Ecosystem Health 

Science-Based Solutions

Environment Environnement

Canada Canada Canada Science-Based Solutions

Solutions fondées sur la science

Canadian Environmental Quality Guidelines

for Nonylphenol and its Ethoxylates

(Water, Sediment, and Soil)

Report No. 1-3
Prepared and published by

National Guidelines and Standards Office

Environmental Quality Branch

Environment Canada

Ottawa

August, 2002

ISSN 1497-2689

ISBN 0-662-32230-4

Cat. no. En1-34/4-2002E

Scientific Supporting Document

(Water, Sediment, and Soil)

Canadian Environmental Quality

Guidelines for Nonylphenol and its

Ethoxylates

Report No. 1-3

© Her majesty the Queen in Right of Canada, represented by the Minister of the Environment, 2002.

All rights reserved. Reproduction authorized if source is acknowledged. The reproduction must be

presented within its proper context and must not be used for profit.
ii

NOTE TO READERS

The Ecosystem Health: Science-based Solutions series is dedicated to the dissemination of

scientific knowledge, information and tools for monitoring, assessing, and reporting on ecosystem

health to support Canadians in making sound decisions. Documents published in this series

include the scientific basis, methods, approaches and frameworks for environmental quality

guidelines and their implementation; monitoring, assessing, and rehabilitating environmental

quality in Canada; and, indicator development, environmental reporting and data management.

Issues in this series are published ad libitum.

This particular issue provides the scientific supporting information and rationale for the

development of Canadian Environmental Quality Guidelines for nonylphenol and its ethoxylates.

The information in this document is current as of April 2001. Minor revisions and editorial

changes have been made for publication in 2002. For additional information regarding these

guidelines, please contact:

Environment Canada Phone: 819-953-1550

Environmental Quality Branch Fax: 819-953-0461

National Guidelines and Standards Office ceqg-rcqe@ec.gc.ca

351 Saint-Joseph Boulevard http://www.ec.gc.ca/ceqg-rcqe

Hull, Quebec K1A 0H3

Canadian Water and Sediment Quality Guidelines are approved by the Water Quality Task Group

of the Canadian Council of Ministers of the Environment (CCME), and Canadian Soil Quality

Guidelines are approved by the CCME Soil Quality Guidelines Task Group. Environment Canada

acts as the federal representative and serves as the technical secretariat to both of these Task

Groups. These guidelines are included as a 2002 update to the Canadian Environmental Quality

Guidelines, which was published by the CCME in October of 1999. For CCME publications,

please contact:

CCME Documents Phone: 1-800-805-3025

c/o Manitoba Statutory Publications Fax: 204-945-7172

200 Vaughan Street spccme@gov.mb.ca

Winnipeg, MB R3C 1T5 http://www.ccme.ca

This scientific supporting document is available in English only. Factsheets are also available in

English under the title Canadian Environmental Quality Guidelines (CCME 1999). Ce document

scientifique du soutien n’est disponible qu’en anglais avec un résumé en français. Des feuillets

d’information sont aussi disponibles en français sous le titre Recommandations canadiennes pour

la qualité de l’environnement (CCME 1999).

Reference listing:

Environment Canada. 2002. Canadian Environmental Quality Guidelines for Nonylphenol and its

Ethoxylates (Water, Sediment, and Soil). Scientific Supporting Document. Ecosystem Health:

Science-based Solutions Report No. 1-3. National Guidelines and Standards Office,

Environmental Quality Branch, Environment Canada. Ottawa.
iii

TABLE OF CONTENTS

NOTE TO READERS........................................................................................................................ii

TABLE OF CONTENTS...................................................................................................................iii

LIST OF TABLES............................................................................................................................. v

LIST OF FIGURES........................................................................................................................... v

LIST OF APPENDICES....................................................................................................................vi

EXECUTIVE SUMMARY.................................................................................................................vii

RÉSUMÉ.......................................................................................................................................... x

ACKNOWLEDGEMENTS.............................................................................................................. xiii

GLOSSARY OF ACRONYMS........................................................................................................xiv

CHAPTER 1. INTRODUCTION ...................................................................................................... 1

Physical and Chemical Properties ......................................................................................... 2

Production and Uses.............................................................................................................. 6

Sources to the Environment................................................................................................... 8

Water ........................................................................................................................... 8

Sediment...................................................................................................................... 8

Soil............................................................................................................................... 8

Air................................................................................................................................. 9

Analytical Methods ................................................................................................................. 9

CHAPTER 2. CONCENTRATIONS IN CANADA.......................................................................... 11

Surface Waters .................................................................................................................... 11

Sediment.............................................................................................................................. 12

Soil ....................................................................................................................................... 12

Sewage and Sludge ............................................................................................................. 12

Industrial Effluents and Sludge ............................................................................................ 13

CHAPTER 3. ENVIRONMENTAL FATE AND BEHAVIOUR ........................................................ 14

Physical and Chemical Processes....................................................................................... 14

Water ......................................................................................................................... 14

Sediment.................................................................................................................... 15

Soil............................................................................................................................. 16

Biodegradation ..................................................................................................................... 17

Primary Biodegradation ............................................................................................. 17

Ultimate Biodegradation ............................................................................................ 20

Uptake/Elimination Pathways, and Metabolism ................................................................... 21

Bioaccumulation................................................................................................................... 23

Toxicity................................................................................................................................. 29

Mode of Action........................................................................................................... 29

Narcosis .......................................................................................................... 29

Endocrine Disruption....................................................................................... 30

Physical Surfactancy....................................................................................... 32

Toxic Effects .............................................................................................................. 33

Mixtures and the TEQ Approach.......................................................................................... 34

CHAPTER 4. GUIDELINES FROM OTHER JURISDICTIONS .................................................... 37
iv

Water ................................................................................................................................... 37

Sediment.............................................................................................................................. 37

Soil ....................................................................................................................................... 38

CHAPTER 5. DEVELOPMENT OF CANADIAN WATER QUALITY GUIDELINES...................... 41

Freshwater ........................................................................................................................... 41

Toxicity Data .............................................................................................................. 41

Fish ................................................................................................................. 41

Invertebrates ................................................................................................... 41

Plants/Algae.................................................................................................... 41

Guideline Derivation................................................................................................... 42

Marine/Estuarine Water ....................................................................................................... 46

Toxicity Data .............................................................................................................. 46

Fish ................................................................................................................. 46

Invertebrates ................................................................................................... 46

Algae............................................................................................................... 46

Guideline Derivation................................................................................................... 47

Recommended Canadian Water Quality Guidelines ........................................................... 51

Guideline for the Protection of Freshwater Life ......................................................... 51

Guideline for the Protection of Marine Life ................................................................ 51

CHAPTER 6. DEVELOPMENT OF CANADIAN SEDIMENT QUALITY GUIDELINES ................ 52

Toxicity Data ........................................................................................................................ 52

Invertebrates.............................................................................................................. 52

Amphibians ................................................................................................................ 52

Protocol Approaches............................................................................................................ 52

Equilibrium Partitioning Approach........................................................................................ 53

Guideline Derivation................................................................................................... 54

Recommended Canadian Sediment Quality Guidelines...................................................... 55

Freshwater Sediments............................................................................................... 55

Marine Sediments...................................................................................................... 55

CHAPTER 7. DEVELOPMENT OF CANADIAN SOIL QUALITY GUIDELINES........................... 58

Toxicity Data ........................................................................................................................ 58

Microorganisms ......................................................................................................... 58

Plants......................................................................................................................... 59

Invertebrates.............................................................................................................. 59

Mammals ................................................................................................................... 59

Agricultural and Residential/Parkland Land Uses................................................................ 62

Soil Contact................................................................................................................ 62

Nutrient and Energy Cycling Check ................................................................ 64

Soil and Food Ingestion ............................................................................................. 64

Commercial and Industrial Land Uses ................................................................................. 65

Soil Contact................................................................................................................ 65

Nutrient and Energy Cycling Check ................................................................ 67

Groundwater Check ............................................................................................................. 67

Off-site Migration Check ...................................................................................................... 68

Environmental Soil Quality Guideline Derivation (SQGE)..................................................... 68

Recommended Canadian Soil Quality Guidelines ............................................................... 69

CHAPTER 8. AREAS FOR FUTURE RESEARCH....................................................................... 71

Concentrations in Canada.................................................................................................... 71

Fate and Behaviour.............................................................................................................. 71

Toxicity Testing .................................................................................................................... 71

Guideline Derivation and Implementation ............................................................................ 71

CHAPTER 9. GUIDANCE ON IMPLEMENTATION...................................................................... 73
v

General Guidance for all Guidelines .................................................................................... 73

Application of the TEQ Approach to EQGs.......................................................................... 73

Water Quality Guidelines ..................................................................................................... 74

Sediment Quality Guidelines................................................................................................ 74

Soil Quality Guidelines ......................................................................................................... 75

REFERENCES............................................................................................................................... 76

ANNEX I. ECOLOGICAL RELEVANCE OF ENDOCRINE DISRUPTION.................................. 102

ANNEX 2. DATASET REQUIREMENTS FOR DERIVING WATER QUALITY GUIDELINES.... 104

ANNEX 3. SUMMARY OF SEDIMENT QUALITY GUIDELINE PROTOCOL............................. 106

LIST OF TABLES

Table 1. Summary of Canadian Environmental Quality Guidelines for nonylphenol and its

ethoxylates. ....................................................................................................................ix

Tableau 1. Résumé des recommandations canadiennes pour la qualité de l’environnement,

concernant le nonylphénol et ses dérivés éthoxylés .....................................................xii

Table 2. Physical and chemical properties of nonylphenol and select nonylphenol ethoxylates... 5

Table 3. Bioconcentration and bioaccumulation factors for nonylphenol and nonylphenol

ethoxylates in aquatic organisms. ................................................................................ 26

Table 4. Summary of relative toxicity of nonylphenolic and related compounds......................... 36

Table 5. Freshwater quality guidelines for nonylphenol from other jurisdictions......................... 38

Table 6. Freshwater quality guidelines for nonylphenol ethoxylates and ethoxycarboxylates

from other jurisdictions................................................................................................. 39

Table 7. Marine water quality guidelines for nonylphenol from other jurisdictions. ..................... 40

Table 8. Soil and sludge guidelines from other jurisdictions. ...................................................... 40

Table 9. Data used in derivation of guideline for toxicity of nonylphenol to freshwater aquatic

life................................................................................................................................. 43

Table 10. Data used in derivation of guideline for toxicity of nonylphenol to marine aquatic

life................................................................................................................................. 48

Table 11. Primary data available for toxicity of nonylphenol though direct soil contact................ 61

Table 12. Summary of soil quality guideline derivation for nonylphenol. ..................................... 70

Table 13. Summary of the available biological effects and related physicochemical data for

sediment-associated nonylphenol (mg·kg-1 dry weight) in freshwater ecosystems.... 112

Table 14. Summary of the available biological effects and related physicochemical data for

sediment-associated nonylphenol (mg·kg-1 dry weight) in marine ecosystems ......... 114

LIST OF FIGURES

Figure 1. The structure of nonylphenol and its derivatives............................................................. 3

Figure 2. Pathways for biodegradation of nonylphenol polyethoxylates....................................... 19

Figure 3. Select freshwater toxicity data for nonylphenol............................................................. 45

Figure 4. Select marine toxicity data for nonylphenol................................................................... 50

Figure 5. Distribution of nonylphenol concentrations in freshwater sediments that are

 associated with adverse biological effects (l) and no adverse effects (m).................. 57

Figure 6. Distribution of nonylphenol concentrations in marine and estuarine sediments that

 are associated with adverse biological effects (l) and no adverse effects (m)............ 57

Figure 7. Overview of steps leading to the development of final Environmental Soil Quality

Guidelines for nonylphenol........................................................................................... 60

Figure 8. Distribution of soil contact toxicity data used in the guideline derivation for

 agricultural and residential/parkland land uses. ............................................................ 63

Figure 9. Distribution of soil contact effects toxicity data used in guideline derivation for

commerical and industrial land uses. ........................................................................... 66
vi

LIST OF APPENDICES

Appendix I. Toxicity of nonylphenol and its ethoxylates to freshwater life through water

column exposure

a. Toxicity of nonylphenol to freshwater algae and macrophytes through water

 column exposure

b. Toxicity of nonylphenol to freshwater invertebrates through water column

 exposure

 c. Toxicity of nonylphenol to freshwater fish through water column exposure

d. Toxicity of nonylphenol ethoxylates to freshwater algae and macrophytes

 through water column exposure

e. Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water

 column exposure

f. Toxicity of nonylphenol ethoxylates to freshwater fish through water column

 exposure

Appendix II. Toxicity of nonylphenol and its ethoxylates to marine aquatic life through water

column exposure

a. Toxicity of nonylphenol to marine algae through water column exposure

b. Toxicity of nonylphenol to marine invertebrates through water column

 exposure

c. Toxicity of nonylphenol to marine fish through water column exposure

d. Toxicity of nonylphenol ethoxylates to marine algae and plants through

 water column exposure

e. Toxicity of nonylphenol ethoxylates to marine invertebrates through

 water column exposure

f. Toxicity of nonylphenol ethoxylates to marine fish through water column

 exposure

Appendix III. Toxicity of nonylphenol to sediment-dwelling life

a. Toxicity of nonylphenol to freshwater life through spiked-sediment exposure

b. Toxicity of nonylphenol to marine life through spiked-sediment exposure

Appendix IV. Toxicity of nonylphenol and its ethoxylates to terrestrial life

a. Toxicity of nonylphenol to soil microorganisms

 b. Toxicity of nonylphenol to soil-dwelling invertebrates

c. Toxicity of nonylphenol to terrestrial plants

d. Toxicity of nonylphenol to terrestrial vertebrates

e. Toxicity of nonylphenol ethoxylates to terrestrial organisms
vii

EXECUTIVE SUMMARY

This scientific supporting document provides the rationale for the development of Canadian

Environmental Quality Guidelines for nonylphenol (NP) and nonylphenol ethoxylates (NPEs). It

contains a review of technical background information on the chemical and physical properties of

NP and NPEs, a review of sources and emissions in Canada, the distribution and behaviour of NP

and NPEs in the environment, and their toxicological effects on a variety of aquatic and terrestrial

species. This information is used to derive water quality, sediment quality, and soil quality

guidelines for NP and NPEs to protect ecological receptors in each of these environmental

compartments. All of the Canadian Environmental Quality Guidelines in this document are based

on the best available toxicity data at the time of writing. The most recent literature search was

conducted in August 2000, but some information obtained since that date has also been included

in response to reviewers’ comments.

NPEs are a group of nonionic organic chemicals that are used as surfactants, detergents,

emulsifiers, wetting agents, and dispersants in a wide variety of industrial processes and

commercial products. They are primarily released to the environment through sewage effluents or

industrial wastewaters. In sewage treatment systems, and upon entering the environment, NPEs

readily undergo an initial primary degradation which largely involves the loss of ethoxylate groups.

This results in the formation of more refractory degradation products such as nonylphenol (NP),

nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), nonylphenoxy acetic

acid (NP1EC), and nonylphenoxy ethoxy acetic acid (NP2EC).

The major pathway for nonylphenolic substances to enter the environment is through the

discharge of municipal and industrial wastewaters into surface waters. Because they are used as

formulants for some pesticides, nonylphenolic substances may also be deposited on surface

waters through aerial spraying of pesticides, or as runoff from agricultural land or forests which

have been sprayed. Once in the aquatic environment, the more hydrophobic chemicals, such as

NP, will tend to adsorb to particulates and accumulate in bottom sediments. In wastewater

treatment plants, NP will also partition to the sludge, and may then become a contaminant of

terrestrial systems when the sludge is landfilled or applied as a soil amendment to agricultural

soils. Leaching of nonylphenolic substances into groundwater has also been documented.

It has been proposed that nonylphenol and its ethoxylates be designated as “CEPA toxic” under

the Canadian Environmental Protection Act (CEPA 1999). Large quantities of NPEs are used in

Canada each year. Therefore, their degradation products (i.e., NP and short-chain NPEs), which

are more persistent, toxic, and estrogenic, may be released at concentrations that could affect

aquatic and terrestrial life in the receiving environments. In light of these possible impacts, the

development of Canadian Environmental Quality Guidelines (CEQGs) has been deemed

necessary. CEQGs are nationally-endorsed, science-based goals that are recommended as

levels that should result in negligible risk to biota and ecosystem health. These guidelines are

developed according to the procedures described in nationally approved protocols for the

derivation of Canadian Environmental Quality Guidelines (CCME 1991; CCME 1995; CCME

1996).

A summary of the Canadian Environmental Quality Guidelines developed in this document is

provided in Table 1. Guidelines in this document were developed based on data for nonylphenol,

but, as described in Chapter 3, it is recommended that a toxic equivalency (TEQ) approach be

used to apply these guidelines to mixtures of nonylphenolic substances. When comparing an

environmental sample to the guidelines, concentrations of each nonylphenolic substance in the

sample should be multiplied by their respective toxic equivalency factors (TEFs) and then

summed to yield a total concentration in NP TEQ units. Based on the toxicity of nonylphenol

ethoxylates and nonylphenoxy carboxylates relative to nonylphenol, an initial set of TEFs has been

recommended. Future work to develop and validate ethoxamer-specific TEF values is needed,

but, in the interim, it is recommended that the TEFs presented here be applied.
viii

Tissue residue guidelines in aquatic biota for the protection of wildlife consumers of aquatic life

have not been derived due to the lack of available toxicity data for mammalian and avian species.

Sufficient toxicity data were available for the derivation of a full freshwater water quality guideline,

but not for a marine water quality guideline. The freshwater guideline was based on the most

sensitive chronic endpoint from a primary study, which was a 91-day lowest observed effect

concentration (LOEC) for growth reduction in rainbow trout at a nonylphenol concentration of

10.3 μg·L-1 (Brooke 1993a). This concentration was multiplied by a safety factor of 0.1 to obtain a

water quality guideline for nonylphenol and its ethoxylates of 1.0 μg·L-1, expressed in NP TEQ

units, for the protection of freshwater aquatic life. The toxicity data available for marine aquatic

species were sufficient to meet the minimum data requirements for an interim water quality

guideline. The critical study was a 28-day test with Americamysis bahia which had a LOEC for

growth reduction of 6.7 μg·L-1 (Ward and Boeri 1991a). Multiplying this by a safety factor of 0.1,

an interim water quality guideline of 0.7 μg·L-1, expressed in NP TEQ units, is recommended for

the protection of marine (including estuarine) aquatic life. These guidelines, which refer to

concentrations in unfiltered samples, are intended to protect all forms of aquatic life and all

aspects of aquatic life cycles during an indefinite period of exposure to the water column (CCME

1991). Current limits of detection for nonylphenolic substances in water range from 0.01 μg·L-1

(for NP) to 1.6 μg·L-1 (for higher molecular weight NPEs).

For the development of Canadian sediment quality guidelines, there were insufficient data to meet

the minimum requirements for any of the three approaches described in the protocol (i.e., National

Status and Trends Program Approach (NSTP), Spiked-Sediment Toxicity Test Approach (SSTT),

adoption of guidelines from other jurisdictions). Instead, an equilibrium partitioning approach was

used to derive provisional interim sediment quality guidelines (ISQGs) from the Canadian water

quality guidelines. The revision and re-assessment of these provisional ISQGs will be possible

when sufficient data become available to support the use of the modified NSTP or SSTT

approaches as described in the formal protocol (CCME 1995). Based on equilibrium partitioning

calculations, a provisional interim sediment quality guideline of 1.4 mg·kg-1 dry weight, expressed

in NP TEQ units, is recommended for the protection of freshwater aquatic life. For the protection

of marine (including estuarine) life, a provisional interim sediment quality guideline of 1.0 mg·kg-1

dry weight, expressed in NP TEQ units, is recommended. These guidelines are normalized to 1%

total organic carbon, but should be adjusted based on local levels of organic carbon in the

sediments. These guidelines are intended to be protective of all forms of aquatic life and all

aspects of aquatic life cycles during an indefinite period of exposure to bed sediments (CCME

1995). These guidelines refer to total concentrations of nonylphenolic substances in surficial

sediments (i.e., top 5 centimetres), as quantified by extraction using standard analytical methods

(e.g., soxhlet extraction, steam distillation, supercritical fluid extraction) followed by detection with

high pressure liquid chromatography or gas chromatography/mass spectrometry. Current limits of

detection for nonylphenolic substances in sediment range from 0.015 to 0.1 mg·kg-1 dry weight.

There were sufficient soil toxicity data to meet the minimum requirements for a weight of evidence

method for soil contact guideline derivation. However, adequate data were not available for the

derivation of a food ingestion guideline value or a nutrient and energy cycling check. For

agricultural and residential/parkland land uses, the soil contact guideline was calculated as the

25th percentile of the effects and no effects data distribution, divided by an uncertainty factor of 5.

For commercial and industrial land uses, the soil contact guidelines were calculated as the 25th

percentile of the effects data distribution only. Therefore, soil quality guidelines for nonylphenol

and its ethoxylates for agricultural and residential/parkland land use are both recommended at

5.7 mg·kg-1, expressed in NP TEQ units. For both commercial and industrial land uses, soil

quality guidelines of 14 mg·kg-1, expressed in NP TEQ units, are recommended. These

guidelines are optimized for soils within the pH range of 5.8 to 7.5 because the toxicological

studies upon which they are based were conducted within this range. The current limit of

detection for nonylphenol in soil is 0.05 mg·kg-1. A groundwater check of 21 mg·kg-1, expressed in
ix

NP TEQ units, was calculated as the concentration of NP and NPEs in soil that should not be

exceeded in order to protect freshwater aquatic life from possible groundwater contamination.

Table 1. Summary of Canadian Environmental Quality Guidelines for nonylphenol and its

ethoxylates.

Environmental Media Media Type Guideline Type Guideline Value*

Water Freshwater Full 1.0 μg·L-1

Marine Interim 0.7 μg·L-1

Sediment †Freshwater Provisional Interim 1.4 mg·kg-1

Marine Provisional Interim 1.0 mg·kg-1

Soil Agricultural SQGE

‡

 5.7 mg·kg-1

Residential/Parkland SQGE 5.7 mg·kg-1

Commercial SQGE 14 mg·kg-1

Industrial SQGE 14 mg·kg-1

Groundwater Check value 21 mg·kg-1

* all guidelines are expressed in nonylphenol toxic equivalent units

†

 these guideline values are for sediments containing 1% total organic carbon

‡

 environmental soil quality guideline; a full soil quality guideline would also consider protection

for human health (CCME 1996)
x

RÉSUMÉ

Le présent document d’évaluation scientifique présente les principes qui sont à la base de

l’élaboration des Recommandations canadiennes pour la qualité de l'environnement pour le

nonylphénol (NP) et les dérivés éthoxylés du nonylphénol (NPE). Il contient une revue de

l’information technique de base sur les propriétés chimiques et physiques du NP et des NPE, les

sources et les émissions au Canada, la distribution et le comportement des NP et des NPE dans

l’environnement et enfin leurs effets toxicologiques sur diverses espèces aquatiques et terrestres.

Cette information sert à l’élaboration de recommandations pour la qualité de l’eau, des sédiments

et des sols concernant le NP et les NPE, de façon à protéger les récepteurs écologiques dans

chacun de ces compartiments de l’environnement. Toutes les Recommandations canadiennes

pour la qualité de l'environnement présentées dans le présent document sont basées sur les

meilleures données toxicologiques disponibles au moment de sa rédaction. Les recherches de

littérature les plus récentes ont été effectuées en août 2000, mais certains renseignements

obtenus depuis cette date ont également été inclus suite aux commentaires des examinateurs.

Les NPE sont un groupe de substances chimiques, organiques et non ioniques, utilisées comme

tensioactifs, détergents, émulsifiants, agents de mouillage et dispersants dans une vaste gamme

de procédés industriels et de produits commerciaux. Ils sont principalement rejetés dans

l’environnement via les effluents d’eaux usées provenant des égouts ou les eaux usées

industrielles. Les NPE sont soumis à une dégradation primaire initiale impliquant principalement

une perte de groupes éthoxy dans les installations de traitements d’eaux usées et à leur entrée

dans l’environnement. Des produits de dégradation plus réfractaires, comme le nonylphénol (NP),

le nonylphénol monoéthoxylé (NP1EO), le nonylphénol diéthoxylé (NP2EO), l’acide

nonylphénoxyacétique (NP1EC) et l’acide nonylphénoxyéthoxyacétique (NP2EC) sont alors

formés.

Les substances nonylphénoliques pénètrent l’environnement majoritairement par la voie des

rejets d’eaux usées des municipalités et des usines à travers les eaux de surface. Étant donné

qu’elles sont employées comme produits de formulation dans certains pesticides, les substances

nonylphénoliques peuvent également être déposées sur les eaux de surface suivant l’épandage

aérien de pesticides, ou encore, être lessivées à partir de terres agricoles ou forestières traitées

par pulvérisation. Dans le milieu aquatique, les substances chimiques les plus hydrophobes,

comme le NP, auront tendance à être adsorbées sur les particules et à s’accumuler dans les

sédiments de fond. Dans les usines de traitement des eaux usées, une partie du NP sera

également distribuée dans les boues et pourra alors devenir un contaminant des systèmes

terrestres, lorsque ces boues sont utilisées comme remblai ou appliquées comme amendement

du sol sur les terres agricoles. La documentation mentionne le lessivage des substances

nonylphénoliques dans les eaux souterraines.

Il a été proposé que le nonylphénol et ses dérivés éthoxylés soient désignés « toxiques » sous la

Loi canadienne sur la protection de l'environnement (LCPE 1999). De grandes quantités de NPE

sont utilisées chaque année, au Canada. Par conséquent, leurs produits de dégradation (c.-à-d. le

NP et les NPE à courte chaîne), qui sont persistents, toxiques et qui possèdent un effet

oestrogène plus marqués, peuvent être rejetés à des concentrations qui pourraient altérer la vie

aquatique et terrestre dans les milieux récepteurs. À la lumière de ces impacts possibles,

l’élaboration des Recommandations canadiennes pour la qualité de l'environnement (RCQE) s’est

avérée nécessaire. Les RCQE, approuvées à l’échelle nationale et fondées sur des données

scientifiques solides, sont des objectifs recommandés sous forme de concentrations qui ne

devraient entraîner qu’un risque négligeable pour la santé du biote et de l’écosystème. Ces

recommandations ont été élaborées selon les méthodes décrites dans les protocoles approuvés à

l’échelle nationale visant l’établissement des Recommandations canadiennes pour la qualité de

l’environnement (CCME, 1991; CCME, 1995; CCME, 1996).
xi

Les Recommandations canadiennes pour la qualité de l'environnement élaborées dans le présent

document sont résumées dans le tableau 1. Les recommandations figurant dans ce document

sont fondées sur les données relatives au nonylphénol, mais, comme décrit au chapitre 3, il est

conseillé d’utiliser une approche d’équivalent toxique (QÉT) lors de l’application de ces

recommandations à des mélanges de substances nonylphénoliques. En comparant un échantillon

de l’environnement aux recommandations, les concentrations de chaque substance

nonylphénolique présente dans l’échantillon devraient être multipliées par leur facteur

d’équivalence de toxicité (FÉT) respectif, et additionnées pour donner la concentration totale en

unités de QÉT de NP. Un ensemble initial de FÉT, fondé sur la toxicité des dérivés éthoxylés et

carboxylés du nonylphénol, a été recommandé. D’autres travaux seront nécessaires pour obtenir

et valider les valeurs FÉT pour des congénères éthoxylés individuels, mais, d’ici là, il est

recommandé d’appliquer les FÉT présentés ici.

Les recommandations pour les résidus dans les tissus du biote aquatique, visant à protéger les

espèces fauniques consommant les organismes aquatiques, n’ont pas été déterminées en raison

du manque de données de toxicité disponibles pour les mammifères et les oiseaux.

Suffisamment de données de toxicité étaient disponibles pour l’élaboration d’une recommandation

complète concernant la qualité de l’eau douce, mais non pour une recommandation concernant la

qualité de l’eau marine. La recommandation pour l’eau douce a été basée sur l’effet toxicologique

chronique le plus sensible provenant d’une étude primaire de 91 jours, dont la concentration

minimale avec effet observé (CMEO) de 10,3 μg·L-1 (Brooke 1993a) de nonylphénol a entraîné

une réduction de la croissance chez la truite arc-en-ciel. Afin de protéger la vie aquatique en eau

douce, cette concentration a été multipliée par un facteur de sécurité de 0,1 pour obtenir une

recommandation de 1,0 μg·L-1, exprimée en unités de QÉT de NP, pour la qualité de l’eau vis à

vis le nonylphénol et ses dérivés éthoxylés. Les données de toxicité disponibles pour les espèces

aquatiques marines étaient suffisantes pour répondre aux exigences de données minimales pour

élaborer une recommandation provisoire pour la qualité de l’eau. L’étude critique était un essai de

28 jours avec Americamysis bahia démontrant une réduction de croissance associée à une

CMEO de 6,7 μg·L-1 (Ward et Boeri, 1991a). En multipliant cette valeur par un facteur de sécurité

de 0,1, une recommandation provisoire pour la qualité de l’eau de 0,7 μg·L-1 est obtenue, en

unités de QÉT de NP, aux fins de la protection de la vie aquatique marine (incluant la vie

estuarienne). Ces recommandations, qui correspondent à des concentrations dans des

échantillons non filtrés, visent à protéger toutes les formes de vie aquatique et tous les aspects de

leurs cycles de vie pendant une période indéfinie d’exposition à la colonne d’eau (CCME, 1991).

Les limites de détection actuelles des substances nonylphénoliques dans l’eau se situent dans

une plage de 0,01 μg·L-1 (pour le NP) à 1,6 μg·L-1 (pour les NPE de masse moléculaire plus

élevée).

Aux fins de l’élaboration des recommandations canadiennes pour la qualité des sédiments, les

données disponibles étaient insuffisantes pour satisfaire aux exigences minimales de n’importe

laquelle des trois démarches décrites dans le protocole (« National Status and Trends Program »

(NSTP), tests de toxicité des sédiments avec dopage (TTSD) et adoption de recommandations

provenant d’autres jurisdictions). Une approche de partage à l’équilibre a plutôt été adoptée pour

élaborer une version intérimaire des recommandations provisoires pour la qualité des sédiments

(RPQS) à partir des recommandations canadiennes pour la qualité de l’eau. La révision et la

réévaluation de ces RPQS provisoires seront possibles lorsque les données seront suffisantes

pour justifier l’emploi des variantes des méthodes NSTP et TTSD, telles que décrites dans le

protocole officiel (CCME 1995). Basée sur des calculs de partage à l’équilibre, une version

intérimaire de la RPQS de 1,4 mg·kg-1 de poids sec, exprimée en unités de QÉT de NP, fut

déterminée pour la protection de la vie aquatique en eau douce. Pour la protection de la vie

marine (y compris la vie estuarienne), une version intérimaire de la RPQS de 1,0 mg·kg-1 de poids

sec, exprimée en unités de QÉT de NP est proposée. Ces recommandations sont normalisées à

1 % de carbone organique total, mais devraient être ajustées en fonction des concentrations

locales de carbone organique dans les sédiments. Les recommandations visent à protéger toutes

les formes de vie aquatique et tous les aspects de leurs cycles de vie pendant une période
xii

indéfinie d’exposition aux sédiments du lit (CCME, 1995). Ces recommandations correspondent

aux concentrations totales de substances nonylphénoliques dans les sédiments de surface (c.-àd. dans la couche supérieure de 5 cm), mesurées par extraction à l’aide de méthodes analytiques

standardisées (p. ex. extraction au soxhlet, distillation à la vapeur, extraction par fluide

supercritique), suivie de détection par chromatographie liquide à haute pression ou

chromatographie gazeuse/spectrométrie de masse. Les limites de détection actuelles des

substances nonylphénoliques dans les sédiments se situent dans une plage de 0,015 à 0,1

mg·kg-1 de poids sec.

Suffisamment de données de toxicité étaient disponibles en ce qui a trait au sol pour satisfaire

aux exigences minimales d’une méthode fondée sur le poids de la preuve permettant d’élaborer

une recommandation pour le contact avec le sol. Par contre, un manque de données ne

permettait pas l’obtention d’une valeur recommandée concernant l’ingestion par les aliments ni

une vérification du cycle nutritif et énergétique. Dans le cas des utilisations sur les terres à

vocation agricole, résidentielle, ou encore destinées aux parcs, la recommandation pour le

contact avec le sol a été calculée au 25e percentile de la distribution des données avec et sans

effets, valeur par la suite divisée par un facteur d’incertitude de 5. Pour les utilisations sur les

terres à vocation commerciale ou industrielle, les recommandations pour le contact avec le sol ont

été calculées au 25e percentile de la distribution des données avec effets seulement. Par

conséquent, les recommandations pour la qualité du sol dans le cas du nonylphénol et de ses

dérivés éthoxylés utilisés sur les terres à vocation agricole, résidentielle, ou encore destinées aux

parcs sont toutes deux de 5,7 mg·kg-1, en unités de QÉT de NP. Pour les utilisations sur les terres

à vocation tant commerciale qu’industrielle, les recommandations pour la qualité du sol sont de

14,2 mg·kg-1, en unités de QÉT de NP. Ces recommandations sont optimalisées pour les sols

dont le pH se situe entre 5,8 à 7,5, du fait que les études toxicologiques sur lesquelles elles sont

fondées ont été effectuées dans cette étendue. La limite de détection actuelle pour le nonylphénol

dans le sol est 0,05 mg·kg-1. Une valeur de vérification d’eau souterraine de 21,2 mg·kg-1, en

unités de QÉT de NP, a été calculée comme concentration de NP et de NPE du sol à ne pas

dépasser afin de protéger la vie aquatique en eau douce contre une contamination possible par

l’eau souterraine.

Tableau 1. Résumé des recommandations canadiennes pour la qualité de l’environnement,

concernant le nonylphénol et ses dérivés éthoxylés

Milieu environnemental Type de milieu Type de recommandation Valeur recommandée*

Eau Eau douce Complète 1,0 μg·L-1

Eau marine Provisoire 0,7 μg·L-1

Sédiments †Eau douce Provisoire (intérimaire) 1,4 mg·kg-1

Eau marine Provisoire

(intérimaire)

 1,0 mg·kg-1

Sol Agricole RQSE

‡

 5,7 mg·kg-1

Résidentiel/parcs RQSE 5,7 mg·kg-1

Commercial RQSE 14 mg·kg-1

Industriel RQSE 14 mg·kg-1

Eau souterraine Valeur pour vérification 21 mg·kg-1

* toutes les recommandations sont exprimées en unités d’équivalent toxique de NP

†

 ces recommandations sont valables pour des sédiments renfermant 1% de carbone organique

total

‡

 recommandation pour la qualité environnementale du sol; une recommandation complète pour

la qualité du sol tiendrait également compte de la protection de la santé humaine (CCME,

1996)
xiii

ACKNOWLEDGEMENTS

This scientific supporting document for the development of Canadian Environmental Quality

Guidelines for nonylphenol and its ethoxylates was prepared by the National Guidelines and

Standards Office, Environmental Quality Branch of Environment Canada. Authors of this

document include Kelly Potter, Kathie Adare and Sherri Smith (National Guidelines and Standards

Office, Environment Canada). Special thanks are extended to Philippa Cureton, Nicole Davidson,

and Thea Rawn (Commercial Chemicals Evaluation Branch, Environment Canada), and to MarcAndré Roy (Ministère de l’Environnment du Québec) for sharing information and references.

This document incorporates review comments received from various scientists representing

federal and provincial government organizations, academic institutions, and the private sector.

Thanks are extended to all those who provided input, including: Pierre-Yves Caux, Janet McCann,

Elizabeth Roberts, Susan Roe, Doug Spry, and Lise Trudel (National Guidelines and Standards

Office, Environment Canada); Bijan Aidun, Isabelle Guay, and Darrell Taylor (CCME Water

Quality Guidelines Task Group); Ruth Hall, Marius Marsh, Ted Nason, Edwin Yee (CCME Soil

Quality Guidelines Task Group); Nicole Davidson, Lucie Desforges, Dominique Dore, MarieLouise Geadah, Colin Gray, Catherine Jefferson, Céline Labossière, Bill Lee, Paul Mudroch, Thao

Pham, Linda Porebski, Louise Power, Thea Rawn, Chantal Roberge, Danielle Rodrique, Mark

Servos, Cecelia Wong (Environment Canada); Wayne Fairchild (Fisheries and Oceans Canada);

Donald Rodier (U.S. Environmental Protection Agency), Nile Kemble (U.S. Geological Survey);

Sylvia Talmage (Oak Ridge National Laboratory); Vince Kramer, Barbara Losey, Richard Mast,

Ellen Mihaich, Carter Naylor, Charles Staples (Alkylphenols and Ethoxylates Research Council).

Funding for the development of Canadian Environmental Quality Guidelines for Nonylphenol and

its Ethoxylates was provided by Environment Canada (National Guidelines and Standards Office,

Environmental Quality Branch).
xiv

GLOSSARY OF ACRONYMS

AP alkylphenol

APE alkylphenol ethoxylate (as a general class)

APnEO alkylphenol ethoxylate, where n = specific number of EO groups

BEDS Biological Effects Database for Sediments

BOD, COD biological oxygen demand, chemical oxygen demand

CAS Chemical Abstracts Service

CCME Canadian Council of Ministers of the Environment

CEC cation exchange capacity

CEPA Canadian Environmental Protection Act

DTED daily threshold effects dose

DT50, DT95 mean time to 50% dissipation, mean time to 95% dissipation

dw dry weight

ECL, ERL effects concentration low, effects range low

EC50, ED50 median effective concentration, median effective dose

EDC endocrine disrupting chemical

EO ethoxylate group

EQGs Environmental Quality Guidelines

GC/MS gas chromatography/mass spectrometry

HLB hydrophile-lipophile balance

HPLC high pressure liquid chromatography

ISQG interim sediment quality guideline

LC50 median lethal concentration

LOEC, LOEL lowest observed effect concentration, lowest observed effect level

LOAEL lowest observed adverse effect level

NE no effect

NOEC, NOEL no observed effect concentration, no observed effect level

NP nonylphenol

NPEC nonylphenol ethoxycarboxylate or nonylphenoxy acetic acid (as a general class)

NPnEC nonylphenol ethoxycarboxylate, where n = specific number of EO groups

NPE nonylphenol ethoxylate (as a general class)

NPnEO nonylphenol ethoxylate, where n = specific number of EO groups

NPER no potential effects range

NR not reported

NSTP National Status and Trends Program

OECD Organisation for Economic Co-operation and Development

OP octylphenol

OPnEO octylphenol ethoxylate, where n = specific number of EO groups

PEL probable effect level

PSL Priority Substance List

QSAR quantitative structure-activity relationship

SQGE, SQGHH environmental soil quality guideline, human health soil quality guideline

SQGI, SQGSC soil quality guideline for ingestion, soil quality guideline for soil contact

SSTT Spiked Sediment Toxicity Test

WWTP wastewater treatment plant

TEC, TEL threshold effect concentration, threshold effect level

TEF, TEQ toxic equivalency factor, toxic equivalency

TOC total organic carbon

UF uncertainty factor

USEPA United States Environmental Protection Agency

WQG water quality guideline

ww wet weight
1

CHAPTER 1. INTRODUCTION

Nonylphenol (NP) is a synthetic substance (typically an isomeric mixture) that belongs to the class

of organic chemicals known as alkylphenols (APs), and consists of a nine-carbon alkyl chain

attached to a phenol ring (Fig. 1). APs, such as NP, may be used to produce various derivatives,

including a class of nonionic surfactants1 called alkylphenol ethoxylates (APEs). APEs are

composed of a phenol group attached to both an alkyl chain [i.e., -(CH2)nH] and an ethoxylate

chain [i.e., -O(CH2CH2O)nH]. In the case of nonylphenol ethoxylates (NPEs), the alkyl chain

contains nine carbon atoms, and the ethoxylate chain may range in length from 1 to 100

ethoxylate groups.

As nonionic surfactants, NPEs are commonly used in a wide variety of industrial processes, as

well as in cleaning products for residential and commercial use. Upon their release to the

environment, the major degradation product of NPEs is NP. Therefore, NP is both the parent

compound of NPEs during production, and a breakdown product of NPEs upon degradation.

There is concern about the occurrence of NP and NPEs in the Canadian environment for a

number of reasons. NPEs are released through sewage and industrial effluents in large

quantities, and although they undergo a quick initial degradation, their degradation products,

including nonylphenol, are much more persistent. NP and NPEs have shown evidence of toxicity

to both aquatic and terrestrial organisms, including effects on endocrine systems.

Nonylphenol and its ethoxylates are among the substances on the second Priority Substances List

(PSL2) that were required to undergo an environmental risk assessment under the Canadian

Environmental Protection Act (CEPA, Bill C-32, June 1, 1999). The draft PSL2 assessment of

nonylphenol and its ethoxylates has found that nonylphenol is entering the Canadian environment

in quantities or concentrations, or under conditions, that are having, or may have, a harmful effect

on the environment or its biological diversity (Government of Canada 2000a). Therefore, it has

been proposed that nonylphenol and its ethoxylates be designated as “toxic” according to Section

64 of CEPA. An estimated no-effects value (ENEV) of 1.0 μg·L-1 was calculated from available

water toxicity data for the protection of aquatic biota (Servos et al. 2000). Risk management

strategies, that may be developed as a follow-up to the CEPA assessment and the likely

designation of NPEs and NP as toxic, will require the use of targets or benchmarks for ensuring

sustainability of ecosystem health. Therefore, in complement to the CEPA assessment, this

report has been undertaken to develop Canadian Environmental Quality Guidelines (EQGs) to

provide such benchmarks for NP and NPEs.

