilation.

Table 47. Summary of non-cancer guidance values and benchmark
concentrations

Approach Guidance value

or benchmark
concentration (μg/m 3 )

NOAEL with dosimetric conversion from rats to humans 5.6 a

NOAEL without dosimetric conversion from rats to 2.3

humans

Benchmark concentration with dosimetric conversion

from rats to humans

Chronic alveolar inflammation 2 a Impaired lung clearance 3 a Hyperplastic lesions 14 a

Benchmark concentration without dosimetric
conversion from rats to humans

Chronic alveolar inflammation 0.9 Impaired lung clearance 1.2 Hyperplastic lesions 6.3

NOAEL, no-observed-adverse-effect level Normalized for lung surface area in rats and humans; after normalization for lung weight, the benchmark concentration increases by a factor of 4.

Emmelin et al. (1993) studied Swedish stevedores in 15 ports where diesel equipment had been introduced between 1957 and 1963; they also performed a nested case-control study on cases of lung cancer identified between 1960 and 1982. Exposure to diesel exhaust was
estimated on the basis of three indices derived from estimated diesel fuel consumption and years of work after the introduction of diesel equipment in each port.

All four studies showed an increased risk for lung cancer with exposure to diesel exhaust. The relative risks reported ranged from about 1.4 for railroad workers with the longest duration of exposure in both studies of such populations to 1.3-2.4 for bus garage workers, depending on the exposure category. The stevedores also had an increased relative odds ratio for lung cancer with increasing exposure with respect to all three indices of exposure examined, with an increase of three- to sixfold in the highest exposure categories and 1.5-2.7-fold in the lowest exposure category. The point estimates were, however, imprecise and had wide confidence intervals: for all but one exposure category, the lower limit of the 90% confidence intervals presented by the authors included 1.0. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmThe effects of cigarette smoking could be adjusted for in the case-control studies of railroad workers (Garshick et al., 1987) and stevedores (Emmelin et al., 1993). The smoking histories of the railroad workers were obtained from next-of-kin. Various regression models (including both dose and duration of smoking) were examined in detail in order to adjust adequately for the effects of smoking. No matter how it was accounted for, the risk for lung cancer based on work in a job with exposure to diesel exhaust was similar to the unadjusted odds ratio. The study of stevedores (Emmelin et al., 1993) was smaller than that of railroad workers, and in order to reduce the number of strata for analysis workers were stratified only as smoker or nonsmoker. In the two studies in which information on smoking was not available (Garshick et al., 1988; Gustavsson et al., 1990), the analysis was based on a comparison with workers in the same occupational cohort, making confounding by smoking less likely.

The results of most of the other studies summarized in Tables 39-41 support those of the four studies discussed above. The relative risks were generally in the range 1.2-1.9 but did not always achieve statistical significance. Furthermore, exposure to diesel exhaust was less precisely defined in these studies; it was usually based on self-reporting, the report of a surrogate, or a census report of occupation (Table 41). Self-reporting or the report of a surrogate would reflect actual exposure to diesel exhaust or work in a job in which exposure was likely; however, defining exposure in this less precise fashion make it harder to detect an effect of the exposure and
leads to lower relative risks and wider confidence intervals.

In three other studies, job titles obtained from an employer were used to define exposure. In the study of Howe et al. (1983), the job
held at the time of retirement from the railroads was used to categorize exposure, and mortality was assessed for 1965-77. Significantly elevated risks for lung cancer were noted among workers who had probably been exposed to diesel exhaust (relative risk = 1.35) and among those who had possibly been exposed (1.20); however, since only retired workers were studied, actual exposure to diesel exhaust would have been short for many workers. Rushton et al. (1983) and Waller (1981) used job titles within the London Transport organization to indicate potential exposure to diesel exhaust. Neither study showed an elevated risk for lung cancer, but the study of Rushton and coworkers was characterized by a short duration of exposure and follow-up, whereas in the study of Waller, deaths occurring after employment were not included.

The relative risks for lung cancer as a result of exposure to diesel exhaust are generally low, and risks of this magnitude are more susceptible to chance and to the effects of unmeasured confounding factors and imprecision in adjusting for known confounding factors. As discussed above, the elevated risk for lung cancer observed in the four most informative studies is unlikely to be due to confounding by cigarette smoking and is probably due to exposure to diesel exhaust. Other studies, although limited primarily by the exposure ascertainment, support this assessment.

B10.4.1.2 Urinary bladder cancer: occupational exposure

The hypothesis that exposure to diesel exhaust results in cancer of the urinary bladder is based on the fact that individuals who smoke cigarettes, a source of PAHs (as is diesel particulate), excrete mutagenic substances in their urine, and cigarette smoking is
associated with an increased risk for bladder cancer. In a recent study of railroad workers with current exposure to diesel exhaust (median concentration of respirable particulates over a work shift, 54-113 μg/m 3 ; adjusted for smoking), mutagenicity in urine at the end of a shift was not associated with exposure (Schenker et al., 1992).

The evidence that bladder cancer is associated with exposure to
diesel exhaust is based on an association between work in industries or occupations with potential exposure to diesel exhaust. The relative risks reported (some of which are adjusted for smoking) are generally in the range 1.2-2 (Hoar & Hoover, 1985; Silverman et al., 1986), although higher risks have been reported. Exposure was assessed mainly by self-reporting and reporting of occupation at interview. The main occupation in which there was considered to be potential exposure to diesel exhaust was truck driving. Measurements of exposure of truck drivers by Zaebst et al. (1991) indicate that current exposure is likely to be low, but there is no knowledge about past exposure

levels.

Thus, although work in occupations related to motor vehicles has been associated with bladder cancer, the extent of exposure of these
workers to diesel exhaust is unknown.

B10.4.2 Dose-response assessment

B10.4.2.1 Lung cancer

Job category and work in an occupation with exposure to diesel exhaust have been used as indicators of exposure. When years of exposure and derived indices of exposure have been used, an exposure-response relationship has generally been found. Measurements of total and respirable dust in exposed workers do not reflect actual exposure to diesel exhaust particles, however, since there are other sources of dust in an occupational environment. The exposure of only two occupational groups, American railroad workers (Woskie et al., 1988b) and truck drivers (Zaebst et al., 1991), to diesel exhaust particles has been quantified (see section B5.2). Current exposure was measured for both these groups, but past exposure levels could not be determined (Woskie et al., 1988b), so that studies of workers do not 

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B10.4.2.2 Urinary bladder cancer

Years of work in an occupation related to motor vehicles (mainly truck driving) was examined in some, but not all, studies, but no measurements were available to estimate exposure to diesel exhaust in these settings.

B10.4.3 Exposure assessment

The ranges of exposures observed for workers are presented in
sections B5.2 and B10.2.

B10.4.4 Risk characterization

B10.4.4.1 Human lung cancer

Thus, historical measurements of exposure to diesel exhaust are unreliable and exist only for current workers in two industries. A quantitative risk assessment cannot be conducted on the basis of epidemiological data in which job title was used as a surrogate of exposure. Attempts to obtain estimates of risk (presented in section B10.4.1) in the retrospective cohort study of American railroad workers (Garshick et al., 1988) were not successful (see below for discussion). Consequently, there are no human data suitable for estimating unit risk.

B10.4.4.2 Human urinary bladder cancer

The risk for urinary bladder cancer cannot be assessed from the available epidemiological data.

B10.4.4.3 Risk characterization based on studies in experimental
animals

(a) Introduction

Most human tumours arise in the bronchial region, and although the frequency of adenocarcinomas has increased in recent years (Martini, 1993), squamous bronchogenic carcinomas still represent the majority (about 60%) of lung tumours in humans (Auerbach & Garfinkel, 1991; Devesa et al., 1991; Campobosso et al., 1993). The tumours observed in rats are located almost exclusively in the bronchoalveolar regions. This difference raises the question of whether the rat is the appropriate species for extrapolating to humans. Although it may be the wrong model, however, studies of hamsters and monkeys give negative results, so the rat is the most appropriate model from a conservative viewpoint and is therefore used for quantitative risk extrapolation.

The objective is to assess quantitatively the potential risk for lung cancer in humans posed by exposure to diesel exhaust emissions in the ambient air. Ideally, human risk due to exposure to an environmental pollutant should be predicted on the basis of human experience. Although several epidemiological studies are available on bus and railroad workers, these results alone are not adequate to assess the potential risk of cancer in humans because of the lack of reliable information on the conditions of exposure of these workers. The challenge is to assess the risk of exposure to diesel exhaust on the basis of all of the available information, for both experimental animals and humans. In contrast to the sparse human data, there is a rich pool of information on diesel-induced lung tumours in two strains of rat. One approach to integrating this information is to make a quantitative risk assessment on the basis of information from bioassays and on the relevant biological mechanism and then to evaluate these animal-based results against available human experience. This approach was followed in this monograph.

Although, the available experimental data on the possible impact of extractable organic matter and PAHs on the lung tumour response associated with exposure to diesel soot are not strong (Heinrich et al., 1991; Heinrich, 1994; Heinrich et al., 1995), a mathematical approach to assessing the risk of exposure to diesel exhaust should take into account the effects of both particles (carbon core) and extractable organic matter, because: (i) organic compounds include a variety of PAHs and nitroaromatic compounds, many of which are known

to be carcinogenic; (ii) the results of recent studies on inert particles and carbon black in rats (Mauderly et al., 1991; Heinrich, 1994; Heinrich et al., 1994) strongly support the hypothesis that the carbon cores of diesel particles are the component primarily responsible for the induction of lung cancer; (iii) PAHs alone are unlikely to be responsible for the tumour response; (iv) the disproportionately high tumour incidence in animals with heavy exposure coincides with a disproportionate increase in the cumulative lung burden of diesel particles. Although a qualitative description of the biological mechanism of diesel exhaust-induced tumours is plausible, however, the lack of quantitative information on the dynamics of tumour initiation, promotion, and progression vitiates the construction of a biologically based dose-response model.

The crudest dose-response model is obtained by fitting observed dose-response data to a mathematical function that is a monotonic, non-decreasing function of exposure, using the inhaled concentration and observed tumour incidence, without considering information on pharmacokinetics and the mechanism of carcinogenesis. This approach may result in uncertain estimates of risk at low doses, because, while the model may adequately fit the dose-response data for high doses at 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmwhich a tumour response is observed, it may greatly underestimate or
overestimate the risk at low doses.

Most previous risk estimates of diesel exhaust-induced cancer risk were developed before the results of many of the bioassays became available (Pepelko & Chen, 1993). Nevertheless, most of the estimates are similar (within an order of magnitude), except for those calculated on the basis of human data, which are higher than those calculated from other databases. Because of great uncertainties associated with the available human data, it is considered more appropriate to calculate risk on the basis of animal data and then use human data to evaluate their validity.

The approach adopted in this assessment is the linearized multistage model (Anderson et al., 1983), in which burden per lung surface area (milligrams per square centimetre) is used as an equivalent dose in animals and humans. The use of lung (alveolar) surface area is justified by the fact that the lung tumours observed in rats are derived exclusively from epithelial cells in the alveolar region of the lung. Normalization on the basis of lung weight can also be done by using a factor of 4, for increasing the dose or decreasing the risk, for the difference in the ratio of rat:human lung surface area and lung weight. The risk prediction calculated by this approach is presumably conservative (i.e. overpredicts the risk) because the dose-response function is assumed to be non-threshold and linear at

low doses.

An alternative model that incorporates a biological mechanism is constructed using statistically estimated parameters. The model assumes that carcinogenic agents present in the organic fraction act directly on the target cells, primarily via initiation. It is further assumed that the majority of the particles are ingested by macrophages. Particle-laden macrophages are then induced to secrete a variety of mediators (e.g. reactive oxygen species and cytokines), which diffuse to the target cells and induce initiation, proliferation, and conversion of initiated cells to malignant cells. This alternative model can be used only as a tool to evaluate the impact of various biological assumptions and not for risk prediction, because the parameters of the model are not measured in the laboratory.

(b) Approaches to quantification of human risk from exposure to
diesel exhaust

Several issues must be addressed in estimating the risk of exposure to diesel exhaust, including the critical target site, the fraction of exhaust responsible for tumour induction, the availability of dosimetric methods for accurately extrapolating dose from that for experimental animals chronically exposed to high concentrations of exhaust to that of humans exposed at ambient concentrations, and the most suitable low-dose extrapolation model.

The critical target organ was considered to be the lung. Although Iwai et al. (1986) reported induction of both malignant lymphomas and lung tumours in rats exposed to diesel exhaust, the lung was the only target site in other experimental studies of this species. Although potentially carcinogenic agents present in diesel exhaust may be absorbed from the lungs, enter the bloodstream, and be transported systemically, no data are available to evaluate this possibility. Organic compounds adsorbed on particles may also reach systemic targets via the gastrointestinal tract, as particles deposited in the conducting airways are cleared rapidly to the oropharynx and swallowed. A considerable volume of particles is also likely to be ingested as a result of grooming when the whole animal is exposed (Wolff et. al., 1982); however, because the half-times for the elution of organic compounds from particles are considerably longer than their passage through the gastrointestinal tract, the fraction absorbed is expected to be small. In any case, there is little evidence for any systemic effects of diesel exhaust.

The site of action in the lungs is assumed to be the epithelial lining of the alveoli and small airways. According to Mauderly et al. (1987), inflammation and tumours appear to arise from this tissue. Although interstitial events (e.g. fibrosis) have been suggested to be associated with induction of lung tumours by particles (Kuschner, 1968), no data are available to support this view with respect to
diesel exhaust.

Accurate extrapolation of dose from studies of animals exposed to high concentrations of exhaust to humans exposed to ambient concentrations requires a variety of adjustments for species differences in deposition efficiency and respiratory ventilation rates. As the normal retention half-times in the alveolar region are several times longer in humans than rats (Chan et al., 1981; Bohning et al., 1982), the lung burden of humans may be underestimated if this difference is not taken into account. The high exposure concentrations used in some experimental studies, however, result in greatly slowed or even completely inhibited clearance (Griffis et al., 1983). In order to extrapolate dose accurately from experimental studies to humans, the detailed dosimetric model developed by Yu et al. (1991) was used, which accounts for species differences in respiratory ventilation rates, deposition efficiency, normal particle clearance rates, transport of particles to lung-associated lymph nodes, and lung surface area. It also accounts for inhibition of particle clearance due to lung overload. In this model, dose is estimated in terms of particle concentration per unit of lung surface area.

Two approaches were used to derive unit risk estimates: the linearized, multistage, low-dose extrapolation model and a model based on the biological mechanism discussed above. In the linearized 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmmultistage model, the lung burden of carbon core per lung surface area is used as the dosimetric parameter. A particle-based assessment was considered to be reasonable for two principal reasons: (i) exposure to the vapour phase alone did not result in detectable tumour induction in rats (Brightwell et al., 1986; Iwai et al., 1986; Heinrich et al., 1986a); and (ii) exposure to carbon black, which is similar in composition to the carbon core of whole diesel exhaust but contains only negligible amounts of organic compounds, was about as effective in inducing lung cancer as whole diesel exhaust (Mauderly et al., 1991; Nikula et al., 1994; Heinrich et al., 1995). Although use of the carbon core as a dosimetric parameter implies that it is primarily the insoluble carbon core of diesel particles that is responsible for the carcinogenic effects of diesel exhaust, it can also be viewed simply as a marker of exposure to diesel exhaust, with no biological implication. Similarly, organic compounds could be used as markers of exposure for the dose-response calculation. When this is done, the resulting unit risk estimates (not presented here) are very similar (within 25%) to those calculated with the carbon core as the surrogate of exposure.

The second approach is based on the assumption that, even though the concentration of carcinogenic compounds on diesel particles is low, they nevertheless can act in concert with the particles to induce carcinogenesis. An alternative low-dose extrapolation model was therefore developed that allows for the possibility that various PAHs and nitroaromatic compounds may induce organ-specific adducts that contribute to cell initiation. This model is described in Appendix A1.

Both approaches incorporate the detailed dosimetric model of Yu et al. (1991) to estimate dose at the airway and alveolar surfaces. Risk is based on the assumption that an equivalent concentration (dose) per unit of alveolar surface area results in equivalent risks
in humans and rats.

(c) Evidence to support a carcinogenic effect of the carbon core

The study of Heinrich et al. (1994) on carbon black and coal-tar pitch provides information useful for evaluating the assumptions used to construct the model that reflects current thinking about the mechanism of action of diesel exhaust in inducing lung tumours. One relevant question about diesel exhaust is whether lung tumours are induced by the carbon core alone or in combination with organic compounds.

In this study, female Wistar rats were exposed to carbon black or tar pitch, or both for 10 months (43 weeks) or 20 months (86 weeks) and followed until the end of the study, which lasted for two-and-a-half years. No lung tumours were observed in the control group. For each exposed group, the probability of dying from lung cancer was calculated by a Kaplan-Meier survival analysis. Since these data are used solely for interpretation and not for risk prediction, it is statistically more appropriate to use only data on mortality, excluding those tumours found at terminal sacrifice. Figure 9 shows the probability of dying from lung cancer for the rats that were exposed either to 6 mg/m 3 of carbon black, or to coal-tar pitch containing 50 μg/m 3 of benzo[a]pyrene, or to a combination of the two for 10 months, and then followed until the end of the study. In Figure 10, each plot represents the time at which a tumour occurred. To provide a better visual presentation, the probabilities at these points are connected by straight lines. (Note: It is statistically more appropriate to present these probabilities by a step function rather than by interpolation when the Kaplan-Meier procedure is used.)

Two conclusions can be drawn from Figure 10: (1) Tumours appeared much earlier in animals exposed to coal-tar pitch alone or in combination with carbon black than in animals exposed to carbon black alone. For instance, the first tumour occurred 665 days after the start of exposure to carbon black, 406 days after exposure to coal-tar pitch, and only 310 days after exposure to the combination. (2) Carbon black and coal-tar pitch together caused a higher rate of mortality from tumours than the sum of mortality caused by carbon black and coal-tar pitch separately. The probability of dying from a lung tumour before terminal sacrifice at 912 days was 0.28 for carbon black, 0.52 for coal-tar pitch, and 0.99 for the combination. These two observations suggest that carbon black and coal-tar pitch act on different stages of tumorigenesis. They are consistent with the hypothesis that carbon black can increase the proliferation rates of normal and cells at different stages of differentiation initiated spontaneously or by coal-tar pitch.

When the rate of mortality from tumours among rats exposed to diesel exhaust is compared with that of groups exposed to carbon black, coal-tar pitch, or the combination (Figure 9), the rate among the animals exposed to diesel exhaust is comaprable to that of rats exposd to carbon black for 10 to 20 months. This implies that carbon black alone can induce a tumour response similar to that induced by diesel exhaust. Figure 9 also shows that doubling the duration of exposure to carbon black from 10 to 20 months does not increase the probability of dying from a lung tumour before terminal sacrifice; however, doubling the duration of exposure to coal-tar pitch significantly increases the probability of dying from a lung tumour. Figure 11 shows that the shapes of the curves for age-dependent mortality from cancer for animals exposed to diesel exhaust and coal-tar pitch for 20 months are similar. It should be kept in mind that the PAH content of the atmosphere containing both carbon black and coal-tar pitch was about 1000 times higher than that of diesel engine exhaust. These observations may have profound implications for elucidating the mechanisms of induction of lung tumours by diesel exhaust, but more refined analyses are necessary before any conjecture

can be made. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htm(d) Data available for calculating risk

Seven bioassays have shown lung tumour responses in rats (Brightwell et al., 1986; Heinrich et al., 1986b; Ishinishi et al., 1986; Iwai et al., 1986; Mauderly et al., 1987; Nikula et al., 1994; Heinrich et al., 1995). Four studies were chosen for use in calculating unit risk (Table 48) because they involved multiple exposure groups. Data on time to event (death with or without a tumour) are available in the studies of both Mauderly et al. (1987) and Heinrich et al. (1995) and were used in all of the risk
calculations.

(e) Calculation of unit risks

The unit risk from exposure to an air pollutant is defined as the 95% upper bound of the increased lifetime cancer risk for an individual continuously exposed to a concentration of 1 μg/m 3 in ambient air. Unit risk is a convenient tool for calculating lifetime risk due to exposure to a pollutant when low-dose linearity is assumed. Under this assumption, the risk due to exposure to d μg/m 3 of pollutant can be calculated from u × d if the pollutant has a

unit risk of u /μg per m 3 . This calculation is valid, however, only
when the level of risk is low. The results of unit risk calculations
are summarized in Table 49. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmTable 48. Incidence of lung tumours in rats exposed to diesel engine exhaust

Strain Exposure Dose metric Lung tumour Refe rence (sex) incidence Schedule Concentration Weekly exposure Lung particle burden (mg/m 3 ) (mg/m 3 × h) (mg/cm 2 lung surface) a

Fischer 7 h/d, 0 0 0 2/230 Maud erly et al.
344 5 d/week 0.35 12 6.4 × 10 -5 3/223 (1987)
(f+m) 3.50 122 2.8 × 10 -3 8/222

7.08 248 6.0 × 10 -3 29/227

Wistar 18 h/d, 0 0 0 1/217 Hein rich et al.
(f) 5 d/week, 0.84 76 5.8 × 10 -4 0/198 (1995) 2 years, 2.50 225 5.2 × 10 -3 11/200

< 6 months 6.98 628 1.5 × 10 -2 22/100 follow-up

Fischer 16 h/d, 0 0 0 1/123 Ishi nishi et al.