Under the auspices of the Canadian Council of Ministers of the Environment (CCME), Canada

has been developing environmental quality guidelines for water, sediment, soil, and tissue quality

since the 1980s (CCREM 1987; CCME 1999). Canadian EQGs are nationally endorsed, sciencebased goals that are recommended as levels that should result in negligible risk to biota and

ecosystem health. These guidelines serve as benchmarks or indicators of environmental quality

that can be used nationally for the protection, evaluation, and enhancement of the Canadian

environment and its beneficial uses. Often the Canadian EQGs form the scientific basis for sitespecific criteria, guidelines, objectives, or standards set by the various Canadian provincial and

territorial jurisdictions.

This document describes the development of Canadian EQGs for nonylphenol and its ethoxylates

for the protection of water, sediment and soil quality. These guidelines are numerical

concentrations that are recommended to support and maintain aquatic (both freshwater and

marine), sediment-dwelling (both freshwater and marine), and terrestrial life, and are developed

 

1

 Nonionic surfactants (as opposed to cationic, anionic, or amphoteric surfactants) contain no

ionic constituents and produce electrically neutral colloidal particles in solution (Davidsohn and

Milwidsky 1967).
2

from the available scientific information on the biological effects of nonylphenolic compounds in

the environment. Water, sediment and soil have been identified as the key compartments of the

environment for which Canadian EQGs should be developed as a result of the potential for

impacts on the beneficial uses of the environment from NP and NPEs. Tissue residue guidelines

in aquatic biota for the protection of wildlife consumers of aquatic life have not been derived due

to the lack of available toxicity data for mammalian and avian species. Nationally approved

protocols have been followed in developing Canadian EQGs for the protection of water, sediment

and soil quality (CCME 1991; CCME 1995; CCME 1996).

All of the guidelines that have been developed in this document are based on toxicity data for

nonylphenol; however, they may be applied to mixtures of nonylphenol, nonylphenol ethoxylates

and nonylphenol ethoxycarboxylates (NPECs) with the use of toxic equivalents. In Chapters 3

and 9, guidance is provided on how to account for NPEs and NPECs when implementing the

guidelines.

In addition to describing the development of environmental quality guidelines, this document also

reviews other available scientific information on nonylphenol and its ethoxylates. This discussion

includes a review of the physical and chemical properties of NP and NPEs, sources of these

substances to the environment, analytical methods used to detect NP and NPEs, levels of these

substances found in the various compartments of the Canadian environment, and their

environmental fate and behaviour. Although the EQGs are solely developed using toxicological

data, this additional information is discussed in order to provide important considerations for the

implementation of the guidelines. More detailed information on some of these aspects has also

been summarized and reviewed to support the CEPA PSL2 assessment of NP and NPEs and can

be found elsewhere (see Servos et al. 2000).

Physical and Chemical Properties

Nonylphenol is a generic term used to refer to a group of isomeric compounds, each consisting of

a nine-carbon alkyl chain (-C9H19) attached to a phenol group, with the molecular formula

C15H24O. The various isomers can differ both in the degree of alkyl chain branching, and in the

position on the phenol ring at which the alkyl chain is attached. Most nonylphenol in production is

of the form 4-nonylphenol (i.e., attachment at the para-position) with varied alkyl chain branching.

Very little straight chain nonylphenol (n-nonylphenol) is produced. Commercially produced

4-nonylphenol may also contain small amounts of 2-nonylphenol (i.e., attachment of the nonyl

chain to the ortho-position of the phenol ring), or 2,4-dinonylphenol (i.e., two nonyl chains attached

to a phenol ring) as impurities (European Commission 1999). The Chemical Abstract Service

(CAS) registry number for a mixture of various branched isomers of nonylphenol is 84852-15-3

(which may be followed by an asterisk) and is given the IUPAC name of “4-nonylphenol

(branched)”. Some of the individual branched isomers also have their own specific CAS numbers.

Straight chain nonylphenol goes by the IUPAC name of “nonylphenol” and has the CAS number

25154-52-3. NP may also be referred to as “isononylphenol” (CAS number 11066-49-2) or

“phenol, nonyl-, branched” (CAS number 90481-04-2) (European Commission 1999). In this

document, the term nonylphenol is used to refer to all isomers and is best represented by CAS

number 84852-15-3. Data specific to particular isomers generally have not been differentiated.

The major derivatives of nonylphenol are nonylphenol ethoxylates (NPEs) and nonylphenol

ethoxycarboxylates (or nonylphenoxy acetic acids, NPECs) (Figure 1). Nonylphenol ethoxylates

have the general formula C9H19-C6H4O(CH2CH2O)nH, with n ranging from 1 to 100, but with the

vast majority of commercially produced NPEs containing between 6 and 12 ethoxylate (EO)

groups. Specific NPE oligomers may be reported as NPnEO, where n refers to the mean number

of EO groups in the ethoxylate chain. For example, an oligomer with 9 EO groups is referred to

as NP9EO. The term NP9EO may also refer to a mixture of various oligomers for which the mean

number of EO groups per molecule is 9 (i.e., the mixture may also contain NP8EO, NP10EO,
3

etc.). A mixture containing equal proportions of both NP1EO and NP2EO, for example, would be

given the abbreviation of NP1.5EO.

Nonylphenol ethoxycarboxylates have the general formula C9H19-C6H4O(CH2CH2O)nCH2COOH.

As with NPEs, nonylphenol ethoxycarboxylates with specific ethoxylate chain lengths may be

reported as NPnEC, where n refers to the number of EO groups in the ethoxylate chain.

The basic structures of the three main types of nonylphenolic compounds are shown in Figure 1.

C9H19

OH

C9H19

O(CH2CH2O)nH

C9H19

O(CH2CH2O)nCH2COOH

Nonylphenol Nonylphenol

ethoxylate

Nonylphenol

ethoxycarboxylate

(NP) (NPnEO) (NPnEC)

Figure 1. The structure of nonylphenol and its derivatives.

Nonylphenol is a colourless to pale yellow, or straw-coloured, viscous liquid with a slight

characteristic phenolic odour (Reed 1978; CCOHS 1995; Budavari et al. 1996). The physical and

chemical properties of nonylphenol are listed in Table 2. NP has a molecular weight of 220.3

(Reed 1978). The vapour pressure is 4.55 (± 3.5) x 10-3 Pa at 25°C, and Henry’s Law Constant is

11.02 Pa m3·mol-1 (Romano 1991 as cited in Maguire 1999; European Commission 1999). This

low Henry’s Law Constant suggests that volatilization of nonylphenol from water would not be

significant (Maguire 1999). The pKa (negative logarithm of the acid dissociation constant) of NP is

10.7, indicating that in most natural waters virtually all NP is present in the undissociated form

(Romano 1991 as cited in Maguire 1999).

Nonylphenol has a low solubility in water or dilute aqueous NaOH, but is readily soluble in

acetone, benzene, chlorinated solvents, aniline, heptane, aliphatic alcohols, and ethylene glycol

(Reed 1978; Budavari et al. 1996). Aqueous solubility of nonylphenol (at 20.5oC) is 5.43 mg·L-1,

with only slight variation of this solubility over the temperature range of 2 to 25oC (Ahel and Giger

1993a). Water solubility is increased by alkyl branching (Talmage 1994). There is also a linear

increase in solubility with increasing number of ethoxylate groups. Solubilities (at 20.5oC) for

NP1EO, NP2EO, NP3EO, NP4EO, and NP5EO are 3.02, 3.38, 5.88, 7.65, and 9.48 mg·L-1,

respectively (Ahel and Giger 1993a). NPnEOs are water soluble for values of n Ú 7 (Talmage

1994). Similar trends were observed with octylphenol (OP) and its ethoxylates (OPnEO).

Octylphenol ethoxylates have the same chemical structure as NPEs, except that their alkyl chain

consists of eight carbons, rather than nine.1 The solubility of the OPnEO oligomers were greater

than those of the corresponding NPnEO oligomers. Therefore, the lengths of both the ethoxylate

and alkyl chains of APEs affect their solubility.

 

1

 OP and NP also differ with respect to the branching of their alkyl chains. While NP is always

present as a mixture of isomers with varied branching of the nine-carbon alkyl chain, OP found in

the environment is almost entirely a single isomer with a 1,1,3,3-tetramethylbutyl eight-carbon

chain (Swisher 1987; Bennie et al. 1997, 1998).
4

Nonylphenol ethoxylates have an amphipathic nature, with the alkylphenyl moiety being

hydrophobic (i.e., lipophilic) in nature, and the polyethoxylate chain being hydrophilic. This

characteristic enables NPEs to concentrate at interfaces between non-mixable phases or liquids

(e.g., air and water, oil and water, oil and minerals, biological membranes). Most commercial and

industrial applications of nonylphenol ethoxylates utilize this characteristic surfactant property.

The sorption and solubilization abilities of a surfactant are related to their hydrophile-lipophile

balance (HLB). The higher the molecular weight of the lipophilic component, the more moles of

hydrophilic ethoxylate units are needed to make the surfactant water-soluble (Davidsohn and

Milwidsky 1967). By calculating the percentage of hydrophilic ethoxylate units compared to the

total mass of the surfactant, HLBs for NP5EO, NP8EO, NP9EO, NP14EO, NP17EO, and

NP40EO have been estimated at 10, 12.3, 12.8, 14.9, 15.8, and 17.8, respectively (Beigel et al.

1998). When present in solution at high enough concentrations, surfactants will form colloidalsized clusters, called micelles, in which the hydrophobic tails of the molecules are grouped

together in the centre, with the hydrophilic heads pointing out towards the solvent (Haigh 1996).

The detergency and solubilisation properties of nonylphenol polyethoxylates are due to this

formation of micelles. The concentration at which thermodynamics favour micelle formation is

referred to as the critical micelle concentration. Critical micelle concentration values for NP5EO,

NP8EO, NP9EO, NP14EO, NP17EO, and NP40EO have been estimated at 13, 32, 42, 66, 112,

and 324 mg·L-1, respectively (Beigel et al. 1998). Therefore, the more hydrophobic compounds

(i.e., those with fewer ethoxylate groups and a lower HLB) are more efficient at micelle formation.

Critical micelle concentration values for nonylphenol ethoxylates increase with increasing HLB

(Beigel et al. 1998).
5

Table 2. Physical and chemical properties of nonylphenol and select nonylphenol ethoxylates.

Property Nonylphenol NP1EO NP4EO NP9EO NP40EO

CAS registry number 84852-15-3 -- -- -- --

chemical formula C15H24O C17H28O2 C23H40O5 C33H60O10 C95H184O41

molecular weight (g·mol-1) 220.3 281.4 396.6 616.8 1982.5

freezing point1 (°C) <20 -- -- -- --

melting point1 (°C) 81-83 -- -- -- --

boiling point2 (°C) 295-320, 310,

304, 293-297

-- -- -- --

flash point3 (°C) 146; 300 (open cup),

285 (closed cup)

-- -- -- --

density1 (g·mL-1 at 43°C) 0.933 -- -- -- --

specific gravity4 (at 20°C referred to water at 4°C) 0.95 -- -- -- --

aqueous solubility5 (mg·L-1 at 20.5°C) 5.43 3.02 7.65 water soluble water soluble

pKa 

6

10.7 -- -- -- --

log Kow

7

4.1, 4.2, 4.48, 4.7 4.17, 4.4 4.24, 4.3 4.1 --

vapour pressure6 (Pa at 25°C) 4.55 x 10-3-- -- -- --

Henry’s law constant8 (Pa m3·mol-1) 11.02 -- -- -- --

hydrophile-lipophile balance9-- -- -- 12.8 17.8

1

 Lorenc et al. 1992

2

 as determined by Reed 1978, Lorenc et al. 1992, CCOHS 1995, and Budavari et al. 1996, respectively

3

 as cited in Lorenc et al. 1992 and Chemical Hazard Response Information System 1995, respectively

4

 Reed 1978

5 Ahel and Giger 1993a; Hall et al. 1989

6 Romano 1991 as cited in Maguire 1999

7

 as cited by McLeese et al. 1981, Barber et al. 1988, Freij 1991, Wild and Jones 1992, and Ahel and Giger 1993b (both experimentally

 determined values and calculated estimations)

8

 European Commission 1999

9

 as cited in Beigel et al. 1998
6

Production and Uses

The industrial manufacture of nonylphenol involves alkylating phenol with a mixture of isomeric

nonenes (propylene trimers) in the presence of an acid catalyst. The resulting nonylphenols (with

various isomers of the branched nonyl chain) are recovered by fractional distillation under reduced

pressure (Reed 1978). As of 1997, three Canadian companies were producing nonylphenol:

Huntsman Canada (Guelph, Ontario), Rhone-Poulenc (Mississauga, Ontario), and Stepan

Canada (Longford Mills, Ontario). Total Canadian domestic production of nonylphenol in 1996

(the most recent year for which data are available) was 7700 tonnes, with 5000 tonnes exported

(Camford Information Services Inc. 1997). In 1996, a total of 1200 tonnes of NP were imported

into Canada, all from the United States. Therefore, the total Canadian domestic demand for

nonylphenol in 1996 was 3900 tonnes (Camford Information Services Inc. 1997).

The domestic demand for nonylphenol can be subdivided into its various use applications. In

1989, 1600 tonnes of NP were used to produce ethoxylated pulp mill specialties, 1000 tonnes

were needed for ethoxylated textile specialties, 500 tonnes were used to produce miscellaneous

ethoxylates, another 500 tonnes were used to produce trinonylphenol phosphite, and 900 tonnes

went towards other miscellaneous products (Camford Information Services Inc. 1990). These

other miscellaneous products include the use of nonylphenol as an additive in lube oils, and a

formulant in pesticides (Camford Information Services Inc. 1990). The most notable Canadian

historical example of NP use in pesticides was as a solvent in the Matacil 1.8D formulation of

aminocarb. This insecticide, which contained NP at almost three times the weight of the

aminocarb, was applied aerially to extensive forested areas of Quebec, New Brunswick, and

Newfoundland between 1975 and 1985 to control spruce budworm (Fairchild et al. 1999). Tris(4-

nonylphenyl) phosphite is an antioxidant used in the stabilization of rubber, vinyl polymers,

polyolefins and styrenics (Lorenc et al. 1992). Nonylphenol may also be used as a catalyst in the

curing of epoxy resins, or it may be converted to barium and calcium phenolates which are used

as heat stabilizers in poly(vinyl chloride) (Lorenc et al. 1992). As evidenced in the above statistics

though, the main use (approximately 70%) of nonylphenol is in the production of nonylphenol

ethoxylates.

Nonylphenol ethoxylates are formed through the process of etherification, whereby a basic

catalyst is used to condense nonylphenol with ethylene oxide (Reed 1978):

 

C9H19

OH

C9H19

O(CH2CH2O)nH

+

n CH2 CH2

 O

catalyst

Nonylphenol ethoxylates belong to a class of compounds called alkylphenol ethoxylates (APEs).

Of the nonionic surfactants, APEs are the second largest group, after alcohol ethoxylates, in

commercial production. Since their introduction in the 1940s, APEs have been widely used in

cleaning products and industrial processes (Talmage 1994; White et al. 1994). More than

300 000 tonnes of APEs are produced annually world-wide (Freij 1991; Metcalfe et al. 1996).

Nonylphenol ethoxylates account for about 82% of the APE world market, followed by octylphenol

ethoxylates (15%) and small amounts of dodecylphenol and dinonylphenol ethoxylates (1-2%;

Naylor 1992). In 1990, total use of NPEs in Canada was estimated at 7000 tonnes per year, of

which 4100 tonnes of NPEs were produced domestically, while the remainder was imported in

cleaning product formulations (Metcalfe et al. 1996). In a 1997 survey of 4500 companies

operating in Canada, 189 companies were involved in the production, import, export, shipping,

acquisition, use or release of at least 1 tonne per year of NP and/or NPEs (Servos et al. 2000).

Nonylphenol ethoxylates have a broad range of uses due to their effectiveness as detergents,

emulsifiers, wetting agents, and dispersing agents. There are eleven major Canadian industrial

sectors that use NPEs. These include the pulp and paper industry, textile manufacturing, plastic
7

and elastomer manufacturing, oil extraction and petroleum production, pesticide formulations,

domestic/industrial/institutional cleaners, metal processing, paint and protective coatings, leather

manufacturing, building and construction, and the food and beverage sectors. Smaller amounts

of nonylphenol ethoxylates are also used in spermicides (Metcalfe et al. 1996).

The commercial and industrial applications of NPEs depend partly upon the length of their

ethoxylate chains. Nonylphenol ethoxylates with 4 or 5 ethylene oxide residues per molecule of

nonylphenol are typically used as oil-soluble detergents and emulsifiers, and if sulphonated or

phosphorylated, can be used as anionic detergents, lubricants and antistatic agents (Reed 1978).

NPnEOs with 8 to 12 EO groups are the most commonly used compounds; they are utilized in

high performance detergents (such as for textile scouring) and in industrial cleaning agents for

hard surfaces. Higher ethoxylates (13 to 15 EO groups per molecule), when combined with an oilsoluble anionic surfactant, make excellent emulsifiers for a wide range of solvents and agricultural

pesticides (Reed 1978). NPnEOs with up to 80 ethoxylate groups are effective as dispersants

(Freij 1991).

Directories are available that contain extensive listings of the trade names, manufacturers and

distributors of the various NPE products available (Ash and Ash 1997; Anonymous 1999).

Examples of some commercial preparations of NPEs include the following products: the Alkasurf

CO series manufactured by Rhodia, Inc.; Caloxylate N-9 (NP9EO) manufactured by Pilot

Chemical Co.; the Carsonon N series manufactured by Lonza, Inc.; the Cedepal CO series

manufactured by Stepan Canada Inc.; the Chemax NP series manufactured by Chemax Inc.; the

Delonic NPE series manufactured by DeForest Enterprises, Inc.; the Iconol NP series

manufactured by BASF Corp. Performance Chemicals; the Igepal CO series manufactured by

GAF Corp. (Rhodia, Inc.); Imbentin-N/7A (NP5EO) and Imbentin-N/200 (NP20EO) manufactured

by W. Kolb, AG; Lissapol TN 450 and Lissapol TN XP (both NP8-9EO) manufactured by ICI

Chemicals; the Marlophen NP series manufactured by CONDEA Chemie GmbH; the Norfox NP

series manufactured by Norman, Fox & Co.; the Polystep F series by Stepan Company; the

Surfonic N series manufactured by the Huntsman Corporation; the Tergitol NP series

manufactured by Union Carbide Corporation; the TERIC N series manufactured by Huntsman

Corp. Australia; and TRITON N by Union Carbide Corporation (Ash and Ash 1997; Anonymous

1999; Lee 1999).

Due to concerns about the toxicity and persistence of NPE degradation products, regulatory and

voluntary initiatives have been taken to eliminate many of the uses of NPEs in Europe. In 1986,

Switzerland banned the use of NPEs in laundry detergents for domestic use (WWF Canada

1997). In an ordinance enacted in 1991, Swedish parliament decided to phase out 90% of NP

and NPE use by the year 2000 (WWF Canada 1997). The Cosmetic, Toiletries, Soap and

Detergent Industries Association (SPT) of Denmark have already phased out alkylphenols

completely under a voluntary agreement with the Danish EPA (Metcalfe et al. 1996). Germany,

Norway, the Netherlands, UK, and France are currently discussing a potential ban and phase out

of NPEs in commercial products such as domestic and industrial detergents. In the UK, domestic

use of NPEs in detergents has been phased-out since 1976, and some agrochemical companies

have voluntarily phased-out NPEs in their pesticides since 1992. In 1995, UK wool scourers

volunteered to eliminate discharges (though not use) of APEs by the end of 1996, and several

have switched to the use of alcohol ethoxylates (WWF Canada 1997). As of 1990, German wool

manufacturers had voluntarily phased out the use of NPEs in wool scouring, and in 1997 a

voluntary agreement was drawn up to phase out the use of NPEs in Germany as flocculants in

wastewater purification (WWF Canada 1997).

This widespread European phase-out of NPEs is the result of a 1992 recommendation from the

Paris Commission. Under Paris Commission Recommendation 92/8, all of the North Sea States

and the European Community agreed to phase NPE out of domestic cleaning products by 1995

and out of industrial cleaning products by 2000 (WWF Canada 1997). The Working Group on

Diffuse Sources of the Oslo and Paris Conventions for the Prevention of Marine Pollution is

currently performing a full risk assessment on nonylphenol (Maguire 1999).
8

In the United States, nonylphenol was first placed on the Environmental Protection Agency’s

Inerts List 1 in 1987, identifying it as a pesticide formulant of toxicological concern because of

human health and/or ecological considerations (PMRA 2000). Pesticide products in the U.S.

which contain nonylphenol must state so on the label. In Canada, the Pest Management

Regulatory Agency is proposing regulations to phase out the use of nonylphenol in pesticide

formulations by December 31, 2002 (PMRA 2000).

Sources to the Environment

Water

Almost all releases of nonylphenolic compounds are initially routed to the aquatic environment in

the form of industrial or municipal wastewater effluents. The bulk of industrial releases to the

Canadian environment originate from formulators/distributors of surfactants, and industrial users

of cleaning products, degreasers and detergents (Servos et al. 2000). Other significant industrial

sources include pulp mills and textile mills, as well as the leather manufacturing, oil extraction,

and petroleum production sectors (Metcalfe et al. 1996). Household, industrial, and institutional

cleaning products are the major source of the nonylphenolic content in effluents from municipal

wastewater treatment plants (WWTPs). Although most nonylphenol and nonylphenol ethoxylates

that enter the aquatic environment are introduced via effluent discharge, there is also the potential

for these substances to be released to the environment during production, transportation,

processing, and during the use of the products (Metcalfe et al. 1996).

Due to their use in some pesticides as adjuvants and dispersants, nonylphenol and nonylphenol

ethoxylates may potentially enter the aquatic environment as residues deposited on surface

waters from aerial spraying (Ernst et al. 1980; Sundaram et al. 1980; Holmes and Kingsbury 1980;

Paveglio et al. 1996) or as runoff from fields. Application of pesticides to forests can introduce NP

and NPEs to forest streams and ponds. Application to agricultural fields and orchards is a source

of NP and NPEs to drainage ditches and farm ponds, and application to emergent vegetation

provides a pathway for entry of NP and NPEs to wetlands and intertidal zones.

Sediment

Once introduced to the aquatic environment, NP and NPEs may adsorb to particulates and be

deposited to bottom sediments. Their amphipathic nature enables adsorption to organic matter

through hydrophobic reactions as well as adsorption to the mineral components of sediment

through hydrophilic reactions (John et al. 2000). Movement of NP and NPEs to the sediments

may involve direct adsorption to sediment particles at the surface of the sediment-water interface,

or adsorption to suspended sediment particles in the water column that may eventually settle out.

Once present in the bottom sediments, there is also the possibility for re-entry of NPEs to the

water column through resuspension of sediment, particularly during storms or through dredging.

Soil

The sources of NP and NPEs to the terrestrial environment are much more limited than for the

aquatic environment. Some NP or NPEs may enter the soil through the spraying of pesticides on

agricultural fields, orchards, golf courses and forests (Sundaram et al. 1980). Another pathway is

through the application of sewage sludge or sludge from recycled paper plants to agricultural soils

(Diercxsens and Tarradellas 1987; Marcomini et al. 1989a,b; Kirchmann and Tengsved 1991;

Hawrelak et al. 1999). Disposal of sewage sludges in landfills may also introduce nonylphenolic

compounds into soil (Marcomini et al. 1989a,b; Marcomini et al. 1991).

Despite the strong tendency for adsorption to soil and sediment particles, alkylphenolic

compounds (such as NP and NPEs) present in surface waters or landfills may leach into

groundwaters (Reinhard et al. 1984; Barber et al. 1988; Zoller et al. 1990; Ahel 1991; Ahel et al.

1996).
9

Air

Some authors have stated that due to the low volatility of nonylphenolic substances, they are

generally not contaminants of the air (e.g., Metcalfe et al. 1996). However, recent studies indicate

that in some locations there may be considerable water-to-air volatilization, resulting in high

atmospheric concentrations of nonylphenolic substances (Dachs et al. 1999).

The fate processes that control the movement of NP and NPEs through various environmental

compartments are discussed further in Chapter 3.

Analytical Methods

Over the past few years there have been major improvements in the methods used for sample

extraction and detection of nonylphenol and nonylphenol ethoxylates in environmental samples,

both in terms of ease of analysis, and in sensitivity and precision (Kubeck and Naylor 1990).

Comprehensive discussions of both historical and more recent analytical methods used for

determination of nonylphenolic compounds are provided by Talmage (1994) and Lee (1999). In

this document, however, brief descriptions of only the most up-to-date and currently accepted

methods are presented. For more detailed information on any of the particular methods that are

mentioned below, readers are advised to consult the references given.

Determining concentrations of nonylphenolic compounds in environmental samples involves two

main procedures: extraction/clean-up, and detection. The optimal methods to use for extraction

and clean-up depend upon the type of environmental sample being analyzed, and the particular

oligomer range of interest.

Extraction of nonylphenol and the lower molecular weight nonylphenol ethoxylates (e.g., NP1EO,

NP2EO) from aqueous samples can be accomplished through liquid-liquid extraction techniques

(Bennie et al. 1997, 1998), or through a combination of steam distillation and solvent extraction by

heating the sample to reflux with iso-octane (Kubeck and Naylor 1990) or cyclohexane (Ahel and

Giger 1985). With steam distillation methods, no further clean-up of the sample is required before

detection analysis. Extraction of higher molecular weight NPnEOs requires other methods due to

their lower volatility and their higher polarity and solubility. Successful extraction and

preconcentration of NPEs, as well as NP and NPECs, from water and effluent samples has been

achieved through solid phase extraction (Marcomini et al. 1987, 1990; Kubeck and Naylor 1990;

Blackburn and Waldock 1995; Field and Reed 1996; Lee et al. 1998; Lee and Peart 1998; Sekela

et al. 1999). Solid phase extraction involves passing the sample through a cartridge or disk

containing an adsorbent such as octadecylsilane, graphitized black carbon, or a strong anion

exchange resin (Lee et al. 1998). For example, Kubeck and Naylor (1990) used a dual column

apparatus in which the aqueous sample was deionized by passing through a mixed-bed ionexchange resin, then passed through octadecylsilica, which adsorbed the organic material. The

organics were flushed from the octadecylsilica with warm methanol, the methanol was removed

under a stream of nitrogen, and the remaining residue was redissolved in a

dichloromethane/hexane mixture for detection analysis (Kubeck and Naylor 1990).

For extraction of NP, NP1EO and NP2E0 from solid samples (i.e., soil, sediment, sludge, or

tissue), Soxhlet extraction (usually with dichloromethane) or steam distillation may be used

(Marcomini et al. 1989a, 1989b, 1991; Bennie et al. 1997, 1998). Marcomini et al. (1990) also

used a combination of Soxhlet extraction and elution through an aminosilica minicolumn cartridge

to extract NP and NPEs from marine sediment and macroalgae. A more recent method using

supercritical fluid extraction is a more efficient alternative that may be applied to a wider range of

nonylphenolic compounds (Lee and Peart 1995; Lee et al. 1997).

For sensitive detection and quantification, it is now common practice to use high pressure liquid

chromatography (HPLC) with fluorescence detection. The phenolic ring in nonylphenolic

compounds absorbs ultraviolet light, giving these substances a distinctive fluorescent profile that
10

can be readily detected (Naylor 1992). With normal-phase HPLC, various nonylphenol

ethoxylates can be separated according to their EO chain length on a silica gel matrix using an

organic solvent gradient (Talmage 1994). The separated components are then detected by

fluorescence and quantitated by comparison with external standards (Talmage 1994). Reversedphase HPLC may be used to separate different alkylphenol ethoxylates by their alkyl chain (i.e.,

into nonylphenol, octylphenol, and dodecylphenyl ethoxylates, etc.) (Talmage 1994). Gas

chromatography/mass spectrometry (GC/MS) is also widely used for analysis. Extracts containing

NP may be analyzed by GC/MS with either electron ionization or chemical ionization (Wahlberg et

al. 1990; Clark et al. 1992; Blackburn and Waldock 1995; Bennie et al. 1997; Bennett and

Metcalfe 1998). Determination of NPECs is also possible though GC/MS, although they must be

derivatized to their volatile esters prior to analysis (Field and Reed 1996). It appears that of the

NPEs, only the lower molecular weight compounds can be adequately determined using GC/MS.

Wahlberg et al. (1990) were able to quantify NP1EO, NP2EO and NP3EO by first derivatizing

them with pentafluorobenzoyl chloride. Clark et al. (1992) found that NPEs with low to moderate

degrees of ethoxylation could be determined by GC/MS if present at high concentrations. Clark et

al. (1992) also found that NP3-8EO could be detected at the parts per trillion level by using

particle-beam liquid chromatography/mass spectrometry.

Although analytical techniques are constantly improving, the analysis of environmental samples

for nonylphenolic substances still remains a complicated procedure. Often environmental

samples will contain other alkylphenolic surfactants which must be separated from NP and NPEs.

Samples may also contain hundreds of different isomers and ethoxamers of NPEs, requiring the

use of several different analytical techniques for the detection of all.

Using various combinations of the analytical methods described above, sensitive analyses may be

conducted with low limits of detection. The detection limits discussed below were determined

through the analysis of environmental samples, and therefore represent concentrations that are

environmentally relevant and can be realistically determined from field samples. Reported limits

of detection and method detection limits for NP, NP1EO, and NP2EO in water, sewage or

effluents are as low as 0.01, 0.02 and 0.02 μg·L-1, respectively (Bennie et al. 1997, 1998; Rudel et

al. 1998). For larger molecular weight NPEs (i.e., NPnEOs where n = 3 to 17) method detection

limits are higher, at about 1.6 μg·L-1 (Radian Corporation 1990). Detection limits for individual

NPECs in water have been reported between 0.04 and 0.4 μg·L-1 (Field and Reed 1996). For NP,

NP1EO, and NP2EO in sediments, current limits of detection are reported at concentrations as

low as 0.004 mg·kg-1 (Marcomini et al. 2000). Detection limits for NP in soil have been reported at

concentrations as low as 0.05 mg·kg-1 (Jobst 1987). For tissue samples, detection limits for NP

and each of NP1EO to NP3EO have been reported at 0.001 mg·kg-1 and 0.006 mg·kg-1,

respectively (Wahlberg et al. 1990). With refinements to the standard analytical methods, it may

be possible to obtain detection limits lower than those reported here, and in some cases (e.g.,

Bennie 1998; Sekela et al. 1999) environmental measurements have been noted at

concentrations below these detection limits.
11

CHAPTER 2. CONCENTRATIONS IN CANADA

Nonylphenol and nonylphenol ethoxylates are synthetic compounds that do not occur naturally.

Therefore, environmental concentrations can be attributed solely to human activities. A

considerable amount of data has been collected on environmental concentrations of

nonylphenolic compounds in North America and Europe for fresh water, marine or estuarine

water, groundwater, freshwater sediment, and marine or estuarine sediment. Many

measurements of these compounds have also been made in sewage effluents, sewage sludge,

and industrial effluents. Considerably less data have been collected on soil concentrations of

nonylphenolic compounds, and the data that do exist focus on soils that have been amended with

sewage sludge.

A comprehensive review and discussion of environmental concentrations of nonylphenol and

nonylphenol ethoxylates, both within Canada and outside of Canada, are provided in the CEPA

PSL2 assessment of nonylphenol and its ethoxylates (Servos et al. 2000). Therefore, the

summary of Canadian environmental concentrations included in this document largely draws upon

the above-mentioned assessment. Data were available for NP and NPE concentrations in

Canadian fresh surface waters, freshwater and marine sediments, sludge-amended soil, raw

sewage, municipal wastewater treatment plant (WWTP) effluent and sludge, and industrial

effluents. No data on concentrations in Canadian marine/estuarine waters or groundwater were

available.

Surface Waters

Canadian freshwater concentrations of NP and NPEs have been reported for various sites in

southern Ontario (largely from rivers, lakes and harbours throughout the Great Lakes - St.

Lawrence River system), for freshwater rivers in Quebec, Prince Edward Island, and Nova Scotia,

and for sites in the Fraser River of British Columbia. Most reported concentrations of NP fall

within the range of <0.01 to 1.7 μg·L-1 (Bennie et al. 1997; Sekela et al. 1999; Sabik et al. 2000;

Rutherford 2001). Agricultural streams monitored in Prince Edward Island in 1999 contained

concentrations of NP as high as 0.25 μg·L-1 (Ernst 2000). The highest measured concentrations

occurred in waters near heavily industrialized areas. For example, nonylphenol concentrations

directly adjacent to an outfall of a secondary municipal wastewater treatment plant treating textile

mill effluent on the Eaton River in Quebec were measured at 94.2 μg·L-1 (Rutherford 2001).

Carey et al. (1981, as cited in Servos et al. 2000) reported NP concentrations as high as

2600 μg·L-1 in the Canagagigue Creek near Elmira, Ontario; however, these measurements were

made following an industrial spill into the small creek, and are not considered representative of

typical levels (Servos et al. 2000). NPEs tend to be more ubiquitous than NP, and also occur at

higher concentrations. Concentrations of NP1EO and NP2EO have been measured in Canadian

fresh waters at <0.02 to 7.8 μg·L-1, and <0.02 to 16.9 μg·L-1, respectively (Bennie et al. 1997;

Rutherford 2001). Unpublished results from an ongoing survey of 8 rivers in Quebec indicate that

the sum of NPE (with 1 to 17 EO groups) ranges between <0.05 and 14.6 μg·L-1 (Berryman 2000).

For the sum of NP1EC and NP2EC, concentrations in Quebec rivers range between <0.05 and

23 μg·L-1 (Berryman 2000). Levels of NP1EC in water from the St. Lawrence River near Montreal

have been reported at 0.014 to 0.51 μg·L-1, and levels of NPnEOs (with n ranging from 7 to 16)

from the same area have been reported to range from below detection limit to 10.3 μg·L-1 (Sabik

et al. 2000). Extremely high levels of NP and NPEs have been measured in the Annapolis River

of Nova Scotia downstream from a textile mill; at 30 metres below the outfall, concentrations of

NP, NP1EO, NP2EO, and the sum of NPEs (with 3 to 17 EO groups) were 644, 180, 44.4, and

186 μg·L-1, respectively (Rutherford 2001). These high levels were attributed to a poorly operating

primary treatment system at the mill (Rutherford 2001).

There is currently a lack of data on all nonylphenolic substances for surface fresh waters in

Alberta, Manitoba, New Brunswick, Newfoundland, Saskatchewan, and the three territories.
12

Sediment

Concentration data for nonylphenolics in Canadian freshwater sediments only exist for the Great

Lakes - St. Lawrence system, and freshwater locations in British Columbia. Reported freshwater

bed sediment concentrations of NP range from below detection limits to 110 mg·kg-1 on a dry

weight basis (Lee and Peart 1995; Bennie et al. 1997; Brewer et al. 1999; Bennett and Metcalfe

1998, 2000; Sabik et al. 2000). Sekela et al. (1999) reported NP concentrations in suspended

solids of the Fraser River, B.C., at 0.021 to 0.087 mg·kg-1 dry weight. Canadian freshwater bed

sediment concentrations of NP1EO and NP2EO fall within the ranges of <0.015 to 75 mg·kg-1 and

<0.015 to 18.0 mg·kg-1 dry weight, respectively (Bennie et al. 1997; Bennett and Metcalfe 2000;

Sabik et al. 2000).

Data on the concentrations of nonylphenolics in Canadian marine sediments are only available for

the Strait of Georgia, British Columbia. Concentrations of NP in surficial marine sediments

ranged from 0.122 to 0.541 mg·kg-1 dry weight (mean = 0.273 mg·kg-1, n = 21) (Shang et al. 1999;

MacDonald and Ikonomou 2001). Surficial marine sediment concentrations of NP1EO and

NP2EO were measured at 0.065 to 0.558 mg·kg-1 dry weight (mean = 0.279 mg·kg-1, n = 21) and

0.034 to 0.198 mg·kg-1 dry weight (mean = 0.069 mg·kg-1, n = 21), respectively. Total

concentrations of NPnEO (3 ≤ n ≤ 19) were measured at 0.29 to 1.42 mg·kg-1 dry weight (mean =

0.78 mg·kg-1, n = 21). These same researchers also collected marine sediment cores and

examined the concentrations of NP and NPEs at various sediment depths. They found that both

the total amount of NPnEO, and the profile of relative concentrations for individual NPnEO

ethoxamers did not change significantly with depth (Shang et al. 1999).

Most of the sediments that have been analyzed for NP and NPEs were collected from locations

near WWTP outfalls or industrialized areas. Therefore, the data reported here likely represent the

upper range of concentrations present in Canadian sediments.

Soil

Only one study has looked at Canadian soil concentrations of nonylphenolic substances, and that

study only examined soil from a single site that had been amended with sludge. Water

Technology International Corporation (1998a as cited in Servos et al. 2000) measured

2.72 mg·kg-1 of NP in sludge-amended soil, <0.538 mg·kg-1 each of NP1EC and NP2EC, and only

trace amounts of NP1EO, NP2EO, and NPnEO.

Sewage and Sludge

Several researchers have reported concentrations of nonylphenolic substances in the raw

sewage, primary effluent, final effluent, and sludge of various municipal WWTPs across Canada.

Reported concentrations of NP, NP1EO, NP2EO, NPnEO, NP1EC, and NP2EC in raw sewage

fall within the ranges of 0.69 to 155 μg·L-1, 2.8 to 43 μg·L-1, 0.26 to 38 μg·L-1, 98.5 to 1300 μg·L-1,

0.9 to 71.3 μg·L-1, and 1.4 to 35 μg·L-1, respectively (Bennie et al. 1998; Lee and Peart 1998; Lee

et al. 1998; Water Technology International Corporation 1998b as cited in Servos et al. 2000).

Most studies have found that concentrations of NP and NPEs are lower in the final effluent from

municipal WWTPs relative to the incoming raw sewage, with the greatest decreases occurring for

the larger NPEs. Reported concentrations of NP, NP1EO, NP2EO, and NPnEO in final effluents

from Canadian municipal WWTPs are <0.02 to 15.1 μg·L-1, 0.072 to 26 μg·L-1, 0.099 to 21 μg·L-1,

and from below detection to 13.3 μg·L-1, respectively (Lee and Peart 1995; Bennie et al. 1998; Lee

and Peart 1998; Sekela et al. 1999; Water Technology International Corporation 1998b as cited in

Servos et al. 2000). Rogers et al. (1986) also identified NP, NP1EO and NP2EO in municipal

wastewater effluents from Vancouver, B.C.; however, they did not report concentrations.

Sewage treatment appears to have the opposite effect on NPECs, with higher concentrations

occurring in the final effluent relative to the raw sewage. Concentrations of NP1EC and NP2EC in
13

final effluents from Canadian municipal WWTPs have been reported at 1.9 to 703 μg·L-1 and 11 to

565 μg·L-1, respectively (Lee and Peart 1998; Lee et al. 1998; Water Technology International

Corporation 1998b as cited in Servos et al. 2000). Similar observations of elevated levels of

NPECs following secondary treatment have also been observed in several Swiss municipal

WWTPs (Ahel et al. 1987). NP1EC and NP2EC are biodegradation products of NPEs, which

explains their occurrence at higher concentrations in sewage following treatment (Ahel et al.

1987).

Substances that are more hydrophobic, such as NP and the lower NPEs, have a greater tendency

to adsorb to particles and thus accumulate in the sludge during treatment. Concentrations of NP,

NP1EO, and NP2EO in the sludge of Canadian municipal WWTPs have been reported at 8.4 to

850 mg·kg-1, 3.9 to 437 mg·kg-1, and 1.5 to 297 mg·kg-1, respectively (Lee and Peart 1995; Lee et

al. 1997; Bennie et al. 1998; Water Technology International Corporation 1998b as cited in Servos

et al. 2000). More hydrophilic substances, such as the NPECs and the higher NPEs, occur at

lower concentrations in sludge. Reported sludge concentrations for NPnEO, NP1EC and NP2EC

are 9 to 169 mg·kg-1, <0.5 to 25 mg·kg-1, and <0.5 to 38 mg·kg-1, respectively (Lee et al. 1997;

Water Technology International Corporation 1998a,b as cited in Servos et al. 2000).

The concentrations and relative proportions of various NPEs in final effluents and sludges vary

considerably among municipal WWTPs and reflect differences in the type of effluent and sludge

treatment used. The operating efficiencies of individual WWTPs may also affect the levels of

NPEs in final effluent and sludge (Ahel et al. 1994a). Differences in NP and NPE concentrations

among the raw sewages entering municipal WWTPs are likely a function of several factors

including the size of the population served by the WWTP, and the number and types of

commercial or industrial clients contributing to the WWTP.