344 6 d/week b 0.46 44 2.5 × 10 -4 1/123 (1986)
(f+m) 0.96 92 2.0 × 10 -3 0/125

1.84 177 4.2 × 10 -3 4/123

3.72 357 8.8 × 10 -3 8/124

Fischer 16 h/d, 0 0 0 4/250 Brig htwell et al.
344 5 d/week 0.7 56 3.5 × 10 -4 1/112 (1986)
(f+m) 2.2 176 4.2 × 10 -3 14/112

6.6 528 1.3 × 10 -2 55/111

a Calculated from the model of Yu et al. (1991)

b Data on heavy-duty diesel engine exhaust used because those for light-duty engines showed no statistically significant differ ence Particle-based model: The linearized multistage model, which is used as a conservative approach to estimating risk, has the mathematical form P = 1 exp(-Z), where Z is either q 0 + q 1 × d + ...

+ q m × d m of a polynomial concentration d, or ( Q 0 + Q 1 × d + ...

+ Q
m × d m ) × t k , a polynomial of concentration d multiplied by a time

factor, t k , when data on time to event are used. In this case, the lifetime risk is calculated from actuarial life tables using the survival probability of control Fischer 344 rats within the National Toxicology Program, provided by Portier et al. (1986). The range of extrapolation is about three orders of magnitude in the present

assessment. Denote P 0 the lifetime cancer risk at concentration 0.

Because the extra risk (P-P 0 )/(1-P 0 ) is dominated by the linear term

q 1 × d at low concentrations, the 95% upper bound of q1 is used
to represent unit risk.

In extrapolating risk from animals to humans, a dose metric that
induces the same tumour incidence rate in animals and humans must be assumed (i.e. dose equivalence). To calculate equivalent doses, a mathematical model is used to adjust for the dosimetric parameters that determine the lung burden of particulate matter in rats and humans and to correct to dose per unit lung surface area. This dose is considered to be equally potent in inducing lung tumours in animals
and in humans. The model accounts for differences between animals and humans in regional deposition efficiency, particle clearance rates at low doses and at doses that result in impaired clearance, and lung surface area. In these calculations, the mass fraction of organic compounds adsorbed on particles is assumed to be 20%; one-half of the mass is assumed to be composed of slowly eluted organic compounds (half-life, 30 days) and the other half of rapidly eluted organic compounds (half-life, 1.3 h). The remainder is considered to be inorganic carbon. At higher concentrations, particle clearance slows and may even stop, and the lung particle burdens increase continually 

# Pa g e 122 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmduring exposure. The organic constituents, however, are eluted from the particles fairly quickly and reach a steady state even during continued exposure to high concentrations. The lung burdens of organic compounds are therefore less affected by inhibition of clearance by overloading.

In the linearized multistage model, determination of the dose of the carbon core is problematic because the lung burdens after low and high exposures differ drastically over time. This difference suggests that use of a lung burden at a fixed time (e.g. one year after the start of exposure) to represent dose may not be appropriate. In this assessment, the average lung burden is used as the dose at the target organ. The average lung burden is calculated by dividing the area under the curve of lung burden over time by the corresponding period of the experiment for which the curve was calculated. Figure 12 shows a comparison of the lung burden predicted from the model and that observed in the laboratory (Muhle et al., 1994) at specific times. It suggests that the model used to calculate lung burden is adequate.

Table 49. Estimates of unit risk per microgram of particles per
cubic metre of diesel exhaust

Upper 95% confidence limit of cancer Study risk due to exposure to 1 μg/m 3 of diesel particulate matter

3.4 × 10 -5 Mauderly et al. (1987)

1.6 × 10 -5a Ishinishi et al. (1988)

7.1 × 10 -5 Brightwell et al. (1986)

3.4 × 10 -5 Heinrich et al. (1995)

3.4 × 10 -5 Geometric mean of
four studies

If milligrams per lung weight are used instead of milligrams per lung surface as the equivalent dose, the risk estimates are reduced by a

factor of 4.

a Heavy-duty diesel engine

Biologically based model: A second approach to estimating risk
was used because it was considered more desirable to base risk on a biologically based dose-response model. Although the data are at present insufficient to replace the linearized multistage model, which is considered more conservative, the implications of hypothetical mechanisms of cancer induction by diesel particles can nevertheless be investigated. The biological issues considered include the effects on the carcinogenic process of particle-adsorbed organic compounds and of a variety of mediators secreted by particle-laden macrophages. A stochastic model was developed, which:

-- accounts for the possible effects of both the carbon particles and their associated organic compounds:

-- allows evaluation of the contribution to tumour induction of both carbon particles and organic compounds;

-- allows for changing parameters with increasing lung burden;

and

-- assumes that cell proliferation and tumour induction are stochastic. (For instance, it is not appropriate to assume that all cells divide at the same age.)

Unlike the linearized multistage model, this approach does not require a constant dose metric but allows for varying lung burden over time.

The model, which is described in Appendix B10.3, allows for initiation by both the carbon and the organic fraction and for the proliferative effects of the carbon fraction. Although these 

# Pa g e 123 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmmechanisms remain to be proven, it is assumed that carcinogenic agents present in the organic fraction act directly on the target cells, primarily by initiation. It is further assumed that most of the particles are ingested by macrophages. Particle-laden macrophages are then induced to secrete a variety of mediators (e.g. reactive oxygen species and cytokines), which diffuse to the target cells, inducing initiation, proliferation, and conversion of initiated cells to malignant cells.

The results reported by Mauderly et al. (1987) for tumour induction were used to estimate the model parameters. These are based on the development of malignant tumours rather than all tumours as in the first method. This was necessary in order to ensure that the data used to estimate the parameters represented the same biological mechanism. Lung burdens were calculated from the same dosimetry model used in the linearized multistage model.

(f) Results of unit risk calculations

Particle-based model: Unit risks were derived from the linearized multistage approach for the tumour incidences seen in four bioassays (Brightwell et al., 1986; Ishinishi et al., 1986; Mauderly et al., 1987; Heinrich et al., 1995) and the corresponding equivalent doses (Table 48). The resulting unit risk estimates, listed in Table 49, range from 1.6 to 7.1 × 10 -5 /μg particles per m 3 with a geometric

mean of 3.4 × 10 -5 /μg per m 3

.

In these calculations, the relationship between air concentration (micrograms per cubic metre) and lung burden (milligrams) in humans is used to determine the lung burden resulting from lifetime exposure to 1 g/m 3 of diesel exhaust particulate matter. The particle burden in terms of mass per unit lung surface area is then multiplied by the slope derived from the bioassay data. For instance, when the data of Mauderly et al. (1987) are used, the slope of the curve for carcinogenicity in rats (i.e. the upper 95% confidence limit of the linear coefficient in the multistage model), expressed in terms of equivalent dose (micrograms of carbon particulate matter per square centimetre) is 1.7 × 10 -2 /μg per cm 2 . According to the dosimetry model, an air concentration of 1 μg/m 3 of particulate matter corresponds to a mass of 1230 μg of carbon particles per human lung. Because the lung epithelial surface area, including the alveolar region and conducting airways, is assumed to be 627 000 cm 2 , a unit

risk of 3.4 × 10 -5 /μg per m 3 is derived from 1.7 × 10 -2 /μg per cm 2
× 1230 μg/627 000 cm 2

.

Biologically based model: The unit risk estimated by the alternative model, based on the data on malignant tumours from the study of Mauderly et al. (1987), is equal to 1.65 × 10 -5 /μg particles

per m 3 . This is lower than the estimate of 3.4 × 10 -5 /μg per m 3 derived from the same study with the linearized multistage approach. Application of the linearized multistage model only to malignant tumours seen in that study, rather than all lung tumours, resulted in

a unit risk estimate of 1.74 × 10 -5 /μg particles per m 3 (Table 50). Thus, the unit risk estimates obtained by the two approaches are virtually identical. The estimated risks may differ somewhat with increasing doses, because the slopes are not identical at all exposure levels. It should be noted, however, that the unit risk predicted by the alternative model is derived under the assumption that particles continue to exert an effect on cell initiation or proliferation (or both) at low doses. There is considerable uncertainty about the effects of particles at low doses. It has been claimed that particles do not induce initiation or proliferation at low doses (Vostal, 1986). At present, the evidence is inadequate to support or refute this claim. Moreover, even if macrophage overload is required, because of uneven distribution of particles, some macrophages may become overloaded even at low concentrations. Because of this uncertainty, the biologically based model, like the linearized multistage model, does not depart from linearity at low doses; however, if initiation and proliferation do not occur at low doses, the risk may be much

smaller.

In an estimate of unit risk for rats, based on the results of seven studies by inhalation and using linear interpolation and the linearized multistage model, the risks were 7 × 10 -5 /μg of diesel particles per m 3 and 10 × 10 -5 /μg carbon core particles per m 3 (Csicsaky et al., 1993; Pott et al., 1993; Roller & Pott, 1994).

(g) Results and implications of the biologically based model

On the assumption that particles continue to affect cell initiation or proliferation at low doses, the risks calculated with this model are comparable to those obtained with the linearized multistage model. Excess risks due to various exposures are shown in Tables 50 and 51. Those in Table 50 are the risks predicted for humans exposed continuously (24 h/day) from the two models: it is interesting
to note that the results are similar. Table 51 shows the excess risks due to exposure to 2.6 μg/m 3 of diesel particulate emission for 16 h/day on seven days per week and to 15 μg/m 3 for 8 h/day on five days per week. The first concentration was reported by the US Environmental Protection Agency Office of Mobile Sources to be the annual mean exposure of the American population to diesel particulate matter in 1986; the second concentration was reported to be that to which workers are exposed on urban freeways (Carey, 1987). The retention half-time for insoluble particles after exposure to

Table 50. Comparison of excess risk for humans due to continuous exposure to various concentrations of diesel exhaust emissions using two
different models 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmExposure concentration Biologically based (alternative) Linearized (μg/m 3 ) multistage Maximum likelihood Upper 95% model
estimate bound estimate

0.01 7.68 × 10 -8 1.35 × 10 -7 1.71 × 10 -7

0.1 8.12 × 10 -7 1.71 × 10 -6 1.72 × 10 -7
1.0 (unit risk) 8.16 × 10 -6 1.65 × 10 -5 1.74 × 10 -5

100 5.58 × 10 -4 9.63 × 10 -4 1.74 × 10 -4

1000 2.60 × 10 -2 4.22 × 10 -2 3.33 × 10 -2

a Slope - 9.04 per mg/cm 2 of lung surface, using carbon core as dosimetric. Only malignant tumours are used in the calculations.

2.6 μg/m 3 is shown to increase from 296 days for members of the general population with normal respiratory function to 519 days for those with a smoking history of 20 pack-years (Bohning et al., 1982).
This information was used to reduce the alveolar clearance rate for
the dosimetric calculations to that used for other risk calculations. Interactive effects of smoking and diesel exhaust are not considered in the risk calculations owing to lack of data. The studies of carbon black and coal-tar pitch (Heinrich, 1994; Heinrich et al., 1994) indicate that smokers have a higher risk for lung cancer than nonsmokers when they are exposed to diesel exhaust.

The excess lifetime risks shown in Tables 50 and 51 are standardized by the actuarial life-table approach, using the survival probability of control animals in the US National Toxicology Program provided by Portier et al. (1986). This approach gives a weighted average of the probability of cancer occurrence over an entire lifetime, weighted by survival probability.

Table 51. Excess lifetime risk for humans due to exposure to diesel exhaust emissions under various exposure scenarios

Exposure pattern Biologically based model Linearized multistage Maximum Upper 95% model a
likelihood bound
estimate estimate

General population (normal 1.41 × 10 -5 2.44 × 10 -5 respiratory function; nonsmoker): 2.6 μg/m 3

, 16 h/day, seven days per week

General population 2.32 × 10 -5 3.61 × 10 -5 5.38 × 10 -5
(20-pack-year smoker) b : 2.6 μg/m 3 , 16 h/day, seven days per week

Occupationally exposed: 3.12 × 10 -5 5.17 × 10 -5 6.18 × 10 -5
15 μg/m 3 8 h/day, five days per week

a Calculated using carbon core as dosimetric; only malignant

tumours are used.

b In this calculation, smoking affects only lung clearance rate; biological interaction between smoking and exposure to diesel
exhaust is not considered. The retention half-times for insoluble particles increased from 296 days for persons with normal respiratory function to 519 days for 20-pack-year smokers (Bohning et al., 1982).

Some implications of the alternative model are:

(1) At a low exposure concentration, the decrease in diesel-induced initiation results in a greater reduction of risk; that is, the number of initiated cells (cancer risk) is reduced more efficiently with a low than with a high exposure. If only organic compounds induce initiation when the concentration is low, they play a more important role than particles in inducing tumours at low concentrations, whereas the roles are reversed when the concentration is high.

(2) Although cells initiated by diesel exhaust play an important role in cancer induction, either organic compounds or the carbon core alone could induce initiation by increasing their respective proportions. Thus, although initiated cells are important for tumour induction, they may be induced by any agent that initiates tumours (e.g. smoking).

(3) A small change in the rate of proliferation induced by diesel exhaust could disproportionately change cancer risk. As this parameter is assumed to be a function of the dose of carbon core, lung overloading has a significant effect on cancer incidence. In the absence of better information, it is assumed in this assessment that the carbon core continues to have a proliferating

effect at low doses.

These observations suggest that, although the effect of particle overload on cell proliferation is important, initiation by the carbon 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmcore or organic compounds or both is also essential. Although this conclusion is only tentative, because the model parameters are estimated statistically on the basis of bioassays conducted at high concentrations, it does suggest the importance of studying the role of the carbon core and organic compounds in initiation and promotion at low and high exposure concentrations. Does the relative initiation potential of organic compounds and the carbon core differ with concentration? These observations also suggest that a subcohort of workers who were smokers and were exposed to high concentrations of diesel exhaust for a long time would have a greater risk of dying from lung cancer.

(h) Comparison of risk estimates derived from experimental studies and human experience

The bioassay-based risk estimates, which range from 1.6 × 10 -5 to

7.1 × 10 -5 , with a geometric mean of 3.4 x 10-5, can be compared with human experience on the basis of three data sets: those of an epidemiological study conducted on London Transport employees (Waller, 1981) and a subsequent analysis (Harris, 1983) and those on American railroad workers (Garshick et al., 1987, 1988). Although these data cannot be used to calculate unit risk, mainly because of a lack of reliable information on exposure, they can be used to evaluate the validity of unit risk estimates based on the results of experimental
studies.

Attempts have already been made to estimate the potential cancer risk due to exposure to diesel exhaust on the basis of epidemiological data. From the results of the study of London Transport workers, Harris (1983) estimated that the increase in the relative risk for lung cancer associated with exposure to 1 μg/m 3 -year of diesel exhaust

was 1.2 × 10 -4 , with an upper bound of the 95% confidence limit of

4.8 × 10 -4 . (Generally, when data from an epidemiological study with

negative results are used to estimate cancer risk, the upper bound is used to calculate unit risk.) The resulting unit risk estimate is 2 ×

10 -3 , which is about 60 times higher than the mean unit risk estimate derived from bioassays, 3.4 × 10 -5 , and about 30 times higher than the upper end of the range of unit risk estimates from bioassays, 7.1 ×

10 -5 . Therefore, the risk estimate based on bioassays is not inconsistent with that for humans, since 2 × 10 -3 is the estimated upper bound in an epidemiological study with negative results.

McClellan et al. (1989) reported risk estimates based on the study of Garshick et al. (1987), in which lung cancers in railroad workers were evaluated. Assuming exposure to concentrations of 500 and 125 μg/m 3 , the upper bounds of the 95% confidence interval for

lifetime cancer risk were estimated to be 6 × 10 -4 and 2 × 10 -3

, respectively. The lower of the two unit risk estimates is only about one order of magnitude higher than the risk estimates based on bioassays.

An epidemiological study potentially more suitable for quantitative risk assessment was reported by Garshick et al. (1988), which was based on a large number of subjects; a small but significant increase in the rate of mortality from lung cancer was seen in some subcohorts. The US Environmental Protection Agency has supported an effort to derive a unit risk estimate from this study, and data on exposure were estimated by Woskie et al. (1988a,b) and Hammond et al. (1988). These data were analysed in a variety of ways, using relative risk and absolute risk dose-response models and by classifying individuals into various exposure categories, including job, duration of employment, age, and exposure markers. Even though at least 50 analyses were carried out, an adequate dose-response relationship
could not be obtained; these data were therefore not used to estimate
unit risk.

The lack of a dose-response relationship is not totally unexpected, given the low increase in mortality rate in the study of Garshick et al. (1988) and the uncertain estimates of exposure. The data on exposure were derived from air samples collected during a limited period (1981-83) on four small railroads operating in a limited geographical area (northern United States), in which concentrations of respirable particulate matter were measured rather than diesel exhaust per se; the measured concentrations were then adjusted to derive markers of exposure. The data were used to estimate exposure to diesel exhaust of railroad workers throughout the United States 30 or more years earlier. Diesel equipment and working conditions have, however, changed since the 1940s when diesel engines first began to be used in large numbers. Woskie et al. (1988b) summarized anecdotal reports of smoky working conditions in diesel repair shops during the 1950s and 1960s and reported that the limited data available on levels of nitrogen oxide during those periods

indicated high levels of diesel exhaust. By the time samples were collected, however, the smoky conditions would have been largely mitigated by improved ventilation and the advent of less smoky diesel engines. The study of Garshick et al. (1988) gives the relative risks for dying from lung cancer in exposed as opposed to unexposed railroad workers classified into five subcohorts by age in 1959. The risks ranged from 0.96 (95% CI, 0.74-1.33) to 1.45 (1.11-1.89). The highest relative risk, 1.45, which was observed in workers who were 40-45 years old in 1959, was used here to evaluate the validity of unit risk based on biossays results. Assuming that this subcohort of workers was exposed to diesel exhaust for 8 h/day on five days per week from age 35 to age 65, the background mortality rate from lung cancer in this subcohort would be 0.038 (0.63 × 0.06), as the unexposed workers in the same age group had a relative risk of 0.63 and the corresponding lifetime mortality rate from lung cancer in the general American white 

# Pa g e 126 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmmale population is about 0.06. If the lung cancer risk due to 1 μg/m 3 is assumed to be 3.4 × 10 -5 , the concentration of diesel exhaust in the working environment was at least 400 μg/m 3 , calculated as follows: The risk due to 1 μg/cm 2 of particles is 0.017 (which

results in a unit risk of 3.4 × 10 -5 ). For a lower bound of the relative risk of 1.1, a lung burden, d, that satisfies the relationship 0.1 × 0.038 = 0.017 d would be needed; that is, d = 0.22 μg/cm 2 , which is equivalent to an air concentration of 0.4 mg/m 3 by the dosimetry model of Yu et al. (1991). If the highest unit risk estimate derived from Brightwell et al. (1986), 7.1 ×

10 -5 , is used, the required minimal air concentration would be about 0.2 mg/m 3 . These calculations imply that a diesel exhaust concentration of at least 0.2 mg/m 3 was necessary to observe a statistically significant increase in the mortality rate from lung cancer in this study. This concentration appears to be reasonable in the light of the working conditions described by Woskie et al. (1988a).

Although the risk estimates derived from bioassays are lower than those derived from human data and may possibly over-predict risk, since the non-linearity of the human response is not considered in this model, they are not inconsistent, for the following reasons:

(1) The risk estimates based on epidemiological studies are not derived from all of the available data but on only a subset with the highest response. The estimates are therefore higher than
those that would be derived from the whole data set.

(2) When a single data point (i.e. an overall relative risk and an averaged exposure concentration) is used in the calculations, the resulting slope for potency will be overestimated if the dose-response relationship is not linear over all exposure concentrations. Assume, for example, that the response follows the simple multistage model P(d) = 1 - exp[-( q 0 + q d + q 2 d 2 )].

The relative risk, R, at concentration d is R(d) = P(d)/P 0 .

Using this mathematical expression, it is easy to demonstrate that the slope factor calculated from [ P(d) - 1] P 0 / d is
greater at high doses (including the averaged concentration used in the risk calculation) than at low doses where the dose-response function is dominated by q 1 . Epidemiologists
have long had a similar (but not identical) concern about the use of averaged data, since ecological associations are not necessarily consistent with those measured at the individual level (see Cohen, 1994; Greenland & Robins, 1994; Piantadosi, 1994).

(3) Some occupational groups were exposed to considerably higher concentrations of diesel exhaust in the past than presently. For example, the average particle concentration in a Finnish roundhouse was reported to be 2 mg/m 3 (Heino et al., 1978). The lung burdens would thus be greater than those predicted on the basis of current exposures, and the unit risk estimates would be higher

(i) Summary and conclusions

A dosimetric model that accounts for differences between experimental animals and humans in lung deposition efficiency, lung particle clearance rates, lung surface area, ventilation, and the rates of elution of organic chemicals from the particle surface was used to calculate equivalent human doses as particle concentration per unit lung surface area. After dosimetric adjustment, four risk estimates were derived by a linearized multistage model from three bioassays with Fischer 344 rats and one with female Wistar rats, which ranged from 1.6 to 7.1 × 10 -5 /μg particles per m 3 , with a geometric

mean of 3.4 × 10 -5 . This quantitative assessment of the carcinogenic risk due to exposure to diesel engine emissions is reasonable,

because:

-- The estimates are based on several well-designed, well-executed, long-term bioassays.