Industrial Effluents and Sludge

Little data exist for concentrations of nonylphenolic compounds in Canadian industrial effluents

and sludge, but most of the data that do exist are from pulp and paper mills or textile mills.

Pulp mill effluents have been found to contain NP, NP1EO and NP2EO in concentrations of <0.02

to 26.2 μg·L-1, <0.015 to 25.9 μg·L-1, and <0.015 to 35.9 μg·L-1, respectively (Bennie 1998 as cited

in Servos et al. 2000). Unbleached whitewater from a Canadian pulp mill was also reported to

contain NPnEO at a concentration of 310 μg·L-1 (Sithole and Allen 1989). Lee and Peart (1999)

measured concentrations of nonylphenolics in both the effluent and sludge of 19 Canadian (and 1

U.S.) pulp and paper mills that use a variety of process types and waste treatment methods.

Secondary treated effluent contained NP, total NPEs, NP1EC, and NP2EC of <0.1 to 4.32 μg·L-1,

<2 to 71.3 μg·L-1, <1.0 to 10.13 μg·L-1, and <1.0 to 32.32 μg·L-1, respectively (Lee and Peart

1999). Effluents commonly contained NPE oligomers with 10 or fewer ethoxy groups, but

ethoxylates with greater than 10 ethoxy groups were seldom detected. Sludge samples from the

mills contained NP, total NPEs, NP1EC, and NP2EC at concentrations of <0.05 to 120.9 mg·kg-1,

<1 to 90.8 mg·kg-1, <1 to 5.78 mg·kg-1, and <1 to 18.5 mg·kg-1 dry weight, respectively (Lee and

Peart 1999). Generally, the NPE oligomers that occurred at the highest concentrations in the

sludge were NP8EO and NP9EO. Sludge from a Canadian recycled paper plant that was used as

an agricultural soil amendment was found to contain NP, NP1EO and NP2EO at concentrations

as high as 4.61, 1.21, and 0.39 mg·kg-1 dry weight, respectively (Hawrelak et al. 1999).

Textile mill effluents appear to contain considerably greater levels of nonylphenolic substances

than pulp and paper mills, with concentrations of NP, NP1EO, NP2EO, and NPnEO (n=3 to 17)

reported at 1.1 to 13.3 μg·L-1, 0.74 to 260 μg·L-1, 0.64 to 590 μg·L-1, and 50.2 to 8600 μg·L-1,

respectively (Bennie 1998 as cited in Servos et al. 2000).

Analysis of two oil refinery effluent samples did not reveal any detectable concentration of NP

(Bennie 1998 as cited in Servos et al. 2000).
14

CHAPTER 3. ENVIRONMENTAL FATE AND BEHAVIOUR

The occurrence of nonylphenol and its ethoxylates in aquatic or terrestrial ecosystems poses a

potential risk to exposed organisms. The bioavailability and toxicity of nonylphenolic compounds

are dependent on a number of physical and chemical processes that affect the fate and behaviour

of these compounds in the environment. Therefore, understanding the role of these factors is

essential for interpreting toxicity data, developing and using environmental quality guidelines, and

implementing site-specific objectives. This chapter provides a discussion of the factors that

govern the fate and behaviour of nonylphenolic compounds in the environment. The major

processes affecting the persistence, distribution and bioavailability of nonylphenolic compounds in

the environment include biodegradation, adsorption to suspended solids with subsequent

sedimentation, soil particle adsorption, infiltration to groundwater, volatilization, and photolysis.

Aerobic and anaerobic biodegradation of nonylphenolic substances by microorganisms is the

most important factor affecting their fate. Various other biotic interactions are also involved in the

fate and behaviour of nonylphenolic compounds. Uptake and bioaccumulation of nonylphenolic

compounds by higher organisms may occur, as well as biotransformation, metabolism, and

excretion. Nonylphenolic compounds, if available to biota at sufficiently high concentrations, have

the potential to cause toxicity and/or endocrine disruption.

Physical and Chemical Processes

The most important physicochemical process affecting nonylphenolic substances is particle

adsorption. Abiotic degradation is not thought to contribute significantly to the dissipation of

nonylphenolic substances. Laboratory studies have shown that under aseptic conditions

nonylphenol is very stable (Sundaram and Szeto 1981; Corti et al. 1995). Therefore, the effects

due to physicochemical transformations such as hydrolysis and photolysis are thought to be

negligible (Trocmé et al. 1988; European Commission 1999). Volatilization from either water or

soil is also considered an insignificant fate process for nonylphenolic substances (European

Commission 1999). Nonetheless, these processes may occur to a minor degree, and therefore

are discussed below.

Water

Most of the nonylphenol and nonylphenol ethoxylates entering the environment are initially

released to surface waters. Parent nonylphenol polyethoxylates do not appear to be persistent in

natural waters, while some breakdown products (e.g., NP, NP1EO, and NP2EO) are moderately

persistent (Maguire 1999). The mean time to 50% dissipation (DT50) of 4-NP from the water

column was estimated in a mesocosm study to be 0.74 days, and mean time to 95% dissipation

(DT95) was 13.8 days (Heinis et al. 1999). In another study where nonylphenol was aerially

applied to a forested plot containing a stream, nonylphenol residues were found to persist in the

stream water for no more than five days, and a half-life of 2.5 hours was estimated (Sundaram et

al. 1980). While in the aquatic environment, potential abiotic fate processes for nonylphenolic

compounds include photolysis, volatilization and adsorption to suspended solids.

Photolysis, the decomposition of chemicals by the action of radiant energy, is one process by

which nonylphenol and its ethoxylates may be degraded. This process is most effective in the

surface layers of the water column where compounds are exposed to sunlight at higher

intensities. The first order rate constant for sunlight photolysis of nonylphenol in fresh water has

been estimated to range from 0.05 to 0.09 m

2

kW-1h

-1 (Ahel et al. 1994d). In other words, if

exposed to continuous noon, summer sunlight, NP in the surface layer of natural waters would

have a half-life of 10-15 hours (Ahel et al. 1994d). In deeper water layers this half-life would be

considerably longer (e.g., at a depth of 20-25 cm the photolysis rate would be 1.5 times slower

than at the surface). The photochemical oxidation of nonylphenol ethoxylates is significantly

slower than that of NP (Ahel et al. 1994d). Often the photolytic process is aided by the presence

of metals or metallic oxides that can act as photocatalysts. Degradation of alkylphenol

ethoxylates is known to be induced by the catalysts Fe (III) and titania (Ti O2) in the presence of
15

sunlight. The degradation process involves a hydroxyl radical attack on the ethoxylate chain, and,

at a much slower rate, on the benzene ring (Pelizzetti et al. 1989; Brand et al. 1998). Further

degradation involves oxidation to lower molecular weight products such as acidic compounds,

polyethylene glycols, ethylene glycols, and ultimately to carbon dioxide (CO2).

Variable results have been found for the effect of temperature on the rate of nonylphenol

degradation in water. Ahel et al. (1994b) found that concentrations of nonylphenolic compounds

in fresh water tend to decrease with higher temperatures. Schöberl and Mann (1976) found little

effect of temperature on degradation of NP9EO in fresh water, with 33-36% decomposition

occurring at 20-23oC, and 37% at 3-4oC. In contrast, they found a substantial temperature effect

in seawater, with 95% degradation occurring at 20-23oC, and only 15% at 3-4oC. Manzano et al.

(1999) observed significant increases in the rates of both primary biodegradation and

mineralization with increased temperature. They also observed shorter acclimation periods at

higher temperatures. Tanghe et al. (1998) found that degradation of nonylphenol in lab-scale

activated sludge units was almost complete at 28oC, but slowed down to 13-86% elimination

capacity at temperatures of 10-15oC, a finding that is significant considering Canadian conditions.

Marcomini et al. (2000) observed considerable seasonality in the concentrations of nonylphenol

ethoxylates present in the lagoon of Venice, Italy, and attributed this to the dependence of their

biodegradation on temperature.

One potential mechanism by which organic compounds may leave surface waters is through

volatilization. In a recent study, nonylphenol was measured in the atmosphere over the Hudson

River Estuary at concentrations ranging from 0.1 to 68.6 ng·m-3 (Dachs et al. 1999). The authors

concluded from these high concentrations that a considerable amount of water-to-air volatilization

must have occurred. These results are surprising because most of the previous literature has

claimed that volatilization of nonylphenol is not likely to occur under most conditions. The Henry’s

Law Constant for nonylphenol, which describes the likelihood for a chemical to move from surface

water to the air, is 11.02 Pa m3·mol-1 (Reed 1978; European Commission 1999). This low Henry’s

Law Constant indicates that volatilization is not likely to be a significant mechanism for removal of

nonylphenol from water systems and that it is unlikely to be transported very far in the atmosphere

(Maguire 1999; European Commission 1999).

Movement of dissolved compounds from the aqueous phase of the water column to the

particulate phase may occur through adsorption. Studies have shown that nonylphenol in water is

readily adsorbed onto suspended solids (Kvestak et al. 1994; Sekela et al. 1999). Using field data

from the Fraser River in British Columbia, the tendency for nonylphenol to partition from the

aqueous phase to the suspended solid phase has been evaluated based on organic carbon (OC)

content. The sorption partition coefficient (log KOC) for nonylphenol was estimated to be in the

range of 4.7 to 5.6 (Sekela et al. 1999). Partitioning of nonylphenol polyethoxylates between the

dissolved and particulate phases can vary slightly among various oligomers; NP6EO and NP7EO

have shown higher relative tendencies to partition into the particulate phase than any other NPEs

(Kvestak et al. 1994).

Sediment

NP and NPEs adsorbed to suspended particles may settle out of the water column into the

sediments. Nonylphenolic compounds dissolved in the water may also directly adsorb to particles

on the sediment surface. John et al. (2000) suggested that the adsorption of nonylphenol

polyethoxylates to sediment involves a combination of two main processes: hydrophobic

interactions with organic matter in the sediment, and hydrophilic interactions with the mineral

components. These two processes have opposing effects in terms of the ethoxylate chain length.

The hydrophobic interactions increase with decreasing ethoxylate chain length since the lower

ethoxamers have higher octanol-water partition coefficients (see Table 2) and are more

hydrophobic. The hydrophilic interaction involves hydrogen-binding due to the hydration of metal

oxides in the sediment to the ether oxygens of the ethoxylate chain. Therefore, the amount of

adsorption due to hydrophilic interactions increases with increasing ethoxylate chain length.

Partitioning of nonylphenol ethoxylates between the water and sediment phases, and
16

achievement of adsorption equilibrium occurred rapidly under artificial conditions (John et al.

2000). In a littoral enclosure study, some of the nonylphenol applied through subsurface injection

into the water column was found to adsorb to the sediment within two days of the initial

application, as well as adsorbing to the plastic walls of the mesocosm enclosures (Heinis et al.

1999). Nonylphenol also appears to be persistent in sediments; in the same freshwater

mesocosm study, the DT50 of nonylphenol from sediments was estimated at 66.0 days, and the

DT95 was estimated at 401 days (Heinis et al. 1999). This indicates that there is the potential for

carry-over from one year to the next, such that a site which receives regular inputs will have

increasing sediment concentrations of NP over time. More studies are needed though to confirm

the fate of nonylphenol in freshwater sediments under natural conditions.

Persistence has also been observed in marine sediments. NPEs (with 1 to 14 ethoxylate units)

applied to estuarine intertidal plots as part of a herbicide formulation showed decreasing

concentrations in the sediments during the first 14 days post-treatment, but then stabilized and

remained constant even after 119 days (Paveglio et al. 1996). The authors suggest that this

stabilization may have been due to partitioning of the NPEs to sediment with reduced availability

for biodegradation; also, lower temperatures at the later dates may have resulted in reduced

microbial degradation.

Soil

The abiotic fate processes affecting nonylphenol and its ethoxylates in soils, in decreasing order

of importance, include particle adsorption, infiltration to groundwater, and volatilization.

The major route for nonylphenolic compounds to enter terrestrial ecosystems is through the

application of sewage sludge or pulp and paper mill sludge to agricultural soils, or through

disposal of sludge in landfills. There appear to be conflicting reports on the persistence of

nonylphenolic compounds in soil. Some researchers report that under aerobic conditions,

nonylphenolic compounds are not persistent and degradation occurs readily (Wild and Jones

1992; Maguire 1999). For example, concentrations of nonylphenol introduced into agricultural

soils through sludge application decreased by 90% in ten days (Kirchmann et al. 1991) and by

92% in three months (Diercxsens and Tarradellas 1987). In contrast, there is also evidence of

persistence with complete degradation taking considerably longer. In a Swiss study where

digested sewage sludge was applied to an experimental field plot, soil concentrations of NP,

NP1EO and NP2EO decreased rapidly during the first 10 days after application, with half-lives of 7

to 8 days for each compound (Marcomini et al. 1988). This was followed by a transition period of

about 90 days, during which the compounds disappeared at a much slower rate; the half-lives for

NP, NP1EO and NP2EO were 90, 150, and 110 days, respectively. The authors continued to

measure the concentrations of the nonylphenolic compounds until 330 days after the sewage

application, and for the rest of that period, the concentrations remained relatively constant, with

estimated half-lives of >360 days. Thus, a small fraction of the initial NP, NP1EO and NP2EO

was highly persistent, possibly because its strong sorption to the soil made it unavailable for

biodegradation. It is unknown whether this constant residual concentration of NP simply

represents the threshold for biological transformation, and therefore will remain the same, or

whether the soil is at risk of increasing accumulation of NP with further sludge applications

(Marcomini et al. 1988, 1989b). Topp and Starratt (2000) also found that NP underwent a biphasic

mineralization in agricultural soil. During a rapid initial phase, that lasted 10 days, about 30% of

the nonylphenol (radioactively-labelled at one of the phenolic ring carbons) was converted to

carbon dioxide; this was followed by a much slower second phase. Under anaerobic conditions,

such as in landfills, it appears that nonylphenolic compounds are persistent in soil (Maguire 1999).

For example, in a survey of several German sludge-only landfills, concentrations of NP and

NP1EO of 4 and 19 mg·kg-1 dry weight, respectively, were detected in samples of sludge that had

been landfilled 30 years previously (Marcomini et al. 1989b).

Based on physical/chemical properties, Wild and Jones (1992) speculated that nonylphenol in

sludge applied to agricultural soils would likely have high adsorption to soil particles. Various

NPEs have also exhibited high adsorption on soil (Beigel et al. 1998). The relative degree of
17

adsorption among NPE oligomers appears to increase with decreasing number of ethoxylate

units, to a maximum soil affinity at about NP9EO, after which the tendency for adsorption

decreases with decreasing number of ethoxylate units (Beigel et al. 1998). It is suggested that

adsorption increases due to increasing hydrophobicity, but with the shorter ethoxylate chains, that

have a lower critical micelle concentration, there may be a preference for surfactant-surfactant

interactions and micelle formation, rather than surfactant-soil surface interactions (Beigel et al.

1998).

Although the high adsorption of nonylphenolic compounds to soil particles suggests that there

would be a low probability of leaching (Wild and Jones 1992), numerous studies have shown

evidence of infiltration of nonylphenolic compounds into groundwater from municipal landfills (Ahel

1991), from contaminated rivers and wastewater canals (Zoller et al. 1990; Ahel 1991; Ahel et al.

1996), and due to sewage disposal through rapid infiltration (Barber et al. 1988; Rudel et al.

1998). Due to their higher water solubility and mobility, nonylphenol ethoxycarboxylates (i.e.,

NP1EC and NP2EC) are thought to pose a greater risk as groundwater contaminants than other

nonylphenolic compounds (Ahel et al. 1996). Although NP, NP1EO and NP2EO are more

effectively retained in the soil, significant concentrations of these substances have been

measured in groundwater at distances greater than 100 metres from the original source (Ahel

1991; Ahel et al. 1996).

As in water, nonylphenol is not likely to volatilize from soils in significant quantities either. For

example, of the initial 1000 mg·kg-1 nonylphenol that was spiked into a compost-sandstone

mixture, only 0.22% was lost through volatilization over a period of 40 days (Trocmé et al. 1988).

There is currently no information available in the literature on photolysis of nonylphenolic

substances on soils.

Biodegradation

The most important mechanism determining the fate of nonylphenolic compounds in water,

sediment, or soil, is likely biological degradation and transformation (Ahel et al. 1994b). NPEs

undergo a two-stage biodegradation process (refer to Figure 2). Primary biodegradation is the

process by which the molecules of NPE are oxidized or altered sufficiently to remove their

characteristic detergent properties (i.e., surfactancy, formation of foam). Often this results in the

formation of intermediate biodegradation products such as mono- (NP1EO) and di-ethoxylates

(NP2EO), nonylphenoxy acetic acid (NP1EC), nonylphenoxy ethoxy acetic acid (NP2EC), and NP.

Ultimate biodegradation (or total biodegradation) refers to the complete conversion of the

molecules to CO2, H2O, and inorganic salts (Swisher 1987). In sewage treatment processes,

efficient elimination of nonylphenol ethoxylates through primary biodegradation may occur,

however, ultimate biodegradation is limited due to the formation of products that are biorefractory,

i.e., that resist further biodegradation (Ahel et al. 1994a; Maguire 1999).

Primary Biodegradation

Nonylphenol ethoxylates undergo a relatively slow degradation process compared with some

other types of surfactants. Steric effects due to the aryl group prevent the occurrence of central

ether fission, and a branched alkyl chain reduces the likelihood of ω-/ß-oxidation1 (White 1993).

The major route of biodegradation is most likely to be ethoxylate shortening through the stepwise

removal of ethoxy groups (White 1993; Jones and Westmoreland 1998; Maguire 1999).

Therefore, the intermediate degradation products of primary biodegradation are lower NPE

ethoxamers, or NPECs (depending on environmental conditions), and nonylphenol. Under

anaerobic conditions (as in the anaerobic digestion of sludge in sewage treatment plants) the

 

1

 ω-/ß-oxidation is a two-step process which results in the shortening of an alkyl chain. Through

ω-oxidation, the terminal unsubstituted carbon of the alkyl chain is oxidized to form a carboxylic

acid (White 1993). With ß-oxidation, the carboxylic acid is further oxidized at the ß-carbon atom,

resulting in the removal of a 2-carbon fragment (Manahan 1994).
18

intermediate breakdown products include NP1EO and NP2EO; under aerobic conditions (that

would be found in most natural river waters and wastewater effluents) major degradation products

include NP1EC and NP2EC (Ahel et al. 1994b; Maki et al. 1996). Nonylphenol is a degradation

product under both aerobic and anaerobic conditions, although formation of NP appears to be

more likely under anaerobic conditions (Giger et al. 1984). Degradation of the hydrophobic moiety

(i.e., the alkyl chain and phenol group) of NPEs has also been observed, but, it occurs at only half

the degradation rate of the hydrophilic polyethoxylate chain (Jones and Westmoreland 1998).

Biodegradabilities of both NP1.5EO and NP9EO have been assessed using the CO2 Evolution

Modified Sturm Test, as described in the Organisation for Economic Co-operation and

Development (OECD) guideline 301B (Gledhill, 1999). According to the results from these tests,

although the substances are both inherently biodegradable (with >50% mineralization occurring

over the 35-day test period), neither meets the requirements for classification as “readily

biodegradable” (i.e., ≥ 60% CO2 produced within 28 days, with this value being achieved within a

10-day window). Among the higher molecular weight NPEs (i.e., with 8 to 30 EO groups)

individual ethoxamers do not appear to differ significantly in their biodegradabilities (Rudling and

Solyom 1974). Sulphated nonylphenol ethoxylates are less readily biodegraded than nonsulphated NPEs (Pitter and Fuka 1979).

Studies examining biodegradation efficiencies in wastewater treatment plants (WWTPs) using

activated sludge have reported greater than 90% primary degradation of NPEs (Rudling and

Solyom 1974; Lee and Peart 1998). Lee and Peart (1998) examined biodegradation in a southern

Ontario municipal WWTP using secondary treatment with activated sludge. Through aerobic

biodegradation, concentrations of NPEs and NP entering the WWTP in raw sewage were reduced

by more than 90 and 82%, respectively, in the final effluent. The degradation of NPEs resulted in

the formation of carboxylates, as evidenced by 418 and 468% increases in concentrations of

NP1EC and NP2EC, respectively, from the raw influent to the effluent. The median total

alkylphenolic concentrations were reduced by 53%. The authors speculated that much of the

“eliminated” nonylphenolics may have been bound to the sludge, that was used for landfill

applications, thus making the total environmental loading even higher. It has been estimated that

at least 63% of the total molar mass of all nonylphenolic compounds entering WWTPs is released

to the environment; 25% as NP, 19% as NPECs, 11% as lipophilic nonylphenol ethoxylates

(NP1EO + NP2EO), and 8% as higher NPEs (Ahel et al. 1994a). The majority of these

compounds are released through the secondary effluents, except for NP which is primarily

disposed via the digested sewage sludge (Ahel et al. 1994a; Brunner et al. 1988).

Degradation of NPEs has also been observed during the composting of sludges from wool scour

effluents (Jones and Westmoreland 1998). Over a 14 week period, NPE concentrations were

observed to decrease by more than 96%.

Laboratory studies have been conducted to simulate biodegradation processes that would occur

in natural waters. For example, Potter et al. (1999) examined the static die-away of NP18EO

added to estuarine water collected from Tampa Bay, Florida. They observed a lag-period of 0 to

12 days, followed by a rapid primary degradation, with primary degradation complete within 4 to

24 days. The first major degradation product to form was NP2EO, which then decayed to NP2EC,

a more persistent degradation product. No formation of NP was observed, perhaps because the

test conditions were aerobic. Ahel et al. (1994c) also conducted die-away tests by adding NP1EO

and NP2EO to river water containing pre-exposed bacterial cultures. They found that

biotransformation rates were 1.5 times faster when the water was stirred, than when it remained

static. Biotransformation rates were also faster at higher temperatures, with 50-60%
19

C9H19 O - (CH2 - CH2 - O) n- CH2 - CH2 - OH Nonylphenol polyethoxylate

 (NPnEO; n = 0 to 99)

 aerobic anaerobic

 (e.g., water, effluent) (e.g., sludge, soil)

 Gram negative bacteria?

 C9H19 O - (CH2 - CH2 - O) 2 - H Nonylphenol diethoxylate

(NP2EO)

 +

 C9H19 O - CH2 - CH2 - OH Nonylphenol monoethoxylate

(NP1EO)

C9H19 O - CH2 - CH2 - O- CH2 - COOH Nonylphenoxy ethoxy acetic acid

 (NP2EC)

 +

C9H19 O - CH2 - COOH Nonylphenoxy acetic acid

 (NP1EC)

 Yeast?

 C9H19 OH Nonylphenol

Stage 1 PRIMARY (NP)

Carbon dioxide, water, and

Stage 2 CO2 + H2O + inorganic salts ULTIMATE inorganic salts 

Figure 2. Pathways for biodegradation of nonylphenol polyethoxylates.
20

transformation after 12 days at 20oC compared with 10-20% at 4oC. A control sterile sample

showed no evidence of analyte transformation throughout the entire study (Ahel et al. 1994c).

In a few studies, specific microorganisms capable of biodegrading nonylphenolic compounds

have been identified. A strain of the bacteria Pseudomonas sp. isolated from activated sludge of

a municipal WWTP was observed to degrade the EO chain of nonylphenol polyethoxylates to

form NP2EO and NP2EC (Maki et al. 1994). Frassinetti et al. (1996) also isolated two strains of

Pseudomonas sp. and a strain of Xanthomonas sp. capable of degrading NP9EO, however none

of these were able to assimilate NP. Biotransformation of nonylphenol has been observed with a

strain of the yeast Candida maltosa, isolated from the sludge of a plant treating textile industry

wastewater (Corti et al. 1995). The major degradation product was 4-acetylphenol, and the

authors suggest ß-oxidation, followed by decarboxylation, as a possible biodegradation process.

It should be noted, however, that the study by Corti et al. (1995) used straight-chain NP; with

branched-chain isomers of NP, it is likely that ß-oxidation would occur much less readily (see

White 1993).

Microorganisms are more efficient at the biodegradation of nonylphenolic compounds if they have

been previously exposed and given time for adaptation; however, pre-exposure is not necessary

for biodegradation to occur. For example, biotransformation of NP1EO and NP2EO in synthetic

sewage using aerobic bacterial cultures was examined by Ahel et al. (1994c). The bacterial

cultures were isolated from three different locations: the wastewater of a detergent manufacturing

plant, from the water of a polluted river, and from a pristine forest soil in Croatia.

Biotransformation occurred quite rapidly with both the river water and wastewater cultures, while

biotransformation with the forest soil culture was less efficient.

Ultimate Biodegradation

Nonylphenol, NP1EO, NP2EO, NP1EC, and NP2EC are very refractory compounds that are much

less readily degraded than their parent compounds, the nonylphenol polyethoxylates. Partial

biodegradation and biotransformation of these compounds will occur, but as evidenced below,

there are fewer reports of complete degradation (i.e., to CO2, H2O, and inorganic salts) than there

are of primary degradation of NPEs.

Ekelund et al. (1993) examined the biodegradation of 4-nonylphenol in seawater, both with and

without sediment present, through the use of radioactive labelling. Initial degradation in the

seawater alone was very slow, with about 0.06% of the NP degraded per day, but after four weeks

this rate had increased to 1% per day. The authors suggested that this increase was due to

adaptation of the microorganisms to the substrate. In the presence of sediment, the degradation

rate was much quicker, at 1.2% per day from the onset, and remained relatively constant. The

faster rate with sediment can be accounted for by the larger number of microorganisms that would

be available, thus increasing the probability of bacteria able to degrade the NP being present.

This rate did not increase with adaptation (as it did in the water only treatment) because

adsorption to sediment particles likely reduced NP bioavailability. In the presence of sediment,

but under anoxic conditions, the biodegradation rate was reduced by 50% (Ekelund et al. 1993).

Biodegradation of NP, NP1EO, and NP2EO in soil-sludge mixtures and sludge slurries were

examined by Madsen et al. (1998). Three different sludges collected from Danish WWTPs, either

activated or anaerobically digested, were tested. After 28 days, no change in concentration was

observed for NP, NP1EO or NP2EO; however, the analysis did not examine longer-chained

nonylphenol ethoxylates, nor nonylphenoxy carboxylates. As such, it is possible that some

degradation of NP and NP1-2EO did occur, but was masked by the transformation of longerchained NPEs to short-chain NPEs and NP (Madsen et al. 1998). Two of the sludges were also

examined for biodegradation by preparing slurries of the sludges and incubating them in the dark

in closed beakers connected to an oxygen supply, and equipped with stirrers, at 15°C for 14 days.

The pre-conditioned sludge bacteria in the slurry, possibly aided by desorption of sludge-bound

contaminants through stirring, resulted in decreases of 70-80% of the initial levels of NP, NP1EO
21

and NP2EO for both sludges. How much of the decrease in NP and NPEs was due to

mineralization is unknown since there also may have been formation of other degradation

products such as NPECs.

Ultimate biodegradation of NP9EO has been observed in soil under aerobic conditions. During a

nine week experimental period, 57% of the initially applied NP9EO was mineralized, including

breakdown of the benzene ring, as confirmed through HPLC with UV detection (Hughes et al.

1996). Mineralization of nonylphenol has been observed in a variety of soil types, including

agricultural soils of various textures, non-cultivated temperate soils, and soils from the Canadian

tundra (Topp and Starratt 2000). The fact that biodegradation occurred in the arctic soils

indicated that the microorganisms already possessed the biochemistry for nonylphenol

metabolism without prior exposure and adaptation. The authors suggested that exposure of

microorganisms to phenolic substances released by the decomposition of plant matter may have

enabled them to degrade nonylphenol (Topp and Starratt 2000).

Mineralization of nonylphenol by soil bacteria can be inhibited by either an overabundance or a

lack of water content (Topp and Starratt 2000). Temperature has also been observed to affect

rates of nonylphenol mineralization in soil. Although biodegradation will occur over the

temperature range of 4 to 30°C, at lower temperatures the rate decreases, and there is a greater

lag period (Topp and Starratt 2000). Overall, the authors conclude that nonylphenol in landapplied sewage sludge would readily undergo biodegradation under the soil conditions of a normal

Canadian growing season.

Wild and Jones (1992) suggest a half-life of <10 days for nonylphenol in sewage sludge. Data

from Ekelund et al. (1993) indicate a half-life for NP in seawater of 56 days, while Sundaram and

Szeto (1981) measured a freshwater half-life of about 16 days. Marcomini et al. (1988), studying

degradation of NP in soil, measured a half-life of 8 days during the first 10 days after application,

followed by a 90 day period in which the degradation rate slowed to give a half-life of 90 days. An

analysis of data from various sources indicated that half-lives measured from the end of the lag

phase to the beginning of the plateau for the NP degradation curves results in half-lives ranging

from 6 to 26 days for water, soil, and water-sediment slurries (Naylor 2000).

Uptake/Elimination Pathways, and Metabolism

The degree to which a substance bioaccumulates in an organism is dependent on the relative

rates of uptake and elimination. These processes can also have a bearing on the type, and

magnitude, of toxic effect that a substance will have. Metabolism, or biotransformation, of a

substance within an organism will produce metabolites that may be more or less toxic than the

parent substance. Therefore, an understanding of the processes affecting a substance within an

exposed organism will permit a greater understanding of the toxicological risks of that substance

to the organism. A discussion of the physiological and metabolical pathways for NP and NPEs in

fish, invertebrates, plants, and wildlife is provided below.

Numerous researchers have examined uptake and elimination pathways for nonylphenolic

compounds in fish. Uptake occurs primarily through the gills with transport to the various

tissues/organs via the blood (Granmo and Kollberg 1976; Arukwe et al. 2000b). For example,

Granmo and Kollberg (1976) found that after only five minutes of exposure to NP10EO, high

levels of the contaminant were observed in the gill tissue and blood of cod (Gadus morhua).

Smaller amounts of NP10EO were also taken up through intestinal resorption. Uptake of

nonylphenolic compounds is fairly rapid, with tissue concentrations reaching a steady state during

the first 3 to 10 days of exposure (Granmo and Kollberg 1976; Ward and Boeri 1991c). Within a

fish, nonylphenol can concentrate in the viscera at levels 1.6 to 7.1 times higher than in the

carcass (Ward and Boeri 1991c). Once a stable equilibrium has been reached, the highest

concentrations of nonylphenolics in fish can be found in the gall bladder, bile, pyloric caecae,

intestine, and faeces (Granmo and Kollberg 1976; Lewis and Lech 1996; Coldham et al. 1998;

Arukwe et al. 2000b). Intermediate levels of nonylphenolic compounds are found in the liver,

kidney and fat tissue, and comparatively low concentrations can be found in the gills, heart,
22

gonad, muscle, spleen, skin, blood, eye and brain (Granmo and Kollberg 1976; Lewis and Lech

1996; Coldham et al. 1998; Arukwe et al. 2000b).

When fish exposed to nonylphenolic compounds are transferred to clean water, elimination of the

contaminants will occur for all tissues, although there appears to be a longer time lag for the gall

bladder, where concentrations of nonylphenolic substances have been observed to remain stable

for extended periods (Granmo and Kollberg 1976; Coldham et al. 1998). For most of the tissues,

Coldham et al. (1998) found that the elimination process was biphasic and fit a two compartment

model. The highest rate of elimination occurs in the blood and gills (Granmo and Kollberg 1976),

and the lowest rates of elimination occur in the liver, muscle, and skin tissues, as evidenced by

the half-lives (Coldham et al. 1998). In rainbow trout (Oncorhynchus mykiss), the estimated halflife of NP in fat was 20 hours, in muscle it was 19 hours, and in the liver the half-life was six hours

(Lewis and Lech 1996). Arukwe et al. (2000b) found that the half-lives for nonylphenol residues in

the carcass, viscera and muscle of Atlantic salmon (Salmo salar) were all between 24 and 48

hours. For all tissues, rates of elimination are slower than rates of uptake (Coldham et al. 1998).

The whole-body excretion half-life for juvenile Atlantic salmon exposed to technical grade

p-nonylphenol was estimated at four days (McLeese et al. 1981). Some excretion is thought to

occur via the kidney, with high levels of nonylphenolics being observed in the urine (Granmo and

Kollberg 1976). Thibaut et al. (1999) gave rainbow trout a single dose of radioactively labelled

nonylphenol and observed urinary elimination of 1.1% of the radioactivity after 24 hours, and 3.0%

elimination through urine after 48 hours. In contrast, Coldham et al. (1998) suggest that the main

pathway for elimination of nonylphenol follows a biliary/faecal route of excretion. Evidence for this

pathway comes from the high equilibrium concentrations of nonylphenolics that have been

observed in tissues associated with the biliary/faecal system (Granmo and Kollberg 1976; Lewis

and Lech 1996; Coldham et al. 1998; Arukwe et al. 2000b). Ekelund et al. (1990) exposed threespined sticklebacks (Gasterosteus aculeatus) to 14C-labelled NP in a flow-through system with

seawater for 16 days, followed by an elimination period of 32 days. They found that the

sticklebacks reached steady states of bioaccumulation after about ten days and elimination of NP

was quite rapid and almost complete after a few days.

Within the fish tissue, nonylphenol may be biotransformed into various metabolic products.

Researchers have suggested that metabolic degradation of nonylphenol in fish occurs through

oxidation and glucuronic acid formation in bile (Lewis and Lech 1996; Meldahl et al. 1996; Arukwe

et al. 2000b). Through in vitro tests with rainbow trout hepatic microsomes, Meldahl et al. (1996)

also observed metabolism of nonylphenol by hepatic cytochrome P450. Some possible

metabolites of nonylphenol have been tentatively identified in rainbow trout as

9-(4-hydroxylphenyl)-nonanoic acid, 4-hydroxybenzoic acid, 3-(4-hydroxyphenyl) propionic acid,

and 3-(4-hydroxylphenyl)-2-propenoic acid (Thibaut et al. 1999). The process of ω-oxidation

followed by ß-oxidation has been proposed as the most likely biotransformation pathway (Thibaut

et al. 1999).

There are few studies that have investigated uptake and elimination rates in invertebrates.

Ekelund et al. (1990) exposed common blue mussels (Mytilus edulis) and common shrimp

(Crangon crangon) to 14C-labelled NP in a flow-through system with seawater for 16 days,

followed by an elimination period of 32 days. The shrimp reached steady states of

bioaccumulation after approximately ten days, however, with the mussels, steady state was not

reached by the end of the 16 day exposure period. Elimination of NP from the shrimp took about

four days, but was much slower in the mussels which still had significant concentrations present

after 30 days in clean water. McLeese et al. (1980b) also found that common blue mussels have

low uptake rates and have high excretion rates of nonylphenol.

There has been considerably greater investigation into the uptake/elimination and metabolism of

nonylphenol in plants than in invertebrates. Much of the literature has focussed on agricultural

plant species exposed to nonylphenolic compounds through contaminated soil. Based on the

physical and chemical properties of nonylphenol, it has been hypothesized that high retention of

nonylphenol at the root surface of plants would be likely, with only moderate uptake and
23

translocation occurring within the plants (Wild and Jones 1992). Kirchmann and Tengsved (1991)

examined the grains of wheat plants grown in soil contaminated with nonylphenol due to sewage

sludge application. They found no significant increase in nonylphenol concentrations in the

grains, though were uncertain whether this indicated a lack of root uptake and transport to the

grains, or rapid decomposition of the nonylphenol in the soil. Harms (1992) also did not observe

any evidence of nonylphenol in the shoots of tomato plants whose roots were exposed to

nonylphenol. In contrast, translocation of nonylphenol or its metabolites from roots to shoots

within wheat, tomato and garden orache plants has been observed by other researchers (Bokern

et al. 1998).

Bokern et al. (1996) examined the metabolism of radioactively labelled 4-n-nonylphenol in wheat

cell suspension cultures. Metabolic products observed were 4-(hydroxy)- and

4-(dihydroxy)nonylphenols that were glucosylated at the phenolic OH-group and further

glucosylated, glucuronidated, and acylated with acetic acid or malonic acid. Metabolism of

nonylphenol was also examined in cell suspension cultures of Daucus carota (carrot) and Atriplex

hortensis (garden orache) (Bokern and Harms 1997). Radiolabelled nonylphenol was taken up

quickly in both cultures with 93-97% of the radioactivity disappearing from the culture medium

after six hours, and concentrations reaching a steady state after 48 hours. Metabolism of

nonylphenol within the cells occurred quickly. In the carrot cell cultures, 80% of the parent

compound had been metabolized within 24 hours, and in the A. hortensis cells about 50% of the

nonylphenol was metabolized after 30 hours (Bokern and Harms 1997). The authors also looked

at the uptake and metabolism of nonylphenol in 14 different cell cultures (predominantly

agricultural crop species). All of the species took up NP and metabolized it. Metabolites were

ß-glucosides of differently hydroxylated nonylphenol, some of which were further acylated with, for

example, glucuronic acid or malonic acid. Cell cultures of plants from similar taxonomic groups

tended to produce similar metabolites (Bokern and Harms 1997). Distribution of radioactivity

among the various cell wall fractions appeared to be species specific. In the majority of species,

high concentrations were associated with the lignin, though in some species it was associated

with the protein or hemicellulose. Although most plant species seem to readily take up

nonylphenol (at least when exposed in vitro) and metabolize it, no mineralization of nonylphenol to

CO2 appears to occur in plants (Bokern et al. 1998).

Considerably less information exists on the uptake of nonylphenolic compounds by aquatic plants.

In a littoral mesocosm study, nonylphenol applied to the water column was found to partition to

macrophytes, with maximum concentrations of 4-NP observed in macrophytes within 1 to 2 days

of application. The authors estimated the mean DT50 and DT95 for nonylphenol persistence in/on

macrophytes at ten and 189 days, respectively, indicating persistence of nonylphenol in

macrophytes (Heinis et al. 1999).

There is a paucity of data on uptake of nonylphenolic compounds by terrestrial animals. Based on

the physical and chemical properties of nonylphenol, Wild and Jones (1992) have speculated that

there is a moderate chance of transfer of nonylphenol to animal tissues through foliage ingestion,

with less chance of transfer occurring through soil ingestion.

Bioaccumulation

Bioaccumulation is defined as the net accumulation of a substance by an organism as a result of

uptake from all routes of exposure (e.g., water intake, food ingestion, dermal absorption, etc.).

Bioconcentration also refers to net accumulation of a substance, but with respect to aquatic

organisms, and involving only direct uptake from an aqueous solution. Thus, bioaccumulation

includes the process of bioconcentration, but not vice versa. Nonylphenolic compounds have

been evaluated for their potential to bioaccumulate or bioconcentrate both through direct

investigations involving exposure of organisms to these substances under natural or artificial

conditions, and through studies of their physical/chemical properties.

Numerous studies have examined bioaccumulation potential through examination of nonylphenol

concentrations in aquatic biota relative to water concentrations, and nonylphenolic compounds
24

appear to bioaccumulate and bioconcentrate to a low or moderate degree. Several researchers

have observed bioaccumulation of NP and NPEs in common blue mussels when placed in cages

near industrial wastewater outflows or when exposed to NP (or wastewater containing NP and

NPEs) under laboratory conditions (McLeese et al. 1980b; Wahlberg et al. 1990; Granmo et al.

1991a,b). Granmo et al. (1991b) found that the amount of bioaccumulation of NP, NP1EO,

NP2EO, and NP3EO depended on the length of the ethoxylate chain with greater bioaccumulation

of compounds with shorter ethoxylate chains (and therefore greater hydrophobicity). Ethoxylate

chain length appears to affect bioconcentration in aquatic plants and algae as well. In their field

study of the Glatt River, Switzerland, Ahel et al. (1993) found that although nonylphenol was

present in the river water at a lower concentration (3.9 μg·L-1) than NP1EO or NP2EO (24 and

9.4 μg·L-1, respectively), it occurred at higher concentrations (2.5 to 38.0 mg·kg-1 dry weight) in the

resident algae and macrophytes than the other two compounds (0.6 to 4.7 mg·kg-1 dry weight).

For the purposes of comparing bioaccumulation potentials from different studies, or across

different substances and different species, bioaccumulation factors (BAFs) and bioconcentration

factors (BCFs) are calculated. Generally, BAFs are calculated in field studies and BCFs are

determined in laboratory studies. Because BCFs do not include dietary uptake, they tend to

underestimate the bioaccumulation potential of a substance. Table 3 lists BAF and BCF values

that have been determined for NP and NPEs on a variety of aquatic species. In general, BAF and

BCF values are higher for NP than for NPEs (i.e., BAFs and BCFs decrease with increasing

ethoxylation). Considerable variation in BAF and BCF values are observed within taxa, and even

within the same species, which makes it difficult to discern any significant differences in

bioaccumulation between algae, plants, invertebrates, fish, and waterfowl. Under the Toxic

Substances Management Policy (Government of Canada 1995) and under Canada’s Persistence

and Bioaccumulation Regulations (Government of Canada 2000b), a substance is considered

bioaccumulative if its BAF (or BCF) is equal to or greater than 5000. Ahel et al. (1993) reported

BAFs greater than 5000 in the algae Cladophora glomerata. Ekelund et al. (1990) calculated

BCFs greater than 5000 in a crustacean, a mollusc, and a fish when based on lipid

concentrations, but not when based on total wet weight. All other reported BAFs and BCFs fall

below the critical value of 5000 (see Table 3), suggesting that nonylphenolic substances are not

highly bioaccumulative.