-- Epidemiological studies indicate that humans are susceptible to the induction of lung cancer after inhalation of diesel exhaust.

-- Dosimetry modelling, especially to account for inhibition of particle clearance at high doses, has allowed accurate extrapolation of doses from animals to humans.

-- The doses are based on actual concentrations of particulate matter per unit of lung surface area.

-- Use of an alternative model that attempts to account for the possible biological effects of the organic and carbon core fractions did not appreciably change the unit risk estimate.

-- The risk estimates based on the results of bioassyas are not inconsistent with the available human experience.

Nevertheless, a number of uncertainties remain, the most significant of which are:

-- In any interspecies extrapolation there may be an inherent difference in sensitivity to the agent being assessed.

-- The assumption of equivalent sensitivity across species is based on concentration per unit of lung surface area, and use of other 

# Pa g e 127 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmassumptions of dose equivalence may lead to different estimates of risk; however, the estimates should not vary by more than one order of magnitude.

-- Although linearized low-dose extrapolation methods are used, it is still uncertain whether inflammatory cells secrete mediators that induce cancer in lung epithelial cells when the particle burden is smaller than that necessary to inhibit clearance. Even if macrophages are activated at low particle burdens, it is uncertain whether the responses of epithelial cells are linear at very low concentrations; however, non-linear carcinogenic responses have been observed in bioassays.

-- Although the unit risk estimate is corroborated in the alternative model, uncertainty about the response at low doses
remains because the estimates in the alternative model are based on the assumption that particles continue to induce cell initiation and/or proliferation at low doses. The actual risk would be much lower if this assumption does not hold.

The risk at low doses derived from the linearized multistage model may thus be overestimated if particles no longer induce cell initiation and/or proliferation. The model was selected for calculating risk because a conservative model was needed in order to ensure protection of public health and because adequate data were not available to use fully the alternative model constructed for this assessment. Thus, the risk derived from the bioassays should be viewed as hypothetical.

These unit risk estimates should not be used to evaluate the carcinogenic risk of other types of particulate matter present in ambient air, which may have different solubilities, surface areas, and free radical contents, which factors greatly affect carcinogenic
potency.

It could be argued that since the types and location of tumours seen in rats after exposure to particles are different from those found in humans, the experimental data are not relevant for humans. The tumours diagnosed in rats after long-term exposure to carbon black particles and diesel exhaust include benign adenomas and malignant adenocarcinomas, squamous-cell carcinomas, adenosquamous carcinomas, and squamous cysts (Mauderly et al., 1994; see section B7.3.2). After injection into athymic mice, cells from 50-67% of squamous-cell carcinomas and 25-40% of adenocarcinomas were found to grow (Table 35), whereas those from squamous cysts did not (Mauderly et al., 1994).

Squamous cysts have been defined by other authors as benign cystic keratinizing squamous-cell tumours (Kittel et al., 1993; Dungworth et al., 1994; Heinrich et al., 1995) and found not to grow after transplantation (Heinrich et al., 1995). As pointed out by Mohr (1992), there is, however, controversy about the correct terminology of this type of lesion. In 1995, an international group of pathologists re-evaluated lung tissue sections from rats exposed to particles by inhalation and identified four distinct lesions: squamous-cell metaplasia, pulmonary keratinizing cyst, cystic keratinizing epithelioma (considered to be benign), and squamous-cell carcinoma (G. Oberdörster, personal communication).

Regardless of the correct classification of lesions, the important fact is that malignant tumours are induced in rats after long-term inhalation of deisel exhaust and carbon black. The tumour response of human beings may not, however, be the same, either qualitatively or quantitatively. In fact, the response of rats to long-term exposure to high concentrations of particles differs from that of other species, including mice and hamsters. In the absence of sufficient information about specific mechanisms unique to rats, however, there is no justification for excluding data on this species from extrapolations to the human situation (US Environmental Protection Agency, 1986).

With respect to the mechanisms of induction of lung tumours by particles in rats, it has been shown that particles devoid of polyaromatic compounds can increase mutation rates in pulmonary epithelial cells, which is an important step in tumour development via cell transformation. In a recent study in which rats were exposed to carbon black for 13 weeks, a significant influx of inflammatory cells was seen at concentrations of 7 and 50 mg/m 3 but not at 1 mg/m 3 (Oberdörster et al., 1995), and a significant, three- to fourfold increase in the frequency of hprt mutations was seen in alveolar epithelial cells (Driscoll et al., 1995 and in press; see section B7.6). Thus, burdens of particles of low toxicity per se, similar to those reached in long-term studies in rats, can be mutagenic, possibly through the involvement of DNA damaging oxidants derived from inflammatory cells.

Appendix B10.1 Construction of a biologically based (alternative)

model

1. Preliminary considerations

In order to evaluate the effects of various biological assumptions on the assessments of the risk of exposure to diesel exhaust, a mathematical dose-response model must be constructed that takes into account the proposed biological mechanisms. As a significant issue in assessing the risk of diesel exhaust is the effect of lung overloading on tumour induction, the model should have the following properties:

-- It should depend on dose metrics for both organic compounds and 

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-- It should allow for changes in the model parameters with time due to increasing lung burden during exposure.

-- It should view cell proliferation and tumour induction and formation stochastically: it is not realistic to assume deterministic clonal growth. For instance, it should not be assumed that all cells divide at the same age.

It is therefore assumed that a normal cell can be initiated by both organic compounds and the carbon core. The initiation rate is denoted by μ 1 , which is a function of the background rate and that
induced by diesel exhaust (as specified below). Because an initiated cell eventually either dies or enters the cell cycle (including cells in quiescence, G 0 ), it is reasonable to assume that the lifetime of
an initiated cell follows a certain probability distribution. In this model, a cell in G 0 phase is equivalent to one with a certain
probability of a very long lifetime (i.e. in the right-hand tail of the distribution of the cell's lifetime). At the end of its lifetime, it either dies (death) with probability b, divides into two daughter cells (birth) with probability a, or divides into one initiated cell and one malignant cell (second transition) with probability μ 2 ; alpha + ß + μ 2 = 1. Instead of assuming that a single malignant cell
is equivalent to a tumour, as in the model proposed by Moolgavkar & Venzon (1979) and Moolgavkar & Knudson (1981), it is assumed here that a malignant cell has a certain probability of becoming a tumour; this probability is assumed to be dose-dependent, thus allowing for an evaluation of the effect of dose on tumour progression. It should be noted that the proposed model does not exclude the possibility that there may be more than one step in the 'initiation' of a normal cell.
The rate of initiation used in the model should be viewed as a net rate that represents several genetic alterations and repairs.

2. Mathematical model and relationship of parameters to lung burden

A dose-response function P(t:d,D) is constructed for the probability of cancer by time (age) t, which depends on both organic compounds, d, and particles (carbon core), D, and incorporates the biologically based concept discussed above. Because the model parameters that are not observed directly in the laboratory can be estimated statistically only from the results of bioassays at high concentrations, the model should not be considered a real model of diesel-induced carcinogenesis; uncertainty about extrapolation to low
doses remains.

A model with the features presented above was originally proposed by Chen & Farland (1991) and was extended into one with time-variable parameters by Tan & Chen (1992). This model was used as the basis for constructing a biologically based dose-response model. A brief mathematical description is presented in Appendix B10.3.

The data on time to event from Mauderly et al. (1987) were used to estimate model parameters. These data are useful because they contain information on natural mortality and serial sacrifice of animals with and without (malignant) tumours, which is valuable for estimating tumour latency. In order to use this information to calculate maximum likelihood estimates of parameters, an 'E-M' algorithm was derived. In the E-M algorithm, each iteration involves an 'expectation' step (E) and a 'maximization' step (M) (see Appendix B10.2).

(a) Model parameters and notations

The following parameters are incorporated into the dose-response model, which includes the rate of initiation (μ 1 ), the rate of
proliferation (gamma alpha ), the rate of conversion (gammaμ 2 ), and
the probability of progression ( q ). The rate of death of the initiated cells is implicitly defined by gamma(1-μ 2 -alpha). The

parameters are all dose dependent.

D: dose of carbon core in milligrams per square centimetre of lung epithelial surface; varies over time;

d: dose of organic compounds in milligrams per square centimetre of lung epithelial surface;

μ1: dose-related initiation rate (per cell per day); depends on

μ 0 (background rate), d, and D by μ 1 = μ 0 (1 + ad + bD ), where

a and b are parameters to be estimated statistically;

μ2: probability that a malignant cell will be produced by the end
of the lifetime of an initiated cell;

alpha: probability that an initiated cell will divide into two daughter cells by the end of its lifetime;

q: probability that a single malignant cell will develop into a malignant tumour;

gamma: 1/gamma is the mean lifetime of an initiated cell in days; the lifetime ends when the cell goes into mitosis or dies. If it is assumed that the probability that a cell will go into mitosis is about the same as the probability that it will die, the mean cell lifetime can be conveniently interpreted as time to mitosis (i.e. cell turnover time); thus, a shorter cell 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmlifetime implies more frequent cell division. Time to mitosis is a random variable, not a fixed constant as in the assumption of the model of Greenfield et al. (1984), which was used by Cohen & Ellwein (1988) to analyse the results of bioassays to dectect urinary bladder cancer.

N: number of (normal) target cells.

(b) Practical considerations

The E-M algorithm developed in Appendix B10.2 is an elegant procedure that can be used by statistical theory alone to test hypotheses about whether a particular parameter is influenced by organic compounds and the carbon core, individually or together. For instance, it could be postulated that the parameter the reciprocal of which represents mean cell lifetime is given by gamma( d,D i ) = gamma 0 +

gamma 11 d + gamma 12 D i , and then proceed to test the null hypothesis

that gamma 11 = 0, i.e. no effect of organic compounds on the cell
lifetime. This temptation must, however, be resisted, because too many parameters would have to be be estimated. Therefore, the biologically plausible assumption that parameters q and gamma depend only on the lung burden of the carbon core, D, is used.

The study of Mauderly et al. (1987) lasted about 940 days. In order to construct a dose-response model including time-dependent lung burden, the time interval (0-940) is divided into five sub-intervals, each spanning six months, except for the last, which spans 730-940 days. The deposition-retention model developed by Yu et al. (1991), corresponding to a concentration of diesel exhaust emissions in ambient air of milligrams per cubic metre, is used to calculate ( d,D i ), where i = 1, 2, ..., 5; the dose of organic compounds,

d, does not change with time because it reaches a steady state soon after exposure begins; and D i is the lung burden of carbon core
during the i th sub-interval.

The assumptions for the dose-parameters relationship are:

-- The rate of initiation associated with a lung burden { d,D i , i = 1, 2, ..., 5} is given by μ i ( d,D i ) = μ 0 (1 + a * d + b * D i ) for

i = 1, 2, ..., 5. This is the only parameter that is assumed to depend on both d and D.

-- The probability of tumour formation from a malignant cell is assumed to be dependent on lung burden, D, from q ( D i ) = q 0

q 0 D i , i = 1, 2, ..., 5. In order to simplify calculation, the
possibility that q 0 is also dependent on organic compounds, d, is

not considered.

-- The cell lifetime, gamma, is assumed to be related nonlinearly to lung burden, D, from gamma( D i ) = gamma 0 + gamma 1 Log(1 + D i ),

i = 1, 2, ..., 5.

In order to reduce the number of parameters that are to be estimated from the data of Mauderly et al. (1987), some of the background parameters for the dose-response model (μ 0 , q 0 , and

gamma 0 ) are estimated from the historical control rates for Fischer
344 rats in the US National Toxicology Program (reconstructed from Portier et al., 1986). The dose-related parameters are then estimated with the E-M algorithm, which is described in Appendix B10.2. The parameters estimated for the model resulting from the tumour response data of Mauderly et al. (1987) and the corresponding dosimetric parameters (Table 52) are given in Table 53.

Table 52. Dosimetric parameters (milligrams per square centimetre of lung surface) used in modelling

Exposure d D 1 D 2 D 3 D 4 D 5

concn (mg/m 3 )

0.35 2.5 × 10 -6 6.2 × 10 -5 8.8 × 10 -5 9.0 × 10 -5 9.0 × 10 -5 9.0 × 10 -5

3.50 3.6 × 10 -5 7.5 × 10 -4 2.4 × 10 -3 3.9 × 10 -3 5.3 × 10 -3 6.3 × 10 -3

7.08 7.3 × 10 -5 2.0 × 10 -3 5.5 × 10 -3 8.6 × 10 -3 1.1 × 10 -2 1.4 × 10 -2

d, organic compounds; D i , i = 1, 2, ..., 5, are average lung burdens of carbon core over

five time intervals. Values calculated from the retention model of Yu et al. (1991)

Table 53. Maximum likelihood estimates for model parameters

Parameter Estimate

μ 1.033 × 10 -7

a 1.103 × 10 4

b 3.214 × 10 2

μ 2 7.907 × 10 -7

q 0 1.035 × 10 -1

q 1 5.332 × 10 -2

gamma 0 1.662 × 10 -2 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmgamma 1 2.647 × 10 -2

alpha 5.443 × 10 -1
N (given) 8.80 × 10 7

For definitions of parameters, see text. Background parameters μ 0 , q 0 , and gamma 0 are estimated separately from historical control data from
the US National Toxicology Program. The number of target cells, N, is assumed to be 10 times the number of type II cells in mice, which is given by Kauffman (1974). It is not essential that N be given accurately because Nμ 0 appears as a single term in the model; the
estimated μ 0 will compensate for the underestimation or overestimation

of N.

Appendix B10.2 E-M algorithm

The E-M algorithm, derived below, was used to calculate maximum likelihood estimates of the parameters for the alternative model. The data used were taken from Mauderly et al. (1987) and include the time when an animal died, naturally or at sacrifice, with or without (malignant) tumours. The theory of the algorithm is given by Dempster et al. (1977).

Assume that the distinct times at which animals died are t 1

< t 2 < ...< t m . The observations can be classified as follows:

a 1x (i): observed number of natural deaths without tumour at time

t i in treatment group x (The four groups are x = 1, 2, 3, 4.)

a 2x (i): observed number of natural deaths with tumour at time

t i in treatment group x

b 1x (i): series sacrificed at time t i without tumour in

treatment group x

b 2x (i): series sacrificed at time t i with tumours in

treatment group x.

Let T d represent the time an animal died and T the time a

tumour developed.

alpha

x (i) = Pr T d = t i | T d > t i , T > t i ,, x } (conditional probability

of death without tumour)

ß

x ( i | u ) = Pr T d = t i | T d > t i , T Epsolin ( t u-1 , t u ], x } (related

to deaths with tumours)

Define

and

S

x

(t) = Pr T > t | x} = exp[- t0 integral t h(x)dx ].

The function S
x (t) is the probability of being tumour-free by

time t.

The exact forms of the hazard function, h(x), and S
x (t) are
given in Appendix B10.3.

Let

a 2x ( i | u ) = number of natural deaths at t i , with a tumour
developing during ( t u-1 , t u ], in treatment group

x, u < i,

b 2x ( i | u ) = number of animals sacrificed at t i , with a tumour
developing during ( t u-1 , t u ], in treatment group

x, u < i.

Then

Let

and 

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Given a 2x ( x ), { a 2x ( i | u ), u = 1, ..., i } is an ( i -1)-dimension multinomial with parameter { a 2x ( i ), P
x ( i | u ), u = 1, ..., i }.

Thus,

E [ a 2x ( i | u ) | a 2x ( i )] = a 2x ( i ) P
x ( i | u ).

Similarly, { b 2x ( i | u ), u = 1, ..., i }, is an ( i - 1)-dimension multinomial with parameters { b 2x ( i ), Q
x ( i | u ), u = 1, ..., i }, and

E [ b 2x ( i | u ) | b 2x ( i )] = b 2x ( i ) Q

x ( i | u ).

It can be shown that the likelihood function is proportional to

where

Let

Let

be a vector of parameters in function S;

alpha

x = [alpha

x (1), alpha

x (2), ..., alpha

x ( m )], and

ß

x ( u ) = [ß

x (1 | u ), ß

x (2 | u, ..., ß

x ( m | u )]

be vectors of parameters related to conditional probabilities of death with and without tumours. These parameters and those in theta

x were estimated by the E-M algorithm described below.

The M step:

Given initial values a 2x ( i | u ) and b 2x ( i | u ), estimate

The E step:

Given the estimated values of alpha

x ( i ), ß

x ( i ), and theta

x from the M

step, compute P
x ( i | u ) and Q

x ( i | u ) and obtain estimates of

a 2x ( i | u ) and b 2x ( i | u ) by

With the estimated values of a 2x ( i | u ) and b 2x ( i | u ) available
from the E step, go back to the M step and repeat the process until
the estimates are stable.

Appendix B10.3 A tumour growth model

A tumour growth model with piece-wise constant parameters taken from Tan & Chen (1992) is an extension of a stochastic model developed by Chen & Farland (1991). The biological justification of this model is similar to that of the two-stage model proposed by Greenfield et al. (1984), which was used by Cohen & Ellwein (1988) to analyse urinary bladder tumour occurrence. The two models differ, however, with respect to their mathematical formulations; the one adopted in this report is a stochastic model, whereas the other is a deterministic model and does not allow for estimation of parameters because it does not have complete mathematical expression.

Although its most general form is not used here because of lack of data, the stochastic model of Chen & Farland (1991) has two 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmdesirable features: (i) it allows for any cell growth distribution (e.g. Gompertz), rather than only exponential distribution as in other models; and (ii) it incorporates the birth and death of tumour cells, rather than assuming that a tumour is born once a single tumour cell occurs, as did Moolgavkar & Venzon (1979) and Moolgavkar & Knudson (1981). Therefore, if information on cell lifetime distribution and the progression of tumour development is available, a reasonably
realistic model can be constructed.

For completeness, a brief description of the model is presented here. The following notations are required:

N(t): number of normal (target) cells at time t,

μ 1: rate of initiation, and
contour the probability density function for the lifetime of an integral (t): an initiated cell.

At the end of its lifetime, an initiated cell either divides (mitosis) or dies (programmed or nonprogrammed death). If it enters mitosis, it either divides either into two initiated cells with probability a or into one initiated cell and one malignant cell with probability μ 2 . At the end of a cell's lifetime, the probability of
it dying is ß = 1 - alpha - μ 2 . A similar set-up (to allow for any
cell lifetime distribution) can be made for a malignant cell; however, we confined ourselves to a simpler version, assuming that the lifetime of a malignant cell follows an exponential distribution. Thus, we assume that a malignant cell follows a simple birth-death process; it can either divide into two malignant cells at a rate alpha

m or die

at a rate ß

m .

When the parameters are constant over time (age), the hazard function is given by

h(t) = μ 1 μ 2 0 integral t N(s)m ( t - s ) ds

where

where y 1 < y 2 are two real roots of alpha y 2 - (alpha + ß + μ 2 q ) y + ß = 0; alpha + ß + μ 2

= 1; q = 1 - ß

m /alpha

m ; A(t) = / 0 integral t a(x) d x, where a(t) =
contour integral( t )/[1 - F(t) ] is the hazard function of the cell lifetime and F(t) is the cumulative function of contour integral (t).

Two cases of special interest are a(t) = gamma when an exponential
distribution is assumed and a(t) = exp(-gamma t ), when the Gompertz
distribution is assumed.

When an exponential distribution (i.e. a(t) = gamma or A(t) =

gamma t ) and q = 1 are assumed, the model is equivalent to the model of Moolgavkar & Venzon (1979) and Moolgavkar & Knudson (1981). A special case that may be more appropriate than the exponential distribution is that when the Gompertz distribution is assumed (i.e. A(t) = [1 - exp(-gamma t )]/gamma).

For the model with time-dependent parameters, assume that the study begins at time t 0 . Divide the time scale ( t 0 , t ] into

k sub-intervals L j = ( t j-1 , t j ], j = 1, 2, ... k- 1 and L k = ( t k-1 , t k ]

where t k = t . (Note that these sub-intervals may not be the same as
those defined by deaths or sacrifice previously.) The parameters that vary over sub-intervals ( t i-1 , t i ], i = 1, 2, ..., k are μ 1j , alpha j , ß j , μ 2j , N, and those parameters related to contour integral( t ). The
hazard functionis given by

where

and

where y 1j < y 2j are two real roots of a alpha y 2 - (alpha j + ß j +

μ 2j q j ) y + ß j = 0; alpha j + ß j + μ 2j = 1; q j = 1 - ß mj /alpha mj , j = 1,

2, ..., k.

When an exponential distribution (i.e. A j ( t ) = gamma j ( t ) and

q j = 1) is assumed, the model is equivalent to the model of
Moolgavkar & Venzon (1979) and Moolgavkar & Knudson (1981), with piece-wise constant parameters. A special case that may be more appropriate than the exponential distribution is that when the 

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/gamma j ).

In the alternative model for exposure to diesel exhaust, in which the total time is divided into five (i.e. k = 5) sub-intervals, Tan & Chen (1992) showed that, under the assumption of exponential cell lifetime distribution, the tumour-free distribution function, S
x ( t ),

can be written:

where s j = t j if j < k and s j = t if j = k , and

where,

W I = [alpha + ß + μ 2 q ) 2 - 4alphaß]  ́

Z I = alpha - ß - μ 2 q , and

Delta I ( s,t ) = gamma i ( t - s ) if both s and t are in the same closed

sub-interval [ t i-1 , t i ] and

if s epsilon L i , t epsilon L j with t j < t j .