Based on their physical and chemical properties, little bioaccumulation and bioconcentration of

nonylphenolic compounds are likely to occur. The potential for a contaminant to bioaccumulate is

largely affected by its hydrophobicity, and therefore is often evaluated based on its octanol-water

partition coefficient (Kow). Estimates of the log Kow for nonylphenol of 4.7 (Barber et al. 1988),

4.48 (Ahel and Giger 1993b), 4.2 (McLeese et al. 1981), and 4.1 have been made (Wild and

Jones 1992). Partition coefficients between n-octanol and water have also been determined for

several of the smaller NPEs. The log Kows for NP1EO, NP2EO and NP3EO are estimated at

4.17, 4.21, and 4.20, respectively (Ahel and Giger 1993b). According to the Toxic Substances

Management Policy (Government of Canada 1995) and Canada’s Persistence and

Bioaccumulation Regulations (Government of Canada 2000b), in the absence of BAF or BCF

data, a substance may be considered as bioaccumulative if it has a log Kow value equal to or

greater than 5. As estimates of the log Kow for nonylphenolic substances all fall below 5, this

reinforces the conclusion based on BAF values that these substances are not bioaccumulative.

Partition coefficients (Khw) have also been determined for the partitioning of nonylphenolics

between n-hexane and water. Log Khws for NP, NP1EO, and NP2EO were estimated at 3.7, 3.4,

and 3.3, respectively (Ahel and Giger 1993b). The compounds generally had a lower solubility in

hexane than in octanol. With both organic solvents, the difference of a single ethoxylate group

had a negligible influence, with NP1EO and NP2EO having almost equal lipophilicity (Ahel and

Giger 1993b). Although neither log Kow nor log Khw values have not been calculated for the higher

NPEs, they are known to be less hydrophobic than nonylphenol and the lower molecular weight

NPEs and would therefore have lower potential for bioaccumulation as well.

Substances that bioaccumulate or bioconcentrate may also have the potential to biomagnify.

There is no evidence in the current literature to suggest that NP or NPEs biomagnify. For
25

example, Ahel et al. (1993) examined concentrations of NP, NP1EO and NP2EO in biota, at

several trophic levels, of the Glatt River, Switzerland. For each of these substances,

bioaccumulation factors in the algae Cladophora glomerata were significantly higher than the

BAFs in three fish species (Squalius cephalus, Barbus barbus, and Oncorhynchus mykiss).

Concentrations of nonylphenolic compounds detected in the tissues of a mallard duck (Anas

boscas) were not significantly different from concentrations observed in fish tissues. Because the

concentrations of NP and NPEs did not increase with increasing trophic level of organisms, the

authors concluded that the compounds did not biomagnify.
26

Table 3. Bioconcentration and bioaccumulation factors for nonylphenol and nonylphenol

ethoxylates in aquatic organisms.

Chemical Species Water

type

BCF BAF Tissue Reference

Plants/Algae

NP Cladophora glomerata

(algae)

fresh NR 10000 *

6600-7700

dw Ahel et al. 1993

Fontinalis antipyretica

(macrophyte)

fresh NR 1100 dw Ahel et al. 1993

Potamogeton crispus

(macrophyte)

fresh NR 600 dw Ahel et al. 1993

NP1EO Cladophora glomerata

(algae)

fresh NR 200 *

3500-5000

dw Ahel et al. 1993

Fontinalis antipyretica

(macrophyte)

fresh NR 40 dw Ahel et al. 1993

Potamogeton crispus

(macrophyte)

fresh NR 50 dw Ahel et al. 1993

NP2EO Cladophora glomerata

(algae)

fresh NR 500 *

1000-1800

dw Ahel et al. 1993

Fontinalis antipyretica

(macrophyte)

fresh NR 60 dw Ahel et al. 1993

Potamogeton crispus

(macrophyte)

fresh NR 200 dw Ahel et al. 1993

Invertebrates

NP Crangon crangon

(common shrimp)

marine 90,110 NR based on

total ww

Ekelund et al. 1990

7500,5500 NR lipidbasedEkelund et al. 1990

Mytilus edulis

(common blue mussel)

marine 1.4-7.9

(K1/K2 = 10)

NR NR McLeese et al. 1980b

NR 340 NR Granmo et al. 1991b

2740,4120 NR based on

total ww

Ekelund et al. 1990

169300,

216600

NR lipidbasedEkelund et al. 1990

NP1EO Mytilus edulis

(common blue mussel)

marine NR 170 NR Granmo et al. 1991b

NP2EO Mytilus edulis

(common blue mussel)

marine NR 100 NR Granmo et al. 1991b

NP3EO Mytilus edulis

(common blue mussel)

marine NR 60 NR Granmo et al. 1991b
27

Table 3 (continued). Bioconcentration and bioaccumulation factors for nonylphenol and

nonylphenol ethoxylates in aquatic organisms.

Chemical Species Water

type

BCF BAF Tissue Reference

Fish

NP Barbus barbus

(barb)

fresh NR 92 muscle,

gut, liver,

heart, roe

(dw)

Ahel et al. 1993

Cyprinus carpio

(carp)

fresh 2.5-3.3

0.9-2.2

NR NR CITI 1992 as cited in

Staples et al. 1998

Gasterosteus

aculeatus

(3-spined stickleback)

marine 1200,1300 NR based on

total ww

Ekelund et al. 1990

17800, 16700 NR lipidbasedEkelund et al. 1990

Lepomis macrochirus

(bluegill sunfish)

fresh 220 NR ww Brooke 1993b

NR 87 ww Liber et al. 1999a

Oncorhynchus mykiss

(rainbow trout)

fresh 21-29 NR carcass Lewis and Lech 1996

90-110 NR viscera

minus gall

bladder/

bile

Lewis and Lech 1996

NR 224 muscle,

gut (dw)

Ahel et al. 1993

Pimephales promelas

(fathead minnow)

fresh 271, 344 NR whole fish Ward and Boeri

1991c

741 NR ww Brooke 1993b

Salmo salar

(Atlantic salmon)

marine 75 NR NR McLeese et al. 1981

190 NR NR McLeese et al. 1981

235 NR NR McLeese et al. 1981

280 NR NR McLeese et al. 1981

Squalius cephalus**

(chub)

fresh NR 246 muscle,

gut, liver,

gills (dw)

Ahel et al. 1993

NP1EO Barbus barbus

(barb)

fresh NR 31 muscle,

gut, liver,

gills, roe,

heart

(dw)

Ahel et al. 1993

Oncorhynchus mykiss

(rainbow trout)

fresh NR 155 muscle,

gut (dw)

Ahel et al. 1993

Squalius cephalus* *

(chub)

fresh NR 263 muscle,

gut, liver,

gills (dw)

Ahel et al. 1993
28

Table 3 (continued). Bioconcentration and bioaccumulation factors for nonylphenol and

nonylphenol ethoxylates in aquatic organisms.

Chemical Species Water

type

BCF BAF Tissue Reference

NP2EO Barbus barbus

(barb)

fresh NR 88 muscle,

gut, liver

(dw)

Ahel et al. 1993

Oncorhynchus mykiss

(rainbow trout)

fresh NR 162 muscle,

gut (dw)

Ahel et al. 1993

Squalius cephalus* *

(chub)

fresh NR 73 muscle,

gut, liver,

gills (dw)

Ahel et al. 1993

Wildlife

NP Anas boscas

(mallard duck)

fresh NR 308 muscle

(dw)

Ahel et al. 1993

NP1EO Anas boscas

(mallard duck)

fresh NR 88 muscle

(dw)

Ahel et al. 1993

NP2EO Anas boscas

(mallard duck)

fresh NR 37 muscle

(dw)

Ahel et al. 1993

NR = not reported

ww = wet weight

dw = dry weight

* the authors (Ahel et al. 1993) note that due to various sources of uncertainty these calculated

BAFs for Cladophora glomerata are only intended to provide an approximation of the order of

magnitude and should not be interpreted as exact bioaccumulation factors

** the more commonly accepted name for this species is Leuciscus cephalus
29

Toxicity

Mode of Action

Although extensive research has been conducted on the toxic effects of nonylphenolic

compounds, much less is known about their actual mode of toxicity. It has been suggested that

the mode of toxicity for nonylphenol and the lower chain length NPEs and NPECs is narcosis,

while toxicity from longer chain length NPEs may be due to physical surfactant effects (Fay et al.

2000; Servos et al. 2000). It is also likely that the mode of toxicity will depend on the dose, with

high concentrations above the critical micelle concentration resulting in more effects due to

physical surfactancy. Nonylphenolic substances may also act as estrogen agonists to disrupt

endocrine functions.

Narcosis

Narcosis is a nonspecific, reversible mode of toxic action in which the presence of hydrophobic

organic chemicals causes a disruption of cellular activity, possibly through binding to proteins in

the cell membrane, or through “swelling” of the lipid portion of cell membranes (Rand 1995).

Numerous researchers have observed changes in the cellular membranes of organisms exposed

to NP and NPEs, thus giving support to the theory that these substances exert a narcotic-type

effect. For example, increased cell membrane permeability occurs in plants exposed to

nonylphenol (Silcox and Holloway 1986). Linsel et al. (1990) found that NPEs caused membrane

depolarization affecting electron transport across the plasmalemma of the aquatic plant Egeria

densa. Octylphenol ethoxylates, that likely have a similar mode of toxicity to their nonylphenol

counterparts, are thought to act on aquatic macrophytes at the cellular membrane (Caux and

Weinberger 1993). It has been proposed that at low surfactant concentrations (i.e., below the

critical micelle concentration) the surfactant molecules mimic membrane phospholipids and

intercalate in the phospholipid bilayer of the cell membrane causing compression of the lipid

constituents, thereby inducing fatty acid desaturate enzymes to render membranes more fluid

(Caux and Weinberger 1993). In unicellular algae, nonylphenol concentrations greater than

1000 μg·L-1 cause membrane disorganization including crennulation, vesiculation and pinocytotic

invaginations of the nuclear membrane, as well as cytoplasm friability and distortion of flagellae

indicative of microtubular damage (Weinberger and Rea 1981; Weinberger and Greenhalgh

1984). Tree seedlings grown in aqueous solutions of nonylphenol display excessive swelling of

cell mitochondria, loss of chloroplast integrity, and convolution of the plasmalemma at

concentrations of 1000 μg·L-1 (Weinberger and Greenhalgh 1984). Massive K

+

 ion leakage from

the roots of tree seedlings also occurs at NP concentrations of 1500 μg·L-1 (Weinberger and

Greenhalgh 1984).

There are indications that nonylphenolic compounds may alter the permeability of cellular

membranes in animals. For example, tissues of saltwater mussels became swollen when

exposed to a nonylphenol polyethoxylate. The authors concluded that the swelling was likely

caused by some nonspecific damage to the cell membranes that could have affected the

osmoregulatory capacity of the tissues (Braaten et al. 1972). At a very high concentration

(63,000 μg·L-1) NP10EO also caused an increase in ion permeability in the gill epithelial cells of a

rainbow trout perfused head (Pärt et al. 1985). A decreased oxygen diffusion capacity was

measured in the gills, possibly because the altered permeability caused swelling of the epithelium

(Pärt et al. 1985). There was also an increase in the vascular resistance of the gills that may have

been caused by an inhibition of the adrenergic dilatation of the gill vasculature by the surfactant

(Pärt et al. 1985).

Nonylphenol also causes vasodilation in the vascular smooth muscle of rats by inhibiting Ca2+

influx through L-type Ca2+ channels in the smooth muscle cells (Ruehlmann et al. 1998). The

sarcoplasmic reticulum (Ca2+ - Mg2+)-ATPase may be another specific target site of nonylphenol

action in animal cells (Michelangeli et al. 1990). At high concentrations (5 to15 μM), nonylphenol

inhibits the ATPase activity, while at low concentrations (1 to 3 μM) it increases ATPase activity
30

(Michelangeli et al. 1990). Nonylphenol may, therefore, affect other P-type ATPase

electroenzymes such as the plasma membrane H

+

-ATPase of plants and fungi (Karley et al.

1997). It appears that NP may partition into the mitochondrial membranes of fungi and induce

ionophoretic uncoupling of respiration (Karley et al. 1997). This process would restrict the

cytosolic ATP supply, thus indirectly inhibiting the plasma membrane H

+

-ATPase that is essential

in maintaining cytosolic ionic homeostasis (Karley et al. 1997). Weinberger and Greenhalgh

(1984) found that NP at 6.25 μg·L-1 causes disruption in the normal sequence of ATP production

in birch and pine seedlings.

QSAR modelling also supports the theory that acute toxicity of APEs follows a narcotic-type

mechanism that is likely to be intermediate with respect to the nonpolar and polar narcosis

syndromes (Schüürmann 1991). Although information is not available for NP itself, QSAR

modelling has shown that other APs, such as 4-propylphenol and p-tert-pentylphenol, also have a

mode of action that is between nonpolar (type I) and polar (type II) narcosis (Veith and Broderius

1990).

Endocrine Disruption

There is considerable evidence to suggest that nonylphenol is an endocrine disrupting chemical

(EDC) with estrogenic properties. The discovery was first made unintentionally when Soto et al.

(1991), who were working with human MCF7 breast tumour cells, found that the use of a new

brand of polystyrene centrifuge tubes resulted in cell proliferation. They were able to extract the

substance being released from the polystyrene using methanol, purify it through flash

chromatography and reverse-phase HPLC, and identify it by GC-MS as nonylphenol. Further

tests showed that the nonylphenol induced progesterone receptors, as well as cell proliferation in

the MCF7 cells, and it induced mitotic activity in rat endometrium - all indicators of estrogenic

activity (Soto et al. 1991).

It has since been determined that NP, NP2EO, and NP1EC mimic the effects of 17ß-estradiol by

binding to the estrogen receptor, thereby acting as an estrogen agonist (White et al. 1994).

Moreover, these compounds will competitively displace 17ß-estradiol from binding to the estrogen

receptor, suggesting that they have intrinsic estrogenic activity (White et al. 1994). Estrogen

receptor competitive binding assays have been conducted with rainbow trout hepatocyte cultures

to ascertain relative potencies of the various nonylphenolics. Relative potencies of nonylphenol

compared to 17ß-estradiol have been measured at 1.0 x 10-3 (Tremblay et al. 1995) and 2.5 x 10-4

(Tremblay and Van Der Kraak 1998). Estrogenicity of NP and NP2EO is similar, while NP1EC

shows slightly greater estrogenicity (White et al. 1994). White et al. (1994) found that APEs with

more than 3 EO groups had little or no estrogenicity. This seems to indicate that the larger NPEs

that are used commercially are unlikely to act as estrogen agonists in their parent form.

Numerous studies have since examined the estrogenicity of nonylphenol in fish through

vitellogenin production. Vitellogenin is a lipoglycophosphoprotein that is normally produced in the

liver of sexually mature female fish under estrogenic control (Jobling and Sumpter 1993;

Bjerregaard et al. 1998). Vitellogenin secreted by the liver is transported via the blood to the

ovaries where it forms the major constituent of the yolk in oocytes. Male fish possess the gene for

vitellogenin, but normally do not express it due to low concentrations of estrogen in their systems.

Treatment of male fish with estrogen, or chemicals with estrogen-like activity, will induce

vitellogenin production and blood concentrations of vitellogenin will increase rapidly. Because this

vitellogenin response is directly related to the binding of chemicals to the estrogen receptor, it may

be used as a biomarker for estrogenicity (Bjerregaard et al. 1998).

Following intraperitoneal injections of nonylphenol (at concentrations ranging from 10 to

200 mg·kg-1 body weight), elevated vitellogenin levels (as much as several hundred-fold and as

high as 8000 mg·L-1) have been detected in the blood plasma of various fish species, including

rainbow trout (Christiansen et al. 1998a), flounder (Platichthys flesus; Bjerregaard et al. 1998),

Atlantic salmon (Yadetie et al. 1999), and eelpouts (Zoarces viviparus; Christiansen et al. 1998b).

Elevation of vitellogenin levels from control levels of 30 mg·L-1 to 40000 mg·L-1 of plasma have
31

also been observed in eelpouts exposed to nonylphenol (at 1000 μg·L-1) through the water column

(Korsgaard and Pedersen 1998). The increase in vitellogenin level observed in the plasma shows

a dose-dependent relationship with the amount of nonylphenol administered (Bjerregaard et al.

1998; Korsgaard and Pedersen 1998; Yadetie et al. 1999).

Arukwe et al. (1997b) have suggested that eggshell zona radiata proteins (Zrp) are even more

sensitive than vitellogenin as biomarkers of estrogenic activity by nonylphenol in fish. Like

vitellogenin, Zrp are produced in the liver in response to estrogen stimulation, and are transported

to the ovary where they form eggshells, while vitellogenin forms the yolks. Juvenile Atlantic

salmon injected intraperitoneally with nonylphenol have shown significantly elevated levels of Zrp

as well as vitellogenin (Arukwe et al. 1998; Yadetie et al. 1999).

Estrogenicity of nonylphenolic compounds has also been investigated through in vitro assays in

which vitellogenin induction in fish hepatocytes has been assessed. Jobling and Sumpter (1993)

assessed the estrogenic capacities of NP, NP1EC, NP2EO, NP9EO, and NP40EO using an in

vitro bioassay with cultures of male rainbow trout hepatocytes. Treatment with NP2EO and

NP1EC resulted in a vitellogenin production 40-50 times higher than that of controls with mean

ED50s of 17.27 μM (5326 μg·L-1) and 15.25 μM (4916 μg·L-1), respectively. NP and NP9EO

caused an 18-20 fold increase in vitellogenin production with mean ED50s of 16.15 μM

(3558 μg·L-1) and 82.31 μM (50800 μg·L-1), respectively. All of these substances became cytotoxic

at concentrations between 50 and 200 μM, with NP being more toxic than the ethoxylates. The

concentration of vitellogenin found in the NP40EO treatment was not significantly different from

the control. The authors noted that the potency of the polyethoxylate compounds seems to

decrease with increasing length of the ethoxylate chain. When NP was added to rainbow trout

hepatocytes with Tamoxifen (which inhibits binding to the estrogen receptor site), the vitellogenin

production was significantly lower than when NP was added alone. This result further confirms

that NP also acts via the estrogen receptor in a manner similar to estradiol itself (Jobling and

Sumpter 1993).

Rainbow trout hepatocytes were treated with estradiol in both the presence and absence of

nonylphenol to assess the effects on induction of vitellogenin mRNA and estrogen receptor mRNA

(Flouriot et al. 1995). No synergistic, additive, or repressive effects were observed. Knudsen et

al. (1998) also found no evidence of synergism in the induction of zona radiata proteins or

estrogen receptor modulation when octylphenol and estradiol were administered simultaneously to

rainbow trout through intraperitoneal injections. When five estrogenic chemicals (nonylphenol,

octylphenol, o,p’-DDT, Aroclor 1221, and bisphenol A) were administered together to rainbow trout

hepatocytes, a greater induction of vitellogenin was observed when compared with the effects of

each chemical when administered singly (Sumpter and Jobling 1995).

Aside from vitellogenesis, other estrogenic effects have also been observed in fish.

Intraperitoneal injection of male eelpouts with 10 and 100 μg·g-1 body weight per week of

nonylphenol resulted in reduced gonadosomatic indices and severe histological effects on the

testicular structure (Christiansen et al. 1998b).

In a study comparing historical pesticide application data with recreational catch data from Atlantic

Canada, epidemiological evidence showed a relationship between use of Matacil 1.8D (an

insecticide formulated with NP) and a decline in catches of Atlantic salmon, Salmo salar (Fairchild

et al. 1999). Similar declines were not observed with the application of other Matacil that did not

contain NP, indicating that NP was the causal agent for the reduced fish abundance. Because the

timing of the pesticide spraying coincided with the final stages of salmon smoltification, the

authors speculated that endocrine disruption of smoltification may have led the lower catches

(Fairchild et al. 1999). Laboratory studies with juvenile Atlantic salmon have also indicated that

NP may affect the smolting and osmoregulatory processes of anadromous fish when they are

exposed to seawater (Madsen et al. 1997; Brown et al. 1998).
32

Endocrine disruption and estrogenic effects of NP, NP2EO, NP1EC and NP2EC have also been

observed in a variety of organisms other than fish. Exposure to nonylphenol has been found to

increase the production of cypris major protein in larvae of the marine barnacle Balanus amphitrite

(Billinghurst et al. 2000). Cypris major protein is required for successful larval settlement and

juvenile development.

Immature female rats showed dose-dependent increases in uterine mass, increased protein and

DNA contents of the uterus, and increased levels of the uterine peroxidase enzyme when injected

intraperitoneally with nonylphenol at concentrations greater than 1 mg per animal (Lee and Lee

1996). This estrogenic response due to nonylphenol exposure was approximately 1x10-3 to 2x10-3

times as potent as that elicited by estradiol. Administration of an estrogen antagonist blocked the

effects of NP, confirming that the action of nonylphenol in rats is through the estrogen receptor

(Lee and Lee 1996). The authors also found that there is direct competition between NP and

estradiol. Because NP is less efficient than estradiol, its presence reduces the estrogenic effects

of estradiol. Chapin et al. (1999) examined effects of NP administered through the diet on several

generations of rats. Effects indicative of an estrogenic response included vaginal opening at a

younger age when exposed to NP at 650 mg·kg-1 food, and increased estrous cycle length at an

NP concentration of 2000 mg·kg-1 food.

Using a recombinant yeast strain, Routledge and Sumpter (1996) examined the ability of

nonylphenolic compounds to stimulate transcriptional activity of the human estrogen receptor

(hER). NP, NP1EC, NP2EC and NP2EO were all found to produce dose-dependent elevations of

estrogenic activity. This yeast estrogen screening assay was repeated by Metcalfe et al. (2001),

confirming the estrogenic responses to NP, NP1EO and NP2EO. With NP1EC and NP2EC,

however, Metcalfe et al. (2001) observed no estrogenic response. Similar results were obtained

in assays with Japanese medaka (Oryzias latipes) in which a mixture of NP1EO and NP2EO was

observed to alter gonadal development, while no evidence of estrogenicity was observed for a

mixture of NP1EC and NP2EC (Metcalfe et al. 2001). The contradiction between these two

studies indicates that further work is needed to confirm whether or not NPECs produce estrogenic

responses.

White et al. (1994) found that NP, NP2EO and NP1EC were estrogenic in mammalian (mouse,

human) and avian (chicken) cells. Lutz and Kloas (1999) compared the estrogenic binding

affinities for several compounds by examining the competitive displacement of estradiol in

cytosolic fractions from the livers of the amphibian Xenopus laevis. They found that NP had a

higher binding affinity than octylphenol. Fifty percent displacement of 18.9 nM estradiol occurred

at 7424 μg·L-1 NP and at 16153 μg·L-1 OP.

Besides acting as an estrogen agonist, it appears that nonylphenol may also be capable of

interacting with androgen receptors, though with a lower affinity than for estrogen receptors (Gray

and Kelce 1996). Both NP and NPEs have been found to reduce the elimination of glucose- and

sulphate-conjugated testosterone metabolites while increasing the elimination of oxido-reduced

derivatives of testosterone and increasing metabolic androgenization in daphnids (Baldwin et al.

1997, 1998).

A brief discussion of the ecological relevance of endocrine disruption, and whether such endpoints

should be used in the development of environmental quality guidelines, is presented in Annex I.

Physical Surfactancy

Apart from interactions with cellular membranes and membrane transport proteins, or endocrinal

effects (as discussed in the next section), the only other mode of toxicity for nonylphenol

mentioned in the literature deals with its physical surfactancy. For insects with an aquatic larval

stage, such as mayflies and caddisflies, decreased adult emergence has been observed during

and immediately after NP application to water. The decreased emergence has been attributed to

the surfactant properties of NP that may have rendered the insects “wettable”, thus preventing

them from escaping the surface tension (Schmude et al. 1999). Effects due to physical
33

surfactancy are most likely to occur if concentrations of NP or NPEs are sufficiently high for

micelles to form. Typical environmental concentrations in Canada are generally below critical

micelle concentrations for these substances, and therefore this mode of toxicity is not likely to

occur often (see the list of critical micelle concentrations in Chapter 1 and the discussion of

Canadian environmental concentrations in Chapter 2).

Toxic Effects

Toxicity of nonylphenol and nonylphenol ethoxylates has been observed in diverse biological taxa

including bacteria, fungi, protozoans, algae, aquatic macrophytes, terrestrial plants, crustaceans,

insects, molluscs, oligochaetes, annelids, rotifers, nematodes, collembolans, fish, amphibians,

and mammals. Researchers have also determined the toxicity of NP and NPEs in many of these

organisms at various developmental stages. Observed toxic effects include mortality, growth

inhibition, reduced fecundity, reduced germination, reduced abundance, reduced egg viability,

reduced larval settlement, reduced emergence, reduced hatch rate, delayed maturity, reduced

photosynthesis, immobilization, lethargy, loss of equilibrium, membrane disruption, induced

vitellogenesis, zona radiata induction, modified sex ratios, induction of testis-ova, and changes in

soil nutrient cycling (e.g., nitrification, nitrogen-fixation by bacteria).

Comprehensive lists of all fresh water, marine water, sediment, and soil toxicity data found in the

literature have been compiled in Appendices I, II, III, and IV, respectively. Within each of these

appendices, the data have been organized under broad taxonomic headings (e.g.,

microorganisms, plants, invertebrates, fish, amphibians, etc.). The data have also been grouped

by compound (i.e., NP, NPEs, NPECs). Toxicity data relevant to the derivation of the water,

sediment, and soil quality guidelines are discussed within Chapters 5, 6 and 7, respectively.

Below is a brief summary of the major, over-arching trends in the toxicity data.

Comparing LC50 and EC50 values across the various taxonomic groupings, there appears to be a

great deal of overlap. Toxic NP concentrations are quite comparable for most invertebrates and

fish, while plants and algae, in general, are slightly less sensitive. Comparing across media, NP is

approximately one hundred times more toxic in water (either freshwater or marine) than it is in

sediment or soil. The bulk of toxic responses in water fall within the range of 0.1 to 10 ppm (i.e.,

100 to 10000 μg·L-1; see Appendix I and II) while most toxic responses in sediment and soil are

within the range of 10 to 1000 ppm (i.e., 10 to 1000 mg·kg-1 dry weight; see Appendix III and IV).

This difference likely reflects the lower bioavailability of the compounds in soil and sediment due

to particle adsorption. There are large differences in toxicity among the various nonylphenolic

ethoxamers. For example, LC50s for freshwater exposure of Japanese killifish, Oryzias latipes, to

NP, NP1EO, NP6.4EO, NP9EO, and NP16.6EO are 1400, 3000, 5400, 12000, and 110000 μg·L-1,

respectively (Yoshimura 1986). In general, toxicity decreases with increasing ethoxylation.

It appears that the concentrations of NP required to elicit estrogenic effects are comparable to

concentrations at which other chronic toxic effects are observed. Estimates of NP chronic toxicity

in freshwater fish, for conventional endpoints, range from 6.0 μg·L-1 (Oncorhynchus mykiss

91-day NOEC growth; Brooke 1993a) to 460 μg·L-1 (Oryzias latipes 17-day LC50 embryotoxicity;

Gray and Metcalfe 1997). For estrogenicity endpoints, the chronic toxicity estimates are of a

similar magnitude, ranging from 10 μg·L-1 (Oryzias latipes 3-month NOEC for testis-ova induction;

Gray and Metcalfe 1997; also Oncorhynchus mykiss 72-hour LOEC vitellogenin induction; Lech et

al. 1996 ) to 100 μg·L-1 (Oryzias latipes 3-month LOEC for sex ratio change; Gray and Metcalfe

1997). These data indicate that guidelines for nonylphenolic substances set at concentrations

that protect against conventional toxicity endpoints are likely to protect against most endocrine

disrupting effects as well.
34

Mixtures and the TEQ Approach

Nonylphenol, nonylphenol ethoxylates, and nonylphenol ethoxycarboxylates typically occur

together in the aquatic environment as mixtures. Therefore, for the purpose of assessing

acceptable levels of these substances in water, sediment or soil, it is more useful to develop

guidelines that consider their combined effects. This can be accomplished through the use of a

toxic equivalency (TEQ) approach.

The TEQ approach is used to characterize the toxicity of a mixture of compounds by expressing

the toxicities of each individual compound in common terms, and summing them. This method is

commonly applied to PCBs, dioxins, and furans, and has previously been used in the

development of Canadian Environmental Quality Guidelines for these chemicals (Environment

Canada 2000a,b). Application of the TEQ approach requires that all components of a mixture

have a common mode of toxicity such that their effects are additive. Nonylphenolic substances

meet this requirement. At typical environmental concentrations, and for conventional toxicity

endpoints, the mode of toxicity for nonylphenol and for at least the lower ethoxylate chain length

NPEs and NPECs is narcosis (see previous discussion). If these substances are all acting

through narcosis, then additivity of their effects (as opposed to synergism or antagonism) is most

likely (Servos et al. 2000).

To apply a TEQ approach, toxic equivalency factors (TEFs) are determined for each individual

compound in a mixture by expressing the potency of each compound relative to the most toxic

compound in the mixture. For PCBs, dioxins, and furans, TEFs are measured relative to the

highly toxic 2,3,7,8-TCDD (van den Berg et al. 1998). Each compound’s concentration in the

mixture is then multiplied by its TEF to obtain its toxic equivalent (TEQ) for that sample, relative to

2,3,7,8-TCDD. The overall toxicity of the mixture is then calculated by summing the TEQs for all

compounds contained in the mixture.

We propose that a similar TEQ approach be applied towards mixtures of nonylphenolic

substances. We are not recommending the incorporation of nonylphenolic substances into the

same TEQ system used for PCBs, dioxins, and furans; the mode of toxicity for these two groups

of substances are different, so their toxicities cannot be compared (Roe et al. 2000; Environment

Canada 2000a). Rather we are suggesting that a separate set of nonylphenol TEF values (NP

TEFs) be used. Several researchers have studied the relative toxicity of various nonylphenol

ethoxylates and found that toxicity increases with decreasing length of the ethoxylate chain (e.g.,

Maxwell and Piper 1968; Macek and Krzeminski 1975; Yoshimura 1986; Miossec and Bocquéné

1986; Hall et al. 1989). Therefore the degradation products of NPEs (e.g., NP, NP1EO, NP2EO)

are more toxic than the parent compounds. In the application of a TEQ approach to these

compounds then, potency should be expressed relative to nonylphenol, the most toxic substance.

Relative toxicity values, or NP TEFs, for various nonylphenolic compounds were calculated for the

CEPA assessment of nonylphenol (Servos et al. 2000) and are presented in Table 4. These

values were derived based on a broad dataset that included both acute and chronic studies on a

range of vertebrate and invertebrate species. Reported toxic concentrations for the various

nonylphenolic compounds were matched up against similar endpoints for nonylphenol with the

same species, and, where possible, from the same laboratory; then a relative toxicity ratio was

calculated. From the resulting list of relative toxicity values for each group of compounds, a mean

relative toxicity value (TEF) was calculated, with more weight given to those studies deemed to be

of higher quality.

These TEFs represent an initial attempt to define the relative toxicity of groups of nonylphenolic

compounds, rather than for each individual ethoxamer. Future work to develop and validate more

specific TEF values is needed, but, in the interim, it is recommended that these more general

TEFs be applied. Also, TEF values for NPEs containing 3 to 8 ethoxylate groups were not

estimated in the CEPA assessment because there have been very few toxicity tests conducted

with these substances. Instead, we have provided our own estimate. As toxicity is known to

generally decrease with increasing number of ethoxylate groups, the relative toxicity of NP3EO-
35

NP8EO likely lies somewhere between the TEFs of 0.5 (for NP2EO) and 0.005 (for NPnEO where

nÚ9). Therefore, for NPEs with 3 to 8 EO groups, until sufficient toxicity data is available to

estimate relative potencies, it is recommended that a conservative TEF of 0.5 be adopted, with

the caveat that the use of this TEF may overestimate the toxicity of a mixture.

The total concentration of NP and NP-equivalents in a sample can be calculated using the

equation:

Total concentration = ∑ (Cx x TEF)

where,

Cx = concentration of each nonylphenolic compound

TEF = toxic equivalency factor for each particular nonylphenolic compound.

In the assessment of a mixture of nonylphenolic compounds, the environmental concentrations of

NPEs and NPECs should be expressed in terms of NP equivalents, and the sum of these

concentrations should then be compared against the guidelines developed in this document.

The NP TEF values determined in the CEPA assessment (Servos et al. 2000) were based on

toxicity to aquatic species which reside in the water column, but they may also be applied towards

the soil and sediment guidelines for NP and NPEs. Narcosis appears to be a common mode of

toxicity for nonylphenolic substances across all taxonomic groups (see earlier discussion under

“Modes of Toxicity”), so the NP TEF values should be applicable to plants, invertebrates, fish,

mammals, etc., regardless of whether they are water-, sediment-, or soil-dwelling species.

If octylphenol, octylphenol ethoxylates, or octylphenol ethoxycarboxylates are present in

environmental samples, it is recommended that these substances also be considered in the

calculation of total NP-equivalents. OP and OPEs are very similar in structure and behaviour to

NP and NPEs, and likely act through a similar toxic mode of action. Servos et al. (2000) have

suggested TEF values relative to NP for OP, OP1EC and OP2EC at 1.0, 0.005, and 0.005,

respectively. We would recommend that for OPnEOs, the same TEF values for the

corresponding NPnEO analogs could be used.

The TEQ approach has not been previously applied towards NP and NPEs, so the guidelines

currently available from other jurisdictions, as described in Chapter 4, are either based solely on a

single ethoxamer (e.g. NP or NP9EO), or they are based on the straight sum of NP and NPE

concentrations with no regard for relative toxicities among the ethoxamers. The Canadian

Environmental Quality Guidelines for NP and NPEs, developed in Chapters 5, 6, and 7, are

expressed on a TEQ basis.
36

Table 4. Summary of relative toxicity of nonylphenolic and related compounds.

Chemical Toxic equivalency factors (TEFs)

relative to NP

NP 1

NP1EO 0.5

NP2EO 0.5

NPnEO (3 Û n Û 8) 0.5*

NPnEO (n Ú 9) 0.005

NP1EC 0.005

NP2EC 0.005

OP 1

OPnEO (1 Û n Û 8) 0.5*

OPnEO (n Ú 9) 0.005*

OP1EC 0.005

OP2EC 0.005

Adapted from Servos et al. (2000) except where otherwise noted.

* conservative estimation by the authors of this document
37

CHAPTER 4. GUIDELINES FROM OTHER JURISDICTIONS

Water

Freshwater

Tables 5 and 6 provide summaries of existing freshwater guidelines, objectives, criteria,

standards, and environmental concern levels for nonylphenol (NP) and nonylphenol ethoxylates

(NPEs), respectively, from various jurisdictions both within and outside of Canada.

In Canada, the Quebec provincial government has established interim water quality criteria for

both nonylphenol (CAS 25154-52-3) and nonylphenol ethoxylates (CAS 9016-45-9). For

nonylphenol, the interim criteria (developed in 1996) for the protection of aquatic life from acute

and chronic toxicity are 6.8 μg·L-1 and 0.3 μg·L-1, respectively. The interim criteria for nonylphenol

ethoxylates (established in 1991) for the protection of aquatic life from acute and chronic toxicity

are 470 μg·L-1 and 21 μg·L-1, respectively (MEF 1998). These criteria have recently undergone

review, and new criteria are scheduled for release in 2000 (Guay 2000). For nonylphenol, new

acute and chronic final criteria will be 29 μg·L-1 and 6.0 μg·L-1, respectively. For NP9EO,

provisional acute and chronic criteria will be 280 μg·L-1 and 12 μg·L-1, respectively. For NP1EC,

provisional acute and chronic criteria will be 100 μg·L-1 and 4 μg·L-1, respectively. It was decided

that there is currently insufficient data available to derive criteria for NP1EO, NP1.5EO, NP2EO,

NP10EO, and NP2EC.

The province of Ontario has an interim provincial water quality objective for nonylphenol of

0.04 μg·L-1. This Ontario objective was based on the 48h LC50 of 100 μg·L-1 for Daphnia pulex

(EPS 1981 as cited in NRCC 1982), that was then divided by a highly conservative uncertainty

factor of 2332 due to the scarcity of toxicological data at the time (OMOE 1991). Many more

aquatic toxicity data are now available.

Outside of Canada, Denmark has an environmental quality standard for freshwater concentrations

of NP and NPEs combined. Australia and New Zealand have an environmental concern level for

nonylphenol in fresh water, and the U.S.A. has developed draft numbers for both acute and

chronic ambient aquatic life water quality criteria for nonylphenol.

Marine/Estuarine

A summary of existing marine and estuarine water quality guidelines for NP from other

jurisdictions are provided in Table 7. There are currently no jurisdictions in Canada with marine

water quality guidelines for NP or NPEs. Australia and New Zealand have an environmental

concern level for NP in marine waters, while the U.S.A. has both acute and chronic ambient

aquatic life water quality criteria for NP in marine waters.

Sediment

There are no jurisdictions within Canada, nor outside of Canada, which have guidelines for NP or

NPEs in either freshwater or marine sediment.
38

Soil

No guidelines for nonylphenol in soil currently exist for other jurisdictions within Canada. Outside

of Canada, only Denmark has a soil quality criterion for nonylphenol in soil (Table 8). Denmark

also has a guideline for the total concentration of NP, NP1EO, and NP2EO that may be present in

sludge which is intended to be applied to agricultural soils. Sweden also has a guideline on the

allowable concentration of NP in sludge used as an agricultural fertilizer (see Table 8).

Table 5. Freshwater quality guidelines for nonylphenol from other jurisdictions.

JURISDICTION GUIDELINE (μg·L-1) LEVEL OF PROTECTION REFERENCE

Ontario, Canada 0.04 interim provincial water quality

objective for the protection of

aquatic life

OMEE 1994

Australia /

New Zealand 1

0.2 environmental concern level See footnote

Quebec, Canada 20.3 interim water quality criteria

for the protection of aquatic

life (chronic exposure)

MEF 1998

Denmark 1.0

(NP and NPEs)

environmental quality

standard

Renner 1997

U.S.A. 35.9

(4-d average)

ambient aquatic life water

quality criteria (draft stage)

Brooke and

Thursby 2000

Quebec, Canada46.0 water quality criteria for the

protection of aquatic life

(chronic exposure)

Guay 2000

Quebec, Canada 26.8 interim water quality criteria

for the protection of aquatic

life (acute exposure)

MEF 1998

U.S.A. 327.9

(1-h average)

ambient aquatic life water

quality criteria (draft stage)

Brooke and

Thursby 2000

Quebec, Canada429.0 water quality criteria for the

protection of aquatic life

(acute exposure)

Guay 2000

1 Australian and New Zealand Environment and Conservation Council, Agriculture and Resource

 Management Council of Australia and New Zealand 1999

2

 These criteria have recently undergone review, and will likely be updated in 2000 (Guay 2000).

3

 These average concentrations should not be exceeded more than once every three years,

 on average.

4

 These criteria are still at the draft stage and have not yet been officially approved or

 implemented (Guay 2000).
39

Table 6. Freshwater quality guidelines for nonylphenol ethoxylates and ethoxycarboxylates from

other jurisdictions.

JURISDICTION GUIDELINE

(μg·L-1)

CHEMICAL LEVEL OF PROTECTION REFERENCE

Denmark 1.0 NP and NPEs environmental quality

standard

Renner 1997

Quebec, Canada 24.0 NP1EC provisional water quality

criteria for the protection

of aquatic life (chronic

exposure)

Guay 2000

Quebec, Canada 212.0 NP9EO provisional water quality

criteria for the protection

of aquatic life (chronic

exposure)

Guay 2000

Quebec, Canada 121.0 NPEs interim water quality

criteria for the protection

of aquatic life (chronic

exposure)

MEF 1998

Quebec, Canada 2100.0 NP1EC provisional water quality

criteria for the protection

of aquatic life (acute

exposure)

Guay 2000

Quebec, Canada 2280.0 NP9EO provisional water quality

criteria for the protection

of aquatic life (acute

exposure)

Guay 2000

Quebec, Canada 1470.0 NPEs interim water quality

criteria for the protection

of aquatic life (acute

exposure)

MEF 1998

1

 These criteria have recently undergone review, and will likely be updated in 2000 (Guay 2000).

2 These criteria are still at the draft stage and have not yet been officially approved or

 implemented (Guay 2000).
40

Table 7. Marine water quality guidelines for nonylphenol from other jurisdictions.

JURISDICTION GUIDELINE (μg·L-1) LEVEL OF PROTECTION REFERENCE

Australia /

New Zealand 1

1.2 environmental concern level See footnote

U.S.A.21.4

(4-d average)

ambient aquatic life water

quality criteria (draft stage)

Brooke and

Thursby 2000

U.S.A.26.7

(1-h average)

ambient aquatic life water

quality criteria (draft stage)

Brooke and

Thursby 2000

1

 Australian and New Zealand Environment and Conservation Council, Agriculture and Resource

 Management Council of Australia and New Zealand 1999

2

 These average concentrations should not be exceeded more than once every three years,

 on average.

Table 8. Soil and sludge guidelines from other jurisdictions.

JURISDICTION GUIDELINE

(mg·kg-1)

CHEMICAL LEVEL OF PROTECTION REFERENCE

Denmark 0.01 NP soil quality criterion Jensen et al. 1997

Denmark 110 4-NP, NP1EO

and NP2EO

guideline for application of

sludge to agricultural soils

European

Commission 1999

Sweden 50 NP allowable dry weight

concentration in sludge for

use as agricultural fertilizer

Bennie et al. 1998

1

 Danish Ministerial Order No. 823, Sept. 16, 1996; guideline takes effect as of June 2000
41

CHAPTER 5. DEVELOPMENT OF CANADIAN WATER QUALITY GUIDELINES

Canadian Water Quality Guidelines (WQGs) are developed using the methods outlined in “A

protocol for the derivation of water quality guidelines for the protection of aquatic life” (CCME

1991; CCME 1999). This protocol was created with the intent that guidelines be protective of all

forms of aquatic life at all life stages during an indefinite period of exposure in the water column.

A brief summary of the protocol is provided in Annex 2, including a definition of primary data.

Freshwater

Toxicity Data

A large number of studies describing the toxic effects of nonylphenol and its ethoxylates on

freshwater aquatic life exist in the scientific literature. A comprehensive list of all freshwater

toxicity data for nonylphenol that was consulted has been compiled in Appendix I. Rather than

giving an exhaustive review of these studies, a brief description of the toxicity data relevant to the

derivation of Canadian Water Quality Guidelines is provided below. This information reflects the

data requirements set forth in the protocol (CCME 1991) and is presented by taxonomic groups.

Fish

From the primary toxicity dataset for nonylphenol (Table 9), the most sensitive freshwater fish

species is the rainbow trout, Oncorhynchus mykiss, with a 91-day LOEC for growth of 10.3 μg·L-1

(Brooke 1993a). A comparable value of 14 μg·L-1 was reported as the 33-day LOEC for survival

with fathead minnows, Pimephales promelas (Ward and Boeri 1991b). The other primary toxicity

endpoints are generally an order of magnitude higher. For example, 28-day and 96-hour LOECs

for survival have been reported for Lepomis macrochirus at 126 and 211 μg·L-1, respectively, and

for P. promelas at 193 and 230 μg·L-1, respectively (Brooke 1993b). Median lethal concentrations

(96h LC50s) for P. promelas are cited at 128 μg·L-1 (Brooke 1993a) and 135 μg·L-1 (Holcombe et

al. 1984), and values of 209 and 221 μg·L-1 are cited for L. macrochirus and O. mykiss,

respectively (Brooke 1993a).

Invertebrates

The most sensitive primary data for a freshwater invertebrate is the 96h LC50 of 20.7 μg·L-1 for

Hyalella azteca (Brooke 1993a). England and Bussard (1995) also report a 96h LC50 of

170 μg·L-1 and a 96h EC50 for immobilization of 150 μg·L-1 for H. azteca. For Daphnia magna,

concentrations of 84.8, 100, and 215 μg·L-1 have been reported for a 48h immobilization EC50

(Brooke et al. 1993a), a 21-day LC50 (Brooke et al. 1993a), and a 21-day LOEC for reproduction

(Comber et al. 1993), respectively. Numerous primary datapoints exist for the midge, Chironomus

tentans, including 14-day EC50s of 95 μg·L-1 for both total adverse effects and larval growth

(England and Bussard 1993), a 14-day LC50 of 119 μg·L-1 (England and Bussard 1993), a 20-day

LOEC for survival of 91 μg·L-1 (Kahl et al. 1997), and a 14-day LOEC for larval growth of

150 μg·L-1 (England and Bussard 1993). Primary data is only available for two non-arthropod

invertebrates. Brooke (1993a) reported 96h LC50s of 342 and 774 μg·L-1 for the annelid

Lumbriculus variegatus and the snail Physella virgata, respectively.

Plants/Algae

A limited amount of primary data is available for toxicity of nonylphenol to algae and aquatic

macrophytes. The macrophyte Lemna minor exhibits reduced frond production after 96 hours of

exposure to nonylphenol at a concentration of 2080 μg·L-1 (Brooke 1993a). The green algae

Selenastrum capricornutum experiences growth reduction after 96 hours of exposure to

nonylphenol concentrations as low as 190.0 μg·L-1, and the corresponding median effective

concentration (EC50) is 410 μg·L-1 (Ward and Boeri 1990a).
42

Guideline Derivation

Primary data which were selected as acceptable for use in guideline derivation (i.e., that met the

screening criteria as described in CCME 1991 and in Annex 2) are shown in Table 9. This

dataset is sufficient to meet the minimum requirements for derivation of a full guideline for

nonylphenol in fresh water.

According to the protocol, water quality guidelines for a chemical are preferably derived from the

chronic study that gives the most sensitive non-lethal lowest-observable-effect level (LOEL) for an

ecologically relevant response. The study must have demonstrated a clear dose-response

relationship, and the LOEL must be statistically significant..

A chronic toxicity test with rainbow trout, Oncorhynchus mykiss, by Brooke (1993a) was selected

as the critical study. This study produced the most sensitive acceptable endpoint, a 91d LOEC for

reduced growth in rainbow trout at 10.3 μg·L-1. Rainbow trout were exposed, under flow-through

conditions, to two replicates each of five concentrations (ranging from 6 to 114 μg·L-1) of

nonylphenol, plus a control. Each replicate originally contained 100 embryos, and after

approximately 45 days, 15 fry from each replicate were randomly chosen for completion of the

assay. The total length of the exposure period was 91 days. Temperature, dissolved oxygen,

total alkalinity, total hardness, pH, and conductivity were measured regularly through the exposure

period and remained within acceptable levels (Brooke 1993a). Water samples from the exposure

chambers were collected and analyzed twice per week, throughout the duration of the test.

Concentrations of nonylphenol were anaylzed using HPLC. One-way analysis of variance and

Dunnett’s test were used to determine NOECs and LOECs. In this study, appropriate

experimental methods and conditions were used, and all relevant information was reported in the

article. Growth is an ecologically relevant response, and the LOEC reported was statistically

significant.

Because of possible variations in toxicity among species, among endpoints, and between lab and

field exposures, it is recommended that a safety factor be used. Therefore, the value of

10.3 μg·L-1 (from Brooke 1993a) was multiplied by a safety factor of 0.1 (CCME 1991) to yield

1.03 μg·L-1. Rounding off, the freshwater guideline value is 1.0 μg·L-1. The effects diagram in

Figure 3 shows the guideline value and the distribution of the primary toxicity data that was used

to derive it.
43

Table 9. Data used in derivation of guideline for toxicity of nonylphenol to freshwater aquatic life.

Species

(Common name)

Life cycle

stage

Duration Endpoint Effect Concentration

(μg·L-1)

Reference

PRIMARY DATA

Algae/Macrophytes

Lemna minor

(duckweed)

2-frond

stage

96h LOEC frond

production

2080 Brooke 1993a

Selenastrum

capricornutum (algae)

4-7 days 96h LOEC growth

(cell counts)

190 Ward and Boeri

1990a

4-7 days 96h EC50 growth

(cell count)

410 Ward and Boeri

1990a

Invertebrates

Chironomus tentans larva 20d LOEC survival 91 Kahl et al. 1997

(midge) 2

nd instar

larva

14d EC50 lethargy,

paleness,

reduced size

95 England and

Bussard 1993

2

nd instar

larva

14d EC50 larval growth 95 England and

Bussard 1993

2

nd instar

larva

14d LC50 mortality 119 England and

Bussard 1993

2

nd instar

larva

14d LOEC larval growth 150 England and

Bussard 1993

Daphnia magna neonate 48h EC50 immobilization 84.8 Brooke 1993a

(water flea) neonate 21d LC50 mortality 100 Comber et al.

1993

neonate 21d LOEC reproduction 215 Brooke 1993a

Hyalella azteca juvenile 96h LC50 mortality 20.7 Brooke 1993a

(amphipod) juvenile 96h EC50 lethargy/

immobility

150 England and

Bussard 1995b

juvenile 96h LC50 mortality 170 England and

Bussard 1995b

Lumbriculus variegatus

(annelid)

adult 96h LC50 mortality 342 Brooke 1993a

Physella virgata

(snail)

adult 96h LC50 mortality 774 Brooke 1993a

Fish

Lepomis macrochirus

(bluegill sunfish)

11-13

weeks

28d LOEC survival 126 Brooke 1993b

juvenile 96h LC50 mortality 209 Brooke 1993a

13-15

weeks

96h LOEC survival 211 Brooke 1993b

Oncorhynchus mykiss

(rainbow trout)

embryo 91d LOEC growth 10.3 Brooke 1993a

45 days 96h LC50 mortality 221 Brooke 1993a
44

Table 9 (continued). Data used in derivation of guideline for toxicity of nonylphenol to freshwater

aquatic life.

Species

(Common name)

Life cycle

stage

Duration Endpoint Effect Concentration

(μg·L-1)

Reference

Pimephales promelas

(Fathead minnow)

embryo 33d LOEC survival 14 Ward and Boeri

1991b

25-35

days

96h LC50 mortality 128 Brooke 1993a

31-35

days

96h LC50 mortality 135 Holcombe et al.

1984

4 weeks 28d LOEC survival 193 Brooke 1993b

4 weeks 96h LOEC survival 230 Brooke 1993b

LOEC = lowest observed effect concentration

LC50 = median lethal concentration

EC50 = median effective concentration

Note: The critical study used in the guideline derivation is shaded.
45

Vertebrates Invertebrates

Acute Chronic

Toxicity

information

Species Toxicity 

endpoint

Concentration (μg·L-1)

Toxicity endpoints:

Canadian Water Quality Guideline: 

1.0 μg·L-1

primary critical value Invertebrates Plants Vertebrates

Canadian Guideline

100 101 102 103 104

P. promelas

P. promelas

P. promelas

O. mykiss

L. macrochirus

L. macrochirus

P. virgata

L. variegatus

H. azteca

H. azteca

H. azteca

D. magna

L. minor

S. capricornutum

S. capricornutum

P. promelas

P. promelas

L. macrochirus

O. mykiss

D. magna

D. magna

C. tentans

C. tentans

C. tentans

C. tentans

C. tentans

96-h LOEC

96-h LC50

96-h LC50

96-h LC50

96-h LOEC

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h EC50

96-h LC50

48-h EC50

96-h LOEC

96-h LOEC

96-h EC50

28-d LOEC

33-d LOEC

28-d LOEC

91-d LOEC

21-d LOEC

21-d LC50

14-d LOEC

14-d LC50

14-d EC50

14-d EC50

20-d LOEC

Figure 3. Select freshwater toxicity data for nonylphenol.
46

Marine/Estuarine Water

Toxicity Data

A large number of studies describing the toxic effects of nonylphenol and its ethoxylates on

marine aquatic life exist in the scientific literature. A comprehensive list of all marine toxicity data

for nonylphenol that was consulted has been compiled in Appendix II. Rather than giving an

exhaustive review of these studies, a brief description of the toxicity data relevant to the derivation

of Canadian Water Quality Guidelines is provided below. This information reflects the data

requirements set forth in the protocol (CCME 1991) and is presented by taxonomic groups.

Fish

From the nonylphenol toxicity dataset, three acceptable (secondary) chronic toxicity studies were

available for marine fish. A 21-day LOEC of 24.5 μg·L-1 was reported for increased

hepatosomatic index in flounder, Platichthys flesus (Allen et al. 1999). A 21-day LOEC of

100 μg·L-1 was reported for increased plasma concentrations of vitellogenin in eelpout, Zoarces

viviparus (Korsgaard and Pedersen 1998). A 10-day LOEC of 2203 μg·L-1 for the induction of

sublethal abnormalities in embryos of the killifish, Fundulus heteroclitus, was also reported (Kelly

and Di Giulio 2000). Six acceptable acute toxicity endpoints have been reported for marine fish.

The 96h LC50 for sheepshead minnow, Cyprinodon variegatus, has been estimated at

310.0 μg·L-1 (Ward and Boeri 1990c) and 142.0 μg·L-1 (Lussier et al. 2000). For inland silversides,

Menidia beryllina, a 96h LC50 of 70.0 μg·L-1 has been cited (Lussier et al. 2000). A 96h LC50 of

17 μg·L-1 has been measured in winter flounder, Pleuronectes americanus (Lussier et al. 2000).

In F. heteroclitus, 96h LC50s have been reported for embryos and for 2-week larvae at 5441 and

209 μg·L-1, respectively (Kelly and Di Giulio 2000).

Invertebrates

The lowest reported endpoint for a marine invertebrate came from a secondary study by

Billinghurst et al. (1998) in which the 48h LOEC for reduced settlement in larvae of the barnacle

Balanus amphitrite was 0.06 μg·L-1. Several studies indicate that the mysid Americamysis bahia

(recently renamed from Mysidopsis bahia) is also a sensitive marine invertebrate. Ward and

Boeri (1991a) reported reductions in the growth of A. bahia after a 28-day exposure to

nonylphenol concentrations as low as 6.7 μg·L-1 and reduced reproduction and survival were both

observed at concentrations as low as 9.1 μg·L-1. Ward and Boeri (1990b) also reported a 96h

LC50 for A. bahia of 43.0 μg·L-1, and Lussier et al. (2000) reported a 96h LC50 of 60.6 μg·L-1 for this

same species. Slightly higher 96h LC50s of 59.4, 61.6, 71.0, and >195 μg·L-1 are reported for the

grass shrimp Paleomonetes vulgaris, the amphipod Leptocheirus plumulosus, the lobster

Homarus americanus, and the mudcrab Dyspanopeus sayii, respectively (Lussier et al. 2000).

Lussier et al. (2000) also reported a 48h EC50 of 37.9 μg·L-1 for inhibition of fertilization in the coot

clam Mulinia lateralis. From a 53-day study on the copepod Tisbe battagliai, a LOEC of

41.0 μg·L-1 has been reported for egg production, offspring survival, and intrinsic rate of increase

(Bechmann 1999). For this same species, the 96h LC50s for nauplii and for adults are

31-62 μg·L-1 and 500-1000 μg·L-1, respectively (Bechmann 1999). For larvae of the Pacific oyster,

Crassostrea gigas, exposure to 100 μg·L-1 resulted in both a 48h LOEC for various deformities,

and a 72h LOEC for delayed development (Nice et al. 2000).

Algae

The marine alga Skeletonema costatum has a 96 hour EC50 for growth inhibition of 27.0 μg·L-1

(Ward and Boeri 1990d). This is the only acceptable toxicity study available for marine algae or

plants.
47

Guideline Derivation

Primary data that were selected as acceptable for use in guideline derivation (i.e., that met the

screening criteria as described in CCME 1991 and in Annex 2) are shown in Table 10. These

data are insufficient to meet the minimum requirements for derivation of a full guideline for the

protection of aquatic marine life. Three additional primary studies are needed for a full guideline.

Of these studies, two must include chronic endpoints for two different temperate, marine fish

species. The third study must have a chronic endpoint for a temperate invertebrate that belongs

to a class other than the Crustacea.

When the minimum data requirements for derivation of a full guideline cannot be met, it may still

be possible to derive an interim guideline. Secondary studies, as well as primary studies, may be

used to meet the minimum requirements for an interim guideline. Studies which were considered

of secondary quality, and therefore could be used to meet the minimum data requirements for an

interim guideline, are also listed in Table 10. Using both the primary and secondary studies, there

are sufficient data to meet the minimum requirements for deriving an interim guideline for

nonylphenol.

As for freshwater, marine water quality guidelines are preferably derived from the chronic study

that gives the most sensitive nonlethal LOEL (CCME 1991). The LOEL is then multiplied by a

safety factor of 0.1 to arrive at the final guideline value.

The LOEC for reduced settlement in barnacles, Balanus amphitrite, was the most sensitive

endpoint reported (48h LOEC at 0.06 μg·L-1; Billinghurst et al. 1998). However, this study was of

secondary quality, involved a short exposure time, and used an unconventional endpoint. Also,

this LOEC value was two orders of magnitude lower than any other endpoints reported in the

literature. This suggests that either larval settlement is much more sensitive than other endpoints,

or that this particular LOEC is an outlier that is not reproducible. Unfortunately, there have been

no other studies conducted on barnacle larval settlement with exposure to nonylphenol with which

to compare these results. The LOECs reported for larval settlement of Balanus amphitrite

exposed to cadmium and phenol (Wu et al. 1997) are higher than the LOECs for many other

endpoints that have been reported for these substances in marine aquatic life (see CCME 1999).

This suggests that larval settlement is not a particularly sensitive endpoint. Due to the suspicion

cast by these various sources of uncertainty, confidence in the study by Billinghurst et al. (1998)

was not high enough to base the guideline value on this LOEC.

The chronic assay with the mysid, Americamysis bahia, by Ward and Boeri (1991a) was selected

as the critical study. This study reported a 28-day LOEC for reduced growth at 6.7 μg·L-1 (Ward

and Boeri 1991a). The study was evaluated as being of primary quality because appropriate

experimental methods and conditions were used, and all relevant information was reported. The

test protocol used was based on U.S. EPA test guidelines. The study involved exposure of

mysids, under flow-through conditions, to five concentrations of nonylphenol (ranging from 3.9 to

21 μg·L-1), plus a dilution water control and solvent control (acetone). Eight replicates of each test

concentration were used, with five mysids per replicate. The total length of the exposure period

was 28 days. Temperature, dissolved oxygen, pH, and salinity were measured and recorded daily

throughout the exposure period and remained within acceptable levels. Water samples from the

test vessels were sampled weekly, with nonylphenol concentrations anaylzed by HPLC with

fluorescence detection. One-way analysis of variance and Dunnett’s test were used to test for

treatment effects. Growth is an ecologically relevant response, and the LOEC reported was

statistically significant.

Therefore, the LOEC value of 6.7 μg·L-1 (from Ward and Boeri 1991a) was multiplied by a safety

factor of 0.1 (CCME 1991), and rounded off, to obtain the interim guideline of 0.7 μg·L-1 for the

protection of marine life against the adverse effects of nonylphenol. The effects diagram in Figure

4 shows the interim guideline value and the distribution of the toxicity data that was used to derive

it.
48

Table 10. Data used in derivation of guideline for toxicity of nonylphenol to marine aquatic life.

Species

(Common name)

Life cycle

 stage

Duration Endpoint Effect Concentration

(μg·L-1)

Reference

PRIMARY DATA

Algae

Skeletonema

costatum

(algae)

NR 96h EC50 growth

inhibition

(cell count)

27 Ward and Boeri

1990d

Invertebrates

Americamysis

bahia (mysid)

neonates 28d LOEC mean total

length

6.7 Ward and Boeri

1991a

neonates 28d LOEC survival 9.1 Ward and Boeri

1991a

neonates 28d LOEC number of

young per

surviving

female

9.1 Ward and Boeri

1991a

neonates 96h LC50 mortality 43 Ward and Boeri

1990b

post-larvae

1d

96h LC50 mortality 60.6 Lussier et al. 2000

Leptocheirus

plumulosus

(amphipod)

adult 96h LC50 mortality 61.6 Lussier et al. 2000

Paleomonetes

vulgaris

(grass shrimp)

2d larvae 96h LC50 mortality 59.4 Lussier et al. 2000

Tisbe battagliai

(copepod)

nauplii 53d LOEC egg

production,

offspring

survival,

intrinsic rate

of increase

41 Bechmann 1999

Fish

Cyprinodon

variegatus

(sheepshead

minnow)

juveniles 96h LC50 mortality 142 Lussier et al. 2000

juveniles 96h LC50 mortality;

lethargy,

bloating,

loss of

equilibrium

310 Ward and Boeri

1990c

Menidia beryllina

(inland silversides)

juveniles 96h LC50 mortality 70 Lussier et al. 2000
49

Table 10 (continued). Data used in derivation of guideline for toxicity of nonylphenol to marine

aquatic life.

Species

(Common name)

Life cycle

 stage

Duration Endpoint Effect Concentration

(μg·L-1)

Reference

SECONDARY

DATA

Invertebrates

Balanus amphitrite

(barnacle)

cyprids

(larvae)

48h LOEC reduced

settlement

0.06 Billinghurst et al.

1998

Crassostrea gigas

(Pacific oyster)

larvae 48h LOEC deformity 100 Nice et al. 2000

larvae 72h LOEC delayed

development

100 Nice et al. 2000

Dyspanopeus

sayii

(mudcrab)

4

th and 5th

stage zoea

96h LC50 mortality > 195 Lussier et al. 2000

Homarus

americanus

(American lobster)

first stage

zoea

96h LC50 mortality 71 Lussier et al. 2000

Mulinia lateralis

(Coot clam)

embryolarval48h EC50 inhibition of

fertilization

37.9 Lussier et al. 2000

Tisbe battagliai

(copepod)

<1 day

nauplii

96h LC50 mortality 31-62 Bechmann 1999

adult 96h LC50 mortality 500-1000 Bechmann 1999

Fish

Fundulus

heteroclitus

(killifish)

2 wk larvae 96h LC50 mortality 209 Kelly and Di Giulio

2000

embryos 10d LOEC sublethal

abnormalities

2203 Kelly and Di Giulio

2000

embryos 96h LC50 mortality 5441 Kelly and Di Giulio

2000

Platichthys flesus

(flounder)

adults 21d LOEC increased

hepatosomatic

index

24.5 Allen et al. 1999

Pleuronectes

americanus

(winter flounder)

2d larvae 96h LC50 mortality 17 Lussier et al. 2000

Zoarces viviparus

(eelpout)

pregnant

females

21d LOEC increased

plasma

levels of

vitellogenin

100 Korsgaard and

Pedersen 1998

LOEC = lowest observed effect concentration

LC50 = median lethal concentration

EC50 = median effective concentration

NR = not reported

Note: The critical study used in the guideline derivation is shaded.
50

Vertebrates Invertebrates

Acute

Toxicity

information

Species Toxicity

endpoint

Concentration (μg·L-1)

F. heteroclitus

F. heteroclitus

C. variegatus

C. variegatus

M. beryllina

P. americanus

T. battagliai

T. battagliai

D. sayii

C. gigas

C. gigas

H. americanus

L. plumulosus

A. bahia

A. bahia

P. vulgaris

M. lateralis

B. amphitrite

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

48-h LOEC

72-h LOEC

96-h LC50

96-h LC50

96-h LC50

96-h LC50

96-h LC50

48-h EC50

48-h LOEC

Chronic

Invertebrates Plants Vertebrates

Toxicity endpoints:

Canadian Interim Water Quality Guideline: 

0.7 μg·L-1

primary

secondary

critical value Canadian Guideline

10-2 100 102 104

S. costatum

F. heteroclitus

Z. viviparus

P. flesus

T. battagliai

A. bahia

A. bahia

A. bahia

96-h EC50

10-d LOEC

21-d LOEC

21-d LOEC

53-d LOEC

28-d LOEC

28-d LOEC

28-d LOEC

Figure 4. Select marine toxicity data for nonylphenol.
51

Recommended Canadian Water Quality Guidelines

Guideline for the Protection of Freshwater Life

A full guideline of 1.0 μg·L-1, expressed in nonylphenol toxic equivalency units (NP TEQs), is

recommended for the protection of aquatic freshwater life from the adverse effects of nonylphenol

and its ethoxylates.

There were sufficient toxicity data available to derive a full freshwater guideline. This guideline

should be applied to the total concentration (in NP TEQs) of all nonylphenolic substances

occurring in a water sample. The total concentration should be calculated using the nonylphenol

toxic equivalency factors provided in Chapter 3 (Table 4). Further guidance and

recommendations for the implementation of this guideline are provided in Chapter 9.

The freshwater guideline for nonylphenol and its ethoxylates proposed in this document is the

same as the environmental quality standard of 1.0 μg·L-1 that exists in Denmark (see Table 5). It

is lower than both the acute and chronic U.S. criteria for nonylphenol (at 27.9 μg·L-1 and

5.9 μg·L-1, respectively), but is higher than the environmental concern level of 0.2 μg·L-1 proposed

by Australia/New Zealand (see Table 5). It is also higher than Quebec’s interim water quality

criteria for chronic exposure of 0.3 μg·L-1, although recommendations have been made to

increase this value to 6.0 μg·L-1 (Guay 2000). The Ontario provincial objective, at 0.04 μg·L-1, is

more than an order of magnitude lower than the Canadian water quality guideline proposed here.

It should be noted that the Ontario objective was calculated using an extremely conservative

safety factor because at the time of its development, in 1991, there were much fewer toxicity data

available.

Guideline for the Protection of Marine Life

An interim guideline of 0.7 μg·L-1, expressed in NP TEQs, is recommended for the protection of

aquatic marine life from the adverse effects of nonylphenol and its ethoxylates.

This guideline is expressed in nonylphenol toxic equivalency (NP TEQ) units and may be applied

to mixtures of nonylphenol and nonylphenol ethoxylates. The total concentration of nonylphenolic

substances in a water sample (in NP TEQs) should be calculated using the nonylphenol toxic

equivalency factors provided in Chapter 3 (Table 4). Further guidance and recommendations for

the implementation of this guideline are provided in Chapter 9.

The guideline proposed here is lower than both the environmental concern level for nonylphenol of

Australia/New Zealand, at 1.2 μg·L-1, and the U.S. acute and chronic criteria of 6.7 μg·L-1 and

1.4 μg·L-1, respectively (see Table 7).
52

CHAPTER 6. DEVELOPMENT OF CANADIAN SEDIMENT QUALITY GUIDELINES

Canadian Sediment Quality Guidelines are developed using the methods outlined in “Protocol for

the derivation of Canadian sediment quality guidelines for the protection of aquatic life” (CCME

1995, 1999). This protocol was created with the intent that guidelines be protective of all forms of

aquatic life and all aspects of their aquatic life cycles during an indefinite period of exposure to

substances associated with bed sediments.

Toxicity Data

Data on the toxicity of nonylphenol in sediments to aquatic life were only available for

invertebrates and amphibians, and were quite limited. These data are also presented in Appendix

III.

Invertebrates

Toxicity of nonylphenol in sediment has been determined for a single freshwater invertebrate

species, the midge, Chironomus tentans (England and Bussard 1993). The study used spikedsediment toxicity testing to assess both lethal and sublethal effects of nonylphenol on the midge

larvae over a 14 day exposure period. Acceptable spiking procedures were used and a standard

test protocol was followed. For both mortality and growth, NOEC values were reported at

20.1 mg·kg-1 dry weight (England and Bussard 1993). A LOEC of 34.2 mg·kg-1 (dry weight) was

observed for both mortality and growth, with 31% mortality and a growth reduction of 27%

occurring at that concentration.

Two studies have examined the toxicity of nonylphenol to marine invertebrates, both through

spiked-sediment toxicity testing. One study evaluated nonylphenol lethality to the amphipod

Ampelisca abdita (Fay et al. 2000). Standard sediment spiking and test procedures were

followed. A ten day LC50 of 98.7 mg·kg-1 was observed, with 100% mortality occurring at a

concentration of 243 mg·kg-1. The other study evaluated sub-lethal effects of nonylphenol on the

polychaete Capitella sp. I over a 78 day exposure period (Hansen et al. 1999). Sub-lethal

endpoints examined included time to first reproduction, time between reproductive events,

number of broods per reproductive individual, number of eggs per individual and per reproductive

individual, population growth rate, and size at maturity. For all of these endpoints, the NOEC was

52 mg·kg-1 and the LOEC was 174 mg·kg-1 (Hansen et al. 1999). The results of this study,

however, should be treated with caution because standard sediment spiking procedures were not

followed.

Amphibians

Ward and Boeri (1992) used a 30 day spiked-sediment toxicity test to examine toxicity of

nonylphenol to tadpoles of the bullfrog, Rana catesbiana. Standard sediment spiking and test

procedures were followed. Reported endpoints for mortality included a NOEC of 155 mg·kg-1 and

a LOEC of 390 mg·kg-1 with a calculated LC50 of 260 mg·kg-1. The authors also examined sublethal endpoints such as growth, lethargy, haemorrhaging, and occurrence of tail lesions. For all

of these sub-lethal effects, the NOECs were 155 mg·kg-1, the LOECs were 390 mg·kg-1, and

EC50s were calculated at 220 mg·kg-1 (Ward and Boeri 1992).

Protocol Approaches

The protocol relies on the modified National Status and Trends Program (NSTP) approach and

the Spiked Sediment Toxicity Test (SSTT) approach. If sufficient information exists to support

both methods, a full sediment quality guideline may be recommended. If information is only

available to support one of the approaches, then an interim sediment quality guideline is

recommended. In cases where there is insufficient information to derive interim guidelines using

either the modified NSTP approach or the SSTT approach, provisional guidelines may be
53

recommended by adopting guidelines from other jurisdictions. Further discussion of these three

approaches is provided in Annex 3 of this document. At present, insufficient information exists to

derive sediment quality guidelines for nonylphenol using any of the three approaches described in

the protocol.

The protocol (CCME 1995) does not provide guidance on other possible options if there is

insufficient data to meet the minimum data requirements for the NSTP and SSTT approaches,

and if there are no acceptable sediment quality guidelines from other jurisdictions that can be

adopted. One possibility would be to not derive a guideline at all. While acknowledging the need

for toxicity data of the highest quality, we believe that guidelines based on the best available data

and protocol approaches are still more practical, scientifically defensible, and protective of the

environment than the absence of guidelines, or than guidelines that are not risk-based.

Therefore, we are recommending the use of an equilibrium partitioning approach for the

development of provisional Canadian interim sediment quality guidelines for nonylphenol and its

ethoxylates.

Equilibrium Partitioning Approach

The water/sediment equilibrium partitioning approach is used in the United States for development

of sediment quality guidelines (MacDonald et al. 1992) and is also supported by the Ontario

Ministry of the Environment (OMOE 1993). This approach establishes a guideline based on the

concentration of a substance in sediments that ensures that the concentration in the interstitial

water does not exceed the water quality guideline for that chemical at equilibrium (USEPA 1992).

It is based on the assumption that continuous equilibrium exchange of a contaminant between

sediment and interstitial water occurs (MacDonald et al. 1992). For non-polar organic

substances, such as nonylphenol, it is known that concentrations in the interstitial water are

dependent on concentrations in the sediment phase with the partitioning largely controlled by the

total organic carbon (TOC) content of the sediment (MacDonald et al 1992). This appears to be

true for NP and NPEs. Studies have shown that organic content is an important determining

factor in the adsorption of at least the short-chain NPEs to sediments (Urano 1984; John et al.

2000). Johnson et al. (1998) found a positive relationship between amounts of octylphenol sorbed

to sediments and the amount of TOC in the sediments; one would expect NP to behave similarly.

Through spiked-sediment bioaccumulation tests with benthic amphipods, Hecht et al. (2000)

found that higher concentrations of NP moved into the pore water of those sediments containing

lower TOC concentrations. Based on similar studies, Gunnarsson et al. (2000) also found that

pore water appears to be the major route of exposure to NP for benthic amphipods, and they

concluded that the equilibrium partitioning theory is valid for nonylphenol.

As the protocol for the development of Canadian sediment quality guidelines (CCME 1995) allows

for the adoption of guidelines from other jurisdictions that were derived using the equilibrium

partitioning approach, it seems reasonable, and in keeping with the spirit of the protocol, that this

approach could be used to derive sediment quality guidelines based on the Canadian water

quality guidelines for nonylphenol and its ethoxylates. Sediment quality assessment values based

on partitioning are less ecologically relevant and scientifically defensible than effect-based

guidelines that are derived from direct sediment exposure studies (CCME 1995). The equilibrium

partitioning method assumes that sediment-dwelling organisms and water column organisms are

equally sensitive to nonylphenol and that the concentration of NP in sediment, interstitial water,

and benthic organisms is at thermodynamic equilibrium. However, because there are insufficient

data available for derivation of sediment quality guideline values by other methods, the equilibrium

partitioning approach will be used to develop provisional ISQGs.
54

Guideline Derivation

Using the equilibrium partitioning approach, provisional ISQGs were derived from the Canadian

water quality guidelines for nonylphenol and its ethoxylates, developed in Chapter 5, using the

following equation (Di Toro et al. 1991; MacDonald et al. 1992):

ISQG = WQG · Kp Equation 1

Where:

WQG = water quality guideline (in μg·L-1);

Kp = sediment/water partition coefficient (in L·kg-1)

For non-polar organic contaminants such as nonylphenol, the organic carbon content of the

sediment is a major factor controlling the partitioning between water and sediment (MacDonald et

al. 1992). The sediment/water partition coefficient can therefore be related to the organic

carbon/water partition coefficient by the following equation (Di Toro et al. 1991):

Kp = Koc ·foc Equation 2

Where:

Koc = organic carbon/water partition coefficient (in L·kgoc

-1);

foc = fraction of organic carbon

Therefore, by substituting Equation 2 into Equation 1, the ISQG may be calculated as:

ISQG = WQG · Koc · foc Equation 3

Sekela et al. (1999) estimated that the log Koc for nonylphenol ranges from 4.7 to 5.6. Taking the

anti-log of these two values, and then calculating the geometric mean, yields a Koc of

141 254 L·kgoc

-1. The freshwater Canadian WQG for nonylphenol, derived herein, is 1.0 μg·L-1

.

Substituting these values into Equation 3 yielded a freshwater ISQG of 141 mg NP per kg of

organic carbon. This value was adjusted in order to express the guideline as dry weight of

sediment, so that the guideline would be consistent with other Canadian SQGs. Therefore, by

normalizing to 1% TOC (i.e., foc = 0.01), the freshwater ISQG of 1.4 mg·kg-1 dry weight was

derived. It is recommended that the guideline be adjusted, as described in Chapter 9, based on

local levels of TOC to obtain a site-specific objective.

Similarly for marine sediments, an ISQG was derived by substituting into Equation 3 the Koc of

141 254 L·kgoc

-1, and the marine Canadian WQG of 0.7 μg·L-1. This resulted in an ISQG of

99 mg NP per kg of organic carbon. The value was expressed as dry weight of sediment by

normalizing to 1% TOC, to give a marine ISQG of 1.0 mg·kg-1 dry weight. Again, it is

recommended that the guideline be adjusted, as described in Chapter 9, based on local levels of

TOC to obtain a site-specific objective.

Comparing these guideline values with the available sediment toxicity data, it would appear that

they are likely to be protective of most sediment-dwelling organisms. The lowest effect

concentrations observed in freshwater and marine sediment toxicity tests with nonylphenol were

34.2 mg·kg-1 and 174 mg·kg-1, respectively. The guidelines are lower than these effect

concentrations by more than a factor of 20, allowing for a large margin of safety.

It should be noted that because these guidelines were derived using an equilibrium partitioning

approach, they represent sediment concentrations that will be protective of sediment-dwelling

biota when the sediment and interstitial water are in equilibrium. Their use requires the

assumptions that there are no changes in the thermodynamics of the system (i.e., constant

temperature and pressure with no input/output of energy or mass), and that there has been

sufficient time for the NP and/or NPEs to distribute throughout the system (Spacie et al. 1995). It

is possible that non-equilibrium conditions could exist in some cases, such as immediately
55

following a spill, in river channels where the sediments are unstable, or in active dredge disposal

areas (NYSDEC 1999). The USEPA SAB (1992) cautions against the use of guidelines based on

equilibrium partitioning in areas of rapid sediment deposition or erosion. Where partitioning of

nonylphenolic compounds between the sediment and interstitial water is not in equilibrium, it is

possible that the sediment guidelines may not be protective against adverse effects to benthos. 

Recommended Canadian Sediment Quality Guidelines

Freshwater Sediments

Insufficient toxicological data exist for freshwater sediments to develop a freshwater TEL and PEL

using the modified NSTP approach, or to calculate a freshwater SSTT value using the SSTT

approach. There are also no sediment quality guidelines that currently exist in other jurisdictions

that could be adopted. Therefore, a provisional freshwater ISQG was calculated from the

Canadian freshwater WQG for nonylphenol proposed in this document using the equilibrium

partitioning approach.

A provisional interim sediment quality guideline of 1.4 mg·kg-1 dry weight, expressed in

nonylphenol toxic equivalent units (NP TEQs) and normalized to 1% TOC, is recommended for

the protection of freshwater life from the adverse effects of nonylphenol and its ethoxylates.

This guideline is recommended for concentrations of nonylphenolic substances in freshwater bulk

surficial sediments (i.e., top 5 cm). The guideline is expressed in nonylphenol toxic equivalency

(NP TEQ) units and may be applied to mixtures of nonylphenol and nonylphenol ethoxylates. The

total concentration of nonylphenolic substances in a sediment sample (in NP TEQs) should be

calculated using the nonylphenol toxic equivalency factors provided in Chapter 3 (Table 4). The

guideline should be adjusted for site-specific levels of total organic carbon. Further guidance and

recommendations for the implementation of this guideline are provided in Chapter 9.

The distribution of the toxicity data for nonylphenol in freshwater sediments, as compared with the

provisional ISQG is shown in Figure 5. There are no effects concentrations below the provisional

freshwater ISQG; therefore, it is thought to be protective of aquatic organisms.

Marine Sediments

As was the case for freshwater sediments, insufficient toxicological data exist for marine

sediments to develop a marine TEL and PEL using the modified NSTP approach, or to calculate a

freshwater SSTT value using the SSTT approach. Also, there are no marine sediment quality

guidelines that currently exist for nonylphenol in any other jurisdictions. Therefore, a provisional

marine ISQG was calculated from the Canadian marine WQG for nonylphenol proposed in this

document by using the equilibrium partitioning approach.

A provisional interim sediment quality guideline of 1.0 mg·kg -1 dry weight, expressed in NP TEQs

and normalized for 1% TOC, is recommended for the protection of marine life from the adverse

effects of nonylphenol and its ethoxylates.

This guideline is recommended for concentrations of nonylphenolic substances in marine bulk

surficial sediments (i.e., top 5 cm). The guideline is expressed in toxic equivalency (TEQ) units

and may be applied to mixtures of nonylphenol and nonylphenol ethoxylates. The total

concentration of nonylphenolic substances in a sediment sample (in NP TEQs) should be

calculated using the nonylphenol toxic equivalency factors provided in Chapter 3 (Table 4). The

guideline should be adjusted based on site-specific levels of total organic carbon. Further

guidance and recommendations for the implementation of this guideline are provided in Chapter

9.
56

The distribution of the toxicity data for nonylphenol in marine sediments, as compared with the

provisional ISQG is shown in Figure 6. There are no effects concentrations below the provisional

marine ISQG; therefore, it is thought to be protective of aquatic organisms.
57

1 10 100 1000

Nonylphenol (mg·kg-1)

Frequency

SQG = 1.4

0.1

Figure 5. Distribution of nonylphenol concentrations in freshwater sediments that are

associated with adverse biological effects (l) and no adverse effects (m).

The provisionally recommended Canadian freshwater interim sediment quality guideline (SQG) is

indicated by the dotted line.

1 10 100 1000

Nonylphenol (mg·kg-1)

Frequency

SQG = 1.0

0.1

Figure 6. Distribution of nonylphenol concentrations in marine and estuarine sediments

that are associated with adverse biological effects (l) and no adverse effects (m).

The provisionally recommended Canadian marine interim sediment quality guideline (SQG) is

indicated by the dotted line.
58

CHAPTER 7. DEVELOPMENT OF CANADIAN SOIL QUALITY GUIDELINES

Canadian soil quality guidelines are developed using the methods outlined in “A protocol for the

derivation of environmental and human health soil quality guidelines” (CCME 1996). This protocol

was created with the intent that guidelines be protective of terrestrial biota and ensure a healthy

functioning ecosystem capable of sustaining current and possible future uses of a site by

ecological receptors and humans. Human health guidelines have not been developed for

nonylphenol, therefore, this chapter discusses only the environmental health component of the

Canadian soil quality guidelines for nonylphenol. According to the protocol, soil quality guidelines

for each chemical are developed for four different land use categories: agricultural,

residential/parkland, commercial, and industrial. A flow chart illustrating the steps that were taken

in developing the environmental soil quality guidelines for nonylphenol is provided in Figure 7.

The primary procedure for derivation of environmental soil quality guidelines is through the use of

direct soil contact toxicological data. A second procedure based on soil and food ingestion is also

applied in the case of agricultural land uses (if sufficient data is available), with adoption of the

lowest of the two values as the guideline for agricultural land use.

Toxicity Data

The first stage in development of environmental soil quality guidelines is the compilation of

toxicity data that is evaluated based on the acceptability of experimental design, laboratory

procedures, and documentation of relevant experimental conditions and parameters. Studies that

provide high quality data are classified as “accepted” and this toxicity data may be used in deriving

the guideline. Studies in which key information is missing, or in which experimental methods or

conditions are questionable, are classified as “consulted” and are not used in the derivation of the

guideline value, though their results may be taken into consideration. A comprehensive list of all

soil toxicity data that was consulted has been compiled in Appendix IV. Studies were excluded

from use because of one or more of the following reasons:

1. the test was not conducted using soil or artificial soil (e.g., conducted in nutrient media

or on agar)

2. the test soil was amended with sewage sludge or a mixture of toxicants, such that

effects could not be conclusively attributed to nonylphenol

3. important details of the experimental procedure were not described (e.g., number of

NP concentrations tested, analytical methodology used, etc.)