B11. RECOMMENDATIONS

B11.1 Recommendations for the protection of human health

Diesel exhaust contributes to the total effect of combustion products on the environment. The data reviewed in this monograph support the conclusion that inhalation of diesel exhaust is of concern with respect to both neoplastic and non-neoplastic diseases. The particulate phase appears to have the greatest effect on health, and both the particle core and the associated organic materials have biological activity, although the gas-phase components cannot be disregarded. The following actions are recommended for the protection

of human health.

-- Diesel exhaust emissions should be controlled as part of the overall control of atmospheric pollution, particularly in urban
environments.

-- Emissions should be controlled strictly by regulatory inspections and prompt remedial action.

-- Urgent efforts should be made to reduce emissions, specifically of particulates, by changing exhaust train techniques, engine design, and fuel composition.

-- In the occupational environment, good work practices should be encouraged, and adequate ventilation must be provided to prevent excessive exposure.

B11.2 Recommendations for the protection of the environment

Too few data are available on which to base specific suggestions with regard to diesel exhaust, except as part of the general control
of emissions.

B11.3 Recommendations for further research

The following recommendations are intended to help reduce the uncertainty associated with assessing the risks of exposure to diesel

exhaust for human health.

-- Research is required on methods for determining concentrations of diesel particulates in the presence of fine particles from other sources, in order to improve assessments of the exposures of occupational groups and the general population indoors and outdoors. The quality and quantity of emissions from engines that are not properly maintained or tuned should be investigated as part of these studies.

-- The effects of diesel exhaust emissions on lung clearance should be studied in animals and humans at concentrations including those likely to be encountered by humans.

-- The mechanisms involved in the etiology of tumours induced in rats exposed to particulate must be investigated by all available techniques.

-- The relative roles of the gas and particulate phases of diesel 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmexhaust in causing adverse effects after short- and long-term exposure should be investigated.

-- Epidemiological investigations into the effects of diesel exhaust should be continued in order to assess issues of dose and latency in human populations with respect to lung cancer; and longer-term studies should be conducted in populations exposed to diesel exhaust with respect to noncarcinogenic effects.

-- Further steps should be undertaken to improve the combustion and exhaust emission characteristics of diesel engines.

B12. PREVIOUS EVALUATIONS BY INTERNATIONAL BODIES

The carcinogenic risks for human beings were evaluated by a working group convened by the International Agency for Research on Cancer in 1988 (International Agency for Research on Cancer, 1989b).
The conclusions were:

'There is sufficient evidence for the carcinogenicity in experimental animals of whole diesel engine exhaust.

'There is inadequate evidence for the carcinogenicity in experimental animals of gas-phase diesel engine exhaust (with particles removed).

'There is sufficient evidence for the carcinogenicity in experimental animals of extracts of diesel engine exhaust particles.

'There is limited evidence for the carcinogenicity in humans of diesel engine exhaust.

'There is limited evidencefor the carcinogenicity in humans of engine exhausts (unspecified as from diesel or gasoline engines).

'Overall evaluation

'Diesel engine exhaust is probably carcinogenic to humans (Group2A).'

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PARTIE A COMBUSTIBLE DIESEL

A1. RESUME

A1.1 Identité, propriétés physiques et chimiques et méthodes d'analyse 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmLe combustible diesel est un mélange complexe d'alcanes normaux ramifiés ou cycliques (de 60 à plus de 90% de volume; longueur de la chaîne hydrocarbonée, généralement comprise entre C 9 et C 30 ), de composés aromatiques, surtout des alkyl benzènes, (5 à 40% en volume) et de petites quantités d'alcènes (0 à 10% en volume) obtenu lors de la distillation fractionnée du pétrole, à partir du distillat moyen qui correspond à la fraction gazole. Le combustible diesel peut contenir, à des concentrations de quelques parties par million, du benzène, du toluène, de l'éthylbenzène, du xylène et des hydrocarbures aromatiques polycycliques (HAP), en particulier du naphtalène et certains de ses dérivés méthylés. La teneur en soufre du combustible diesel dépend de l'origine du pétrole et du procédé de raffinage utilisé. Dans un certain nombre de pays, elle est soumise à réglementation et se situe en général entre 0,05 et 0,5% en poids. On utilise un certain nombre d'additifs pour modifier la viscosité, la conservation et la combustion, pour différencier les différents produits et également pour satisfaire aux spécifications commerciales. A la température ambiante, le combustible diesel est en général modérément volatile, légèrement visqueux, inflammable et se présente sous la forme d'un liquide brun à odeur de kérosène. L'intervalle d'ébullition est habituellement de 140°C à 385°C (plus de 588°C pour le carburant destiné aux moteurs marins); à 20°C, la densité est de 0,87-1,0 g/cm 3 et la solubilité dans l'eau de 0,2-5 mg/litre. La qualité et la composition du combustible diesel influent considérablement sur l'émission de polluants par les moteurs diesel. Les conditions d'allumage (exprimées au moyen de l'indice de cétane), la densité, la viscosité et la teneur en soufre sont des variables importantes. Les spécifications du combustible diesel commercial varient fortement d'un pays à l'autre.

Les différents types de mazout destinés au chauffage ou de kérosène pour avions à réaction produits lors du raffinage du pétrole peuvent avoir une composition analogue à celle du combustible diesel, mais avec des additifs différents. Les données biologiques relatives à ces mélanges ont donc été prises en considération lors de l'évaluation toxicologique et écotoxicologique.

En raison de la complexité du mélange, il n'existe pas de méthode d'analyse spécifique pour le combustible diesel et les techniques utilisées pour la plupart des études d'impact dur l'environnement ne se prêtent qu'au dosage des hydrocarbures totaux. Elles consistent tout d'abord à effectuer une extraction par solvant, puis à éliminer les hydrocarbures d'origine naturelle et enfin à procéder au dosage par gravimétrie, spectrophotométrie infrarouge ou chromatographie en phase gazeuse. Ni la méthode gravimétrique, ni la spectrophotométrie infrarouge ne donnent de renseignements qualitatifs ou quantitatifs utiles et ne peuvent donc être utilisées que pour un premier tri. La chromatographie en phase gazeuse couplée à des techniques de détection telles que l'ionisation de flamme ou la spectrométrie de masse, est la méthode classique d'analyse des échantillons prélevés dans l'environnement. Il existe de nombreuses autres méthodes pour la recherche et le dosage des divers hydrocarbures présents dans le
combustible diesel.

A1.2 Sources d'exposition humaine et environnementale

Le combustible diesel est produit par raffinage du pétrole brut. Afin de satisfaire aux spécifications techniques relatives au rendement des moteurs, ces combustibles sont généralement mélangés; l'adjonction ultérieure d'additifs en améliore également l'adaptation à diverses utilisations particulières. Ces combustibles sont largement utilisés dans les transports. Les plus volatils d'entre eux, qui présentent une faible viscosité, sont destinés aux moteurs tournant à haut régime, les combustibles plus lourds étant réservés aux transports ferroviaires et maritimes. Une grande partie des véhicules lourds de transport routier sont mus par des moteurs diesel. Les cars à moteur diesel destinés au transport des voyageurs se répandent de plus en plus en Europe et au Japon (10 à 25%) alors qu'en Amérique du Nord, la proportion de cars assurant les transports en commun est d'environ 1 à 2%, et a même une légère tendance à se tasser. On utilise également le combustible diesel pour alimenter les chaudières et les moteurs fixes, qu'il s'agisse de moteurs à pistons, de turbines à gaz, de pompes d'oléoducs, de compresseurs, de générateurs de vapeur pour centrales thermiques, de brûleurs, et d'installations de chauffage industriel par convection ou circulation d'eau.

La demande mondiale de combustible diesel a régulièrement progressé au cours des 5 dernières années. En 1985, les chiffres de consommation étaient les suivants: environ 170 000 kilotonnes par an en Amérique du Nord, environ 160 000 kilotonnes par an, gazole y compris, dans l'Union européenne, environ 46 000 kilotonnes par an en Australie, au Japon et en Nouvelle-Zélande, soit l'équivalent de 1062 kilotonnes par jour. En 1990, la demande mondiale a été estimée à environ 1110 kilotonnes par jour.

On ne dispose d'aucun renseignement sur les émissions qui se produisent lors de la production des combustibles diesel; toutefois, il semble que cette source soit d'importance secondaire, étant donné que le raffinage s'effectue en vase clos. Si des émissions doivent se produire, c'est surtout au cours du stockage et du transport. Ces combustibles peuvent également être répandus dans l'environnement par suite de déversements accidentels ou dans des stations service lorsque l'on fait de plein des véhicules. L'atmosphère et l'hydrosphère sont les compartiments du milieu les plus fortement affectés par ces décharges accidentelles. Il peut y avoir contamination du sol par du combustible diesel à la faveur d'accidents et ce type de contamination pose également un problème dans les gares de triage.

Parmi les nombreuses techniques utilisables pour nettoyer les sols contaminés par du combustible diesel, on peut citer l'excavation, 

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A1.3 Transport, distribution et transformation dans
l'environnement

On ne dispose que très rares données sur la destinée du combustible diesel dans l'environnement mais on suppose que son mode de distribution et de transformation est comparable à celui des huiles lourdes destinées au chauffage comme le No 2 qui a été bien étudié. En cas de déversement dans l'eau, il se forme presque immédiatement une nappe de mazout. Les constituants polaires et ceux dont la masse
moléculaire est relativement faible se dissolvent et s'éliminent de la nappe par lessivage; les constituants volatils s'évaporent en surface et il y a un début de dégradation microbienne. Par attaque chimique et biologique, la composition de la nappe se modifie. Ces processus dépendent de la température; les déversements qui se produisent en milieu arctique conduisent à des nappes plus durables que sous les climats tempérés. En milieu marin, la plupart des composés aromatiques de masse moléculaire relative faible se dissolvent dans la phase aqueuse, mais les alcanes normaux, les alcanes ramifiés, les cycloalcanes et les composés aromatiques restantes peuvent demeurer dans les sédiments pendant plus d'un an.

Bien qu'on ne dispose d'aucune donnée sur la photo-oxydation du combustible diesel dans l'air et l'eau, on sait que les composants qui s'évaporent subissent une décomposition photochimique. Ainsi, on a montré que l'huile lourde No 2 subissait une photo-oxydation rapide en milieu aqueux dans les conditions naturelles.

Les différents constituants du combustible diesel sont intrinsèquement biodégradables, mais à des degrés et à des vitesses variables. Les alcanes normaux ainsi que les dérivés

n- alkylaromatiques et les molécules aromatiques simples en

C 10 -C 22 , sont les plus facilement dégradables. Les petites

molécules se métabolisent en général rapidement. Les n- alcanes à longue chaîne sont plus lentement dégradés en raison de leur hydrophobicité et du fait qu'ils sont visqueux ou solides à la température ambiante. Les alcanes ramifiés et les cycloalcanes sont relativement résistants à la décomposition biologique et les hydrocarbures aromatiques polycycliques, franchement résistants. La vitesse globale de dégradation des hydrocarbures est limitée par la température, la teneur en eau et en oxygène, le pH, la présence de nutriments inorganiques et la versatilité métabolique microbienne.

Les algues unicellulaires peuvent fixer et métaboliser les hydrocarbures aliphatiques et aromatiques mais on connaît mal l'ampleur de ce phénomène. Contrairement aux microorganismes qui utilisent les hydrocarbures du pétrole comme source de carbone, le métabolisme animal a généralement tendance à oxyder et à conjuguer les produits pour les transformer en substances plus solubles et donc plus faciles à excréter. Toutes les espèces animales étudiées sont capables de fixer des hydrocarbures du pétrole. On sait que les hydrocarbures aromatiques polycycliques, le pétrole brut et les produits pétroliers raffinés induisent les enzymes du cytochrome P450 et chez de nombreuses espèces de poissons de mer et d'eau douce, on constate qu'il y a accroissement du métabolisme des hydrocarbures.

On n'a que peu de données sur la bioaccumulation du combustible diesel dans les conditions du laboratoire mais il est largement prouvé, par l'étude des nappes de mazout et d'autres études de laboratoire consacrées à ce genre de produits, en particulier le No 2, que les organismes aquatiques concentrent les hydrocarbures. Le coefficient de partage du combustible diesel entre le n- octanol et l'eau est égal à 3,3-7,06, ce qui incite à penser que son potentiel de bioaccumulation est élevé; quoi qu'il en soit, de nombreux composés de
faible masse moléculaire relative sont facilement métabolisés et la bioaccumulation effective des produits de masse moléculaire relative plus élevée est limitée par leur faible solubilité dans l'eau et les dimensions importantes de leur molécule. Il en résulte donc que la bioaccumulation peut en réalité être faible.

On a constaté qu'après des déversements de combustible diesel, le poisson pouvait devenir inconsommable. On ne dispose d'aucune donnée sur la bioamplification du combustible diesel.

On ne possède non plus aucune donnée expérimentale sur le déplacement du combustible diesel à travers le sol, encore qu'il ait été avancé qu'il existait une corrélation directe entre ce déplacement et la viscosité cinématique du produit. Le déplacement du kérosène dans un sol dépend de la teneur en eau et de la nature de ce sol.

A1.4 Concentrations dans l'environnement et exposition humaine

Les divers types de combustible diesel étant constitués de mélanges complexes, on n'en n'a pas mesuré la concentration dans l'environnement. On peut mettre en évidence la présence de leurs divers constituants dans presque tous les compartiments du milieu, même s'il n'est pas possible d'en vérifier l'origine. Lorsqu'il y a exposition de la population générale, elle se produit dans les stations service ou par suite de déversements.

Il peut y avoir exposition professionnelle au combustible diesel à la faveur d'un grand nombre d'activités. Du fait de leur faible volatilité, ces combustibles ne devraient produire qu'une vapeur assez ténue à la température normale mais si l'on opère à température élevée, la concentration peut augmenter sensiblement.

A1.5 Effets sur les mammifères de laboratoire et les systèmes d'épreuve in vitro 

# Pa g e 165 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmLes combustibles diesel présentent une faible toxicité aiguë après administration par voie orale, percutanée ou respiratoire. Chez toutes les espèce étudiées (souris, lapin, rat, cobaye) on a obtenu pour la DL 50 par voie orale une valeur > 5000 mg/kg de poids
corporel. En application cutanée, on obtient une valeur de la DL 50 également > 5000 mg/kg de poids corporel chez la souris et le lapin, mais des valeurs > 2000 mg/kg de poids corporel ont été relevées pour certains types de kérosène et de distillats moyens, selon le protocole d'application et la limite inférieure de la dose. Chez des rats exposés par la voie respiratoire, on a obtenu une valeur de la CL 50 d'environ 5 mg/litre, sauf dans le cas d'un distillat moyen de première distillation pour lequel on a obtenu une valeur de 1,8 mg/litre.

Chez des lapins badigeonnés avec du combustible diesel à des doses allant jusqu'à 8000 μl/kg de poids corporel et par jour et chez
des souris traitées dans les mêmes conditions avec des doses quotidiennes atteignant 40 000 mg/kg de poids corporel, on a observé une acanthose et une hyperkératose dues à une grave irritation. Les lapins se sont révélés plus sensibles que les souris. Chez la souris, l'inhalation de combustible diesel a provoqué des effets neurodépresseurs à des concentrations allant jusqu'à 0,2 mg/litre; en revanche ces effets n'ont pas été observés chez des rats exposés à des doses allant jusqu'à 6 mg/litre. Chez les rats, on a observé une réduction du poids corporel et du poids du foie.

Après inhalation de doses de combustible diesel allant jusqu'à 1,5 mg/litre dans des conditions de subchronicité, des souris, des rats et des chiens n'ont pas présenté de signes sensibles de toxicité cumulative. Le syndrome néphropathique spécifique observé chez les rats mâles est lié à l'accumulation intrinsèque d'inclusions hyalines
dans les tubules rénaux.

Les seuls effets de l'exposition à long terme ont été des ulcérations après application cutanée de combustible diesel à des souris (doses quotidiennes: 250 ou 500 mg/kg de poids corporel) et une modification importante du poids des organes après inhalation par des rats de ce même produit à la dose de 1 ou 5 mg/litre. Dans les deux études, on a constaté une réduction du poids corporel moyen.

Les divers types de combustible diesel se révèlent légèrement à fortement irritants pour la peau du lapin. Ils ne produisent pas
d'irritation oculaire mais dans le cas d'un certain nombre de kérosènes, on a fait état d'un léger effet irritant. Il n'y a pas de
sensibilisation cutanée.

Le combustible diesel et les carburéacteurs (kérosène) ne se sont révélés ni embryotoxiques, ni tératogènes lors de deux études effectuées sur des rats à qui l'on avait fait inhaler ces produits aux doses de 100 ou 400 ppm ainsi que dans une autre étude au cours de laquelle des rats avaient reçu par gavage des doses quotidiennes de ces produits allant jusqu'à 2000 mg/kg de poids corporel. Dans la dernière étude, on a observé une réduction du poids des foetus.

Les épreuves effectuées sur Salmonella typhimurium n'ont pas permis de prouver de manière nette l'existence d'une activité mutagène. Des résultats positifs ont bien été observés chez S. typhimurium ainsi que sur des cellules lymphomateuses de souris
mais leur caractère contradictoire les a fait considérer comme équivoques. Les tests de génotoxicité effectués sur des souris

in vivo (induction de la formation de micronoyaux ou d'aberrations chromosomiques) ont également donné des résultats équivoques ou négatifs.

Les combustibles diesel présentent un faible pouvoir cancérogène au niveau cutané. Dans l'état actuel de la recherche, on ne peut déterminer si l'activité cancérogène de ces produits est due à leur génotoxicité ou aux lésions chroniques qu'ils induisent dans le derme.

A1.6 Effets sur l'homme

Il peut y avoir exposition non professionnelle aux combustibles diesel lors du remplissage manuel de citernes. C'est principalement lors de déversements accidentels que la peau peut se trouver fortement exposée à ces produits, encore que pour une brève durée.

A la suite de contacts directs avec le peau on a observé de l'anurie, une insuffisance rénale, des symptômes gastro-intestinaux ainsi qu'une hyperkératose cutanée. Des pneumopathies d'aspiration d'origine toxique ont été observées par suite de l'ingestion accidentelle de combustible diesel. Après inhalation, on peut observer

une toux grasse persistante. Lors d'une étude cas-témoins portant sur des hommes exposés à du combustible diesel, on a constaté qu'ils courraient un risque accru de cancers du poumon, autres que les adénocarcinomes; on a également observé une association positive avec le cancer de la prostate, encore que le risque ait été plus important dans le groupe soumis à une exposition 'non substantielle' que dans celui qui était soumis à une exposition 'substantielle'.

Lors d'une étude transversale portant sur des ouvriers d'une usine exposés à des carburéacteurs, on a constaté qu'ils étaient plus fréquemment sujets à des étourdissements, des maux de tête, des nausées, des palpitations, une sensation d'oppression thoracique et une irritation oculaire que les témoins non exposés. La concentration moyenne pondérée par rapport au temps de la vapeur dégagée par le combustible au niveau de la zone de respiration avait été estimée à 128-423 mg/m 3

. 

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leur milieu naturel

Le combustible diesel est plus toxique que le pétrole brut pour les animaux et les plantes aquatiques. L'écotoxicité du combustible diesel est généralement attribuée à la présence de composés aromatiques solubles, mais les hydrocarbures aliphatiques insolubles peuvent également jouer un rôle. Parmi les composés aromatiques, les dérivés monocycliques sont les moins toxiques, leur toxicité aiguë augmentant avec la masse moléculaire jusqu'aux composés tétra- ou pentacycliques, encore que ces derniers soient peu solubles dans l'eau de mer. Certains animaux, comme les poissons et les oiseaux, peuvent avoir le corps enduit de produit, avec des effets toxiques parfois

mortels.

On a étudié en laboratoire le combustible diesel et notamment ses fractions solubles dans l'eau, les dispersions huile-eau et le pétrole microencapsulé. Il apparaît que le combustible diesel ne réduit pas sensiblement la croissance des cultures d'algues vertes du genre Euglena gracilis, mais à faible concentration (0,1%), il inhibe presque complètement la croissance de Scenedesmus quadricauda. Le diesel léger (0,05%) stimule la croissance, la photosynthèse et la synthèse de la chlorophylle a chez Chlorella salina, mais il en inhibe légèrement la respiration. A concentration plus élevée, le taux de croissance et la photosynthèse sont fortement réduits. Une exposition de longue durée inhibe la croissance des algues benthiques

comme Ascophyllum nodosum et Laminaria digitata. Chez les algues bleues, la photosynthèse est réduite par les fractions aromatiques et asphaltiques, mais pas par la fraction aliphatique.

Le combustible diesel est fortement toxique pour les daphnies, pour les larves de chironomidés et pour le mollusque Viviparus bengalensis (un gastéropode). A la concentration de 0,1 ml/litre, il a provoqué la mort de copépodes du genre Tigriopus californicus, en l'espace de cinq jours.

Les moules du genre Mytilus edulis, accumulent le combustible diesel, avec pour conséquence une réduction marquée de leur rythme d'alimentation et de croissance; leur reproduction souffre également d'une exposition de longue durée à ce produit. La CE 50 relative au frai des moules exposées pendant 30 jours à du combustible diesel, a été trouvée égale à 800 μg/litre. La CL 50 de gazole microencapsulé a
été égale à 5000 μg/litre pour des moules en maturation exposées pendant 30 jours. Le gazole s'est révélé plus toxique pour les larves que pour les jeunes moules: il avait un effet nocif sur la croissance des larves à la dose de 10 μg/litre.

Des crabes d'eau douce (Barytelphusa cunicularis) exposés à des concentrations sublétales de combustible diesel pendant des durées allant jusqu'à 96 heures, ont généralement réduit leur consommation d'oxygène, en particulier aux concentrations les plus faibles et pour des durées d'exposition allant jusqu'à 8 heures. Lorsque l'exposition se prolongeait, la consommation d'oxygène était égale ou supérieure à
celle des animaux témoins.

Lors d'épreuves de 96 heures visant à évaluer la toxicité aiguë du produit sur des alevins de salmonidés dans des conditions statiques, on a constaté que le combustible diesel était plus toxique

pour Onchorhychus gorbuscha (CL 50 : 32-123 mg/litre), que pour

O. kisutch (CL 50 : 2186-3017mg/litre) ou pour la truite arc-en-ciel
O. mykiss (CL 50 : 3333-33 216 mg/litre) quel que soit le type

d'eau.

Le seuil de détection des réactions comportementales de la morue
( Gadus marhua L.), exposée à du combustible diesel dans de l'eau de mer, a été trouvé égal à 100-400 ng/litre. Un poisson de l'antarctique, Pagothenia borchgrevinki, a résisté pendant des durées allant jusqu'à 72 heures à la fraction hydrosoluble non diluée du combustible diesel, tout en présentant cependant des signes de

stress.

Dans le cas des oiseaux, il peut y avoir contamination externe ou interne par les produits pétroliers. Le combustible diesel supprime l'hydrophobicité du plumage et peut être ingéré lorsque l'oiseau lisse ses plumes. Du combustible diesel et du mazout administrés par gavage à des canards à la dose de 2 ml/kg de poids corporel, a provoqué au bout de 24 heures une stéatose, une inflammation extrême des poumons, une infiltration graisseuse du foie et une dégénérescence hépatique.

L'administration de combustible diesel ou de mazout à la dose de 1 ml/kg a également entraîné une grave irritation des voies digestives et une néphrose toxique. A doses plus élevées, on a observé une hypertrophie des surrénales (due principalement à une hyperplasie du tissu cortical), une chute du taux de la cholinestérase plasmatique, de l'ataxie et des tremblements. Jusqu'à 20 ml/kg de poids corporel, la contamination n'a pas été mortelle pour les oiseaux en bonne santé
mais la DL 50 s'abaissait à 3-4 ml/kg de poids corporel lorsque le

combustible diesel ou le mazout était administré à des oiseaux
stressés.

En cas de déversement de combustible diesel, le zooplancton se révèle extrêmement vulnérable aux constituants dispersés ou dissous du pétrole, mais moins aux nappes d'huile flottante. Pour les organismes aquatiques, la nocivité de ces produits peut se manifester de diverses manières: mortalité directe (oeufs de poisson, copépodes et plancton), contamination externe (chorions des oeufs de poisson ou cuticules et appendices buccaux des crustacés), contamination tissulaire par des constituants aromatiques, développement anormal des embryons de poisson et perturbation du métabolisme. 

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Il peut y avoir exposition de la population générale au combustible diesel et à d'autres distillats moyens sur les lieux et dans les circonstances suivants: dans les stations service, lors de déversements accidentels, lors de la manipulation de ces combustibles et lors de l'utilisation de pétrole lampant pour la cuisine ou le chauffage. Il peut y avoir exposition des travailleurs à ces produits lors de la manipulation et du transvasement du combustible dans les terminaux, les citernes et les stations services; lors de la fabrication, de la réparation, de l'entretien et de l'essai des
moteurs diesel et autres matériels; lors de l'utilisation de combustible diesel pour le nettoyage ou comme solvant ou encore lors de la manipulation des prélèvements de routine au laboratoire. En raison de sa faible volatilité, le produit ne devrait pas émettre de vapeurs très denses à la température ambiante, encore que dans un espace confiné et à température élevée, la vapeur puisse être plus
concentrée.

Lors de la manipulation normale du combustible diesel, l'exposition aux vapeurs est minime. L'effet le plus probable sur la
santé humaine consiste en une dermatite de contact. Le combustible diesel est irritant pour la peau mais il ne semble pas irriter la muqueuse oculaire. Des effets toxiques aigus au niveau rénal peuvent s'observer après exposition cutanée, mais on ignore quels peuvent être les effets à long terme d'une absorption percutanée de faibles
concentrations.

L'ingestion de combustible diesel peut avoir des effets toxiques qui se traduisent quelquefois par une régurgitation et une aspiration pouvant provoquer une pneumonie chimique, comme dans le cas de tout hydrocarbure entre certaines limites de viscosité.

Chez des rongeurs exposés par la voie respiratoire à du combustible diesel à des concentrations allant jusqu'à 0,2 mg/litre, on a observé un effet neurodépresseur dans le cas de souris, mais pas chez des rats, mêmes aux concentrations les plus élevées. Chez des rats mâles, une exposition subchronique par la voie respiratoire à divers distillats a produit une néphropathie spécifique à alpha2-microglobulines; cette observation n'est pas considérée comme transposable à l'homme.

Les combustibles diesel ne se sont révélés ni embryotoxiques ni tératogènes chez les animaux exposés par la voie orale ou par la voie respiratoire.

Rien n'indique de façon nette qu'il existe une activité mutagène chez les bactéries, mais les résultats d'autres tests de génotoxicité
in vitro et in vivo sont plutôt équivoques.

Un étude cas-témoins portant sur des ouvriers exposés à du combustible diesel incitent à penser qu'il existe un risque accru de cancers du poumon n'appartenant pas au type épithélium glandulaire ainsi que de ce cancer de la prostate. Il n'a pas été possible d'établir de relation dose-réponse pour l'un ou l'autre de ces cas. En raison du petit nombre d'études disponibles, du petit nombre de cas et par voie de conséquence, de l'étendue de l'intervalle de confiance, il n'est pas possible de tirer la moindre conclusion de ces données au sujet de la cancérogénicité du produit pour l'homme. Chez la souris, du combustible diesel administré par voie intradermique a présenté une faible activité cancérogène. En raison de l'absence d'une activité génotoxique avérée, il est possible d'invoquer un mécanisme non génotoxique pour ces cancers; il pourrait s'agir par exemple d'une irritation dermique chronique caractérisée par la répétition de lésions cutanées entraînant une hyperplasie de l'épiderme.

A1.9 Evaluation des effets sur l'environnement

Il peut y avoir pollution de l'environnement par libération accidentelle à grande échelle de combustible diesel, comme cela peut se produire lors catastrophes affectant des réservoirs ou encore en cas de fuites d'oléoduc ou, à plus petite échelle, lorsqu'il y a contamination du sol aux alentours d'une usine ou d'un garage. Dans l'eau, le combustible diesel s'étale presque immédiatement, les composants polaires de masse moléculaire relative peu élevée se dissolvent et disparaissent pas lessivage, les constituants volatils s'évaporent et la décomposition microbienne s'amorece. L'élimination

des différents constituants dépend de la température et des conditions climatiques. La composition chimique de la nappe varie dans le temps: une fois répandues dans l'eau, certaines fractions s'évaporent et subissent une décomposition photochimique; les fractions lourdes, transportées par les particules qui se déposent, parviennent jusqu'aux
sédiments du fond; dans le sol les constituants du combustible diesel migrent à des vitesses variables, en fonction de la nature de ce sol.

Les divers constituants du combustible diesel sont intrinsèquement biodégradables mais leur vitesse de biodécomposition dépend largement des conditions physiques et climatiques et de la composition microbiologique du milieu.

Des organismes aquatiques et en particulier les mollusques, accumulent les hydrocarbures à des degrés divers mais ceux-ci peuvent être éliminés par passage dans l'eau claire. Le combustible diesel peut subir une bioaccumulation; en revanche on ne dispose d'aucune donnée sur une bioamplification éventuelle.

Tout déversement de combustible diesel a un effet nocif immédiat sur l'environnement, qui se traduit par une mortalité notable pour la 

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Les organismes aquatiques qui survivent aux déversements de combustible diesel peuvent cependant subir une contamination externe par ces produits et en accumuler dans leurs tissus: le stress qui en résulte se traduit par un développement anormal et une modification du
métabolisme

PARTIE B GAZ D'ECHAPPEMENT DES MOTEURS DIESEL

B1. RESUME

B1.1 Identité, propriétés physiques et chimiques et méthodes d'analyse

Les gaz d'échappement des moteurs diesel contiennent des centaines de composés chimiques qui sont émis en partie dans la phase gazeuse proprement dite et en partie dans la phase particulaire. Les principaux produits gazeux de la combustion sont le dioxyde de carbone, l'oxygène, l'azote et la vapeur d'eau: on trouve également du monoxyde de carbone dixoyde de soufre, des oxydes d'azote et des hydrocarbures et leurs dérivés. La phase gazeuse de la fraction hydrocarbonée contient également un faible pourcentage en poids de benzène et de toluène. Les autres constituants des gaz d'échappement sont des hydrocarbures aromatiques polycycliques (HAP) de faible masse
moléculaire relative.

L'échappement des moteurs diesel est principalement caractérisé par l'émission de particules dans une proportion environ 20 fois supérieure à celle des moteurs à essence. Ces particules sont composées de carbone élémentaire, de dérivés organiques adsorbés provenant du combustible et de l'huile lubrifiante, de sulfates formés à partir du soufre contenu dans le combustible et de dérivés métalliques à l'état de traces. La granulométrie globale de ces particules les situe en dessous du micron, entre 0,02 et 0,5 μm. Le vieillissement peut conduire à une agglomération qui les amène à un diamètre maximal de 30 μm. Les particules émises ont une aire superficielle importante. Les composés organiques constituent généralement 10 à 30% des particules totales mais avec des moteurs mal conçus et mal entretenus cette proportion peut atteindre 90%. Dans cette fraction on trouve également des hydrocarbures aromatiques polycycliques de masse moléculaire élevée, entre autres, sous forme oxygénée et nitrée, à des concentrations de l'ordre de plusieurs parties par million.

Pour mesurer les émissions des véhicules on travaille en régime transitoire ou stationnaire. L'échantillonnage peut se faire à partir de gaz d'échappement dilués ou non dilués. Il est difficile d'obtenir
des échantillons débarrassés de tout artéfact car les constituants peuvent subir des réactions chimiques, être adsorbés ou désorbés, ou encore soumis à une condensation ou à une diffusion. Les hydrocarbures aromatiques polycycliques d'importance toxicologique qui sont présents dans les particules émises par les moteurs diesel sont généralement dosées selon la procédure suivante: extraction au Soxhlet, purification et fractionnement puis analyse finale par chromatographie liquide à haute performance ou chromatographie en phase gazeuse couplée à la spectométrie de masse.

B1.2 Sources d'exposition humaine et environnementale

Les gaz d'échappement des moteurs diesel proviennent essentiellement des véhicules à moteur; parmi les autres source on peut citer les installations fixes, le matériel de traction
ferroviaire et les machines de navires. Les émissions de ces véhicules à moteurs diesel ont été bien décrites mais les résultats obtenus pour chaque catégorie particulière ne sont souvent pas comparables en raison de différences concernant certains paramètres comme le cycle de fonctionnement, le type de moteur et la composition du combustible. Les divers constituants sont émis dans les proportions suivantes: dioxyde de carbone, environ 1 kg/km; monoxyde de carbone, oxydes d'azote, hydrocarbures gazeux totaux et particules 0,1-20 g/km; dérivés aliphatiques, alcools, aldédhydes, hydrocarbures aromatiques légers et hydrocarbures aromatiques polycycliques, quelques microgrammes/km. Dans un certain nombre de pays, il y a réglementation des émissions de monoxyde de carbone, d'oxyde d'azote, d'hydrocarbures gazeux totaux et de particules.

En principe, il n'y a aucune différence entre les moteurs de faible ou de forte puissance pour ce qui est de la nature et de la quantité des émissions, encore que les véhicules lourds aient tendance à dégager une quantité relativement plus importante de particules. Les émissions dépendent du cycle de fonctionnement du moteur (régime transitoire ou régime stationnaire), du type et de l'état du moteur (injection ou aspiration, entretien, kilométrage total) et de la composition du combustible (teneur en soufre, teneur en composés aromatiques, volatilité); le réglage du moteur jour un rôle très important.

Le dégagement de particules augmente en raison inverse du rapport air/combustible, mais en raison directe de la charge et de la température. Les moteurs vieillissants, soumis à une utilisation intense, libèrent plus de particules que les moteurs récents, qui ont peu de kilométrage, probablement du fait de la plus grande consommation d'huile lubrifiante. Dans le cas des véhicules légers à moteur diesel, il y a également une corrélation entre l'émission de particules et la teneur en soufre du combustible, car la formation de sulfates métalliques accroît la masse des particules. Dans le cas des véhicules lourds, ce type de corrélation n'a pas encore été établi. 

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Les hydrocarbures aromatiques polycycliques, oxygénés ou non, émis par les moteurs diesel et les moteurs à allumage commandé, sont de nature similaire. Des hydrocarbures aromatiques polycycliques oxygénés ou nitrés sont émis à raison de quelques microgrammes/km, mais la concentration effective de ces composés n'est pas connue avec certitude car il peut y avoir décomposition ou formation lors de l'échantillonnage. Les émissions de HAP augmentent avec la charge et

la température ainsi qu'avec l'âge du moteur, probablement du fait d'une consommation plus importante d'huile lubrifiante. Les émissions d'HAP dépendent également de la technique d'injection utilisée dans le moteur: ces émissions sont proportionnelles au rapport air/combustible dans les moteurs à injection directe mais la situation est inversée dans le cas des moteurs à injection indirecte. La teneur en dérivés aromatiques et la volatilité du combustible sont directement liées aux
émissions de HAP. En cas de mauvais fonctionnement de certains éléments du moteur, en particulier des injecteurs, il y a augmentation de l'émission des principaux constituants des gaz d'échappement. On ne possède guère de données sur la contribution des moteurs diesel aux émissions totales de produits de combustion d'origine artificielle.

Les émissions des moteurs diesel peuvent être réduites moyennant une meilleure conception du moteur, la pose de pièges à particules (pièges oxydants) et de convertisseurs catalytiques. Alors que les pièges à particules éliminent la suie et les composés organiques solubles adsorbés sur les particules, les convertisseurs catalytiques réduisent principalement la teneur en oxyde de carbone et hydrocarbures gazeux. En pratique, il est difficile de régénérer les pièges à particules. Les convertisseurs catalytiques nécessitent
l'utilisation de combustibles à faible teneur en soufre car le soufre empoisonne les centres actifs du catalyseur.

B1.3 Transport, distribution et transformation dans l'environnement

C'est principalement l'atmosphère qui est affectée par les émissions de moteurs diesel. L'hydrosphère et la géosphère peuvent être contaminées indirectement par des dépôts secs ou humides. La destinée environnementale des divers constituants des gaz d'échappement des moteurs diesel est en général bien connue: les particules se comportent comme les molécules de gaz non réactives pour ce qui est de leur transport mécanique dans l'atmosphère; elles peuvent être transportées sur de longues distances et même pénétrer dans la stratosphère. On pense que leur vitesse d'élimination est faible, ce qui fait qu'elles séjournent plusieurs jours dans l'atmosphère. Avec le temps, ces particules peuvent s'agglomérer, et leur vitesse de chute augmentant, la quantité restante aéroportée diminue. Le carbone élémentaire présent dans des particules émises par les moteurs diesel peut catalyser la formation d'acide sulfurique par oxydation du dioxyde de soufre. Les constituants organiques adsorbés à la surface des particules de carbone élémentaire peuvent subir un certain nombre de transformations et de rections chimiques avec d'autres composés atmosphériques ou par exposition à la lumière
solaire.

B1.4 Concentrations dans l'environnement et exposition humaine

Etant donné que les gaz d'échappement des moteurs diesel sont des mélanges complexes contenant des composés très divers, on ne peut pas définir de 'concentration dans l'environnement'. On devrait pouvoir mettre en évidence la présence de ces divers constituants dans tous les compartiments de l'environnement, encore qu'il ne soit pas généralement possible d'en déterminer l'origine. On connaît la concentration dans l'environnement de la plupart de ces constituants.

C'est le plus probablement dans les rues très passantes ou dans les parkings, en particulier souterrains, que la population générale a le plus de chances d'être exposée aux émissions des moteurs diesel. Il est difficile d'identifier les sources de pollution et on calcule généralement la contribution des gaz d'échappement des moteurs diesel à la pollution totale due à la circulation des véhicules à moteur, sur la base des facteurs d'émission et du pourcentage de véhicules à moteur diesel dans le parc automobile général. Le niveau d'exposition de la population générale et des travailleurs aux particules émises par les moteurs diesel est toxicologiquement significatif.

Les concentrations quotidiennes moyennes ambiantes de particules à proximité des voies de communication sont de 8-42 μg/m 3 . On a calculé que les concentrations annuelles moyennes de particules étaient de 5 à 10 μg/m 3 dans les zones urbaines et < 1,5 μg/m 3 dans les zones rurales d'Allemagne et de 1 à 2 μg/m 3 dans les zones urbaines et 0,6-1 μg/m 3 dans les zones rurales des Etats-Unis d'Amérique. Ces concentrations sont directement corrélées à la densité de la circulation automobile et diminuent à mesure qu'on s'éloigne des

routes.

Il peut être également difficile de déterminer l'origine de la pollution sur les lieux de travail, en particulier dans les mines où la quantité totale de poussière est élevée. Le dosage du carbone particulaire a permis de déterminer l'exposition spécifique aux
émissions de moteurs diesel sur les lieux de travail et il en ressort que les travailleurs seraient exposés à des concentrations de particules de l'ordre de 0,04 à 0,134 mg/m 3 pour les camionneurs et de 0,004 à 0,192 mg/m 3 pour les cheminots. On a également eu recours au dosage des particules respirables totales et des particules totales en suspension pour évaluer l'exposition professionnelle.

B1.5 Cinétique et métabolisme chez les animaux de laboratoire et 

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B1.5.1 Dépôt

Les particules émises par les moteurs diesel, dont le diamètre aérodynamique médian massique est égal à environ 0,2 μm, sont quelque peu filtrés par le nez et leur aptitude au dépôt dans les poumons n'est que légèrement supérieure à la valeur minimale trouvée pour les

particules de diamètre aérodynamique médian massique égal à environ 0,5 μm. Autrement dit, lorsque les particules de suie émises par les moteurs diesel sont inhalées, 10 à 15% d'entre elles se déposent au niveau des alvéoles chez le rat et le cobaye, le dépôt au niveau des
alvéoles étant d'environ 10% chez l'homme.

B1.5.2 Rétention et élimination des particules

L'élimination des particules par l'ascenseur muco-ciliaire est pratiquement complète au bout de 24 heures. L'élimination à long terme des particules déposées dans la zone alvéolaire a fait l'objet de plusieurs études sur des rats qui avaient été exposés par la voie respiratoire à deux types de particules: les particules émises par un moteur diesel et les particules servant de référence. Dans le cas des témoins chez lesquels la charge pulmonaire était faible (< 1 mg par poumon), on a obtenu des temps de demi-élimination de l'ordre de 60 à 100 jours, alors que chez les rats présentant une charge pulmonaire allant de 1 à 60 mg/poumon, le temps de demi-élimination était de l'ordre de 100 à 600 jours. Dans plusieurs études, on a constaté que les effets observés étaient dus à une surcharge en particules, laquelle a été décrite chez diverses espèces et pour un certain nombre de matériaux particulaires. Ce phénomène s'observe généralement lorsque la vitesse de dépôt des particules de faible solubilité et de faible toxicité aiguë reste supérieure à leur vitesse d'élimination pendant une très longue durée. Chez l'homme, le temps de demi-élimination alvéolaire normal est de plusieurs centaines de jours, c'est-à-dire plus long que chez le rat.

On a mis au point un modèle pulmonaire dosimétrique sur la base des données de dépôt et de rétention des particules diesel chez le rat après inhalation de longue durée et des mêmes données chez l'homme. Ce modèle peut être utilisé pour prévoir la rétention des particules diesel et des composés organiques adsorbés dans les poumons de personnes d'âges divers. On ne dispose d'aucune donnée sur la manière dont la rétention de différents composés évolue après une exposition prolongée aux gaz d'échappement de moteurs diesel.

B1.5.3 Rétention et élimination des hydrocarbures aromatiques polycycliques adsorbés sur la suie de moteurs diesel

Les hydrocarbures aromatiques polycycliques présents dans la suie émise par les moteurs diesel adhèrent fortement à la surface des particules. Environ 50% des HAP adsorbés sur les particules sont éliminés par les poumons dans l'espace d'une journée, mais le temps de demi-rétention de la fraction restante s'est révélé égal à 18-36 jours. Des études portant sur du 3 H-benzo[ a ]pyrène et du

14 C-nitropyrène ont montré que lorsqu'ils sont associés à des particules, les HAP sont sensiblement plus longs à être éliminés des poumons que leurs homologues libres.