The primary data selected as acceptable for use in guideline derivation are shown in Table 11.

Effective concentration data in this table were considered to be biologically significant in addition

to statistically significant.

A brief description of the toxicity data relevant to the derivation of Canadian Soil Quality

Guidelines is provided below. This information reflects the data requirements set forth in the

protocol (CCME 1996) and is presented by taxonomic groups.

Microorganisms

A number of studies have looked at the toxic effects of nonylphenol on microorganisms in artificial

nutrient media, but only toxicity tests conducted with soil are described here. The effects of

nonylphenol on the respiration of soil microorganisms, as measured through CO2 evolution, have

been studied by two sets of researchers. In a 40 day test, Trocmé et al. (1988) observed no

effect of CO2 evolution when nonylphenol was present in the soil at a concentration of

100 mg·kg-1, but at 1000 mg·kg-1 there was a significant reduction in CO2 evolution of 64%. In a

three month assay, Kirchmann et al. (1991) observed no effect on CO2 evolution at a nonylphenol

concentration of 10 mg·kg-1. At 500 mg·kg-1 Kirchmann et al. (1991) did see an effect, but in

contrast with the results of Trocmé et al. (1988), the effect was an increase in CO2 evolution by

26%. Kirchmann et al. (1991) also examined effects of nonylphenol on the nitrogen-cycling of

microorganisms. No effects on nitrification were observed at a nonylphenol concentration of
59

10 mg·kg-1. With 500 mg·kg-1, no effect on nitrogen mineralization occurred, but a 35% decrease

in nitrification was observed.

Plants

Toxicity studies have been conducted with a wide range of plants, or plant cell cultures, grown in

culture medium (see Appendix IVc). These studies cannot be used in the derivation of a soil

quality guideline, and therefore will not be discussed. Nonylphenol toxicity tests using soil have

been conducted for five crop species. From a 14 day test with the turnip, Brassica rapa, an EC5

for growth reduction of 10 mg·kg-1 was calculated (Günther and Pestemer 1992). Other reported

endpoints are one to two orders of magnitude higher. In 21 day tests with nonylphenol, soybean

(Glycine max), sunflower (Helianthrus rodeo), and sorghum (Sorghum bicolor) all showed no

growth effects at 100 mg·kg-1, but had EC50s for growth reduction at 1000 mg·kg-1 (Windeatt and

Tapp 1986). EC50s for growth reduction have also been reported for lettuce (Lactuca sativa) at

559 mg·kg-1 after seven days of exposure, and at 625 mg·kg-1 after 14 days of exposure

(Hulzebos et al. 1993).

Invertebrates

Toxicity tests with nonylphenol have been conducted for three soil-dwelling invertebrates. From a

28 day exposure of the springtail Folsomia candida, Madsen et al. (1998) reported that the EC50

for reproduction falls between 16 and 30 mg·kg-1, and the median lethal concentration is greater

than 30 mg·kg-1. Researchers have also studied survival and reproduction in another species of

springtail, Folsomia fimetaria. In 21 day exposures with this second species, Krogh et al. (1996)

reported an LC10 of 75.1 mg·kg-1 and a LC50 of 151.3 mg·kg-1. For reproduction in F. fimetaria,

Krogh et al. (1996) reported a 21 day EC10 of 23.6 mg·kg-1, while Holm (as cited in Jensen et al.

1997) reported EC10s of 27 mg·kg-1 and 48 mg·kg-1. The EC50s for reproduction in F. fimetaria

were reported at 65.5 mg·kg-1 (Krogh et al. 1996) and 39 mg·kg-1 and 59 mg·kg-1 (Holm, as cited

in Jensen et al. 1997). The effects of nonylphenol on the earthworm Apporectodea calignosa in a

21 day exposure was also studied (Krogh et al. 1996). The median lethal concentration occurred

at greater than 40 mg·kg-1. The median effective concentration for growth reduction was

23.9 mg·kg-1. The authors calculated the EC10 and EC50 for effects on reproduction as

3.44 mg·kg-1 and 13.7 mg·kg-1, respectively.

Mammals

The only mammals which have been tested for nonylphenol toxicity through ingestion are

laboratory rats. In a multigenerational study, Chapin et al. (1999) examined the effects of

nonylphenol administered through dosed food on Sprague Dawley rats (Rattus norvegicus). At a

diet concentration of 650 mg·kg-1 (i.e., a dose of 30-108 mg·kg-1 body weight) vaginal opening at

an earlier age was observed in the F1 generation. Significant effects observed at a diet

concentration of 2000 mg·kg-1 (i.e., a dose of 100-360 mg·kg-1 body weight) included increased

relative kidney weights and decreased adult ovary weights in the F1 generation, and increased

estrous cycle length in both the F1 and F2 generations. Laws et al. (2000) reported LOECs for

increased uterine weight and earlier vaginal opening both at a diet concentration of 50 mg·kg-1,

and disruption of estrous cyclicity at a diet concentrations of 100 mg·kg-1. A LOEC of 75 mg·kg-1

body weight per day for increased uterine weight in ovariectomized female rats has been reported

by Odum et al. (1999).
60

 AGRICULTURAL RESIDENTIAL/PARKLAND COMMERCIAL & INDUSTRIAL

 SOIL SOIL AND FOOD SOIL SOIL

 CONTACT INGESTION CONTACT CONTACT

 Calculate TEC Calculate DTED* Calculate TEC Calculate ECL

 ̊

 WOE LOEC ME WOE LOEC ME WOE LOEC 

Method Method Method Method Method Method Method Method

  ̧  ̧  ̧

 Nutrient and Energy Nutrient and Energy Nutrient and Energy

 Cycling Check* Cycling Check* Cycling Check*

  ̊  ̊  ̊

 SQGSC SQGI* SQGSC SQGSC

  ̧  ̊  ̧  ̧

 Take lowest of

 two values

 SQGE SQGE SQGE

  ̧  ̧  ̧

 Groundwater Groundwater Groundwater

 Check* Check* Check*

  ̧  ̧  ̧

Figure 7. Overview of steps leading to the development of final Environmental Soil Quality Guidelines for nonylphenol.

TEC = threshold effects concentration; DTED = daily threshold effects dose; ECL = effects concentration low;

WOE = weight of evidence; LOEC = lowest observed effect concentration; ME = median effects;

SQGSC = soil quality guideline for soil contact; SQGI = soil quality guideline for soil and food ingestion;

SQGE = final environmental soil quality guideline

 ̧ Indicates steps used in the derivation of guidelines for nonylphenol.

 ̊ Indicates values that were not calculated in the derivation of guidelines for nonylphenol due to insufficient data.

* Indicates steps that are not mandatory for deriving a guideline if insufficient data is available.
61

Table 11. Primary data available for toxicity of nonylphenol though direct soil contact.

Species

(Common name)

Duration Endpoint Effect Concentration

(mg·kg-1)

Reference

Plants

Brassica rapa

(turnip)

14d EC5 growth 10 Günther and

Pestemer 1992

Glycine max

(soybean)

21d NOEC growth 100 Windeatt and Tapp

1986

21d EC50 growth 1000 Windeatt and Tapp

1986

Helianthrus rodeo

(sunflower)

21d NOEC growth 100 Windeatt and Tapp

1986

21d EC50 growth 1000 Windeatt and Tapp

1986

Lactuca sativa

(lettuce)

7d EC50 growth 559 Hulzebos et al. 1993

14d EC50 growth 625 Hulzebos et al. 1993

Sorghum bicolor

(sorghum)

21d NOEC growth 100 Windeatt and Tapp

1986

21d EC50 growth 1000 Windeatt and Tapp

1986

Invertebrates

Apporectodea 21d EC10 reproduction 3.44 Krogh et al. 1996

calignosa 21d EC50 reproduction 13.7 Krogh et al. 1996

(earthworm) 21d EC50 growth 23.9 Krogh et al. 1996

21d EC10 mortality >40 Krogh et al. 1996

21d EC50 mortality >40 Krogh et al. 1996

Folsomia candida 28d EC50 reproduction 16-30 Madsen et al. 1998

(springtail) 28d LC50 survival >30 Madsen et al. 1998

Folsomia fimetaria 21d LC10 survival 75.1 Krogh et al. 1996

(springtail) 21d LC50 survival 151.3 Krogh et al. 1996

21d EC10 reproduction 23.6 Krogh et al. 1996

21d EC10 reproduction 27 Holm*

21d EC10 reproduction 48 Holm*

21d EC50 reproduction 65.5 Krogh et al. 1996

21d EC50 reproduction 39 Holm*

21d EC50 reproduction 59 Holm*

 * As cited in Jensen et al. 1997

NOEC = no observed effect concentration

ECx = concentration at which effects are observed in x% of the population

LCx = concentration that is lethal to x% of the population
62

Agricultural and Residential/Parkland Land Uses

Soil Contact

There are slight differences in the derivation procedures for soil contact guidelines (SQGSC)

among land uses such that a higher level of protection is provided for agricultural and

residential/parkland soils than for commercial and industrial soils. For agricultural and

residential/parkland land use, derivation of the threshold effect concentration (TEC) due to direct

soil contact may proceed by one of three possible methods (weight of evidence method, lowest

observed effect concentration method, median effects method), depending on the toxicity data

available. A nutrient and energy cycling check is performed (when sufficient data are available)

and compared with the TEC. The lower of these two values is adopted as the soil quality

guideline for soil contact.

The preferred approach for derivation of the soil contact guideline is to use a weight of evidence

method that considers a range of acceptable endpoints. The minimum requirements for this

approach are 10 data points from at least three studies, with at least two soil invertebrate species

and at least two plant species represented. According to Section 7.5.2.2 of the protocol (CCME

1996), the geometric mean should be used when multiple data are available for the same

endpoint with the same species. For the nonylphenol data, geometric means have been applied

to the three EC10 values and the three EC50 values reported for reproduction in springtails (Krogh

et al. 1996; Holm as cited in Jensen et al. 1997). The soil toxicity data for nonylphenol satisfied

the minimum data requirements with a total of 20 data points (after geometric means were taken

for multiple data) from 6 studies representing 5 plant species and 3 soil invertebrate species (see

Table 11).

For agricultural and residential/parkland land uses, if the minimum data requirements are met,

then the TEC may be calculated by taking the 25th percentile of the frequency distribution (i.e., the

“no potential effects” range) and dividing it by an uncertainty factor (if deemed necessary).

Therefore,

TEC = NPER ÷ UF

where,

TEC = threshold effects concentration (mg·kg-1 soil)

NPER = no potential effects range (25th percentile of distribution)

UF = uncertainty factor (if needed)

Using rank percentile and linear reqression methods, the 25th percentile of the soil contact toxicity

data was calculated as 17.15 mg·kg-1 soil (Figure 8).

Although there were ample data available representing a wide range of taxonomic groups, the

use of an uncertainty factor was deemed necessary. There was a preponderance of median

lethal and median effective concentration data that may have introduced some bias into the

dataset. Also, more than 25% of the data below the 25th percentile were definitive effects data

(i.e., LCx and ECx data, as opposed to LOECs and NOECs). Therefore, as recommended in the

protocol (CCME 1996), an uncertainty factor of 3 was chosen.

Thus,

TEC = 17.15 ÷ 3 = 5.72 mg·kg-1 soil ≈ 5.7 mg·kg-1 soil
63

y = 38.25 [Log(x)] - 22.22

R2 = 0.95

0

10

20

30

40

50

60

70

80

90

100

1 10 100 1000

Nonylphenol Concentration (mg·kg-1)

Rank Percentile

25th percentile

17.15

Figure 8. Distribution of soil contact toxicity data used in the guideline derivation for agricultural and residential/parkland land uses.

 The no potential effects range (i.e., the 25th percentile of the distribution) is indicated by the dotted line.
64

Nutrient and Energy Cycling Check

Once a soil contact TEC has been calculated, it is then checked against a microbial (nutrient and

energy cycling) value. Microorganisms are an essential component of healthy, functioning soils.

The ability of soil to support plant life depends on the coordinated activities of microorganisms that

mediate processes such as decomposition, respiration, and organic nutrient cycles. The rates of

these processes may be affected by contaminants, therefore, the microbial check provides an

additional tool for assessing the toxicological impact of soil-associated contaminants on terrestrial

life.

The microbial check may be calculated by weight of evidence, LOEC, or median effects methods,

depending on available data. Preferred endpoints are nitrogen fixation and nitrification, although

carbon cycling and nitrogen mineralization data may also be considered.

The data that were available for microbial responses to nonylphenol are presented in Appendix

IVa. Acceptable endpoints included a NOEC and a LOEC for nitrification, a NOEC for nitrogen

mineralization, and two NOECs and two LOEC for respiration (Trocmé et al. 1988; Kirchmann et

al. 1991). These data were insufficient to meet the minimum requirements for the weight of

evidence, LOEC, or median effects approaches, so a nutrient and energy cycling check could not

be calculated. It appears though, on the basis of the acceptable LOEC values that are available

for microbial processes (ranging from 500 to 1000 mg·kg-1), that microorganisms are generally

less sensitive to nonylphenol than other terrestrial organisms. Hence, it is unlikely that omission

of this check will result in a lack of protection for microbial processes. To calculate the nutrient

and energy cycling check using the LOEC method, at least one more study reporting a LOEC

value would be required. For the weight of evidence approach, several studies reporting median

effects data would be required, preferably with endpoints of nitrogen fixation and nitrification.

As a nutrient and energy cycling check could not be calculated, the soil contact TEC of

5.7 mg·kg-1 was adopted as the soil quality guideline for soil contact (SQGSC) for both agricultural

and residential/parkland land uses.

Soil and Food Ingestion

The soil quality guideline for soil and food ingestion (SQGI) applies only to agricultural land use.

Derivation of the soil and food ingestion guideline first involves calculating the daily threshold

effects dose (DTED). A minimum of three studies is required, at least one of which is an oral

avian study, and two of which are oral mammalian studies (no more than one laboratory rodent

study, and preferably including an ungulate study). The DTED is calculated as the lowest

observed adverse effect level (LOAEL) from the most sensitive species divided by an uncertainty

factor. The soil quality guideline for ingestion is then calculated such that the ingestion of soil and

plants growing on the soil does not result in the most sensitive species receiving more than 75%

of the DTED. More detailed information on these calculations is provided in the protocol (CCME

1996).

There is a lack of data on the toxicity of nonylphenol to grazing livestock and wildlife through

ingestion. The few studies that have been conducted have all examined effects on laboratory

rodents (Appendix IVd).

The minimum data requirements cannot be met for calculating a soil quality guideline for ingestion

(SQGI). Rough calculations based on the data available suggest that an SQGI likely would not be

lower than the SQGSC; however, due to the high uncertainty involved, no conclusions concerning

ingestion can be made at this time.
65

Commercial and Industrial Land Uses

Soil Contact

For commercial and industrial land use, the effects concentration low (ECL) due to direct soil

contact may be calculated using either the weight of evidence method or LOEC method,

depending on the toxicity data available. A nutrient and energy cycling check is performed (when

sufficient data are available) and the resulting microbial value is compared with the ECL.

As with the agricultural and residential/parkland land uses, the weight of evidence approach was

used because the minimum data requirements for this method were met. For commercial and

industrial land uses, if the minimum data requirements are met, then the ECL may be calculated

by taking the 25th percentile of the “effects” data distribution only (rather than the 25th percentile of

the entire data distribution, as is done for the NPER). At this percentile, called the effects range

low (ERL), it is expected that some effects will be incurred by soil-dependent biota, but not at the

level of median lethality in the population(s). Less stringent protection levels are required than for

residential/parkland and agricultural land because the primary land use activities of commercial

and industrial lands are not directly dependent on the need to sustain a high level of ecological

processes. For this same reason, it is not desirable to achieve lower estimates of the ECL using

uncertainty factors.

Therefore, the effects concentration low (ECL) for nonylphenol was calculated as

ECL = ERL

where,

ECL = effects concentration low (mg·kg-1 soil)

ERL = effects range low (25th percentile of effects data distribution, in mg·kg-1 soil)

Using ranked percentile and linear reqression methods, the 25th percentile of the soil contact

effects data was calculated as 14.16 mg·kg-1 soil (Figure 9).

Thus,

ECL = 14.16 mg·kg-1 soil ≈ 14 mg·kg-1 soil
66

y = 34.863 [Log(x)] - 15.126

R2 = 0.94

0

10

20

30

40

50

60

70

80

90

100

1 10 100 1000

Nonylphenol Concentration (mg·kg-1)

Rank Percentile

25th percentile

14.16

Figure 9. Distribution of soil contact effects toxicity data used in guideline derivation for commercial and industrial land uses.

 The effects range low (i.e., the 25th percentile of the effects data distribution) is indicated by the dotted line.
67

Nutrient and Energy Cycling Check

Once a soil contact ECL has been calculated, it is then checked against a microbial (nutrient and

energy cycling) value. The microbial check may also be calculated by either the weight of

evidence method or the LOEC method, depending on available data. Preferred endpoints are

nitrogen fixation and nitrification, although carbon cycling and nitrogen mineralization data may

also be considered. If the microbial value is lower than the ECL, then significant adverse effects

may be experienced by microorganisms controlling nutrient and energy cycling processes. In this

case, the geometric mean of the two values is taken and used as the soil quality guidelines for soil

contact (SQGSC) for commercial and industrial land use. However, if the microbial value is above

the ECL, then a level of effects consistent with the objective of this procedure is expected and the

ECL is adopted directly as the SQGSC for commercial and industrial land use.

The data that were available for microbial responses to nonylphenol are presented in Appendix

IVa. These data were insufficient to meet the minimum requirements for either the weight of

evidence method or the LOEC method. To calculate the nutrient and energy cycling check using

the LOEC method, at least one more study reporting a LOEC value would be required. For the

weight of evidence approach, several studies reporting median effects data would be required,

preferably with endpoints of nitrogen fixation and nitrification.

As a nutrient and energy cycling check could not be calculated, the soil contact ECL of 14 mg·kg-1

was adopted as the soil quality guideline for soil contact (SQGSC) for both commercial and

industrial land uses.

Groundwater Check

A major concern with soil contamination is that it can lead to contamination of groundwater. Soil

quality guidelines for all four classes of land use must be designed to account for possible

transfers to groundwater such that concentrations of a substance in groundwater do not exceed

the water quality guidelines for the protection of freshwater aquatic life. To prevent such an

exceedence, a groundwater check is calculated based on equilibrium partitioning between solid

and aqueous phases. The applicability of equilibrium partitioning to nonylphenol is discussed in

Chapter 6. For non-ionic organics, such as nonylphenol, the following equation (CCME 1996) is

used:

Y = DF · Cwa · (Kd + θm)

where,

Y = concentration of nonylphenol in soil (mg NP·kg-1 dry soil)

DF = dilution factor = 50 (default)

Cwa = water protection value = Canadian water quality guideline (mg·L-1)

Kd = distribution coefficient

θm = soil moisture content = 0.1 (default)

The distribution coefficient (Kd) is directly related to the organic matter content and can be derived

using the sorption partition coefficient (Koc) and the organic fraction of dry soil (foc) as described by

the equation:

Kd = Koc · foc

A default value of 0.3% (or 0.003) is used for foc (CCME 1996). The log Koc for nonylphenol has

been estimated to range from 4.7 to 5.6 (Sekela et al. 1999), so, a mean Koc can be determined

by taking the anti-log of these two values and then calculating the geometric mean.

Therefore,
68

 Koc = 141 254 L·kgoc

-1

 Kd = 423.8 L·kgsoil

-1

For the protection of freshwater aquatic life, a water quality guideline of 1.0 μg·L-1 (or

0.001 mg·L-1) was developed for nonylphenol in Chapter 5. Therefore:

Y = (50) (0.001) (423.8 + 0.1)

= 21.2 mg NP·kg-1 dry soil

The groundwater check indicates that for the protection of freshwater aquatic life, the level of

nonylphenol in soil should not exceed 21 mg·kg-1.

Off-site Migration Check

Contaminated soil can be transferred from one property to another through erosion by water and

wind with subsequent deposition. This transfer of contaminants can result in unacceptable

degradation of an adjacent property that was previously uncontaminated, or used for a more

sensitive land use. Therefore, an off-site migration check was calculated to determine the

concentration of nonylphenol in soil from an industrial site that could, through erosion,

hypothetically result in levels on an adjacent residential/parkland property that exceed the

residential/parkland SQGE. The check value was calculated using the following equation (CCME

1996):

Ci = (14.3 · SQGRP) - (13.3 · BSC)

where,

Ci = concentration of nonylphenol in the eroded soil (mg NP·kg-1)

SQGRP = soil quality guideline for residential/parkland land use

BSC = background concentration of nonylphenol in the receiving soil (mg NP·kg-1)

The soil quality guideline for soil contact for residential/parkland land uses is 5.7 mg·kg-1, and the

background concentration of nonylphenol is assumed to be zero. Substituting these values into

the equation yields the following off-site migration check value:

Ci = 82 mg NP·kg-1

The off-site migration check indicates that for the protection of adjacent properties, the level of

nonylphenol in soil on industrial sites should not exceed 82 mg·kg-1.

Environmental Soil Quality Guideline Derivation (SQGE)

The final environmental soil quality guideline for agricultural land use is determined by taking the

lower of either the soil contact guideline (SQGSC) or the soil ingestion guideline (SQGI), if both are

available. If there are not enough data to calculate a soil ingestion guideline, then the soil contact

guideline is taken as the final environmental guideline; however, the final environmental guideline

cannot be derived if there is insufficient data for the soil contact guideline. For all other soil uses,

the soil contact guideline is used as the final environmental soil quality guideline.

Because no data were available on toxicity of nonylphenol through ingestion of food or soil, the

SQGSC of 5.7 mg·kg-1 will be taken as the final SQGE for agricultural land use. For the other three

land uses, the SQGSC is used as the SQGE also. Therefore, the SQGEs for residential/parkland,

commercial, and industrial land uses are 5.7 mg·kg-1 soil, 14 mg·kg-1 soil, and 14 mg·kg-1 soil,

respectively. The groundwater check value was higher than all of the SQGE values, indicating that

these guidelines are sufficiently low to protect against contamination of groundwater. The off-site

migration check value was higher than the SQGE for industrial land uses, indicating that this
69

guideline is sufficiently low to protect against degradation of adjacent lands through deposition of

eroded soil.

Recommended Canadian Soil Quality Guidelines

The final Canadian Soil Quality Guidelines for a substance are determined as the lower of the

environmental soil quality guideline (SQGE) and the human health soil quality guideline (SQGHH)

for each land use. Because human health soil quality guidelines have not been developed for NP

and NPEs, the environmental guidelines will be taken as the final Canadian Soil Quality

Guidelines for nonylphenol and its ethoxylates. A summary of the soil quality guideline values for

each land use type is provided in Table 12. These guidelines are expressed in nonylphenol toxic

equivalency units (NP TEQs) and may be applied to mixtures of nonylphenol and nonylphenol

ethoxylates. The total concentration of nonylphenolic substances in a soil sample (in NP TEQs)

should be calculated using the nonylphenol toxic equivalency factors provided in Chapter 3 (Table

4). Further guidance and recommendations for the implementation of these guidelines are

provided in Chapter 9.

The Canadian environmental soil quality guidelines recommended for agricultural and

residential/parkland land use are higher than the Danish soil quality criterion for nonylphenol (see

Table 8). The Danish criterion was based on the 21 day EC50 for reduced reproduction in

earthworms at 13.7 mg·kg-1 as determined by Krogh et al. (1996). This endpoint, which

represented the lowest toxicity value in their data set, was divided by an application factor of 1000

and rounded off to 0.01 mg·kg-1. It is due to this extremely conservative application factor that the

Danish soil criterion is so much lower than the Canadian soil guideline. Both Sweden and

Denmark have also set limits for NP in sludge used as agricultural fertilizer (Table 8). The Danish

guidelines refer to the total concentration of 4-NP, NP1EO and NP2EO in sludge (European

Commission 1999; Danish Ministerial Order No. 823, Sept. 16, 1996). The higher guideline

values for sludge, compared with soil, reflect the fact that the nonylphenol concentrations in the

sludge will be effectively diluted through the application of the sludge over a large area of soil. 
70

Table 12. Summary of soil quality guideline derivation for nonylphenol. *

Land use

Guideline

(mg·kg-1)

 Agricultural Residential/

Parkland

Commercial Industrial

Final Canadian SQG 5.7 5.7 14 14

SQGE 5.7 5.7 14 14

SQGSC 5.7 5.7 14 14

Nutrient and energy cycling NC NC NC NC

SQGI NC NA NA NA

Groundwater check

(freshwater aquatic life)

21 21 21 21

Off-site migration check NA NA NA 82

SQGHH NC NC NC NC

* All guideline values are expressed in toxic equivalent (TEQ) units.

SQGE = soil quality guideline for environmental health; SQGSC = soil quality guideline for soil

contact; SQGI = soil quality guideline for soil and food ingestion; SQGHH = soil quality guideline

for human health

NC = not calculated; NA = not applicable
71

CHAPTER 8. AREAS FOR FUTURE RESEARCH

The guidelines developed in this document are based on the best available data. In the process

of deriving these guidelines, however, it was noted that there are certain areas of study in which

limited data are available for nonylphenol and its ethoxylates. This chapter highlights some of the

gaps in our knowledge of these substances with the hope that it will provide stimulation and

direction for further research.

Concentrations in Canada

Historically, concentrations of NP and NPEs have not been routinely monitored in the Canadian

environment. Consequently, there is limited data concerning the distribution and levels of these

substances that occur in Canada. In particular, there is either no data, or very limited data, that

exist for Canadian marine and estuarine waters, marine sediments, groundwaters, soils, and

industrial effluents and sludges. There is also a lack of data on levels of NPECs and NPnEOs

(n≥3) in freshwater sediments.

Fate and Behaviour

There is a need for further studies into the fate and behaviour of NP and NPEs in the

environment. In particular, more field studies on the fate of these substances in sediments are

needed because most work to date has been conducted in a laboratory setting. There does not

appear to have been any work done on the effects of photolysis on NP/NPEs present on soil

surfaces. There also has been no research into the effects of varying pH on the aquatic toxicity or

bioavailability of nonylphenol. Further investigation into the biological fate of NP and NPEs is also

needed. Specifically, the data available on uptake and elimination rates in invertebrates, uptake

by aquatic plants, and uptake by terrestrial animals are limited.

Toxicity Testing

An area of research that has been largely unexplored is the toxic effects of NP and NPEs on

mammalian and avian consumers of aquatic life. There are very limited toxicity data available for

sediment-dwelling organisms. Toxicity testing has also not been conducted for terrestrial wildlife

and grazing livestock.

The toxicity data that does exist for aquatic algae, plants, invertebrates and vertebrates is largely

for NP, NP1-2EO, NP9-10EO, and, to a lesser degree, NP1-2EC. There is a need for more

toxicity data on other NPE oligomers, particularly those containing between 3 and 8 ethoxylate

groups.

Further investigation into the mode of toxicity for these substances would also be useful. At

present, the exact mechanisms of toxicity via narcosis are unclear. There is also some

controversy over the exact mechanisms involved in the endocrine disrupting effects produced by

these substances.

Guideline Derivation and Implementation

When more toxicity data becomes available, it may be possible to upgrade the status of the

marine interim WQG developed in this document. A full marine WQG could be derived if more

chronic toxicity data becomes available for marine fish and for marine invertebrates other than

crustaceans.

An unconventional approach (equilibrium partitioning) was used to derive the sediment quality

guidelines in this document due to a lack of sediment toxicity data. In the future, if more sediment

toxicity data becomes available for nonylphenol it may be possible to derive guidelines which are

based on approaches in the CCME protocol (CCME 1995). In order to use the NSTP approach,
72

12 more entries in the effect data set and 3 more entries in the no-effect data set would be

required for the freshwater SQG; 11 more entries in the effect data set and 6 more entries in the

no-effect data set would be required for the marine SQG. Alternatively, the freshwater SQG could

be derived using the SSTT approach if three more studies on sediment-resident invertebrates are

conducted, including at least one study on a benthic amphipod, and at least one partial or full lifecycle test that reports ecologically relevant sub-lethal endpoints. A marine SQG could be derived

using the SSTT approach if at least two more studies on sediment-resident invertebrates are

conducted, at least one of which is a partial or full life-cycle test with sub-lethal endpoints.

The soil quality guidelines in this document were developed without the calculation of either a soil

and food ingestion guideline, or a nutrient and energy cycling check. The soil and food ingestion

guideline could be calculated in future if at least one oral avian study was conducted, and at least

one oral mammalian study was conducted with a mammal other than a laboratory rodent, but

preferably with an ungulate species. To calculate a nutrient and energy cycling check, at least

one more study reporting a LOEC value would be required, or several studies reporting median

effects data for endpoints such as nitrogen fixation and nitrification.

This document recommends the use of TEF values that were developed by Servos et al. (2000).

Further work is needed to verify and refine these TEF values. For example, a single TEF is

currently given for the entire group of NPEs ranging from NP9EO and up. It may be possible to

develop oligomer-specific TEFs if variations in toxicity are known to occur for different ethoxylate

chain lengths. Due to a lack of toxicity data for NPEs with 3 to 8 EO groups, a conservative TEF

value was estimated from the other TEFs. Further work is need to develop TEFs for this group of

ethoxamers.

At present, we are recommending that OP, OPEs, and OPECs be included in the TEQ approach

for nonylphenolic substances. Further investigation is needed to confirm whether these

substances should be grouped together, or whether they warrant separate guidelines. As with

NPEs, further refinement and validation of the TEF values for OPEs is needed.
73

CHAPTER 9. GUIDANCE ON IMPLEMENTATION

General Guidance for all Guidelines

The legislative authority for implementation of Canadian Environmental Quality Guidelines (EQGs)

lies with each provincial and territorial jurisdiction (with the exception of federal lands). The

provincial and territorial governments may develop their own science-based criteria, guidelines,

objectives and standards, but often the Canadian EQGs form the scientific basis for these (CCME

1999). Canadian EQGs may also form the basis for federal use-specific guidelines and

objectives.

The Canadian EQGs developed in this document for nonylphenol should be used along with all

other relevant information in making practical and informed decisions regarding water, sediment,

and soil quality. It is widely recognized that no single assessment tool can always be used to

predict whether adverse biological effects will occur as a result of contaminant exposure. Rather,

the appropriate use of several different tools provides the most useful information (Chapman

1995). Apart from EQGs, other scientific tools available to assist in the protection and

management of water, sediment, or soil quality include other assessment values (such as the PEL

for sediment), biological assessments, or guidelines for other media.

It should also be noted that socio-economic or technological factors that may influence the

application of Canadian EQGs are not considered in their development process. However, such

factors may play a varying role in the application of EQGs (and/or in the development of sitespecific objectives) within the decision-making framework of different jurisdictions and programs

(Gaudet et al. 1995).

Negligible risk to biota, their functions, or any interactions that are integral to sustaining the health

of ecosystems and the designated resource uses they support, should result when nonylphenol

(and other nonylphenolic substances) are present in the ambient environment at the

concentrations recommended by the Canadian EQGs developed in this document. It should be

noted, however, that for ecosystems of superior quality, impairment to these guideline

concentrations is not advocated.

Additional general information on the use of Canadian EQGs is available in the introductory

chapter of the publication Canadian Environmental Quality Guidelines (CCME 1999). More

specific guidance, relevant to the guidelines for nonylphenol and its ethoxylates, is provided

below.

Application of the TEQ Approach to EQGs

When comparing environmental samples to the guidelines developed in this document, it is

important that concentrations of NP and all NPEs and NPECs in the sample are measured,

converted to nonylphenol toxic equivalents (NP TEQs), and then summed. It is also

recommended that OP, OPEs and OPECs, if present in the sample, be included in the calculation

of the total NP TEQ. Toxic equivalency factors (TEFs) are provided in Table 4 of Chapter 3. The

following example illustrates the proper use of the TEQ approach and its application towards the

Canadian Environmental Quality Guidelines for nonylphenol and its ethoxylates.

Consider a water sample collected from a freshwater stream that has been analysed for all

possible nonylphenolic substances. The sample was found to contain 0.1 μg·L-1 of NP, 1.0 μg·L-1

of NP1EO, 0.2 μg·L-1 of NP3EO, and 2.0 μg·L-1 of NP1EC. The TEF values for toxicity of NP,

NP1EO and NP1EC are 1, 0.5, and 0.005, respectively, and we will assume a relative potency of

0.5 for NP3EO (see Table 4). Therefore, using the equation

Total concentration = ∑ (Cx x TEF),
74

and plugging in the concentrations from our water sample, we get:

Total concentration = (0.1 μg·L-1 x 1) + (1.0 μg·L-1 x 0.5) + (0.2 μg·L-1 x 0.5) + (2.0 μg·L-1 x 0.005)

 = 0.71 μg·L-1

Therefore, our water sample contains 0.71 μg·L-1 of nonylphenolic substances, as expressed in

NP TEQ units. This total concentration 0.71 μg·L-1 is the value that should then be compared

against the nonylphenol Canadian Water Quality Guideline for the protection of freshwater aquatic

life (i.e., 1.0 μg·L-1). In this example then, the water sample does not exceed the freshwater

guideline for nonylphenol and its ethoxylates and adverse effects to aquatic life are not expected

to occur in this water.

These same NP TEF values should be used to determine total NP TEQs for sediment and soil

samples before comparing them to the respective guideline values. Narcosis is a common mode

of toxicity for nonylphenolic substances across all taxonomic groups (see earlier discussion under

“Modes of Toxicity” in Chapter 3), so the NP TEF values should be applicable to plants,

invertebrates, fish, mammals, etc., regardless of whether they are water-, sediment-, or soildwelling species.

Water Quality Guidelines

The toxicological data upon which these guidelines are based were conducted within the pH

ranges of 5.6 to 8.9 and 7.3 to 9.6 for fresh and marine waters, respectively. Most natural waters

will have pH values within these ranges. However, site-specific considerations may be needed for

those locations where the water pH falls outside of these ranges. In particular, these guidelines

may not be sufficiently protective for highly alkaline waters. The acid dissociation constant (pKa)

of nonylphenol is 10.7, indicating that in most natural waters NP will remain predominantly in the

undissociated form. In waters with a pH greater than 10.7, the dissociated form, which is more

soluble and may differ in bioavailability or toxicity, will be present. It is currently unknown whether

pH has any effect on the aquatic toxicity or bioavailability of NP and NPEs.

The presence or absence of sensitive species at a local site is another factor that might play a

role in the development of site-specific objectives.

These guidelines refer to concentrations in unfiltered samples.

Sediment Quality Guidelines

The interim sediment quality guidelines presented here represent the concentrations of

nonylphenol or nonylphenol equivalents in sediments below which adverse effects in sedimentdwelling aquatic biota are not expected to occur. Although these interim sediment quality

guidelines are considered provisional at this time, they should not be used differently than they

would be if they were full sediment quality guidelines. The revision and re-assessment of these

provisional ISQGs will be possible when sufficient data become available to support the use of the

modified NSTP or SSTT approaches as described in the formal protocol (CCME 1995).

Using the equilibrium partitioning approach, the provisional interim sediment quality guidelines

were normalized to 1% TOC (foc = 0.01) in order to express them in terms of sediment dry weight.

In jurisdictions where the percentage of organic carbon in sediments is known to be consistently

higher or lower than 1%, local estimates of TOC may be used to set site-specific sediment

objectives. This is done by multiplying the ISQG by the percentage of organic carbon. For

example, for freshwater sediments with 2% TOC, the freshwater ISQG of 1.4 mg·kg-1 would be

multiplied by 2 to obtain a site-specific objective of 2.8 mg·kg-1. With lower levels of organic

carbon there will be less binding of contaminants to the sediments, and consequently greater

bioavailability. Therefore, it is important that the foc value used is based on the lowest sediment

organic carbon concentration that might possibly occur, such that all aquatic life in the designated
75

jurisdiction will be protected. It is not recommended that foc values outside of the range 0.002 to

0.12 be used in deriving site-specific objectives. For sediments that contain less than 0.2% or

greater than 12% TOC, other factors that the equilibrium partitioning method does not account for

may influence the contaminant partitioning (USEPA SAB 1992). Further generic guidance

regarding the implementation of Canadian sediment quality guidelines is provided in CCME

(1995).

Soil Quality Guidelines

The toxicological studies upon which these guidelines are based did not report soil pH in many

cases; however, where reported, the studies were conducted within the pH range of 5.8 to 7.5.

Therefore, site-specific consideration may be required for soils that fall outside of this pH range.

In particular, these guidelines may not be sufficiently protective for sites with highly alkaline soils

(i.e., with a pH greater than 10). The high acid dissociation constant of nonylphenol (pKa = 10.7)

indicates that in most natural environments NP will remain predominantly in the undissociated

form. Extremely high pHs may create dissociated forms that are less readily sorbed than the

parent nonylphenol, and much more mobile in soil. In these cases, the groundwater check as

utilized in this document will not be applicable (see CCME 1996), and the development of more

conservative site-specific guidelines than presented here may be necessary to ensure adequate

protection.
76

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ANNEX I. ECOLOGICAL RELEVANCE OF ENDOCRINE DISRUPTION

Environmental guidelines must be based on toxicity data that have some ecological relevance.

Conventional toxicity endpoints, such as mortality, growth inhibition, and reduced reproductive

output are widely acknowledged as ecologically-relevant effects that can cause impacts at

organizational levels higher than the individual (i.e., population, community and ecosystem levels).

There has recently been much debate over the ecological relevance of effects caused by

endocrine disrupters. Endocrine disrupting chemicals (EDCs) work at various biochemical levels,

such as affecting the synthesis of hormones, hindering movement of hormones through the blood

by hormone transporting proteins, upsetting the metabolism of hormones, or by directly interfering

with cellular hormone receptors (Bjerregaard et al. 1998). However, even though an EDC may

cause biological and physiological changes within an organism, whether or not these changes

affect whole organism performance in terms of survival, reproduction or growth will depend on the

hormone affected, the degree of impairment, the timing of the exposure in terms of the organism’s

life stage, and the relative sensitivity of the particular species (Van Der Kraak 1998).

Evidence of endocrine disruption, in various forms, has been extensively documented in

vertebrates such as fish, marine mammals, predatory birds, and even alligators (Fry and Toone

1981; Reijnders 1986; Brouwer et al. 1989; Munkittrick et al. 1991, 1992; Jobling and Sumpter

1993; Purdom et al. 1994; Fry 1995; Guillette et al. 1994, 1995, 1996; Harries et al. 1997). There

has been less documentation of endocrine disruption in invertebrates species; most cases have

involved daphnids or marine molluscs (Taylor et al. 1999). Although there is a wealth of data

describing the effects of endocrine disrupters at the cellular and biochemical levels, very few

studies have investigated the long-term ramifications of these effects at the population or

community level (Hutchinson and Matthiessen 1999). In some cases, although population-level

changes have been observed, the effects have not been linked to a specific chemical.

Ankley and Giesy (1998) have developed a weight of evidence approach for conclusively

demonstrating ecologically relevant effects of an EDC. First, there must be documentation of

effects in individuals and of adverse effects in populations. Second, there must be a connection

between the effects observed in populations and those observed in individuals. Third, a plausible

mode of action must be identified that is consistent with the effects observed in the individuals.

Fourth, there must be positive identification of specific contaminants operating through this mode

of action. Fifth, the exposure metrics must be reasonable in terms of individual-level effects

occurring via the mode of action under consideration. And finally, there should be evidence that

the population or individuals will recover upon removal of the chemical stressor.

For NP, short-chain NPEs and NPECs, there is extensive documentation of effects at the

individual level, and the mechanism of action is generally understood. However, how these

effects on individuals may translate into adverse effects at the population level is not well

understood.

In the case of fish, to determine the significance for populations of endocrine disrupter effects

such as vitellogenin induction and intersex states, it will first be necessary to establish their

impacts on individual fitness (Taylor et al. 1999). It is important to establish possible links

between the vitellogenin response and ecologically important parameters that are relevant to

population dynamics, such as semen quality, testicular development, histological changes and

offspring sex ratio and development (Bjerregaard et al. 1998). A few studies have examined the

effects of xenobiotically-induced vitellogenesis on the individual fitness of fish. Carragher and

Sumpter (1991) found that when they induced vitellogenin synthesis in juvenile rainbow trout with

17ß-estradiol, there was a significant decrease in the amount of calcium in the fish scales, and a

significant increase in plasma calcium levels (possibly for incorporation into vitellogenin). The

authors suggested that at higher demands for calcium due to vitellogenesis, the scales could

become loosened, thus compromising the fish’s defenses. Therefore, there may be a link

between vitellogenesis and fish survival rates (Carragher and Sumpter 1991). Herman and
103

Kincaid (1988) found that when rainbow trout underwent prolonged vitellogenin induction due to ßestradiol exposure, mortality rates increased, and necrosis and damage to the kidney and liver

tissues were severe. Induction of vitellogenin in male fish has also been correlated to decreases

in testicular growth and serum testosterone levels (Jobling et al. 1996; Folmar et al. 1996). These

effects might have a bearing on reproductive success.

In their multigenerational study with rats, Chapin et al. (1999) observed increased estrous cycle

length in those individuals exposed to NP. This estrogenic response could have adverse effects

at the population level because lengthened estrous cycles have been associated with reduced

fecundity (Chapin et al. 1997).

It seems likely that these estrogenic effects of NP and short-chain NPEs and NPECs could result

in population-level effects. However, at this point such effects are largely speculative and actual

documentation of population-level effects in the field are needed.

In some cases, population-level effects have been observed due to NP, but further research is

needed to confirm that these effects are the result of endocrine disruption. For example, adverse

effects of NP on the smoltification of juvenile Atlantic salmon (Salmo salar) have been observed in

laboratory studies (Madsen et al. 1997; Brown et al. 1998), and resultant decreases in population

sizes have been documented in the field (Fairchild et al. 1999). Although the authors suspect

endocrine disruption, the mechanism by which NP affects smoltification is unknown.

In an attempt to address the current data gaps concerning the ecological impacts of EDCs, there

has been a call for more multigenerational toxicity tests with suspected endocrine disrupters

(Taylor et al. 1999). Studies examining population parameters such as offspring survival and

recruitment are needed to assess the effects of long-term exposure to EDCs. The OECD is

currently working to develop and validate test methods for assessing the potential of new and

existing chemicals for endocrine disruption (Taylor et al. 1999).

Endocrine disruption endpoints were considered in the development of the guidelines in this

document. In the aquatic toxicity dataset, the range of concentrations at which EDC effects were

observed were comparable with the range of concentrations for more conventional endpoints.

The lowest primary toxicity data available for both the freshwater and marine Water Quality

Guidelines used conventional endpoints, so, there was no need to decide whether a guideline

could be based on endpoints related to endocrine disruption, and whether these endpoints are

ecologically relevant.

If future research concludes that EDC effects due to NP and short-chain NPEs do result in

ecologically relevant impacts on populations, then there may be a need to re-examine the toxic

equivalency factors that are recommended in this document. Servos et al. (2000) calculated

relative estrogenicity values for various alkylphenolic compounds, relative to NP. These relative

estrogenicity values for NP1EO, NP2EO, NPnEO (n≥9), NP1EC, NP2EC, OP, OP1EC and

OP2EC are 0.67, 0.67, 0, 0.63, 0.63, 4.1, 0.63 and 0.63, respectively. These relative

estrogenicities differ from the TEF values based on conventional toxicity endpoints. For example,

although NPECs have a low relative conventional toxicity, with TEFs of 0.005, they are

considerably more estrogenic, with relative estrogenicity values estimated at 0.63. Octylphenol,

though similar to NP in toxicity, is about four times more estrogenic. Therefore, environmental

samples which do not exceed the guidelines based on conventional toxicity TEFs, could still

presumably cause adverse effects due to estrogenic effects, particularly if they contain significant

quantities of OP, OPECs, or NPECs. In future, this is an issue that will need to be resolved as

more and more information on EDCs is coming to light.
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ANNEX 2. DATASET REQUIREMENTS FOR DERIVING WATER QUALITY GUIDELINES

Water quality guidelines are based on a single, ecologically-relevant, toxicity endpoint for the most

sensitive life stage of the most sensitive aquatic species investigated. To determine the

concentration that represents the most sensitive and protective toxicity endpoint, toxicological

data are evaluated according to a number of screening criteria. The screening criteria (e.g.,

acceptable experimental design, laboratory procedures, documentation of relevant experimental

conditions and parameters) have been described in detail in the protocol (CCME 1991; CCME

1999). Toxicity studies are classified as primary, secondary, or unacceptable. Only primary

studies may be used in the actual derivation of the WQG. Secondary studies, however, may be

used to help meet the minimum toxicological data set requirements.

Studies which were identified as being of primary quality, and therefore could be used in guideline

derivation, met all of the following criteria:

1. flow-through or renewal test conditions were used (rather than static)

2. the study was conducted under controlled laboratory conditions rather than in the field

3. water parameters, such as temperature, pH, and hardness, were measured and

reported

4. sufficient replication was used

5. more than a single concentration of NP was tested

6. concentrations of NP in the water were measured (i.e., did not just report nominal

concentrations)

7. conventional endpoints with documented ecological relevance were used

8. dose-dependent concentration responses were observed

9. experimental conditions and/or analytical methods were described sufficiently to

evaluate the validity of the study (note: data obtained from published abstracts,

citations in other articles/reports, or from articles published in foreign languages were

considered unacceptable)

The following discussion summarizes the minimum data set requirements for development of

freshwater and marine WQGs.

Freshwater

For a full freshwater life guideline, the toxicological data set of primary studies must minimally

include the following:

• at least three studies on three or more freshwater fish species resident in North

America, including at least one cold-water species (e.g., trout) and one warm-water

species (e.g., fathead minnow), and at least two must be chronic (partial or full lifecycle) studies

• at least two chronic (partial or full lifecycle) studies on two or more invertebrate

species from different classes, one of which includes a planktonic species resident in

North America (e.g., daphnid).

• at least one study of a freshwater vascular plant or freshwater algal species resident in

North America (note: for highly phytotoxic variables, four acute and /or chronic studies

on nontarget freshwater plant or algal species are required)

The primary data available for freshwater toxicity due to nonylphenol met all of the minimum

dataset requirements.
105

Marine

The minimum requirements for a full marine guideline are the same as those for freshwater life

except that the studies must include temperate marine fish, invertebrates and plants/algae. Also,

the invertebrate studies must include two species from different classes, but it is not necessary

that one of those species be planktonic.

The marine toxicity dataset for nonylphenol was unable to meet these minimum requirements,

and therefore a full marine guideline could not be derived.

When the minimum data requirements for derivation of a full guideline cannot be met, it may still

be possible to derive an interim guideline. The minimum data set requirements for an interim

marine life guideline are:

• at least two acute and/or chronic studies on two or more marine fish species, one of

which is a temperate species

• at least two acute and/or chronic studies on two or more marine invertebrate species

from different classes, one of which is a temperate species

Secondary studies, as well as primary studies, may be used to meet the minimum requirements

for an interim guideline.

Studies which were considered of secondary quality, and therefore could be used to meet the

minimum data requirements for an interim guideline, met the same criteria as the primary studies,

except for one or more of the following characteristics:

1. test conditions were static (rather than flow-through or renewal)

2. only nominal (rather than measured) concentrations of NP in water were reported

3. ecologically relevant endpoints other than survival, reproduction or growth were

reported

There were sufficient marine toxicity data to meet the minimum requirements for the derivation of

an interim guideline.
106

ANNEX 3. SUMMARY OF SEDIMENT QUALITY GUIDELINE PROTOCOL

There are three approaches to deriving sediment quality guidelines that are recommended in the

CCME protocol: the National Status and Trends Program Approach, the Spiked-Sediment Toxicity

Test Approach, and adoption of guidelines from other jurisdictions (CCME 1995). The data

available for nonylphenol did not support the use of any of these three approaches, and therefore

they were not used in the final derivation of Canadian Sediment Quality Guidelines for

nonylphenol and its ethoxylates. This annex provides a brief summary of the three protocol

approaches and includes a discussion of the nonylphenol sediment toxicity data that would be

needed to support the use of these guideline derivation methods.

The Modified National Status and Trends Program (NSTP) Approach

The NSTP approach to deriving sediment quality assessment values was originally developed by

Long and Morgan (1990). This approach was adopted, with some modifications, for use in

developing sediment quality guidelines in Canada (CCME 1995) and is therefore referred to in this

document as the modified NSTP approach. Modifications to this approach include the separate

evaluation of information for freshwater and marine systems, an expansion of the information

originally compiled, and the use of derivation procedures that consider all of the compiled

information (see also Long and MacDonald 1992; MacDonald 1994; CCME 1995; and MacDonald

et al. 1996 for more detailed explanations of these modifications).

The NSTP approach primarily relies on the use of field-collected data, in which chemical mixtures

occur, to derive sediment quality assessment values (Long & Morgan 1990; Long 1992; Long and

MacDonald 1992; MacDonald 1994; CCME 1995; Long et al. 1995). Data for individual studies

are evaluated to establish an association between the concentration of each of the chemicals

measured in the sediments and any adverse biological effect observed. Cause-and-effect

relationships between the concentration of individual chemicals and the observed adverse

biological effect cannot be inferred from this evaluation.

Other sediment chemical and biological data may also be used, including models of equilibrium

partitioning in sediments, sediment quality assessment values from other jurisdictions (e.g., those

derived using the apparent effects threshold approach and the screening level concentration

approach), spiked-sediment toxicity tests, and field studies (including acute and chronic toxicity

results from sediment bioassays and analyses of benthic community composition). Candidate

studies are critically evaluated according to a number of screening criteria to ensure that high

quality data sets are used. These screening criteria (e.g., appropriate sediment handling

procedures, acceptable toxicity test procedures) have been described in more detail elsewhere

(MacDonald 1994; CCME 1995; Long et al. 1995; Environment Canada 1996).

Acceptable data is sorted by sediment type (i.e., freshwater versus marine) and arranged in

ascending order of the chemical's concentration to produce separate ascending data tables, or

guideline derivation tables, for individual chemicals and sediment types (CCME 1995;

MacDonald 1994; Long et al. 1995). These tables summarize the compiled information that

associates a chemical's concentration with either adverse effects or no adverse effects on aquatic

organisms (also referred to as the effect data set and the no-effect data set, respectively).

Concentrations are expressed as the total concentrations of chemicals in sediments on a dryweight basis. Many of the previous Canadian sediment quality guidelines were derived based on

ascending data tables produced from a large central database called the Biological Effects

Database for Sediments (BEDS). However, the guideline derivation tables for nonylphenol did not

originate from the BEDS database, but instead were independently compiled.

The guideline derivation tables for individual chemicals (e.g., nonylphenol; Tables 13 and 14)

consist of a number of entries, each of which belongs to either the effect data set (concentrations

associated with an adverse biological effect) or the no-effect data set (concentrations not
107

associated with an adverse biological effect), that are sorted according to ascending chemical

concentrations. Entries in the guideline derivation tables were designated as being associated

with an effect (an asterisk in the 'Hit' column) if an adverse biological effect was reported. These

effects included acute or chronic toxicity observed during a controlled spiked-sediment test,

apparent effect thresholds (concentrations above which specific biological effects would always be

expected), and predicted toxicity based on equilibrium partitioning theory (which determines a

sediment chemical concentration that ensures the concentration of the chemical in the interstitial

water does not exceed the water quality guideline for that chemical at equilibrium). An entry was

also assigned an asterisk (effect descriptor) if concordance was apparent between the observed

biological response and the measured chemical concentration in a field (i.e., co-occurrence)

study. Concentrations of individual chemicals reported in field studies were considered to be

associated with the observed toxic response (i.e., concordance was apparent) if the mean

concentration at sites at which significant adverse effects were observed was a factor of two or

more greater than the mean concentration at sites at which effects were not observed (i.e., at

toxic versus non toxic sites) (Long et al. 1995). A factor of two was chosen to ensure that the

difference in the response (i.e., adverse effect) was associated with a significant difference in the

chemical concentration. For each chemical, all of the entries designated by an asterisk, as

described above, are collectively referred to as the effect data set.

All entries in the guideline derivation tables other than those designated by an asterisk are

collectively referred to as the no-effect data set, and are represented by those entries for which

chemical concentrations were not associated with adverse biological effects. These entries

include those associated with non-toxic, reference, or control conditions (i.e., no effects; NE).

Individual entries in the guideline derivation tables consist of the chemical concentration in the

sediment, an indication of whether this concentration was part of the effect data set (i.e., *) or the

no-effect data set (i.e., NE), location of the study, analysis type (or approach used), test duration,

toxicological end-point measured, species and life stage tested, information on the characteristics

of the sediments if available (e.g., concentrations of TOC, physical classification of the

sediments), and the study reference. In most cases, information for individual entries

(i.e., chemical concentrations and sediment chemical and physical characteristics) represents the

means of several samples. Standard deviations of these means are provided wherever possible.

The following discussion summarizes the information contained in the freshwater and marine

guideline derivation tables for nonylphenol (Tables 13 and 14).

Guideline Derivation Tables for Nonylphenol

The freshwater guideline derivation table for nonylphenol (Table 13) contains 25 entries, all of

which are results from spiked-sediment toxicity tests (SSTT). The freshwater sedimentassociated nonylphenol concentrations in the table range from 2.34 to 680 mg·kg -1 dw. A number

of adverse biological effects are associated with nonylphenol concentrations ranging from 34.2 to

680 mg·kg -1 dw.

Chironomids (Chironomus tentans) and frogs (Rana catesbiana) are the only freshwater taxa for

which toxicological information was obtained and included in the freshwater guideline derivation

table. Effects on survival and growth are the most common indicators of adverse biological

effects measured for sediment bioassays with laboratory-spiked sediments. Other chronic

toxicological endpoints tested include lethargy, haemorrhaging, and tail lesions.

Information on total organic carbon (TOC), pH of the sediment and/or overlying water column,

water temperature, conductivity, chemical oxygen demand (COD), biological oxygen demand

(BOD), cation exchange capacity (CEC), and the physical classifications of the bed sediments is

also included for each entry in the freshwater guideline derivation table, when available (Table 13).

The mean TOC content of sediments ranges from 0.05 to 1.27%. The mean proportions of sand,

silt, and clay in the freshwater sediments represented in the ascending data table range from 26.4

to 99.4%, <0.7 to 52%, and <0.7 to 21.6%, respectively.
108

The marine guideline derivation table (Table 14) contains 23 entries, all of which are results from

spiked-sediment toxicity tests (SSTT). Concentrations of sediment-associated nonylphenol

contained in the table range from 14 to 243 mg·kg-1 dw (Table 14). Adverse biological effects

associated with nonylphenol in marine sediments occur at concentrations of 98.7 to 243 mg·kg-1

dw.

Only two taxa, the amphipod Ampelisca abdita and the polychaete Capitella sp., are represented

in the marine guideline derivation table. Most of the observed adverse biological effects were

measurements of various reproductive effects, both at the individual and population level. Size of

the organisms at maturity and survival were other endpoints examined.

Information on total organic carbon (TOC), temperature, and salinity were also included for each

entry in the marine guideline derivation table (Table 14). The mean TOC content of sediments

was 7.4%. The grain size composition of the sediments was not reported in either study.

Derivation of TELs and PELs

As is described in CCME (1995), the derivation procedures used to calculate two assessment

values in the original NSTP approach (Long and Morgan 1990) were modified to consider both the

effect and no-effect data compiled in the guideline derivation tables. In the modified NSTP

approach, the lower value, referred to as the threshold effect level (TEL), represents the

concentration below which adverse biological effects are expected to occur rarely. The upper

value, referred to as the probable effect level (PEL), defines the level above which adverse

effects are expected to occur frequently. Concentrations that fall in the range between the TEL

and the PEL are occasionally expected to be associated with adverse biological effects. The

definition of these ranges (also referred to as the minimal, possible, and probable effect ranges) is

based on the assumption that the potential for observing toxicity of sediment-associated

chemicals increases with increasing chemical concentrations (Long et al. 1995).

Minimum toxicological data requirements have been set to ensure that the TELs calculated from

the guideline derivation tables provide adequate protection of aquatic life and that these values

are supported by a weight of evidence for a broad range of sediment types and characteristics.

For a given guideline derivation table, both the effect data set and the no-effect data set must

contain at least 20 entries (CCME 1995). In the case of nonylphenol, the freshwater guideline

derivation table contained 8 entries in the effect data set and 17 entries in the no-effect data set.

For marine sediments, there were 9 entries in the effect data set and 14 entries in the no-effect

data set. Therefore, both the freshwater and marine datasets contained insufficient data to derive

TELs or PELs for nonylphenol. To derive a freshwater sediment quality guideline using the NSTP

approach, 12 more entries in the effect data set and 3 more entries in the no-effect data set would

be required. For derivation of a marine sediment quality guideline using the NSTP approach, 11

more entries in the effect data set and 6 more entries in the no-effect data set would be required

to meet the minimum data requirements.
109

The Spiked-Sediment Toxicity Test (SSTT) Approach

The SSTT approach, which involves an independent evaluation of information from spikedsediment toxicity tests (that are included in the guideline derivation tables described above), is a

complementary procedure to the modified NSTP approach for estimating the concentration of a

chemical in sediments below which adverse effects are not expected to occur (CCME 1995). In

contrast to the modified NSTP approach, which is used to derive TELs and PELs using

information on associations between chemical concentrations in sediments and effects on

sediment-associated organisms, the SSTT approach is used to derive SSTT values using data

from controlled laboratory tests in which organisms are exposed to sediments that have been

spiked with known concentrations of a chemical. Such studies provide quantifiable cause-andeffect relationships between the concentration of a chemical in sediments and the observed

biological response (e.g., survival, reproductive success, growth). Spiked-sediment toxicity tests

are generally used to evaluate the toxic effect of a single chemical, or a specific mixture of

chemicals, to exposed organisms. They may also be used to determine the extent to which

environmental conditions modify the bioavailability of a chemical, and ultimately the response of

organisms exposed to the spiked-sediments.

Data from spiked-sediment toxicity tests must be evaluated and determined to be acceptable

before it can be used in the SSTT approach (CCME 1995). In acceptable studies, information on

the analytical chemistry of the sediments and overlying water (particularly characteristics that

might influence bioavailability of sediment-associated chemicals) should be reported for both the

beginning and end of the test. Variables that should be measured in the overlying water include

pH, dissolved oxygen, total suspended solids, suspended and dissolved organic carbon, and

water hardness or salinity. In the sediment, reported variables should include total organic

carbon, particle size distribution, acid volatile sulphide, pH, redox conditions, and sediment type.

Generally accepted laboratory practices of exposure and environmental controls should be

employed and tests that follow standardized guides or protocols are preferable. Information on

the health and survival of the test species for at least a week prior to the test, and repeated

evaluation of the condition of the test species throughout the test, should be reported. Where

significant mortalities have occurred prior to the test period, the data should not be used.

Concentrations of the chemical in sediment must be measured – nominal concentrations are not

acceptable. Also, appropriate statistical procedures should be used and reported in detail.

Further discussion of data evaluation in provided in the protocol (CCME 1995).

Minimum toxicological data requirements have been set for the SSTT approach to ensure that

SSTT values derived using spiked-sediment toxicity information provide adequate protection to

aquatic organisms (CCME 1995). Data requirements include at least four independent studies on

two or more sediment-resident invertebrate species that occur in North American waters. For

freshwater sediments, at least one species must be a benthic crustacean and one must be a

benthic arthropod (other than a crustacean). For marine sediments, at least one species must be

a benthic amphipod. SSTT values can be derived from studies conducted on sensitive species

(e.g., fish, aquatic plants, protozoa, fungi, bacteria) provided that the minimum data set

requirements are met. An additional requirement is that at least two of the studies must be partial

or full life cycle tests that consider ecologically-relevant endpoints (e.g., growth, reproductive

success, developmental effects). In addition to these requirements, the procedures used to

generate spiked-sediment toxicity data must be evaluated and determined to be appropriate

before they can be used to develop SSTT values. Although methods (e.g., spiking procedures,

equilibration periods) for spiked-sediment toxicity tests are currently not standardized

(Environment Canada 1995), information provided in individual studies (e.g., equilibrium time,

stability of concentrations over the test duration, responses in control treatments) can be used to

evaluate the influence of the use of specific methods on test results.

Two independent studies have examined the toxicity of nonylphenol to North American freshwater

organisms in spiked-sediment toxicity tests (Appendix IIIa). These studies evaluated nonylphenol

toxicity (lethality) to the midge, Chironomus tentans (England and Bussard 1993), and to the
110

amphibian, Rana catesbiana (Ward and Boeri 1992). England and Bussard (1993) also assessed

sub-lethal endpoints (i.e., growth and reproduction) for Chironomus tentans, and Ward and Boeri

(1992) examined sub-lethal endpoints for Rana catesbiana such as growth, lethargy,

haemorrhaging, and occurrence of tail lesions. With only two independent studies, insufficient

data are available to derive an SSTT value for freshwater sediments. At least three additional

independent studies on North American freshwater sediment-resident invertebrates, at least one

of which is a benthic amphipod, are required. Also, at least one of the studies must be a partial or

full life-cycle test that reports ecologically relevant sub-lethal endpoints (e.g., growth, reproduction,

developmental effects) in order to fulfill the minimum data requirements for the derivation of a

freshwater SSTT value.

Two independent studies examined the toxicity of nonylphenol to a marine organism in spikedsediment toxicity tests (Appendix IIIb). One study evaluated nonylphenol toxicity (lethality) to the

amphipod Ampelisca abdita (Fay et al. 2000). The other study evaluated sub-lethal effects of

nonylphenol on the polychaete Capitella sp. (Hansen et al. 1999). Sub-lethal endpoints examined

included time to first reproduction, time between reproductive events, number of broods per

reproductive individual, number of eggs per individual and per reproductive individual, population

growth rate, and size at maturity. These data, however, are insufficient to derive an SSTT value

for marine sediments. At least two additional independent studies on North American marine

sediment-resident invertebrates are required. Also, at least one of the studies must be a partial or

full life-cycle test that reports ecologically relevant sub-lethal endpoints (e.g., growth, reproduction,

developmental effects) in order to fulfill the minimum data requirements for the derivation of a

marine SSTT value.

Adoption of Guidelines from Other Jurisdictions

When insufficient information is available to derive ISQGs using either the modified NSTP or

SSTT approaches, sediment quality assessment values from other jurisdictions are evaluated

and, if appropriate, adopted provisionally as ISQGs (CCME 1995). This adoption process gives

preference to biological-effects based sediment quality assessment values that have been derived

using methods that are philosophically consistent with the protocol’s guiding principles.

Guidelines that have been derived using effects-based approaches are considered to be the most

ecologically relevant and scientifically defensible. Using this approach, the lowest of the

guidelines that incorporate data on the effects of sediment-associated substances on sedimentdwelling organisms is selected. For organic contaminants, such as nonylphenol, guidelines

derived using the equilibrium partitioning approach can also be considered for adoption.

No sediment quality guidelines for NP or NPEs, either for freshwater or marine sediments,

currently exist in any other jurisdiction. Therefore, the approach of adopting sediment guidelines

from another jurisdiction cannot be used to develop Canadian sediment quality guidelines.
111
112

Table 13. Summary of the available biological effects and related physicochemical data for

NP

(mg·kg-1)

Hit Analysis

Type

Test

Type

Endpoint Measured Species Life

Stage

TOC

(%)

pH

sediment

2.34 NE SSTT 14d not toxic (7% mortality) Chironomus tentans (freshwater midge) larvae 1.27 5.5

2.34 NE SSTT 14d not toxic (13% growth increase) Chironomus tentans (freshwater midge) larvae 1.27 5.5

4.79 NE SSTT 14d not toxic (11% mortality) Chironomus tentans (freshwater midge) larvae 1.27 5.5

4.79 NE SSTT 14d not toxic (10% growth increase) Chironomus tentans (freshwater midge) larvae 1.27 5.5

9.51 NE SSTT 14d not toxic (8% mortality) Chironomus tentans (freshwater midge) larvae 1.27 5.5

9.51 NE SSTT 14d not toxic (13% growth increase) Chironomus tentans (freshwater midge) larvae 1.27 5.5

20.1 NE SSTT 14d not toxic (10% mortality) Chironomus tentans (freshwater midge) larvae 1.27 5.5

20.1 NE SSTT 14d not toxic (6% growth reduction) Chironomus tentans (freshwater midge) larvae 1.27 5.5

34.2 * SSTT 14d toxic (31% mortality) Chironomus tentans (freshwater midge) larvae 1.27 5.5

34.2 * SSTT 14d toxic (27% growth reduction) Chironomus tentans (freshwater midge) larvae 1.27 5.5

36 NE SSTT 30d not toxic (0% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

36 NE SSTT 30d not toxic (5% sublethal effects) Rana catesbiana (bullfrog) tadpole 0.05 NR

36 NE SSTT 30d not toxic (6% growth reduction) Rana catesbiana (bullfrog) tadpole 0.05 NR

57 NE SSTT 30d not toxic (0% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

57 NE SSTT 30d not toxic (0% sublethal effects) Rana catesbiana (bullfrog) tadpole 0.05 NR

57 NE SSTT 30d not toxic (7.5% growth reduction) Rana catesbiana (bullfrog) tadpole 0.05 NR

155 NE SSTT 30d not toxic (10% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

155 NE SSTT 30d not toxic (10% sublethal effects) Rana catesbiana (bullfrog) tadpole 0.05 NR

155 NE SSTT 30d not toxic (25% growth reduction) Rana catesbiana (bullfrog) tadpole 0.05 NR

220 * SSTT 30d toxic (sublethal effects and growth) Rana catesbiana (bullfrog) tadpole 0.05 NR

260 * SSTT 30d toxic (50% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

390 * SSTT 30d toxic (85% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

390 * SSTT 30d toxic (100% sublethal effects -

lethargy, haemorrhaging, tail

lesions)

Rana catesbiana (bullfrog) tadpole 0.05 NR

390 * SSTT 30d toxic (29% growth reduction) Rana catesbiana (bullfrog) tadpole 0.05 NR

680 * SSTT 30d toxic (100% mortality) Rana catesbiana (bullfrog) tadpole 0.05 NR

TOC = total organic carbon

Temp. = temperature

Cond. = conductivity measured in μMhos·cm-1

COD = chemical oxygen demand

BOD = biological oxygen demand

CEC = cation exchange capacity

NE = no effect

* = effect

d = days

NR = not reported
113

sediment-associated nonylphenol (mg·kg-1 dry weight) in freshwater ecosystems.

pH

water

Temp.

(°C)

Cond. COD

(mg·L-1)

BOD

(mg·L-1)

CEC

(meq·L-1)

Sand

(%)

Silt

(%)

Clay

(%)

Area Reference

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

7.2-8.5 19-21 320 352.5 9.2 23.9 26.4 52 21.6 Laboratory England and Bussard 1993

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992

6.9-8.3 17.1-18.8 740 620 μg/g 45 μg/g 2.3 meq/100g 99.4 <0.7 <0.7 Laboratory Ward and Boeri 1992
114

Table 14. Summary of the available biological effects and related physicochemical data for

NP

(mg·kg-1)

Hit Analysis

Type

Test

Type

Endpoint Measured Species Life

Stage

TOC

(%)

pH

sediment

14 NE SSTT 78d not toxic (1% decrease in time to

first reproduction)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (7% increase in time

between reproductive events)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (2% decrease in number

of broods per reproductive

individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (12% increase in number

of eggs per reproductive

individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (25% decrease in number

of eggs per individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (11% increase in size at

maturity)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

14 NE SSTT 78d not toxic (6% increase in

population growth rate)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (0% change in time to

first reproduction)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (3% increase in time

between reproductive events)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (2% increase in number

of broods per reproductive

individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (3% decrease in number

of eggs per reproductive

individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (3% decrease in number

of eggs per individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (16% decrease in size at

maturity)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

52 NE SSTT 78d not toxic (0% change in population

growth rate)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

98.7 * SSTT 10d toxic (50% mortality) Ampelisca abdita (amphipod) juvenile NR NR

174 * SSTT 78d toxic (17% increase in time to first

reproduction)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (46% increase in time

between reproductive events)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (44% decrease in number of

broods per reproductive individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (78% decrease in number of

eggs per reproductive individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (76% decrease in number of

eggs per individual)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (33% decrease in size at

maturity)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

174 * SSTT 78d toxic (24% decrease in population

growth rate)

Capitella sp. I (polychaete) larvae to

adult

7.4 NR

243 * SSTT 10d toxic (100% mortality) Ampelisca abdita (amphipod) juvenile NR NR

TOC = total organic carbon

Temp. = temperature

NE = no effect

* = effect

d = days

NR = not reported
115

sediment-associated nonylphenol (mg·kg-1 dry weight) in marine ecosystems.

Temp.

(°C)

Water Salinity

(g·L-1)

Sand

(%)

Silt

(%)

Clay

(%)

Area Reference

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 29-32 NR NR NR Laboratory Fay et al. 2000

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 26-27 NR NR NR Laboratory Hansen et al. 1999

20 29-32 NR NR NR Laboratory Fay et al. 2000
116

APPENDIX I

Toxicity of nonylphenol and its ethoxylates to freshwater life

through water column exposure
APPENDIX 1a: Toxicity of nonylphenol to freshwater algae and macrophytes through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hard.

ness**

Temp.

(°C)

Evaluation

Rank

Reference

Ceratophyllum

demersum

(coontail)

NR 21d S, N bleaching EC100 6250 6.8 NR NR 23 U Weinberger and Greenhalgh

1984

Chlamydomonas

reinhardii

NR 1h S, N membrane

disruption

NR 10 NR NR NR 22 U Weinberger & Rea 1981

(flagellate algae) NR 1h S, NR inhibition of

photosynthesis

ICF100 500-750 6.7 NR NR 23 U Moody et al. 1983

NR 21d S, N growth

(biomass)

LOEC 1000 NR NR NR 23 U Weinberger & Greenhalgh 1984

NR 21d S, N algistasis LOEC 6250-1000 NR NR NR 23 U Weinberger & Greenhalgh 1984

9d old

culture

24h S, N ATP reduction EC75

EC20

6250

2500

NR NR NR 23 U Weinberger & Greenhalgh 1984

Chlamydomonas segnis

(algae)

4 life

cycle

phases

24h S, N algistatic or

algicidal

NR 2500 6.8 NR NR 23 2 Weinberger et al. 1987

NR 24h NR,NR mortality LC50 1500 NR NR NR NR U Weinberger 19841

NR 24h NR,NR inhibition of

photosynthesis

NR 500-750 NR NR NR NR U Weinberger 19841

Chlorella pyrenoidosa

(algae)

NR 24h S, N mortality LC50

LC100

1500

25000

6.8 NR NR 22 U Weinberger & Rea 1981, 1982

Lemna minor

(duckweed)

2-4

fronds

4d R, N growth (dry

matter

production)

LOEC 125 NR NR NR 20 U Prasad 1989

2-4

fronds

6d S, N growth (frond

production)

LOEC 500 NR NR NR 20 U Prasad 1989

2-4

fronds

4d S, N growth (dry

matter

production)

LOEC 500 NR NR NR 20 U Prasad 1989

NR 7d S, N growth (new

frond

production)

LOEC 1250 5.6 NR NR 25 U Weinberger and Iyengar 1983

NR 7d S, N growth (fresh

and dry mass)

LOEC >1250

>2500

5.6 NR NR 25 U Weinberger and Iyengar 1983

NR 7d S, N chlorophyll

content

LOEC 1600 5.6 NR NR 25 U Weinberger and Iyengar 1983

2-frond

stage

96h F, M frond production NOEC

LOEC

MATC

901

2080

1369

8.0-8.7 6.5-8.5 108-210 22.8-

24.2

1 Brooke 1993a

NR 7d S, N photosynthesis

(CO2 fixation)

LOEC 2500 5.6 NR NR 25 U Weinberger and Iyengar 1983
APPENDIX 1a (continued): Toxicity of nonylphenol to freshwater algae and macrophytes through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Lemna minor

(duckweed)

NR 24h S, N ATP content

measurement

LOEC 2500 5.6 NR NR 25 U Weinberger and Iyengar 1983

NR 24h NR, NR total ATP LC50 2500 NR NR NR NR U Weinberger 19841

NR 96h S, N growth inhibition EC50 5500 5.6 NR NR 25 U Weinberger and Iyengar 1983

NR 21d S, N bleaching EC100 6250 6.8 NR NR 23 U Weinberger and Greenhalgh

1984

Potamogeton sp. and

Chara sp. primarily

(macrophytes)

NR 21d R, M survival NOEC

LOEC

MATC

243

>243

>243

7.7 7.3 122 19.5 1 Liber et al. 1999b

Salvinia molesta

(aquatic fern)

2-4

fronds

9d S, N growth (frond

production)

LOEC 2500 NR NR NR 20 U Prasad 1989

Scenedesmus

subspicatus

(green algae)

NR 72h NR,NR growth EC50 1300 NR NR NR NR U Hüls 19962

Selenastrum

capricornutum

(green algae)

4-7d 96h S, M growth

(cell counts)

EC50

NOEC

LOEC

MATC

410

92

190

132

7.4-8.9 NR NR 23.2-

23.7

1 Ward & Boeri 1990a

NR 96h S, M cell production NOEC

LOEC

MATC

694

1480

1013

7.3-7.7 NR 16 25.2-

26.0

1 Brooke 1993a

NR = not reported

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

ICF100 = concentration causing 100% inhibition of fluorescence response

1

 As cited in Servos et al. 2000

2

 As cited in OECD 1997 (as cited in Servos et al. 2000)
APPENDIX Ib. Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Crustacea

Acanthocyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Acroperus sp.

(cladoceran)

NR 92d R, M abundance NOEC

LOEC

23

76

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Alona sp

(Cladocera)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Bosmina sp.

(cladoceran)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Calanoida

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

MATC

5

23

11

7.8 8.97 118 17.3 U O'Halloran et al. 1999;

Liber et al. 1999b

Ceriodaphnia dubia

(water flea)

NEO 7d R, N survival NOEC

LOEC

MATC

3

10

5.5

8.4-8.8 6.9-8.0

(85-101%

saturation)

134 -

166

24-25 2 England 1995

NEO 7d R, N total adverse

effects of

mortality and

delayed

maturity

EC50 27 8.4-8.8 6.9-8.0

(85-101%

saturation)

134 -

166

24-25 1 England 1995

NR 96h F, NR NR EC50 69 NR NR NR NR U Weeks et al. 1996

NR 7d NR,NR NR EC50 99 NR NR NR NR U Weeks et al. 1996

NEO 7d R, N reproduction

(time to first

brood, #

young/ adult

reprod. days

NOEC

LOEC

MATC

100

300

170

8.4-8.8 6.9-8.0

(85-101%

saturation)

134 -

166

24-25 1 England 1995

NR 7d F, NR reproduction MATC 134 NR NR NR NR U Weeks et al. 1996

NR 7d NR,NR mortality LC50 258 NR NR NR NR U Weeks et al. 1996

NR 96h NR,NR mortality LC50 276 NR NR NR NR U Weeks et al. 1996

NR 7d NR,NR NR MATC 276 NR NR NR NR U Weeks et al. 1996)

NEO 7d R, N mortality LC50 >300 8.4-8.8 6.9-8.0

(85-101%

saturation)

134 -

166

24-25 2 England 1995
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Ceriodaphnia dubia

(water flea)

NEO 96h R, N mortality, total

adverse

effects

LC50

EC50

both >300 8.4-8.8 6.9-8.0

(85-101%

saturation)

134 -

166

24-25 2 England 1995

NR 48h S, N mortality LC50 470 (mean of

340, 710,

430, 410)

8 NR NR 25 U Ankley et al. 1990

Ceriodaphnia sp.

(water flea)

NR 92d R, M abundance NOEC

LOEC

23

76

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Chydorus sp.

(Cladocera)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Cladocera

(taxa combined)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Copepoda

(taxa combined)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Daphnia galeata mendotae

(cladoceran zooplankton)

NEO 30d R, N deformity LOEC 10 NR NR NR 20 2 Shurin & Dodson

1997

NEO 30d R, N sex

determination

(more female

offspring)

LOEC 50 NR NR NR 20 U Shurin & Dodson

1997

Daphnia magna

(water flea)

NEO 21d R, M reproduction NOEC 24 8.25 NR 180 20 1 Comber et al. 1993

NR chronic NR, NR reproduction

rate

NOEC

LOEC

MATC

24

39

31

NR NR NR NR U Naylor 1995

NEO 21d R, M length of

parent

NOEC 39 8.25 NR 180 20 1 Comber et al. 1993

NEO 21d R, N fecundity MATC 71 8 NR NR 22 2 Baldwin et al. 1997

NR 48h NR, NR acute toxicity NOEC

LOEC

MATC

77

160

110

NR NR NR NR U Naylor 1995

NEO 48h R, M mortality EC50 84.8 7.66-8.04 6.3-8.2 175.6-

195.1

21-23 1 Brooke 1993a

10d adult

females

48h S, N inc. androgen

(from inhibition

of testost'one

metab. elim.)

LOEC 100 8 NR NR 22 U Baldwin et al. 1997

NR 21d NR,NR reproduction NOEC 100 NR NR NR NR U Hüls 19922
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Daphnia magna

(water flea)

NEO 21d R, M reproduction NOEC

LOEC

MATC

116

215

157.9

8.14-8.65 7.5-9.6 156-

188

19.7-21.8 1 Brooke 1993a

NEO 21d R, M survival of

parent

(mortality)

LC50 100 8.25 NR 180 20 1 Comber et al. 1993

9-10d

adult

females

96h R, N immobilization

or mortality

NOEC 0.617 μM

(136 μg·L-1)

7.8-8.4 >60%

saturation

227 20 2 Gerritsen et al. 1998

NR 48h NR,NR mortality LC50 140 NR NR NR NR U Hüls 19922

NR 48h S, N immobilization EC50 180 NR NR NR NR U Bringmann and Kühn

1982

NEO 48h NR, NR mortality LC50 190 NR NR NR 20 or 25 U Naylor 1995

NEO 48h S, M immobilization EC50 190 8.25 NR 180 20 1 Comber et al. 1993

NEO 96h R, N immobilization

or mortality

NOEC 1.37 μM

(302 μg·L-1)

7.8-8.4 >60%

saturation

227 20 2 Gerritsen et al. 1998

NR 48h NR,NR mortality LC50 440 NR NR NR NR U Monsanto 1985b1

NEO 48h NR, NR reproduction EC50 440 NR NR NR 20 or 25 U Naylor 1995

NEO 48h S, N immobilization EC50

NOEC

LOEC

440

250

500

7.5-8.7 6.7-8.4 264-

320

22.1 U Calvert & Adams

1981

Daphnia pulex

(water flea)

JUV 48h S, N mortality LC50 140, 176,

190

NR NR 27.1 23-27 U Ernst et al. 1980

Diacyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Eucyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Harpacticoida

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Hyalella azteca

(amphipod)

JUV (2

mm)

96h F, M mortality LC50 20.7 7.69-8.01 7.6-8.8 46-60 23.7-25.6 1 Brooke 1993a

JUV (2

mm)

96h F, M immobilization EC50 20.7 7.69-8.01 7.6-8.8 46-60 23.7-25.6 1 Brooke 1993a

NR 96h F, NR NR EC50

MATC

150

123

NR NR NR NR U Weeks et al. 1996

JUV 96h F, M lethargy and

immobility

EC50 150 7.9 - 8.7 1.4 - 8.0 152 -

158

21 1 England & Bussard

1995

JUV 96h F, M mortality LC50 170 7.9 - 8.7 1.4 - 8.0 152 -

158

21 1 England & Bussard

1995
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Kurzia sp.

(cladoceran)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Macrocyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Mesocyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Ostracoda

(ostracod)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Paracyclops sp.

(copepod)

nauplii to

adult

92d R, M abundance NOEC

LOEC

5

23

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Pleuroxus sp.

(Cladocera)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Simocephalus sp.

(cladoceran)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al. 1999

Insecta

Chironomini

(chironomid)

NR 21d R, M survival NOEC

LOEC

MATC

243

>243

>243

7.7 7.3 122 19.5 1 Liber et al. 1999b;

Schmude et al. 1999

Chironomus tentans

(freshwater midge)

LAR (24h

to 4th

instar)

20d R, M survival NOEC

LOEC

42

91

7.73 7.04 NR 23 1 Kahl et al. 1997

LAR-ADT 53d R, M emergence,

reproduction,

sex ratio

NOEC 45 7.73 7.04 NR 23 2 Kahl et al. 1997

LAR (2nd

instar)

14d F, M total adverse

eff. (lethargy,

small size,

pale)

EC50 95 7.6-8.6 0.5-7.5 130-

154

19-21 1 England and Bussard

1993

LAR (2nd

instar)

14d F, M larval weight

(growth)

EC50

LOEC

NOEC

MATC

95

150

76

107

7.6-8.6 0.5-7.5 130-

154

19-21 1 England and Bussard

1993

LAR (2nd

instar)

14d F, M mortality

(survival)

LC50

LOEC

NOEC

MATC

119

150

76

107

7.6-8.6 0.5-7.5 130-

154

19-21 1 England & Bussard

1993

LAR 96h NR,NR mortality LC50

MATC

119

107

NR NR NR NR U Weeks et al. 1996,

Naylor 1995

NR 96h F, NR NR EC50

MATC

160

97

NR NR NR NR U Weeks et al. 1996
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Ophiogomphus sp.