B1.5.4 Métabolisme

Déposé sur des particules émises par un moteur diesel, du benzo[ a ]pyrène a subi une métabolisation oxydative au niveau du poumon et dans des cultures cellulaires de macrophages pulmonaires aboutissant à la formation de phénols, de diols et de quinones substituant le noyau benzo[ a ]pyrène. Du nitropyrène adsorbé sur des particules diesel a été métabolisé en acétylaminopyrène-phénol après inhalation. Chez des rats exposés à des émissions de moteur diesel on a constaté, dans les poumons et les cellules de type II, un accroissement des adduits de l'ADN par rapport aux témoins. Le métabolisme oxydatif de certains composés organiques qui conduit à des époxydes pourrait être responsable de la formation des adduits, mais il est vrai qu'on ne les trouve qu'après exposition à des particules. Certains dérivés organiques présents dans les émissions de moteur diesel se sont révélés capables de former des adduits avec l'ADN; toutefois le noyau carboné lui-même (sans composés organiques extractibles) peut également conduire à la formation d'adduits, en provoquant des lésions chroniques au niveau des cellules épithéliales.

B1.6 Effets sur les mammifères de laboratoire et les systèmes d'épreuve in vitro

Les quelques données disponibles incitent à penser que les émissions de moteur diesel ne présentent qu'une faible toxicité aiguë. Des souris auxquelles on avait administré par voie intratrachéale, des particules émises par un moteur diesel, sont mortes des suites d'une oedème du poumon. La DL 50 était égale à environ 20 mg/kg de poids
corporel. Des particules du même type, extraites au méthanol, n'ont entraîné aucune mortalité jusqu'à une concentration d'environ 33 mg/kg de poids corporel. Chez le hamster, on a obtenu une DL 50 de 1280 mg/kg de poids corporel après administration par voie intrapéritonéale. On ne dispose d'aucune donnée concernant l'exposition par la voie respiratoire.

Après exposition de rats, de cobayes et de chats pendant environ quatre semaines à des émissions de moteur diesel ayant une teneur en particules de 6 mg/m 3 , on a observé une altération de la fonction pulmonaire, et notamment une augmentation de 35% de la résistance ventilatoire chez les cobayes et une diminution de 10% de la capacité vitale chez les chats. Du point de vue histopathologique, on a observé 

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se situaient au niveau des bronchioles terminales et elles ont augmenté de volume lors de la période de récupération postérieure à l'expérience en raison de la fixation des macrophages (séquestration).

Après avoir inhalé pendant de longues périodes des émissions de moteur diesel à des concentrations atteignant 4 mg/m 3 , des rats, des souris, des hamsters, des chats et des singes n'ont pas présenté de chute spectaculaire de leur poids corporel ni de réduction de leur longévité. Les effets toxiques liés à la dose qui ont été observés chez toutes les espèces après une longue période d'inhalation étaient les suivants: augmentation du poids des poumons pouvant aller jusqu'à 400%; inflammation pulmonaire objectivée par des paramètres biochimiques (marqueurs enzymatiques cytoplasmiques, collagène) et cytologiques (augmentation des neutrophiles polynucléaires); perturbation de la mécanique pulmonaire; accroissement du nombre de macrophages chargés de particules avec accumulations focales (séquestration) dans des conditions de surcharge; enfin, altérations prolifératives des cellules épithéliales et début de fibrose.

D'après les données limitées dont on dispose sur la toxicité des émissions de moteurs diesel pour la fonction de reproduction et le développement, on peut penser qu'il n'y a pas d'effets toxiques déterminants. Dans la plupart des expériences, on n'a pas constaté d'effets chez des souris, des rats, des hamsters, des lapins ou des singes; toutefois, après injection par voie intrapéritonéale, on a observé chez des souris qui avaient reçu de particules émises pas des moteurs diesel, des anomalies touchant les spermatozoïdes et constaté une certaine embryotoxicité chez des hamsters auxquels on avait
administré des extraits d'émissions de ces mêmes moteurs.

La plupart des épreuves de génotoxicité in vitro ont été réalisées avec des extraits d'émissions de moteurs diesel plutôt qu'avec les émissions totales elles-mêmes et l'on a obtenu des réactions positives en l'absence d'activation métabolique. Autrement dit, il semble que ces effets génotoxiques soient indépendants de la présence d'HAP. Environ 50% des études in vivo ont donné des résultats négatifs; les seuls résultats positifs obtenus consistaient en échanges de chromatides soeurs dans le cas des émissions totales et des extraits organiques, et dans la présence de micronoyaux dans le cas des extraits organiques.

On n'a généralement pas observé d'effets immunotoxiques après
inhalation d'émissions de moteurs diesel; toutefois on a observé dans une étude une augmentation du titre des immunoglobulines anti-ovalbumine et deux autres expériences ont mis en évidence un
accroissement de la sensibilité aux infections chez des souris.

Des études effectuées sur des rats incitent à penser que l'inhalation de gaz d'échappement de moteurs diesel affecte l'état comportemental et neurophysiologique.

Lors d'études sur la cancérogénicité des émissions de moteur diesel, au cours desquelles des rats étaient exposés à ces émissions par la voie respiratoire, on a constaté que la phase gazeuse (c'est-à-dire dépourvue de particules) n'était pas cancérogène. Toutes les études sur des rats qui ont été validées ont montré que les émissions de moteurs diesel avaient un effet cancérogène lorsque la concentration en particules était supérieure à 2 mg/m 3 , soit l'équivalent d'une exposition continue à une concentration d'environ
1 mg/m 3 . Aucun effet n'a été observé en revanche chez les hamsters et les souris. Lors d'études au cours desquelles on a pratiqué une instillation intratrachéale, on a constaté que les particules présentes dans les émissions de moteurs diesel et le noir de fumée produisaient des tumeurs; en outre on a observé l'existence d'une corrélation entre l'aire superficielle des particules carbonées et leur activité tumorigène.

L'inhalation, pendant une longue période, de noir de fumée pratiquement dépourvu d'hydrocarbures aromatiques polycycliques à ces concentrations, a également provoqué des tumeurs pulmonaires chez le

rat.

On ne sait pas avec certitude si la cancérogénicité des émissions
de moteurs diesel est due à un mécanisme faisant intervenir ou non des réactions au niveau de l'ADN (ou à un mécanisme mixte). Différents modèles ont été proposés pour élucider la cancérogénicité des
émissions de moteurs diesel.

B1.7 Effets sur l'homme

Les émissions de moteurs diesel contribuent à la pollution globale de l'air. Les études toxicologiques, qu'elles cherchent à mettre en évidence des effets aigus ou des effets chroniques, ne permettent pas d'assigner un rôle particulier à ces particules mais il apparaît qu'elles pourraient être en partie responsables d'un certain nombre d'effets attribués à la pollution de l'air.

Les émissions de moteur diesel ont des caractéristiques qu'un certain nombre de personnes trouvent agressives, en particulier à fortes concentrations. Les symptômes observés après exposition de brève ou de longue durée à ce type d'émissions ont été décrits dans un certain nombre d'études et de rapports concernant des personnes exposées de par leur profession. Les effets aigus consistent en irritation des muqueuses oculaires et nasales et l'on a observé dans les cohortes professionnelles une augmentation de la fréquence des symptômes respiratoires; toutefois on ignore qu'elle est la contribution exacte des particules émises par les moteurs diesel à ce 

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d'asthme.

Lors d'une étude contrôlée au cours de laquelle huit volontaires non fumeurs, en bonne santé, ont été exposés à des émissions diluées de moteurs diesel, pendant 60 minutes dans une chambre fermée, on a constaté une réduction du taux de phagocytose chez des macrophages alvéolaires recueillis par lavage broncho-alvéolaire.

Un certain nombre d'études transversales et longitudinales portant sur des ouvriers longtemps exposés de part leur profession à des émissions de moteur diesel, ont révélé une altération de la fonction pulmonaire et une augmentation de la prévalence des symptômes respiratoires, toutefois la brièveté des épisodes d'exposition limite la portée de ces études. Aucune surmortalité n'a été constatée lors d'études de cohorte au cours desquelles on s'est efforcé d'étudier des décès par maladie cardio-vasculaire ou accident vasculaire cérébral, imputables à l'exposition des émissions de moteurs diesel.

Lors d'un certain nombre d'études épidémiologiques on a tenté d'établir des relations entre certains cancers du poumon et de la vessie et une exposition professionnelle à des émissions de moteurs diesel. Seules les études jugées utiles pour l'évaluation des effets cancérogènes de ces émissions ont été prises en considération dans la présente monographie. Celles qui sont les plus intéressantes pour ce qui est du cancer du poumon, concernent des cheminots, des personnes travaillant dans des garages de cars et des débardeurs qui constituent des cohortes dont les membres sont effectivement exposés aux émissions de moteur diesel. Les quatre études les plus informatives font toutes état d'un risque accru de cancer du poumon, avec un risque relatif allant de 1,4 pour les cheminots, et de 1,3-2,4 pour les ouvriers des garages (selon le type d'exposition), jusqu'à un risque trois à six fois plus élevé pour les débardeurs (en fonction du mode d'évaluation de l'exposition, mais avec un large intervalle de confiance). Il a été possible de tenir compte du tabagisme dans une étude cas-témoins sur des cheminots et dans une étude sur des débardeurs. Dans les deux cas, cette correction pour tenir compte du tabagisme n'a eu aucune influence sur l'effet de l'exposition aux émissions diesel. Dans les trois études pour lesquelles on n'a pas pu tenir compte du tabagisme, l'analyse était basée sur des comparaisons entre les différents sous-groupes de ces cohortes, de sorte que l'effet de confusion créé par le tabagisme avait moins de chance d'être gênant qu'en cas de comparaison avec des groupes extérieurs.

Plusieurs études cas-témoins ont été menées afin d'examiner la relation pouvant exister entre les cancers de la vessie et une exposition supposée à des émissions de moteurs diesel. On a constaté un accroissement du risque, en particulier pour le chauffeurs de
camion; toutefois toutes ces études souffrent d'une caractérisation insuffisante de l'exposition. De la sorte, il n'est pas possible d'affirmer qu'une exposition aux émissions de moteurs diesel entraîne un risque accru de cancer vésical.

B1.8 Effets sur les autres êtres vivants au laboratoire et dans
leur milieu naturel

Les effets des émissions de moteur diesel n'ont été abordés que dans une seule étude, portant sur des algues vertes.

B1.9 Evaluation des risques pour la santé humaine

L'évaluation des risques de cancer et de maladies non malignes a été menée conformément au modèle d'évaluation des risques établi par l'Académie nationale des sciences des Etats-Unis (National Research Council, 1983). Ce modèle d'évaluation comporte quatre étapes: (1) identification du danger; (2) évaluation de la relation dose-réponse; (3) évaluation de l'exposition et (4) caractérisation du risque.

On estime que les travaux les plus intéressants sont les études épidémiologiques de longue durée dans lesquelles l'exposition est bien définie et le suivi supérieur à 20 ans. Quatre études portant sur le cancer du poumon chez des personnes professionnellement exposées satisfont à ces critères. Il apparait que le risque relatif de cancer du poumon lié à une exposition à des émissions de moteur diesel est généralement faible et qu'il est influencé par les circonstances, les
effets des facteurs de confusion non évalués et la difficulté de tenir suffisamment compte des facteurs de confusion reconnus. D'autres études, où l'exposition est définie avec une moindre précision, corroborent les conclusions des études précédentes. Globalement, on estime que les émissions de moteurs diesel sont probablement cancérogènes pour l'homme; toutefois on ne dispose d'aucune donnée quantitative permettant d'évaluer le risque.

B.1.9.1 Effets non néoplasiques

Pour caractériser le risque, on a procédé de deux manières: premièrement une concentration sans effets nocifs observables que l'on divise par un coefficient d'incertitude; deuxièmement, une concentration de référence. Dans les deux cas, on a utilisé un modèle dosimétrique élaboré qui permet de réduire l'imprécision due à l'extrapolation interspécifique des doses.

La dose de particules sans effet pour l'homme a été estimée à
0,139 mg/m 3 . La valeur guide pour la population générale calculée d'après le modèle dosimétrique a été établie à 5,6 μg/m 3 , la valeur calculée sans recours au modèle étant de 2,3 μg/m 3

.

La méthode basée sur l'utilisation d'une concentration de référence prend en compte l'ensemble de la relation exposition-réponse 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmde préférence aux données ponctuelles fournies par les études d'inhalation, comme c'est le cas dans la méthode de la dose sans effets observables. On a identifié trois paramètres biologiques sensibles: l'inflammation alvéolaire chronique, la diminution

d'efficacité de l'ascenseur mucociliaire et les lésions pulmonaires hyperplasiques. Les concentrations de référence calculées à partir du même modèle dosimétrique que dans le cas de la méthode de la dose sans effets observables, étaient de 0,9 à 2 μg/m 3 pour l'inflammation, de 1,2 à 3 2 μg/m 3 pour la dégradation de l'ascenseur muco-ciliaire et de 6,3 à 14,2 μg/m 3 pour les lésions hyperplasiques.

B1.9.2 Effets néoplasiques

Pour évaluer le risque d'une exposition aux émissions de moteur diesel, on a eu recours à un modèle linéarisé en plusieurs étapes. Comme les résultats des études épidémiologiques avaient été jugés insuffisants pour une évaluation quantitative du risque unitaire, on a eu recours à des données provenant de plusieurs études d'inhalation à long terme chez des rats, études qui montraient l'existence d'une cancérogénèse à partir de 2 mg/m 3

.

On a ainsi établi par le calcul que le risque unitaire était
de 3,4 × 10 -5 μg/m 3 (moyenne géométrique de quatre estimations). Un autre modèle de type biologique a fourni une valeur similaire pour le risque unitaire, en prenant pour hypothèse que les particules émises par les moteurs diesel agissent à faibles concentrations sur la prolifération cellulaire ou l'initiation du processus de
transformation.

B1.10 Evaluation des effets sur l'environnement

On ne dispose pas d'informations suffisantes pour évaluer les effets spécifiques des émissions de moteurs diesel. La combustion du
carburant diesel devrait avoir des effets similaires à celle des autres combustibles fossiles, effets qui sont liés à la consommation de ce type de combustible.

PARTE A DIESEL

A1. RESUMEN

A1.1 Identidad, propiedades físicas y químicas y métodos
analíticos

El diesel es una mezcla compleja de alcanos normales, ramificados y cíclicos (60%-> 90% en volumen; hidrocarburos de longitud de cadena comprendida entre C 9 y C 30 ); compuestos aromáticos, especialmente
alquilbencenos (5%-40% en volumen), y pequeñas cantidades de alquenos (0%-10% en volumen) obtenidos a partir de la fracción de gasóleo del destilado medio durante la separación del petróleo. El diesel puede contener benceno, tolueno, etilbenceno, y xilenos e hidrocarburos aromáticos policíclicos (HAP), sobre todo naftaleno y derivados del mismo con metilos sustituyentes, a niveles de partes por millón. El contenido de azufre de los combustibles diesel depende del origen del crudo y del proceso de refino. En un cierto número de países está reglamentado por ley, y suele estar comprendido entre 0,05% y 0,5% en peso. Se utilizan aditivos para influir en el flujo, el almacenamiento y la combustión del diesel, para diferenciar productos y para satisfacer las especificaciones comerciales. A temperatura ambiente, los combustibles diesel suelen ser líquidos de color marrón moderadamente volátiles, ligeramente viscosos, inflamables y de olor parecido al del queroseno. Los márgenes de ebullición están comprendidos por lo general entre 140°C y 385°C (> 588°C en el caso del diesel marino); a 20°C la densidad es de 0,87-1,0 g/cm 3 , y la hidrosolubilidad de 0,2-5 mg/litro. La calidad y composición del diesel influye considerablemente en los contaminantes emitidos por los motores diesel. Otras variables importantes son el proceso de encendido (expresado en términos del índice de cetano), la densidad, la viscosidad y el contenido de azufre. Las especificaciones del diesel comercial varían considerablemente de un país a otro.

Los combustibles de calefacción y algunos combustibles de aviación a base de queroseno producidos durante el proceso de refino tienen a veces una composición parecida a la del diesel, aunque con distintos aditivos. Por ello, a la hora de evaluar la toxicidad y la ecotoxicidad se han tenido también en cuenta los datos biológicos

sobre estas mezclas.

Debido a la complejidad de las mezclas diesel, no existe un método analítico específico para ellas, y las técnicas analíticas empleadas en la mayoría de las evaluaciones ambientales sólo permiten determinar la mezcla total de hidrocarburos de petróleo. Los métodos consisten en una extracción previa con solventes, un proceso de limpieza para eliminar los hidrocarburos naturales, y la posterior detección mediante gravimetría, espectroscopía infrarroja o cromatografía de gases.

Ni la gravimetría ni la técnica infrarroja proporcionan
información cualitativa o cuantitativa de utilidad sobre los contaminantes, por lo que sólo pueden usarse con fines de cribado.

El método utilizado habitualmente para analizar muestras ambientales es la cromatografía de gases unida a técnicas de detección tales como la ionización por conductor y la espectrometría de masas. Existen otros muchos métodos para analizar hidrocarburos particulares
en los combustibles diesel.

A1.2 Fuentes de exposición humana y ambiental 

# Pa g e 174 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmLos combustibles diesel se producen por refinación del petróleo bruto. Para satisfacer las especificaciones técnicas en cuanto al rendimiento, normalmente se combinan varios componentes para obtenerlos; la ulterior inclusión de aditivos mejora sus propiedades para usos específicos. Los combustibles diesel son muy utilizados con fines de transporte. En los motores de alta velocidad se deben emplear los más volátiles y de menor viscosidad, y los de cadena más larga se reservan para los motores diesel de ferrocarriles y barcos. Una gran parte del transporte pesado por carretera está propulsado por motores
diesel.

Los automóviles para pasajeros propulsados por motores diesel son cada vez más frecuentes en Europa y el Japón (10%-25%), mientras que en América del Norte el porcentaje de ese tipo de automóviles es de aproximadamente 1%-2%, con una ligera tendencia a la baja. El diesel es utilizado en motores fijos y en calderas, como por ejemplo motores de pistones, turbinas de gas, bombas de oleoductos, compresores de gas, unidades de procesamiento de vapor de centrales de energía eléctrica, quemadores e instalaciones industriales de calefacción de espacios y de agua.

En los últimos cinco años la demanda mundial de combustibles
diesel ha aumentado constantemente. En 1985 se consumieron las siguientes cantidades: unas 170 000 kilotoneladas al año en América del Norte; unas 160 000 kilotoneladas al año, gasóleos incluidos, en la Unión Europea; y aproximadamente 46 000 kilotoneladas anuales en Australia, el Japón y Nueva Zelandia, lo que equivale a un total de 1062 kilotoneladas diarias. En 1990 se notificó que la demanda mundial ascendía aproximadamente a 1110 kilotoneladas al día.

No se dispone de información sobre las emisiones a que da lugar la producción de combustibles diesel; no obstante, al parecer esta fuente tiene una importancia secundaria, dado que el proceso de
refinación se lleva a cabo en sistemas cerrados. Las emisiones se producen sobre todo durante el almacenamiento y el transporte. Se libera diesel como resultado de derrames y durante el reabastecimiento de los vehículos en las gasolineras.

La atmósfera y la hidrosfera son los compartimentos ambientales más gravemente afectados. En algunos accidentes se produce contaminación del suelo por combustibles diesel, problema que también afecta a las playas ferroviarias. Entre las numerosas técnicas empleadas para limpiar los suelos contaminados por diesel, cabe citar la excavación, los métodos biológicos y el confinamiento.

A1.3 Transporte, distribución y transformación en el medio

ambiente

Apenas hay datos sobre el destino ambiental de los combustibles diesel, pero se considera que los mecanismos de su distribución y transformación son comparables a los de combustibles de calefacción que han sido bien estudiados, como por ejemplo el fuel-oil No 2. Los derrames de diesel en agua se extienden casi de inmediato para formar una 'capa aceitosa'. Los componentes polares y de bajo peso molecular relativo se disuelven y se separan por lixiviación de dicha capa, mientras que los componentes volátiles se evaporan a partir de la superficie; además da comienzo la degradación microbiana. La degradación química y biológica altera la composición del derrame. Estos procesos dependen de la temperatura; así, los derrames que se producen en el Artico duran más que los que se producen en zonas de clima templado. En entornos marinos, la mayoría de los agentes aromáticos de bajo peso molecular relativo se disuelven en la fase acuosa, pero los alcanos ramificados, los cicloalcanos y los restantes compuestos aromáticos primarios pueden persistir en los sedimentos
durante más de un año.

No se dispone de información sobre la fotooxidación de los combustibles diesel en el agua y el aire, pero se sabe que los componentes del petróleo evaporados se degradan fotoquímicamente. Se ha demostrado que el fuel-oil No 2 se fotooxida rápidamente en el agua
en condiciones ambientales.

Los distintos componentes del diesel son de por sí biodegradables, en diversa medida y a distinta velocidad. Los

n- alcano, los compuestos n- alquilaromáticos y las moléculas aromáticas simples comprendidas en el margen de C 10 a C 22 son los que más fácilmente se degradan. Las moléculas más pequeñas son metabolizadas por lo general rápidamente. Los n- alcanos de cadena larga se degradan más lentamente, debido a su hidrofobicidad y a que son viscosos o sólidos a temperatura ambiente. Los alcanos ramificados y los cicloalcanos son relativamente resistentes a la degradación biológica, y los HAP son resistentes. La velocidad global de degradación de los hidrocarburos depende de la temperatura, el contenido de agua, el oxígeno, el pH, los nutrientes inorgánicos y la
variabilidad metabólica microbiana.

Las algas unicelulares pueden incorporar y metabolizar hidrocarburos tanto alifáticos como aromáticos, pero no se sabe con seguridad en qué medida ocurre realmente tal cosa en la naturaleza. A diferencia de los microorganismos que utilizan carbonos de petróleo como fuente de carbono, los animales generalmente oxidan y conjugan los productos, generando productos finales que son más solubles y por consiguiente más fáciles de excretar. Todas las especies animales analizadas pueden incorporar hidrocarburos de petróleo. Se sabe que los HAP, el petróleo bruto y los productos refinados derivados del petróleo son inductores de las enzimas del citocromo P450 y aumentan el metabolismo de los hidrocarburos en numerosas especies de peces marinos y de agua dulce.

Hay pocos datos disponibles sobre la bioacumulación de diesel en 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmcondiciones de laboratorio, pero diversos estudios sobre derrames y estudios de laboratorio sobre otros tipos de petróleo, en particular el fuel-oil No 2, han aportado pruebas abundantes de que los organismos acuáticos bioconcentran los hidrocarburos. El coeficiente de reparto n- octanol-agua para el diesel es de 3,3-7,06, valores que indican una alta bioacumulación potencial; no obstante, muchos de los compuestos de bajo peso molecular relativo son metabolizados fácilmente, y la bioacumulación real de los compuestos de mayor peso molecular relativo se ve limitada por su escasa hidrosolubilidad y su gran tamaño molecular. Así pues, la bioacumulación real posiblemente es baja.

Se han descubierto peces envenenados por diesel después de derrames, pero no se dispone de datos sobre la biomagnificación del combustible. No hay datos experimentales sobre el movimiento del diesel a través del suelo, aunque se ha señalado que existe quizá una correlación directa entre el movimiento y la viscosidad cinemática. El movimiento del queroseno a través del suelo depende del contenido de humedad y de las características del suelo.

A1.4 Niveles ambientales y exposición humana

Debido a su carácter de mezclas complejas, no se han determinado los niveles ambientales de los combustibles diesel. Se pueden detectar componentes individuales del diesel en casi todos los compartimentos del medio ambiente, pero no es posible averiguar su origen. La población general puede verse expuesta al diesel en las gasolineras y

de resultas de los derrames.

Hay muchas actividades que acarrean exposición ocupacional al diesel. Dada su escasa volatilidad, en principio estos combustibles sólo suelen generar bajas concentraciones de vapores a temperaturas normales, pero a temperaturas altas las concentraciones pueden llegar a ser importantes.

A1.5 Efectos en mamíferos de laboratorio y en sistemas de pruebas
in vitro

La toxicidad aguda de los combustibles diesel es baja en caso de exposición oral o cutánea o de inhalación. El valor de la DL 50 oral
fue superior a 5000 mg/kg de peso corporal en todas las especies analizadas (ratón, conejo, rata, cobayo). La DL 50 del producto aplicado a nivel cutáneo fue superior a 5000 mg/kg de peso corporal en el ratón y el conejo, si bien se notificaron valores de más de 2000 mg/k de peso corporal para algunos querosenos y destilados medios, con distintos protocolos y dosis límite menores. La CL 50 observada en ratas expuestas por inhalación fue de aproximadamente 5 mg/litro, con excepción de un destilado medio directo para el que se halló un valor de 1,8 mg/litro.

En conejos y en ratones tratados cutáneamente con cantidades diarias de hasta 8000 μl/kg de peso corporal y hasta 40 000 mg/kg de peso corporal respectivamente, se observó la aparición de acantosis e hiperqueratosis como consecuencia de una irritación grave. Los conejos eran más sensibles que los ratones. La inhalación de diesel tuvo efectos neurodepresivos en ratones a concentraciones de hasta 0,2 mg/litro, pero no así en ratas expuestas a concentraciones de hasta 6 mg/litro. En ratas se observó una disminución del peso corporal y del peso del hígado.

No se observaron signos importantes de toxicidad acumulativa en ratones, ratas y perros que habían inhalado hasta 1,5 mg/litro de forma subcrónica. El síndrome de nefropatía específico observado en ratas macho se asocia a una acumulación inherente de gotas hialinas en
los túbulos renales.

Los únicos efectos de las exposiciones a largo plazo fueron la ulceración por aplicación cutánea en ratones (250 ó 500 mg/kg de peso corporal al día) y alteraciones importantes del peso de los órganos tras la inhalación de 1 ó 5 mg/litro en ratas. En los dos estudios el peso corporal medio disminuyó.

Diversos tipos de diesel se revelaron entre leves y gravemente irritantes para la piel del conejo. Los combustibles diesel no son irritantes para el ojo, si bien se ha observado que algunos querosenos tienen un ligero efecto de esa naturaleza. Los combustibles diesel no
causan sensibilización cutánea.

El diesel y los combustibles de avión (queroseno) no tuvieron efectos embriotóxicos ni teratogénicos en dos estudios realizados en ratas expuestas por inhalación a concentraciones de 100 ó 400 ppm y en un estudio en que las ratas recibieron hasta 2000 mg/kg de peso corporal diarios mediante sonda. En este último estudio se observó una disminución del peso fetal.

Las pruebas realizadas con Salmonella typhimurium no aportaron indicios claros de mutagenicidad. Algunos datos positivos obtenidos

con S. typhimurium y en células de linfoma de ratón se consideraron equívocos dada la incoherencia de los resultados. Las pruebas de genotoxicidad realizadas con ratones in vivo (inducción de micronúcleos o aberraciones cromosómicas) también dieron resultados equívocos o negativos.

Se ha observado la inducción de un bajo nivel de carcinogenicidad cutánea por combustibles diesel. En el estado actual de las investigaciones, es imposible discernir si la actividad carcinogénica del diesel se debe a un mecanismo genotóxico o a la aparición de
lesiones cutáneas crónicas.

A1.6 Efectos en el ser humano 

# Pa g e 176 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmUna posible fuente de exposición no ocupacional al combustible diesel es el llenado manual de depósitos de combustible. La fuente principal de exposición cutánea son los derrames accidentales, que dan lugar de inmediato a niveles altos de exposición, si bien de corta
duración.

Como efectos del contacto cutáneo accidental se han notificado anuria, insuficiencia renal, síntomas gastrointestinales e hiperqueratosis cutánea. Se ha observado un cuadro pulmonar tóxico tras la ingestión accidental de diesel y su sucesiva aspiración. Se ha notificado el desarrollo de tos productiva persistente tras la inhalación del producto.

En un estudio de casos y testigos realizado en hombres expuestos al diesel, se observó un aumento del riesgo de cáncer de pulmón
distinto del adenocarcinoma; se detectó asimismo una correlación positiva con el cáncer de próstata, si bien se halló un riesgo mayor para el grupo sometido a una exposición 'no considerable' que para el sometido a una exposición 'considerable'.

En un estudio transversal realizado entre obreros expuestos a combustibles de avión a base de queroseno se observó que los trabajadores presentaban mareos, cefaleas, náuseas, palpitaciones, presión torácica e irritación ocular con mayor frecuencia que los testigos no expuestos. Se calculó que la concentración promedio ponderada por el tiempo del vapor desprendido por el combustible en la zona en que se respiraba era de 128-423 mg/m 3

A1.7 Efectos en otros organismos en el laboratorio y en el medio
ambiente

El diesel es más tóxico que el petróleo bruto para los organismos y las plantas acuáticas. La ecotoxicidad del diesel se atribuye por lo general a compuestos aromáticos solubles, pero es posible que también intervengan hidrocarburos alifáticos insolubles. Entre los compuestos

aromáticos, los monoaromáticos son los menos tóxicos, y su toxicidad aguda aumenta con el peso molecular hasta los compuestos de cuatro a cinco anillos, si bien estos últimos son escasamente solubles en agua de mar. En algunos animales, como por ejemplo peces y aves, el recubrimiento físico de la superficie corporal por el combustible puede ser causa de toxicidad y mortalidad.

Se han llevado a cabo experimentos de laboratorio con combustible diesel, fracciones hidrosolubles, dispersiones de petróleo-agua y petróleo microencapsulado. El diesel no redujo significativamente el crecimiento en cultivo del alga verde Euglena gracilis, mientras que una concentración baja (0,1%) inhibió casi por completo el crecimiento

de Scenedesmus quadricauda. La presencia de diesel a baja concentración (0,05%) estimuló el crecimiento, la fotosíntesis y la
síntesis de clorofila a de Chlorella salina, pero inhibió ligeramente su respiración; a concentraciones superiores el ritmo de crecimiento y la fotosíntesis se vieron considerablemente reducidos. La exposición prolongada inhibió el crecimiento de las algas
bentónicas Ascophyllum nodosum y Laminaria digitata. En algas verdeazuladas, la fotosíntesis se vio reducida por las fracciones aromática y asfáltica, pero no por la fracción alifática.

El diesel tuvo efectos tóxicos agudos en Daphnia, en larvas de quironómidos, y en el molusco Viviparus bengalensis (gasterópodos). Una concentración de 0,1 ml/litro causó la muerte de copépodos de

charca Tigriopus californicus en el término de cinco días.

Los mejillones Mytilus edulis acumulan diesel, presentan índices de alimentación y crecimiento considerablemente reducidos y acusan efectos de toxicidad reproductiva tras la exposición crónica a
dicho combustible. La CE 50 para el desove en mejillones expuestos durante 30 días fue de aproximadamente 800 μg/litro. La CL 50 de gasóleo microencapsulado en mejillones en fase de maduración expuestos durante 30 días al producto fue de unos 5000 μg/litro. El gasóleo fue más tóxico para las larvas que para los ejemplares jóvenes: 10 μg/litro tenían efectos adversos para el crecimiento de las larvas.

Cangrejos de agua dulce (Barytelphusa cunicularis) expuestos a concentraciones subletales de diesel por espacio de hasta 96 horas redujeron por lo general su consumo de oxígeno, sobre todo a exposiciones bajas de hasta ocho horas. A exposiciones más largas el consumo de oxígeno fue similar o superior al de los controles.

En las pruebas de toxicidad aguda de 96 horas llevadas a cabo en salmónidos jóvenes en condiciones estáticas, el diesel tuvo efectos más tóxicos para el salmón rosado Onchorhychus gorbuscha
(CL 50 : 32-123mg/litro) que para el salmón plateado O. kisutch (CL 50 : 2186-3017 mg/litro) o para la trucha arco iris O. mykiss

(CL 50 : 3333-33216 mg/litro), con independencia del tipo de agua.

El umbral de detección de efectos comportamentales en bacalaos
( Gadus morhua L.) expuestos a diesel en agua de mar fue de 100-400 ng/litro. El pez antártico Pagothenia borchgrevinki soportó
una fracción hidrosoluble no diluida de combustible diesel durante 72 horas, si bien mostró signos de estrés.

Las aves se ven afectadas externa e internamente por los petróleos contaminantes. El diesel destruye la impermeabilidad del plumaje de las aves y éstas lo ingieren cuando se limpian las plumas. El diesel y el fuel-oil administrados mediante sonda a razón de 2 ml/kg de peso corporal a patos causó neumonía lipídica, inflamación extrema de los pulmones, infiltración grasa del hígado y degeneración 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmhepática al cabo de 24 horas. La administración de diesel o fuel-oil a dosis de 1 ml/kg causó irritación grave del tracto digestivo y nefrosis tóxica. Dosis mayores causaron aumento de las glándulas suprarrenales (debido sobre todo a una hiperplasia del tejido cortical), disminución de los niveles plasmáticos de colinesterasa, ataxia y temblores. La administración de estos productos a dosis de hasta 20 ml/kg de peso corporal no tuvieron efectos mortales en pájaros sanos, pero la DL 50 del diesel y el fuel-oil administrados a

aves estresadas fue de 3-4 ml/kg de peso corporal.

Tras los derrames de diesel, el zooplancton parece altamente vulnerable a los componentes dispersos y disueltos del petróleo, pero no tanto al petróleo flotante. Los organismos acuáticos pueden verse afectados de varias formas: mortalidad directa (huevos de pescados, copépodos y plancton mixto), contaminación externa por petróleo (corion de huevos de pescados y cutículas y apéndices de alimentación de los crustáceos), contaminación tisular por componentes aromáticos, desarrollo anormal de embriones de peces, y alteración de las tasas
metabólicas.

A1.8 Evaluación de los riesgos para la salud humana

La población general puede verse expuesta al diesel y a otros destilados medios en las gasolineras de resultas de derrames accidentales, durante la manipulación de dichos combustibles y con motivo del uso de queroseno en la cocina o la calefacción domésticas. Los trabajadores pueden verse expuestos al diesel o a otros destilados medios al manipular y descargar el combustible en las terminales, los tanques de almacenamiento y las gasolineras; durante la fabricación, la reparación, el mantenimiento y el ensayo de motores y demás equipo a base de diesel; durante la utilización del diesel como agente limpiador o disolvente; y durante el manejo y el muestreo sistemáticos de combustible diesel en el laboratorio. Debido a su baja volatilidad a temperatura ambiente sólo suelen producirse concentraciones bajas de vapor, si bien en espacios cerrados y a altas temperaturas se pueden alcanzar niveles importantes.

La exposición al vapor desprendido es mínima durante la manipulación habitual del diesel. El efecto más probable en la salud
humana es la dermatitis resultante del contacto cutáneo. El diesel es irritante para la piel, pero al parecer no para el ojo. La exposición cutánea puede dar lugar a efectos tóxicos agudos a nivel renal, pero se desconoce qué efectos tiene la absorción cutánea prolongada de bajas concentraciones.

La ingestión de combustibles diesel tiene efectos tóxicos, y en ocasiones se produce regurgitación y aspiración, posible causa de neumonía química, al igual que puede ocurrir con cualquier hidrocarburo que tenga un determinado grado de viscosidad.

En roedores, la exposición por inhalación a concentraciones de diesel de hasta 0,2 mg/litro tuvo efectos neurodepresivos en ratones, pero no en cambio en ratas a las concentraciones más altas. La exposición subcrónica por inhalación a diversos destilados combustibles indujo una nefropatía específica para la alpha2-microglobulina en ratas macho; se consideró que este efecto era irrelevante para el hombre.

No se observaron efectos embriotóxicos ni teratogénicos en animales expuestos oralmente o por inhalación a combustibles diesel.

No hay indicios claros de una actividad mutagénica en bacterias, y los resultados de otras pruebas de genotoxicidad
in vitro e in vivo fueron ambiguos.

Los resultados de un estudio de casos y testigos realizado entre trabajadores expuestos a diesel llevaron a pensar en un aumento del riesgo de cáncer de pulmón distinto del adenocarcinoma y de cáncer de próstata. En ninguno de los casos se observó una relación exposición- respuesta. En vista de los pocos estudios disponibles, del reducido número de casos y de la consiguiente amplitud de los intervalos de confianza, no es posible extraer conclusión alguna acerca de la carcinogenicidad del diesel para el hombre.

En ratones, la administración cutánea de combustibles diesel fue débilmente carcinógena. Dado que no existe una genotoxicidad clara, es posible que la inducción de cáncer se deba a mecanismos no genotóxicos, por ejemplo a una irritación cutánea crónica caracterizada por ciclos repetidos de lesiones cutáneas, con la consiguiente hiperplasia epidérmica.

A1.9 Evaluación de los efectos en el medio ambiente

El medio puede contaminarse de resultas de la liberación accidental de diesel en gran escala, como la asociada a desastres en depósitos o escapes en oleoductos, o a menor escala como resultado de la contaminación del suelo de las inmediaciones de fábricas o garajes.

En el agua, el diesel se propaga de forma casi inmediata, los componentes polares y de bajo peso molecular relativo se disuelven y se separan por lixiviación, los componentes volátiles se evaporan a partir de la superficie del agua, y da comienzo la degradación microbiana. La intensidad de la degradación depende de la temperatura y de las condiciones climáticas. La composición química de los derrames cambia con el tiempo: después de un vertido al agua, algunas fracciones se evaporan, y los componentes de diesel evaporados se degradan fotoquímicamente; en el sedimento, parece ser que el diesel es transportado en general a los sedimentos del fondo por las partículas que se depositan; en el suelo, los componentes del diesel migran a distintas velocidades, en función del tipo de suelo. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmLos componentes individuales son de por sí biodegradables, pero la velocidad de biodegradación depende estrechamente de las condiciones físicas y climáticas y de la composición de la población
microbiana.

Los organismos acuáticos, en particular los moluscos, bioacumulan hidrocarburos en diversa medida, pero se produce una depuración de los hidrocarburos al transferirlos a agua limpia. Puede darse una bioacumulación de diesel; ahora bien, no hay datos sobre su biomagnificación.

Los derrames de diesel tienen efectos nocivos inmediatos en el medio, donde causan una considerable mortalidad de la biota. Puede producirse una recolonización al cabo de aproximadamente un año, en función de la especie de animal o planta y del contenido químico y físico de los restos del derrame. Los organismos acuáticos que sobreviven a los derrames de diesel pueden verse afectados por la contaminación externa y por la acumulación del producto en los tejidos: un desarrollo anormal y la presencia de tasas metabólicas
alteradas son indicios del estrés resultante.

PARTE B EMISIONES DE GASES DE ESCAPE DE MOTORES DIESEL

B1. RESUMEN

B1.1 Identidad, propiedades físicas y químicas y métodos
analíticos

Las emisiones de gases de escape de los motores diesel contienen cientos de compuestos químicos, que son emitidos en parte en la fase gaseosa y en parte en la fase de materia particulada de los gases de escape. Los principales productos gaseosos de la combustión son los siguientes: dióxido de carbono, oxígeno, nitrógeno y vapor de agua; hay también monóxido de carbono, dióxido de azufre, óxidos de nitrógeno, e hidrocarburos y sus derivados. Se detectan benceno y tolueno en el margen porcentual correspondiente a los compuestos de menor peso en la parte gaseosa de la fracción de hidrocarburos. Otros componentes de los gases de escape son los hidrocarburos aromáticos policíclicos (HAP) de bajo peso molecular relativo.

Una importante característica de las emisiones de estos gases de escape es que la liberación de partículas es 20 veces más intensa que en los vehículos propulsados por gasolina. Las partículas están compuestas de carbono elemental, compuestos orgánicos adsorbidos del combustible y el aceite lubricante, sulfatos derivados del azufre del combustible, y cantidades mínimas de componentes metálicos. La mayor parte de la materia particulada total tiene al parecer dimensiones del orden de la submicra: entre 0,02 y 0,5 μm. Con el paso del tiempo puede producirse aglomeración, hasta un diámetro máximo de 30 μm. Las partículas emitidas presentan una gran superficie. Los compuestos orgánicos representan por lo general un 10%-30% de la materia particulada total, pero los motores mal diseñados o mantenidos pueden generar hasta un 90%. En esta fracción se encuentran también, a concentraciones de partes por millón, HAP, HAP oxigenados y nitro-HAP de alto peso molecular relativo.

Para medir las emisiones de los vehículos se utilizan ciclos de pruebas transitorios y estables específicos. Pueden tomarse muestras de gases de escape diluidos o no diluidos. Es difícil conseguir muestras sin artefactos, ya que los componentes de los gases de escape pueden sufrir reacciones químicas, procesos de adsorción y desorción, y condensación y difusión. Los HAP de mayor interés toxicológico de la materia particulada del diesel se determinan por lo general mediante la extracción de Soxhlet, limpieza y fraccionamiento, seguidos de análisis por cromatografía líquida de alta resolución o cromatografía de gases unida a espectrometría de masas.

B1.2 Fuentes de exposición humana y ambiental

La principal fuente de gases de escape de motores diesel son los vehículos de motor; otras fuentes son los motores diesel fijos, de locomotoras de ferrocarril y de barco. Las emisiones de los vehículos de motor diesel han sido bien caracterizadas, pero a menudo los resultados de los estudios no se prestan a comparación, debido a diferencias que afectan a parámetros tales como los ciclos de pruebas, el tipo de motor y la composición del combustible. Las cantidades emitidas de los diversos componentes son las siguientes: dióxido de carbono, aproximadamente 1 kg/km; monóxido de carbono, óxidos de nitrógeno, hidrocarburos gaseosos totales y materia particulada, 0,1-20 g/km; y compuestos alifáticos, alcoholes, aldehídos, compuestos aromáticos ligeros y HAP, microgramos por kilómetro. Las emisiones de monóxido de carbono, óxidos de nitrógeno, hidrocarburos gaseosos totales y materia particulada están reglamentadas por ley en varios países.

En lo que respecta a la calidad y cantidad de los gases de escape, no suele haber diferencias entre los motores de baja y de gran potencia, si bien los vehículos pesados liberan cantidades relativamente mayores de materia particulada. La naturaleza de los gases de escape depende del ciclo de pruebas (transitorio o estable), de las características del motor (técnicas de inyección y de aspiración, mantenimiento, kilometraje), y de la composición del combustible (contenido de azufre, aromaticidad, volatilidad); el ajuste del motor es muy importante en este sentido.

La liberación de materia particulada aumenta con la disminución de la proporción aire/combustible, el aumento de la carga y el aumento de la temperatura. Emiten más partículas los motores viejos y muy usados que los nuevos y de bajo kilometraje, probablemente debido a un 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmmayor consumo de aceites lubricantes. La emisión de partículas por los vehículos diesel de poca potencia también está correlacionada con el contenido de azufre del combustible, ya que la formación de sulfatos de metal se traduce en un aumento de la masa de partículas; no se ha determinado la emisión de partículas por parte de vehículos de gran potencia. El aumento de la aromaticidad del combustible también conlleva una mayor emisión de partículas.

Los HAP y los HAP oxigenados emitidos por los motores diesel y de encendido por chispa son parecidos cualitativamente. Los HAP oxigenados y nitrados se emiten a niveles del orden de pocos microgramos por kilómetro, pero no se sabe con seguridad qué concentración real alcanzan estos compuestos, ya que pueden descomponerse y formarse durante el muestreo. Las emisiones de HAP aumentan con la carga y con la temperatura, así como con a antigüedad del motor, a causa probablemente de un mayor consumo de aceites

lubricantes. La emisión de HAP depende también de la técnica de inyección del motor: las emisiones de HAP aumentan al mismo tiempo que la proporción aire/combustible en los motores de inyección directa, mientras que disminuyen en los motores de inyección indirecta. La aromaticidad y la volatilidad del combustible guardan relación directa con la emisión de HAP. El mal funcionamiento de los dispositivos del motor, sobre todo del sistema de inyección de combustible, aumenta la emisión de los principales componentes de escape. Se dispone de pocos
datos sobre la contribución de las emisiones de los motores diesel a la liberación total de productos de combustión por el hombre.

Es posible reducir las emisiones de gases de escape producidos por el diesel mejorando el diseño de los motores, los captores de partículas (oxidantes de captores) y los catalizadores. Mientras que los captores de partículas eliminan tanto el hollín como los compuestos orgánicos solubles adsorbidos en las partículas, los catalizadores reducen sobre todo los niveles de monóxido de carbono y de hidrocarburos gaseosos. En la práctica es difícil regenerar los captores de partículas. Los catalizadores exigen combustibles de bajo contenido de azufre, ya que este elemento daña los centros activos del
catalizador.

B1.3 Transporte, distribución y transformación en el medio
ambiente

El compartimento que antes se ve afectado por las emisiones de gases de escape de los motores diesel es la atmósfera. La hidrosfera y la geosfera se ven contaminadas indirectamente por depósitos secos y húmedos. El destino ambiental de los distintos componentes de estos gases de escape es en general bien conocido: las partículas se comportan como moléculas de gas (no reactivas) por lo que se refiere a su transporte mecánico en la atmósfera; pueden ser transportadas a gran distancia y penetrar incluso en la estratosfera.

Se estima que el índice global de eliminación de las partículas diesel es bajo, lo que se traduce en una permanencia de varios días en la atmósfera. Con el tiempo se puede producir una coagulación de las partículas, con lo que aumenta la precipitación de las mismas y disminuye su concentración total en el aire. El carbono elemental de las partículas diesel puede actuar como catalizador de la formación de ácido sulfúrico por oxidación del dióxido de azufre. Los componentes orgánicos adsorbidos en el carbono elemental pueden experimentar varias reacciones físicas y químicas con otros compuestos atmosféricos y durante la exposición a la luz solar.

B1.4 Niveles ambientales y exposición humana

Dado que los gases de escape diesel son una mezcla compleja de gran diversidad de compuestos, por lo general no es posible precisar los 'niveles ambientales'. Los distintos componentes de los gases de escape diesel se pueden detectar en principio en todos los compartimentos del medio ambiente, aunque por lo general no es posible averiguar su origen. Se conocen los niveles ambientales de la mayoría de los distintos componentes.

La exposición de la población general a los gases de escape de motores diesel se produce especialmente en las calles transitadas y en los aparcamientos, sobre todo los subterráneos. La identificación de las fuentes resulta difícil, y la contribución de esos gases a la contaminación total por productos de combustión del tráfico se calcula en general a partir de los coeficientes de emisión y del porcentaje de vehículos propulsados por diesel. Los niveles de exposición de la población general y de los trabajadores a la materia particulada de los motores diesel tienen importancia toxicológica. Las concentraciones ambientales diarias de partículas diesel cerca de las carreteras oscilan como promedio entre 8 y 42 μg/m 3 . Las concentraciones promedio anuales estimadas fueron de 5-10 μg/m 3 en zonas urbanas y < 1,5 μg/m 3 en zonas rurales de Alemania, y de 1-2 μg/m 3 en zonas urbanas y 0,6-1 μg/m 3 en zonas rurales de los Estados Unidos de América. Las concentraciones están directamente relacionadas con la densidad del tráfico y son tanto menores cuanto más lejos están las carreteras.

La identificación de las fuentes también puede revestir dificultad en los lugares de trabajo, sobre todo en las minas, donde la carga total de polvo es elevada. Se han hecho análisis de muestras de carbono para determinar la exposición específica a gases de escape de motores diesel en el lugar de trabajo y, según se ha notificado, los niveles de materia particulada a que están expuestos los trabajadores son del orden de 0,04-0,134 mg/m 3 para los conductores de camiones y de 0,004-0,192mg/m 3 para los ferroviarios. Como medidas de la exposición ocupacional se han utilizado también las partículas respirables totales y las partículas suspendidas totales. 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmB1.5 Cinética y metabolismo en animales de laboratorio y en el

hombre

B1.5.1 Depósito

Las partículas de los gases de escape de motores diesel, con un diámetro aerodinámico mediano de aproximadamente 0,2 μm, son filtradas en cierta medida por la nariz, y la eficiencia de depósito en el pulmón es sólo ligeramente superior al valor mínimo hallado para las

partículas que tienen un diámetro aerodinámico mediano de aproximadamente 0,5 μm. Así, un 10%-15% de las partículas de hollín diesel inhaladas se depositan en la región alveolar de los pulmones de la rata y el cobayo; en el hombre se deposita en la región alveolar un

10%.

B1.5.2 Retención y eliminación de las partículas

La eliminación mucociliar de partículas en las vías respiratorias ciliadas es casi completa en un plazo de 24 horas. La eliminación a largo plazo de las partículas diesel en la región alveolar se ha determinado mediante varios estudios realizados en ratas expuestas por inhalación a partículas diesel o control marcadas. Se notificaron periodos de semieliminación de 60-100 días en testigos con cargas pulmonares bajas (< 1 mg/pulmón), mientras que los periodos de semieliminación aumentaron a 100-600 días en ratas con cargas pulmonares crecientes de 1 a 60 mg/pulmón. En varios estudios se halló que los efectos se debían a una sobrecarga de partículas, fenómeno descrito en diversas especies y con varios tipos de materia particulada. Este hecho se observa por lo general cuando la velocidad de depósito de las partículas de baja solubilidad y baja toxicidad aguda supera su velocidad de eliminación durante un tiempo considerable. El periodo de semieliminación alveolar libre en el hombre es de varios cientos de días, mayor que en la rata.

Se desarrolló un modelo pulmonar dosimétrico a partir de datos relativos al depósito y retención de las partículas diesel en ratas sometidas a inhalación prolongada, así como de datos referentes al depósito y retención de partículas en el hombre. Este modelo puede utilizarse para predecir la retención de partículas diesel y de material orgánico adsorbido en los pulmones de personas de diferentes edades. No se dispone de datos sobre la variación de la retención de distintos componentes tras la exposición prolongada a gases de escape
de motores diesel.

B1.5.3 Retención y eliminación de hidrocarburos aromáticos policíclicos adsorbidos en hollín de diesel

Los HAP del hollín de diesel se adhieren fuertemente a la superficie de las partículas. Aproximadamente un 50% de los HAP adsorbidos en las partículas diesel desaparece de los pulmones en el plazo de un día, pero los periodos de semieliminación de los restantes
HAP fueron del orden de 18-36 días. Estudios realizados con

3 H-benzo[ a ]pireno y 14 C-nitropireno han demostrado que, cuando los HAP están asociados a materia particulada, su eliminación de los pulmones se retrasa significativamente en comparación con los HAP inhalados no asociados a materia particulada.

B1.5.4 Metabolismo

Se ha observado que el benzo[ a ]pireno adsorbido en partículas de gases de escape de motores diesel se metaboliza por oxidación en fenoles, dioles y quinonas de benzo[ a ]pireno en el pulmón y en cultivos celulares de macrófagos pulmonares.

El nitropireno adsorbido en partículas diesel se metabolizó en acetilaminopirenofenol después de su inhalación. Se han hallado más aductos de ADN en los pulmones y las células de tipo II de ratas expuestas a gases de escape de motores diesel que en las testigo. El metabolismo oxidativo de algunos compuestos orgánicos en epóxidos podría ser la causa de la formación de esos aductos, pero únicamente se hallaron aductos tras la exposición a partículas. Se ha demostrado que algunas sustancias orgánicas de los gases de escape de motores diesel dan lugar a la formación de aductos de ADN; sin embargo, el núcleo carbonáceo por sí mismo (sin material orgánico extraíble) también puede inducir aductos, por lesión crónica de las células epiteliales.

B1.6 Efectos en mamíferos de laboratorio y en sistemas de pruebas
in vitro

Los escasos datos disponibles parecen indicar que la toxicidad aguda de los gases de escape de motores diesel es escasa. Ratones tratados intratraquealmente con partículas de tales gases murieron de edema pulmonar; la DL 50 fue de aproximadamente 20 mg/kg de peso
corporal. Partículas extraídas con metanol no tuvieron efectos letales a concentraciones de hasta aproximadamente 33 mg/kg de peso corporal. En hámsters, la DL 50 tras administración intraperitoneal fue de
1280 mg/kg de peso corporal. No hay datos respecto a la exposición por
inhalación.

La exposición de ratas, cobayos y gatos a gases de escape de motores diesel con un contenido de partículas de 6 mg/m 3 durante aproximadamente cuatro semanas alteró la función pulmonar, causando en particular un aumento del 35% de la resistencia al flujo pulmonar en los cobayos y una disminución del 10% de la capacidad vital (flujo espiratorio) en los gatos. El análisis histopatológico reveló un espesamiento focal de las paredes alveolares, un aumento significativo del índice de marcaje de las células de tipo II, y acumulaciones de 

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3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmmacrófagos cargados de partículas. Estas acumulaciones se hallaban cerca de los bronquiolos terminales, y aumentaron de tamaño, debido a la adhesión (secuestro) de macrófagos, durante el periodo ulterior de recuperación.

No se observaron disminuciones importantes del peso corporal ni acortamiento alguno del tiempo de supervivencia en ratones, ratas, hámsters, gatos y monos después de un periodo prolongado de inhalación de concentraciones de hasta 4 mg/m 3 . En todas las especies sometidas a inhalación prolongada de gases de escape de motores diesel se observaron los siguientes efectos tóxicos relacionados con la dosis: aumentos de hasta un 400% del peso pulmonar; inflamación pulmonar cuantificable mediante parámetros bioquímicos (enzimas citoplasmáticas marcadoras, colágeno) y citológicos (aumento de los neutrófilos polimorfonucleares); trastornos de la mecánica pulmonar; aumento del número de macrófagos cargados de partículas con acumulaciones focales (secuestro) en condiciones de sobrecarga; y ulteriores cambios proliferativos de las células epiteliales, con inicio de fibrosis.

La limitada información disponible sobre la toxicidad para la reproducción y el desarrollo lleva a pensar que la inhalación de gases de escape de motores diesel no reviste gran importancia. En la mayoría de los experimentos no se observaron efectos en ratones, ratas, hámsters, conejos o monos; no obstante, tras inyección intraperitoneal se observaron anomalías en el semen de ratones sometidos a partículas de esos gases de escape, y embriotoxicidad en hámsters sometidos a extractos de tales gases.

La mayoría de las pruebas de genotoxicidad in vitro se han llevado a cabo con extractos de gases de escape de motores diesel en lugar de con la mezcla total de tales gases; se detectaron respuestas positivas en ausencia de activación metabólica, es decir: los efectos genotóxicos eran al parecer independientes de los HAP. Aproximadamente la mitad de las investigaciones in vivo arrojaron resultados negativos; los únicos resultados positivos fueron la inducción del intercambio de cromátides hermanas en respuesta a la mezcla total de gases de escape y a extractos orgánicos, y la inducción de micronúcleos en respuesta a extractos orgánicos.

Por lo general no se observaron efectos inmunotóxicos después de la inhalación de gases de escape de motores diesel; sin embargo, en un estudio se halló un mayor título de anticuerpos IgE antiovoalbúmina, y en dos experimentos, una mayor susceptibilidad a las infecciones en ratones. Algunos estudios realizados en la rata parecen indicar que la inhalación de estos gases de escape tiene efectos sobre el comportamiento y el estado neurofisiológico.

En estudios de determinación de la carcinogenicidad de los gases de escape de motores diesel administrados por inhalación a ratas se observó que la fase gaseosa (sin la fracción particulada) no era carcinogénica. En todos los estudios válidos realizados en la rata se halló que los gases eran carcinogénicos a concentraciones de partículas superiores a 2 mg/m 3 , lo que equivale a una exposición

continua de aproximadamente 1 mg/m 3 . No se observaron efectos ni en
el hámster ni en el ratón. En estudios realizados mediante instilación intratraqueal, tanto las partículas de los gases de escape como el negro de humo indujeron la aparición de tumores, y se observó una aparente correlación entre la superficie de las partículas carbonosas y la actividad tumorigénica. La inhalación prolongada de negro de humo prácticamente desprovisto de HAP a concentraciones similares también provocó la aparición de tumores pulmonares en ratas.

No está claro si en los mecanismos de carcinogenicidad de los gases de escape de motores diesel intervienen reacciones del ADN o de otro tipo (o quizá ambas). Se han utilizado diversos modelos para dilucidar la carcinogenicidad de esos gases.

B1.7 Efectos en el ser humano

Los gases de escape de los motores diesel contribuyen a la contaminación atmosférica en general. Aunque en los estudios agudos o crónicos no es posible diferenciar el papel de las partículas diesel, éstas podrían ser parcialmente responsables de los diversos efectos
sobre la salud asociados a la contaminación atmosférica.

Los gases de escape habituales de los motores diesel tienen un olor peculiar, que algunas personas consideran desagradable, sobre todo a altas concentraciones. Los síntomas que siguen a la exposición aguda y crónica a esos gases han sido descritos en estudios y en referencias anecdóticas de personas expuestas en el lugar de trabajo. Se ha observado la aparición de irritación de las membranas mucosas oculares y nasales en respuesta a la exposición aguda a tales gases, así como un aumento de la frecuencia de síntomas respiratorios en cohortes ocupacionales; no obstante, se ignora la contribución de las partículas diesel. No se han detectado efectos sistemáticos a corto plazo en la función pulmonar, pero se han notificado ataques de asma.

En un estudio controlado en el que se expuso a ocho voluntarios sanos no fumadores en una cámara durante 60 minutos a gases de escape de motores diesel diluidos, se detectó una disminución de la fagocitosis en los macrófagos alveolares del líquido de lavado

broncoalveolar.

En algunos estudios transversales y longitudinales realizados entre trabajadores expuestos de forma prolongada en su entorno laboral a esos gases se han hallado decrementos de la función pulmonar y una mayor frecuencia de síntomas respiratorios, pero estos estudios presentan la limitación de haberse realizado con una exposición breve. No se observaron aumentos importantes en estudios de cohortes 

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En varios estudios epidemiológicos se ha evaluado la asociación entre cánceres de pulmón y de la vejiga urinaria y la exposición ocupacional a gases de escape de motores diesel. En esta monografía se incluyen sólo los estudios considerados pertinentes para evaluar los efectos carcinogénicos de esos gases. Los estudios más importantes por lo que se refiere al cáncer de pulmón son los realizados en ferroviarios, trabajadores de garajes de autobuses y estibadores, esto es, cohortes de individuos cuya actividad laboral no deja lugar a dudas sobre la exposición a gases de escape diesel. La totalidad de los cuatro estudios más informativos pusieron de manifiesto un mayor riesgo de cáncer de pulmón, con riesgos relativos que van desde 1,4 para los ferroviarios, pasando por 1,3-2,4 para los trabajadores de garajes de autobuses (según el grado de exposición), hasta 3-6 para los estibadores (según el método empleado para evaluar la exposición, pero con amplios intervalos de confianza). En un estudio de casos y testigos realizados entre ferroviarios y en el estudio de los estibadores fue posible introducir ajustes en función del consumo de tabaco. En los dos casos, el efecto de la exposición al diesel no se vio materialmente alterado por el ajuste realizado en función del consumo de tabaco. En los tres estudios en que no pudo hacerse ese ajuste, el análisis se basó en comparaciones de subgrupos de las cohortes, de manera que la probabilidad de influencia del tabaco como factor de confusión fue menor que al utilizar grupos de comparación

externos.

Se han llevado a cabo varios estudios de casos y testigos para examinar la relación entre el cáncer de vejiga urinaria y la presunta exposición a gases de escape de motores diesel. Se detectó un aumento del riesgo, sobre todo para los conductores de camión; no obstante,
todos estos estudios adolecían de una deficiente caracterización de la exposición. Por ello, no es posible establecer una asociación causal entre la exposición a los gases de escape diesel y un aumento del riesgo de cáncer de vejiga urinaria

B1.8 Efectos en otros organismos en el laboratorio y en el medio
ambiente

El único estudio sobre los efectos de las emisiones de motores diesel propiamente dichas es un trabajo realizado con algas verdes.

B1.9 Evaluación de los riesgos para la salud humana

Se utilizó el paradigma de evaluación de riesgos de la Academia Nacional de Ciencias de los Estados Unidos (National Research Council, 1983) para evaluar los riesgos asociados a variables de valoración relacionadas y no relacionadas con el cáncer. Este proceso consta de los cuatro pasos siguientes: 1) identificación del riesgo, 2) evaluación de la relación dosis-respuesta, 3) evaluación de la exposición, y 4) caracterización del riesgo.

Se considera que los estudios más informativos son los epidemiológicos de larga duración realizados con una exposición bien definida y con seguimiento ulterior (> 20 años). Satisfacían esos criterios cuatro estudios sobre el cáncer de pulmón en personas expuestas en su entorno de trabajo. Los riesgos relativos de cáncer de pulmón asociado a la exposición a gases de escape de motores diesel fueron por lo general bajos y podían haberse debido al azar, a los efectos de factores de confusión no determinados y a la dificultad para corregir con precisión la influencia de los factores de confusión conocidos. Las conclusiones de estos estudios están avaladas por los resultados de otros estudios menos precisos en cuanto a la definición de la exposición. Globalmente se considera que los gases de escape de los motores diesel son probablemente carcinogénicos para el hombre; sin embargo, no se dispone de datos cuantitativos que permitan estimar el riesgo para el ser humano.

B1.9.1 Efectos no neoplásicos

Para caracterizar los riesgos se utilizaron dos métodos generales: el nivel sin efectos adversos observados (NOAEL) dividido por un factor de incertidumbre y una concentración de referencia. En los dos casos se empleó un sofisticado modelo dosimétrico para reducir la incertidumbre asociada a la extrapolación interespecies de las
dosis.

Se calculó que el nivel sin efecto de las partículas de los gases de escape de los motores diesel en el hombre era de 0,139 mg/m 3 . El valor orientativo para la población general calculado a partir del modelo dosimétrico fue de 5,6 μg/m 3 , y el calculado sin el modelo, de 2,3 μg/m 3

.

El método de la concentración de referencia tiene en cuenta la totalidad de la relación exposición-respuesta en lugar de puntos únicos obtenidos en estudios de inhalación, como ocurre con el método
del NOAEL. Se identificaron tres variables de evaluación sensibles: inflamación alveolar crónica, eliminación pulmonar alterada, y lesiones pulmonares hiperplásicas. Las concentraciones de referencia, calculadas mediante el mismo modelo dosimétrico utilizado para el método del NOAEL, fueron de 0,9-2 μg/m 3 para la inflamación, 1,2-3 μg/m 3 para la disminución de la eliminación pulmonar, y
6,3-14 μg/m 3 para las lesiones hiperplásicas.

B1.9.2 Efectos neoplásicos

Se utilizó un modelo polietápico linearizado para hacer estimaciones del riesgo unitario asociado a la exposición a gases de escape de motores diesel. Se consideró que los resultados de los 

# Pa g e 183 of 184 Diesel fuel and exhaust emissions (EHC 171, 1996)

3/8/2011 htt p ://www.inchem.or g /documents/ehc/ehc/ehc171.htmestudios epidemiológicos disponibles eran insuficientes para hacer una estimación cuantitativa del riesgo unitario, por lo que se decidió utilizar datos de varios estudios de inhalación prolongada en los que se había detectado carcinogénesis a concentraciones superiores a 2 mg/m 3 . Se calculó un riesgo unitario de 3,4 × 10 -5 μg/m 3 (media geométrica de cuatro estimaciones del riesgo) para las partículas de los gases de escape. Usando otro modelo biológico se obtuvo un riesgo unitario parecido, en el supuesto de que las partículas diesel influyen en la iniciación y/o proliferación celular a bajas concentraciones.

B1.10 Evaluación de los efectos en el medio ambiente

No se disponía de suficiente información para evaluar los efectos específicos de las emisiones de gases de escape de motores diesel. Los efectos del combustible diesel quemado deben de ser parecidos a los de otros combustibles fósiles y están relacionados con el consumo de

diesel.

See Also: Toxicological Abbreviations 

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