(dragonfly)

NYM

(285 mg)

96h F, M loss of

equilibrium,

posterior of

abdomen

flexed over

back

EC50 596 7.82-8.52 7.7-8.9 50.7-

54.6

21.5-25.0 1 Brooke 1993a

NYM

(285 mg)

96h F, M mortality LC50 >768 7.82-8.52 7.7-8.9 50.7-

54.6

21.5-25.0 1 Brooke 1993a

Tanytarsini

(chironomid)

LAR 21d R, M survival NOEC

LOEC

76

243

7.7 7.3 122 19.5 U Schmude et al. 1999

Mollusca

Anodonta cataractae

(freshwater clam)

15g 144h R, M mortality LC50 1700 NR NR NR 10 U McLeese et al. 1980a

Elliptio complanata

(mussel)

excised

gills

50 min S, N metabolic heat

rate inhibition

LOEC 70000 NR NR NR 30 U Levine and Cheney

2000

excised

gills

50 min S, N respiratory

inhibition

LOEC 220300 NR NR NR 25 U Levine and Cheney

2000

Gastropoda

(snails, mostly

Planorbidae)

NR 21d R, M survival NOEC

LOEC

76

243

7.7 7.3 122 19.5 U Schmude et al. 1999

Physella virgata

(snail)

ADT (476

mg)

96h F, M mortality LC50 774 7.73-8.02 7.8-8.2 50-60 22.8-24.5 1 Brooke 1993a

ADT (476

mg)

96h F, M unable to

retract into

shell with

prodding

EC50 378 7.73-8.02 7.8-8.2 50-60 22.8-24.5 1 Brooke 1993a

Pisidium sp.

(bivalve)

NR 21d R, M survival NOEC

LOEC

MATC

23

76

42

7.7 7.3 122 19.5 U Schmude et al. 1999;

Liber et al. 1999b

Oligochaeta

Naididae

(oligochaete)

NR 21d R, M survival NOEC

LOEC

23

76

7.7 7.3 122 19.5 U Schmude et al. 1999

Tubificidae

(oligochaete)

NR 21d R, M survival NOEC

LOEC

243

>243

7.7 7.3 122 19.5 U Schmude et al. 1999

Annelida

Lumbriculus variegatus

(annelid)

ADT (5

mg)

96h F, M mortality LC50 342 6.63-6.9 7.2-8.5 50.7-

54.6

22.2-25.9 1 Brooke 1993a

ADT (5

mg)

96h F, M inactivity EC50 268 6.63-6.9 7.2-8.5 50.7-

54.6

22.2-25.9 1 Brooke 1993a
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Rotifera

Rotifera

(taxa combined)

NR 92d R, M abundance NOEC

LOEC

MATC

243

>243

>243

7.8 8.97 118 17.3 U O'Halloran et al. 1999;

Liber et al. 1999b

Brachionus sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Colurella sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Conochilus sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Euchlanis sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Keratella sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Lecane sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Lepadella sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Monostyla sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Mytilina sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Notholca sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Platyias sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Pleosoma sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Polyarthra sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Testudinella sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Trichocerca sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

23

76

7.8 8.97 118 17.3 U O'Halloran et al.

1999

Trichotria sp.

(rotifer)

NR 92d R, M abundance NOEC

LOEC

76

243

7.8 8.97 118 17.3 U O'Halloran et al.

1999
APPENDIX Ib (continued). Toxicity of nonylphenol to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Miscellaneous

Macroinvertebrates

(taxa combined)

NR 21d R, M survival NOEC

LOEC

MATC

76

243

136

7.7 7.3 122 19.5 U Liber et al. 1999b

Periphyton

(taxa combined)

NR 92d R, M survival NOEC

LOEC

MATC

243

>243

>243

7.8 8.97 118 17.3 U O'Halloran et al.

1999; Liber et al.

1999b

Tetrahymena pyriformis

(freshwater protozoan)

NR 24h S, N relative growth

rate

EC50 460 NR NR NR 30 U Yoshioka et al. 1985

Zooplankton

(taxa combined)

NR 92d R, M abundance NOEC

LOEC

MATC

76

243

136

7.8 8.97 118 17.3 U O'Halloran et al. 1999;

Liber et al. 1999b

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, PUP = pupae, LAR = larvae, NYM = nymph, NEO = neonates

* duration reported in minutes (min), hours (h), or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Etnier 1985 as cited in Talmage 1994

2

 As cited in Servos et al. 2000
APPENDIX Ic. Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Culaea inconstans

(5-7-spined

stickleback)

NR NR 21d R, M survival NOEC

LOEC

MATC

243

>243

>243

7.7 7.3 122 19.5 1 Liber et al. 1999b

Gasterosteus

aculeatus

(stickleback)

NR NR 96h NR,NR mortality LC50 370 NR NR NR NR U Granmo et al. 1991a

Gila elegans

(bonytail chub)

0.29-

0.52 g

NR 96h S, N mortality LC50 290 8.35 low

values

173 22 2 Dwyer et al. 1995

Ictalurus punctatus

(channel catfish)

JUV (65-

95g)

NR 7d IPS, N vitellogenesis LOEC 237

mg·kg-1 (1)

NR NR NR NR U Nimrod & Benson

1996

hepatocytes

from

adult fish

400-

500g

male 4-6d R, N vitellogenin

induction

LOEC

NOEC

10 μM

(2203

μg·L-1)

1 μM (220

μg·L-1)

NR NR NR NR 2 Monteverdi and

Di Giulio 1999

hepatocytes

from 2 yr

old fish

(300-

400g)

female 5h S, NR inhibition of

estradiol

binding to

estrogen

receptor site

IC50 4531 nM

(998

μg·L-1)

NR NR NR room

temp

U Nimrod and Benson

1997

Lebistes reticulatus

(guppy)

NR NR 24h F, NR NR NR NR NR NR NR NR U Swedmark 1968

Lepomis macrochirus

(bluegill sunfish)

11-13

wks old

NR 28d F, M mortality NOEC

LOEC

59.5

126

6.80-

7.78

4.4-7.4 46.8-

51.2

23.3-

26.0

1 Brooke 1993b

JUV

(9 cm,

12 g)

NR 21d R, M survival NOEC

LOEC

MATC

76

243

136

7.7 7.3 122 19.5 1 Liber et al. 1999a

13-15

wks old

NR 96h F, M mortality NOEC

LOEC

86.5

211

7.51-

7.99

5.2-8.0 49.4-

50.8

25.3-

26.6

1 Brooke 1993b

JUV (<1

year old),

23 mm,

294 mg

NR 96h F, M loss of

equilibrium,

loss of

schooling

ability

EC50 203 7.26-

7.79

7.5-8.2 50.7-

54.6

21.5-

23.8

1 Brooke 1993a
APPENDIX Ic (continued). Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Lepomis macrochirus

(bluegill sunfish)

JUV (<1

yr old), 23

mm, 294

mg

NR 96h F, M mortality LC50 209 7.26-

7.79

7.5-8.2 50.7-

54.6

21.5-

23.8

1 Brooke 1993a

Leuciscus idus

(golden orfe)

NR NR 48h NR,NR mortality LC50 950 NR NR NR NR U Hüls 1987

Oncorhynchus apache

(Apache trout)

0.38-0.85

g

NR 96h S, N mortality LC50 170 8.24 low

values

169 12 2 Dwyer et al. 1995

Oncorhynchus clarki

henshawi

(Lahontan cutthroat

trout)

0.31 g NR 96h S, N mortality LC50 180, 170 8.24 low

values

169 12 2 Dwyer et al. 1995

Oncorhynchus clarki

stomias

(greenback cutthroat

trout)

0.34-0.57

g

NR 96h S, N mortality LC50 150, 156 8.24 low

values

169 12 2 Dwyer et al. 1995

Oncorhynchus mykiss

(rainbow trout)

JUV female 22d,35d F, N growth

inhibition

LOEC 1 6.5 NR 12.5 7-13 U Ashfield et al. 1998

eggs male 12 mo. NR,NR vitellogenin

induction

LOEC 1 NR NR NR NR U Fent et al. 1999

eggs male 12 mo. NR,NR Vg mRNA

induction

LOEC 1 NR NR NR NR U Fent et al. 1999

EMB NR 91d F, M growth NOEC

LOEC

MATC

6

10.3

7.861

6.34-

7.84

7.7-

11.7

39.0-

66.3

8.9-14.4 1 Brooke 1993a

ADT

and

JUV

male

and

female

72h F, N vitellogenin

induction

LOEC 10 NR NR NR 10-15 U Lech et al. 1996

ADT

and

JUV

male

and

female

72h F, N

(calc)

estrogenic

(vitellogenin

induction)

EC50 14.14 NR NR NR 10-15 U Lech et al. 1996

2 yr male 21d F, M elevation of

plasma

vitellogenin

LOEC 20.3 NR NR NR NR 2 Jobling et al. 1996

hepatocytesmale 24h S, N Vg mRNA

induction; rtER

mRNA

induction

NOEC

LOEC

10-7 M

(22 μg·L-1)

10-6 M (220

μg·L-1)

NR NR NR NR U Flouriot et al. 1995
APPENDIX Ic (continued). Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oncorhynchus mykiss

(rainbow trout)

JUV

(sexually

immature

35-50g)

NR 21d R, N vitellogenin

induction

NOEL

LOEL

25

50

8.0-8.4 >90%

saturation

NR 13 2 Tremblay and Van

Der Kraak 1998

2 yr male 21d F, M 500 fold

increase in

vitellogenin

NR 37 NR NR NR NR U Jobling et al. 1996

JUV (46-

174g)

NR 9d IPS, N vitellogenin

induction

NOEC 50 mg·kg-1 NR NR NR NR U Andersen et al. 1999

JUV

(103-

168g)

NR 12d IPR,

NR

vitellogenin

induction

LOEC (only

conc tested)

50 mg·kg-1 NR NR NR NR U Pedersen et al. 1999

JUV

(103-

168g)

NR 12d IPR,

NR

vitellogenin

induction

NOEC (only

conc tested)

50 mg·kg-1 NR NR NR NR U Pedersen et al. 1999

JUV

(100-

200g)

NR NR F, N induction of

vitellogenin

mRNA

significant

effect

100 NR NR NR 10-15 U Ren et al. 1996

45d, 27.2

mm, 241

mg

NR 96h F, M loss of

equilibrium,

poor fright

response

EC50 109 6.53-

6.98

9.3-10.4 54.6-

58.5

11.3-

12.6

1 Brooke 1993a

JUV

(103-

168g)

NR 9d F, M vitellogenin

induction

LOEC 75 NR NR NR NR U Pedersen et al. 1999

JUV

(103-

168g)

NR 9d F, M vitellogenin

induction

NOEC 109 NR NR NR NR U Pedersen et al. 1999

0.27-

1.25 g

NR 96h S, N mortality LC50 190, 250 8.24 low

values

169 12 2 Dwyer et al. 1995

ADT

and

JUV

 (50-

200g)

male

and

female

72h F, N mortality LC50 193.65 NR NR NR 10-15 U Lech et al. 1996

45d, 27.2

mm, 241

mg

NR 96h F, M mortality LC50 221 6.53-

6.98

9.3-10.4 54.6-

58.5

11.3-

12.6

1 Brooke 1993a

JUV NR 96h NR,NR mortality LC50 230 NR NR NR NR U Armstrong and

Kingsbury 1979(2)
APPENDIX Ic (continued). Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oncorhynchus mykiss

(rainbow trout)

42-

60mm,

0.5-2.7g

NR 96h S, N mortality LC50 230 NR NR NR 9 U Holmes and

Kingsbury 1980

15-30d NR 96h NR,NR mortality LC50 230 NR >6 NR 12 U Naylor 1995

EMB NR acute24h exp

+ 48h

post-exp

S, M mortality LC50 484 NR NR 27.1 9.5-28.5 U Ernst et al. 1980

JUV

(1.4g,

5.2 cm)

NR 96h S, N mortality LC50 920, 560 6.2-8.3 4.6-10.6 27.1 15-16 U Ernst et al. 1980

hepatocytes

from

JUV

fish

males 48h S, N vitellogenin

production

ED50 16.15 μM

(3558

μg·L-1)

NR NR NR NR U Jobling and Sumpter

1993

Oryzias latipes

(Japanese medaka)

5-8d old,

larval

swim-up

stage

NR 28d

(& 1 mo.

in clean

water)

F, M sex ratio,

fecundity, egg

viability or

hatchability

NOEC 1.93 NR NR NR 25 2 Nimrod and Benson

1998

hatch to

3 mths

NR 3 mo. R, N induction of

testis-ova

NOEC

LOEC

10

50

7.4-7.8 NR 80-100 20 U Gray & Metcalfe

1997

hatch to

3 mths

NR 3 mo. R, N sex ratio NOEC

LOEC

50

100

7.4-7.8 NR 80-100 20 U Gray & Metcalfe

1997

EMB NR 17d S, N embryotoxicity LC50 460 NR NR NR 25 U Gray and Metcalfe

1997

2 cm,

0.2 g

NR 48h S, NR mortality LC50 1400 7.2 NR 50 16.2 U Yoshimura 1986

Pimephales promelas

(fathead minnow)

ADT NR 42d F, M histologic

lesions

LOEC

NOEC

1.6 - 3.4

0.4 - 0.93

NR NR NR 25-26 2 Miles-Richardson et

al. 1999

EMB NR 33d F, M survival NOEL

LOEL

MATC

7.4

14

10.2

7.1-8.2 6.3-9.7 160-180 23.4-

26.3

1 Ward and Boeri

1991b

early life

stage

NR 30d F, NR NR MATC 10.2 NR NR NR NR U Weeks et al. 1996

EMB NR 33d F, M # embryos

hatched, time

to hatch, time

to first feeding,

total length,

weight

NOEC

LOEC

MATC

for all

endpoints:

23

>23

>23

7.1-8.2 6.3-9.7 160-180 23.4-

26.3

1 Ward and Boeri

1991b
APPENDIX Ic (continued). Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1

)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Pimephales promelas

(fathead minnow)

4 wks old NR 28d F, M mortality NOEC

LOEC

77.5

193

6.91-

7.92

4.4-7.7 45.8-

52.2

23.2-

26.3

1 Brooke 1993b

4 wks old NR 96h F, M mortality NOEC

LOEC

83.1

230

7.50-

7.84

5.0-6.7 46.2-

48.4

23.0-

25.6

1 Brooke 1993b

25-35d

old,

81.6g,

17.1 mm

NR 96h F, M loss of

equilibrium,

poor schooling

and fright

responses

EC50 96 6.77-

7.58

6.6-8.2 53.6-

63.4

21.3-

23.1

1 Brooke 1993a

31-35d,

220 mg

NR 96h F, M loss of

equilibrium

LOEC 98 6.9-7.7 7.4 44.9 24.6 2 Holcombe et al. 1984

25-35d

old,

81.6g,

17.1 mm

NR 96h F, M mortality LC50 128 6.77-

7.58

6.6-8.2 53.6-

63.4

21.3-

23.1

1 Brooke 1993a

31-35d,

220 mg

NR 96h F, M mortality LC50 135 6.9-7.7 7.4 44.9 24.6 1 Holcombe et al. 1984

32d NR 96h F, M mortality LC50 140 7.29 7.1 45.1 24.5 1 Geiger et al. 1985

31-35d NR 96h F, M haemorrhagingLOEC 187 6.9-7.7 7.4 44.9 24.6 2 Holcombe et al. 1984

31-35d NR 96h F, M bodies

swollen with

fluid

LOEC 187 6.9-7.7 7.4 44.9 24.6 2 Holcombe et al. 1984

31-35d,

220 mg

NR 96h F, M lethargic LOEC 187 6.9-7.7 7.4 44.9 24.6 2 Holcombe et al. 1984

0.32-

0.56g

NR 96h S, N mortality LC50 270, 320 8.35 low

values

173 22 2 Dwyer et al. 1995

0.21g, 26

mm

NR 96h S, N mortality LC50

NOEC

300

180

7 9.2 45 22 2 Analytical BioChemistry

Laboratories, Ltd. 1981

Ptychocheilus lucius

(Colorado squawfish)

0.32-

0.34 g

NR 96h S, N mortality LC50 260, 240 8.35 low

values

173 22 2 Dwyer et al. 1995

Salmo salar

(Atlantic salmon)

1 yr,

immatur

e 23g

NR 30d IPR,

N

smoltification

vitellogenesis

sig. effect 3 mg NP

in 100μL

oil

NR NR NR 8-12 U Madsen et al. 1997

1 yr,

immatur

e23g

NR 30d IPR,

N

osmoregulationsig. effect 3 mg NP

in 100μL

oil

NR NR NR 8-12 U Madsen et al. 1997
APPENDIX Ic (continued). Toxicity of nonylphenol to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Sex Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Salvelinus fontinalis

(brook trout)

36-55mm,

0.5-1.4g

NR 96h S, N mortality LC50 145 6.5-7.0 NR 51.6 (3) ambient

stream

temp.

U Holmes and Kingsbury

1980

NR NR 96h NR,NR mortality LC50 145 NR NR NR NR U Armstrong and Kingsbury

19791

Salvelinus namaycush

Walbaum

(lake trout)

54-92mm,

0.6-60g

NR 35d S, N mortality LC50 >40 NR NR NR 12.5 U Holmes and Kingsbury

1980

Xiphophorus

maculatus

(platyfish)

ADT

0.6-1.2 g

males 28d R, N survival NOEC

LOEC

320

640

8.1 NR NR 28 U Kinnberg et al. 2000

ADT

0.6-1.2 g

males 28d R, N growth NOEC 1280 8.1 NR NR 28 U Kinnberg et al. 2000

Xyrauchen texanus

(razorback sucker)

0.31-0.32 g NR 96h S, N mortality LC50 170, 200 8.35 low

values

173 22 2 Dwyer et al. 1995

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h), days (d), or months (mo.)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent), IPS = single intraperitoneal injection, IPR = repeated intraperitoneal injections

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

Critical study for freshwater Canadian Water Quality Guideline appears in bold

(1) Units are in mg·kg-1 because the nonylphenol was administered as an agar suspension that was injected intraperitoneally

(2) As cited in McLeese et al. 1981

(3) In Holmes and Kingsbury (1980) this value was given as 3 GPG (grains per gallon), but has been converted here to mg·L-1 CaCO3 (by multiplying by a factor of 17.2).
APPENDIX Id. Toxicity of nonylphenol ethoxylates to freshwater algae and macrophytes through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Algae

Scenedesmus sp.

(green algae)

NR NP4EO NR NR,NR mortality lethal

threshold

6000 NR NR NR NR U Janicke et al. 1969

Scenedesmus sp.

(green algae)

NR NP6EO NR NR,NR mortality lethal

threshold

10000 NR NR NR NR U Janicke et al. 1969

Selenastrum capricornutum

(green algae)

NR NP6EO 21d S, N growth NOEC 500000 NR NR NR 25 U Nyberg 1988

Scenedesmus sp.

(green algae)

NR NP7EO NR NR,NR mortality lethal

threshold

16000 NR NR NR NR U Janicke et al. 1969

Selenastrum capricornutum

(green algae)

NR NP7EO 96h NR,NR growth EC50 >1000000 NR NR NR NR U Markarian et al. 19891

,

1990

Selenastrum capricornutum

(green algae)

NR NP8EO 48h S, N growth

inhibition

EC50 20000 7.4 NR NR 24 2 Yamane et al. 1984

Selenastrum capricornutum

(green algae)

NR NP9EO 96h NR,NR growth EC50 50000 NR NR NR NR U Lewis 1986

4-7d NP9EO 96h S, M cell count

(growth)

EC50

NOEC

LOEC

MATC

12000

8000

16000

11300

7-7.5 2-5 150 22-25 1 Dorn et al. 1993

NR NP9EO 21d S, N growth NOEC 500000 NR NR NR 25 U Nyberg 1988

NR NP9EO 48h S, N growth

inhibition

EC50 50000 7.4 NR NR 24 2 Yamane et al. 1984

green and blue-green algae NR NP9.5EO NR NR,NR growth

inhibition

NR 20000 NR NR NR NR U Davis & Gloyna 19672

Scenedesmus sp.

(green algae)

NR NP10EO NR NR,NR mortality lethal

threshold

31000 NR NR NR NR U Janicke et al. 1969

Scenedesmus sp.

(green algae)

NR NP20EO NR NR,NR mortality lethal

threshold

125000 NR NR NR NR U Janicke et al. 1969
APPENDIX Id (continued). Toxicity of nonylphenol ethoxylates to freshwater algae and macrophytes through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluatio

n

Rank

Reference

Selenastrum capricornutum

Printz (green algae)

NR NP30EO 21d S, N growth NOEC 500000 NR NR NR 25 U Nyberg 1988

Scenedesmus sp.

(green algae)

NR NP30EO NR NR,NR mortality lethal

threshold

5000000 NR NR NR NR U Janicke et al. 1969

Macrophytes

Egeria densa

(macrophyte)

leaflets

removed

from

plants

NP4EO several

hours

S, NR depolarization

of membrane

potential

LOEC 10000 7.3 NR NR NR U Linsel et al. 1990

Egeria densa

(macrophyte)

leaflets

removed

from

plants

NP20EO several

hours

S, NR depolarization

of membrane

potential

LOEC 100000 7.3 NR NR NR U Linsel et al. 1990

NR = not reported

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Servos et al. 2000

2

 As cited in Talmage 1994
APPENDIX Ie. Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Crustacea

Hyalella azteca

(amphipod)

2-4d mixture of

APEs

96h S, N mortality LC50 450 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

0-2d mixture of

APEs

96h S, N mortality LC50 600 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

6-8d mixture of

APEs

96h S, N mortality LC50 750 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

4-6d mixture of

APEs

96h S, N mortality LC50 850 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

16-18d mixture of

APEs

96h S, N mortality LC50 870 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

10-12d mixture of

APEs

96h S, N mortality LC50 900 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

20-22d mixture of

APEs

96h S, N mortality LC50 1250 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

24-26d mixture of

APEs

96h S, N mortality LC50 1520 7.4-8.1 6.3-8.2 40 25 U Collyard et al. 1994

Ceriodaphnia dubia

(water flea)

NEO NP1.5EO 7d R, M survival and

reproduction

NOEC

LOEC

MATC

285

886

503

8.0 - 8.9 4.0-8.5

(49-106%

saturation

)

140 -

154

23-25 1 England &

Downing 1995

NEO NP1.5EO 7d R, M total adverse

effects

EC50 319 8.0 - 8.9 4.0-8.5

(49-106%

saturation

)

140 -

154

23-25 1 England &

Downing 1995

NR NP1.5EO 7d NR,NR NR EC50 319 NR NR NR NR U Weeks et al. 1996

NR NP1.5EO 7d NR,NR reproduction MATC 503 NR NR NR NR U Weeks et al. 1996

NR NP1.5EO 96h NR,NR NR EC50 626 NR NR NR NR U Weeks et al. 1996

NEO NP1.5EO 96h R, M total adverse

effects -

mortality,

small size,

pale

appearance

EC50 626 8.0 - 8.9 4.0-8.5

(49-106%

saturation

)

140 -

154

23-25 1 England &

Downing 1995

NEO NP1.5EO 7d R, M mortality LC50 886 8.0 - 8.9 4.0-8.5

(49-106%

saturation

)

140 -

154

23-25 1 England &

Downing 1995
APPENDIX Ie (continued). Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Ceriodaphnia dubia

(water flea)

NR NP1.5EO 7d NR,NR mortality LC50 886 NR NR NR NR U Weeks et al. 1996

NEO NP1.5EO 96h R, M mortality LC50 1016 8.0 - 8.9 4.0-8.5

(49-106%

saturation

)

140 -

154

23-25 1 England &

Downing 1995

NR NP1.5EO 96h NR,NR mortality LC50 1016 NR NR NR NR U Weeks et al. 1996

NR NP1.5EO 48h S, N mortality LC50 1040 8 NR NR 25 2 Ankley et al. 1990

Daphnia magna

(water flea)

NEO NP2EO 48h S, NNR mortality LC50 67 NR NR hard 20 U Maki et al. 1998

NEO NP2EO 48h S, NR mortality LC50 148 NR NR hard 20 U Maki et al. 1998

Daphnia magna

(water flea)

NR NP7EO 96h NR,NR mortality LC50 4100 U Markarian et al.

19891

Daphnia magna

(water flea)

NEO NP9EO 7d R, M mortality LC50

NOEC

LOEC

MATC

9000

10000

20000

14000

7-7.5 2-5 150 22-25 1 Dorn et al. 1993

NEO NP9EO

(Igepal

CO-630,

GAF Corp.)

7d R, M carapace

length

(growth)

NOEC

LOEC

MATC

10000

>10000

>10000

7-7.5 2-5 150 22-25 1 Dorn et al. 1993

(also Kravetz et al.

1991)

NEO NP9EO 48h R, M mortality LC50 14000 7-7.5 2-5 150 25 1 Dorn et al. 1993

(also Kravetz et al.

1991)

Daphnia pulex

(water flea)

NR NP9EO 48h NR,NR immobilizationEC50 2870 NR NR NR NR U Salanitro et al.

1988

Daphnia magna

(water flea)

NR NP10EO 24h NR,NR mortality LC50 44200 NR NR NR NR U Unilever Research

Laboratories 19772

Daphnia pulex

(water flea)

NR NP10EO 48h S, NR mortality LC50 12520 7.0-8.0 3.7-7.5 25-40 20-21 U Moore et al. 1987

Daphnia magna

(water flea)

NEO NPnEO 21d R, N fecundity NOEC 5000 8.0 NR NR 22 Baldwin et al. 1998
APPENDIX Ie (continued). Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Daphnia magna

(water flea)

NEO NPnEO 21d,

then

16h

with

testosterone

R, N metabolic

elimination

of

testosterone

EC100 5000 8.0 NR NR 20 U Baldwin et al. 1998

10d

females

NPnEO 64h (16

with

testosterone)

S, N metabolic

elimination

of

testosterone

NOEC 5000 8.0 NR NR 22 U Baldwin et al. 1998

NEO NPnEO 21d R, N reproduction chronic value 5700 8.0 NR NR 22 U Baldwin et al. 1998

NEO NPnEO 48h NR, N mortality LC50 8600 8.0 NR NR 22 U Baldwin et al. 1998

Daphnia magna

(water flea)

NR NP1EC 48h NR,NR mortality LC50

NOEC

14000

11000

NR NR NR NR U Naylor et al. 1997

Ceriodaphnia dubia

(water flea)

NR NP1EC 7d NR,NR reproduction NOEC 2200 NR NR NR NR U Naylor et al. 1997

NR NP1EC 7d NR,NR mortality NOEC 8400 NR NR NR NR U Naylor et al. 1997

NR NP1EC 96h NR,NR mortality LC50 17000 NR NR NR NR U Naylor et al. 1997

Daphnia magna

(water flea)

NEO NP2EC 48h S, NR mortality LC50 141 NR NR hard 20 U Maki et al. 1998

NEO NP2EC 48h S, NR mortality LC50 990 NR NR hard 20 U Maki et al. 1998

Insecta

Culex pipiens

(southern house

mosquito)

PUP NP1EO NR S, NR emergence EC50 80000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP1.5EO NR S, NR emergence EC50 67000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP4EO NR S, NR emergence EC50 6000 -

8000

7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP6EO NR S, NR emergence EC50 6000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968
APPENDIX Ie (continued). Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Culex pipiens

(mosquito)

PUP NP9EO NR S, NR emergence EC50 10000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP9.5EO NR S, NR emergence EC50 7000 -

19000

7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP10EO NR S, NR emergence EC50 19000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Aëdes aegypti

(mosquito)

LAR (4th

stage)

NP11EO 10d NR, NR pupae

mortality

LC50 50000 NR NR NR 25 U van Emden et al.

1974

LAR (2nd

& 3rd

stage)

NP11EO 24h NR, NR mortality LC50

LC100

500000

1000000

NR NR NR 25 U van Emden et al.

1974

EGG NP11EO 10d NR, NR hatch rate EC50 500000 NR NR NR 25 U van Emden et al.

1974

Culex pipiens

(mosquito)

PUP NP15EO NR S, NR emergence EC50 9000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP20EO NR S, NR emergence EC50 28000,

120000,

>200000

7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

 (mosquito)

PUP NP30EO NR S, NR emergence EC50 >400000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968

Culex pipiens

(mosquito)

PUP NP50EO NR S, NR emergence EC50 >400000 7.5-8.0 NR NR 78°F U Maxwell and Piper

1968
APPENDIX Ie (continued). Toxicity of nonylphenol ethoxylates to freshwater invertebrates through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Mollusca

Elliptio complanata

(mussel)

excised

gills

NP1.5EO 50 min S, N metabolic

heat rate

inhibition

LOEC 9000 NR NR NR 30 U Levine and Cheney

2000

Biomphalaria glabrata

(snail)

1.5-2 cm NP11EO 24h NR, NR mortality LC100 23000 NR NR NR 25 U van Emden et al.

1974

Cnidaria

Hydra littoralis

(hydra)

mixture of

budded,

unbudded

NPnEO 11d R, NR growth threshold

concentration

640 NR NR NR NR U Stephenson 1979

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, PUP = pupae, LAR = larvae, NYM = nymph, NEO = neonates

* duration reported in minutes (min), hours (h), or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Servos et al. 2000

2

 As cited in Anonymous 1977 as cited in Talmage 1994
APPENDIX If. Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP1EO 48h S, NR mortality LC50 3000 7.2 NR 50 16.2 U Yoshimura 1986

Oryzias latipes

(Japanese medaka)

1 day posthatchNP1.5EO 90d R, N growth NOEC 100 NR NR NR 22-24 2 Metcalfe et al.

2001

1 day posthatchNP1.5EO 90d R, N sex ratio NOEC 100 NR NR NR 22-24 2 Metcalfe et al.

2001

1 day posthatchNP1.5EO 90d R, N testis-ova NOEC

LOEC

50

100

NR NR NR 22-24 U Metcalfe et al.

2001

Oncorhynchus mykiss

(rainbow trout)

JUV

(females)

NP2EO 22d,35d F, N growth

inhibition

LOEC 1 6.5 NR 12.5 7-13 U Ashfield et al.

1998

2 yr-old

males

NP2EO 21d F, M 200 fold

increase in

vitellogenin

NR 38 NR NR NR NR U Jobling et al.

1996

hepatocytes from

JUV males

NP2EO 48h S, N vitellogenin

production

ED50 17.27 μM

(5326

μg·L-1)

NR NR NR NR U Jobling and

Sumpter 1993

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP3.3EO 48h S, NR mortality LC50 2500 7.2 NR 50 16.2 U Yoshimura 1986

Lepomis macrochirus

(bluegill sunfish)

NR NP4EO 96h S, M mortality LC50 1300 7.1 9.0-5.1 NR 18 U Macek and

Krzeminski 1975

Lepomis macrochirus

(bluegill sunfish)

NR NP5EO 96h S, M mortality LC50 2400 -

2800

7.1 9.0-5.1 NR 18 U Macek and

Krzeminski 1975

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP5EO 48h S, NR mortality LC50 3600 7.2 NR 50 16.2 U Yoshimura 1986

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP6.4EO 48h S, NR mortality LC50 5400 7.2 NR 50 16.2 U Yoshimura 1986

Pimephales promelas

(fathead minnows)

NR NP7EO 48h NR,NR mortality LC50 3200 NR NR NR NR U Markarian et al.

19891
APPENDIX If (continued). Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oncorhynchus mykiss

(rainbow trout)

12-16 cm NP8EO 14d F, N mortality LC50 4250 7.3-7.4 >70%

saturation

290-310 15-15.6 U Calamari and

Marchetti 1973

12-16 cm NP8EO 96h F, N mortality LC50 4700 7.3-7.4 >70%

saturation

290-310 15-15.6 U Calamari and

Marchetti 1973

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP8.4EO 48h S, NR mortality LC50 11600 7.2 NR 50 16.2 U Yoshimura 1986

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP8.9EO 48h S, NR mortality LC50 11200 7.2 NR 50 16.2 U Yoshimura 1986

Carassius auratus

(goldfish)

NR NP9EO 48h NR,NR mortality LC50 18000 NR NR NR NR U Tomiyama 1974

Lepomis macrochirus

(bluegill sunfish)

NR NP9EO 192h F, N mortality LC50 6300 7.1 9.4-8.9 38 21 U Macek and

Krzeminski 1975

NR NP9EO 96h S, M mortality LC50 7900 7.1 9.0-5.1 NR 18 U Macek and

Krzeminski 1975

Leuciscus idus

(golden orfe)

NR NP9EO 48h NR,NR mortality LC50 7000 NR NR NR NR U Fischer 19732

Oncorhynchus mykiss

(rainbow trout)

hepatocytes from

JUV males

NP9EO 48h S, N vitellogenin

production

ED50 82.31 μM

(50800

μg·L-1)

NR NR NR NR U Jobling and

Sumpter 1993

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP9EO 48h S, NR mortality LC50 12000 7 NR 3400 16.2 U Yoshimura 1986

2 cm,

0.2 g

NP9EO 48h S, NR mortality LC50 14000 7 NR 25 16.2 U Yoshimura 1986

Pimephales promelas

(fathead minnow)

1d NP9EO 7d R, M weight at 7d

(growth)

NOEC

LOEC

MATC

1000

2000

1400

7-7.5 2-5 150 22-25 1 Dorn et al. 1993

(also Kravetz et

al. 1991)

NR NP9EO 96h NR,NR mortality LC50 1620 NR NR NR NR U Salanitro et al.

1988

1d NP9EO 7d R, M mortality LC50

NOEC

LOEC

MATC

2900

1800

2000

1400

7-7.5 2-5 150 22-25 1 Dorn et al. 1993

7-27d NP9EO 96h R, M mortality LC50 4600 7-7.5 2-5 150 22-25 1 Dorn et al. 1993

NR NP9EO 96h NR,NR mortality LC50 6600 NR NR NR NR U Williams et al.

19961
APPENDIX If (continued). Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Carassius aurtus

(goldfish)

5-6 cm NP9.5EO 6h S, NR mortality LC50 6900 NR NR 100 20 U Reiff et al. 1979

Rasbora heteromorpha

(harlequin fish)

1.3-3.0

cm

NP9.5EO 96h F, NR mortality LC50 8600 NR NR 20 20 U Reiff et al. 1979

Lepomis macrochirus

(bluegill sunfish)

NR NP9.5EO 96h S, M mortality LC50 7600 7.1 9.0-5.1 NR 18 U Macek and

Krzeminski 1975

Leuciscus idus4

(golden orfe)

5-7 cm NP9.5EO 48h S, NR mortality LC50 4900 NR NR 150 20 U Reiff et al. 1979

5-7 cm NP9.5EO 96h F, NR mortality LC50 7000 NR NR 268 20 U Reiff et al. 1979

5-7 cm NP9.5EO 96h S, NR mortality LC50 11200 NR NR 268 20 U Reiff et al. 1979

Oncorhynchus mykiss

(rainbow trout)

1 yr old,

9.4 cm,

8.3g

NP9.5EO 24h S, NR loss of

equilibrium

EC50 2800 7.5-8.1 >90%

saturation

114 15 U Muller 1980

NR NP9.5EO 96h NR,NR mortality LC50 7500-

12500

4500-

5900

NR NR NR NR U Unilever Research

Laboratories 19773

1 yr old,

9.4 cm,

8.3g

NP9.5EO 24h S, NR mortality LC50 8500 7.5-8.1 >90%

saturation

114 15 U Muller 1980

Salmo trutta

(brown trout)

2.8 or 5.8

cm

NP9.5EO 96h F, NR mortality LC50 1000 NR NR 26-30 15 U Reiff et al. 1979

Carassius auratus

(goldfish)

5cm NP10EO 48h NR, N mortality LC50 5400 NR NR NR NR U Kurata et al. 1977

NR NP10EO 48h NR,NR mortality LC50 13800 NR NR NR NR U Unilever Research

Laboratories 19773

Rasbora heteromorpha

(harlequin fish)

NR NP10EO 48h NR,NR mortality LC50 11300 NR NR NR NR U Unilever Research

Laboratories 19773

Leuciscus idus

(golden orfe)

NR NP10EO 48h NR,NR mortality LC50 7400,

9500

NR NR NR NR U Unilever Research

Laboratories 19773

Oncorhynchus mykiss

(rainbow trout)

perfused

head of

fish 160-

220 g

NP10EO 1h F, N change in gill

resistence;

sodium net

flux

NOEC

LOEC

87.6

(+0.9 Cd)

63000

(+905

Cd)

7 measuredNR 10 U Pärt et al. 1985
APPENDIX If (continued). Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oncorhynchus mykiss

(rainbow trout)

23 days

after

hatching

NP10EO 6h S, N mortality LC50 2100 7.3-7.4 5 240-260 10-14 U Marchetti 1965

fry 25

days after

hatching

NP10EO 6h S, N mortality LC50 2300 7.3-7.4 5 240-260 10-14 U Marchetti 1965

40d old

(fry)

NP10EO 6h S, N mortality LC50 5200 7.3-7.4 5 240-260 10-14 U Marchetti 1965

210d old

(fingerling)

NP10EO 6h S, N mortality LC50 5200 7.3-7.4 5 240-260 10-14 U Marchetti 1965

alevin 12

days after

hatching

NP10EO 6h S, N mortality LC50 5200 7.3-7.4 5 240-260 10-14 U Marchetti 1965

alevin 19

days after

hatching

NP10EO 6h S, N mortality LC50 5200 7.3-7.4 5 240-260 10-14 U Marchetti 1965

3-10 cm NP10EO 96h NR,NR mortality LC50 7500-

11700

NR NR 20-300 10 and

15

U Unilever Research

Laboratories 19773

alevin 6

days after

hatching

NP10EO 6h S, N mortality LC50 22000 7.3-7.4 5 240-260 10-14 U Marchetti 1965

alevin

immediately after

hatching

NP10EO 6h S, N mortality LC50 42000 7.3-7.4 5 240-260 10-14 U Marchetti 1965

perfused

head of

fish 160-

220 g

NP10EO 1h F, N change in gill

resistence &

O2 transfer

factor; Na+

net flux

significantly

different

 from control

63000 7 measuredNR 10 U Pärt et al. 1985

Phoxinus phoxinus

(minnow)

NR NP10EO 48h NR,NR mortality LC50 8800 NR NR NR NR U Unilever Research

Laboratories 19773

Salmo trutta

(brown trout)

NR NP10EO 48h NR,NR mortality LC50 2700 NR NR NR NR U Unilever Research

Laboratories 19773
APPENDIX If (continued). Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Lebistes reticulatus

(guppy)

14d JUV

(0.007g,

0.7cm)

NP11EO 24h NR, NR mortality LC100 52000 NR NR NR 25 U van Emden et al.

1974

Lebistes reticulatus

(guppy)

females

(0.12-

0.38g,

2 cm)

NP11EO 24h NR, NR mortality LC100 57000 NR NR NR 25 U van Emden et al.

1974

Lebistes reticulatus

(guppy)

males

(0.05-

0.08g,

2 cm)

NP11EO 24h NR, NR mortality LC100 64000 NR NR NR 25 U van Emden et al.

1974

Oncorhynchus mykiss

(rainbow trout)

JUV (46-

174g)

NP12EO 9d IPS, N vitellogenin

induction

NOEC 50

mg·kg-1

NR NR NR NR U Andersen et al.

1999

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP13.1EO 48h S, NR mortality LC50 48000 7.2 NR 50 16.2 U Yoshimura 1986

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

NP16.6EO 48h S, NR mortality LC50 110000 7.2 NR 50 16.2 U Yoshimura 1986

Lepomis macrochirus

(bluegill sunfish)

NR NP30EO 96h S, M mortality LC50 >1 x 106

7.1 9.0-5.1 NR 18 U Macek and

Krzeminski 1975

Pimephales promelas

(fathead minnow)

sexually

mature

NP7-11EO 42d F, M histologic

lesions

NOEC 10 NR NR NR 25-26 2 Miles-Richardson

et al. 1999

Oncorhynchus mykiss

(rainbow trout)

JUV

(females)

NP1EC 22d,35d F, N growth

inhibition

LOEC 10 6.5 NR 12.5 7-13 U Ashfield et al.

1998

2yr males NP1EC 21d F, M 100-fold

increase in

vitellogenin

NR 32 NR NR NR NR U Jobling et al. 1996

hepatocytes

from

immature

males

NP1EC 48h S, N vitellogenin

production

ED50 15.25 μM

(4916

μg·L-1)

NR NR NR NR U Jobling and

Sumpter 1993

Pimephales promelas

(fathead minnow)

NR NP1EC 96h NR,NR mortality LC50

NOEC

2000

1000

NR NR NR NR U Williams et al.

19961
APPENDIX If (continued). Toxicity of nonylphenol ethoxylates to freshwater fish through water column exposure

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Hardness**Temp.

(°C)

Evaluation

Rank

Reference

Oryzias latipes

(Japanese medaka)

1d posthatchNP1.5EC 90d R, N growth NOEC 100 NR NR NR 22-24 2 Metcalfe et al.

2001

1d posthatchNP1.5EC 90d R, N sex ratio NOEC 100 NR NR NR 22-24 2 Metcalfe et al.

2001

1d posthatchNP1.5EC 90d R, N testis-ova NOEC 100 NR NR NR 22-24 2 Metcalfe et al.

2001

Oryzias latipes

(Japanese killifish)

2 cm,

0.2 g

sodium 4-

NP2EC

48h S, NR mortality LC50 8900 7.2 NR 50 16.2 U Yoshimura 1986

2 cm,

0.2 g

sodium 4-

NP1EC

48h S, NR mortality LC50 9600 7.2 NR 50 16.2 U Yoshimura 1986

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent), IPS = single intraperitoneal injection, IPR = repeated intraperitoneal injections

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

** hardness reported in mg CaCO3·L-1

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Servos et al. 2000

2

 As cited in Gloxhuber 1974

3

 As cited in Anonymous 1977 as cited in Talmage 1994

4

 Although reported as Idus idus in the original article, this species has since been renamed as Leuciscus idus
APPENDIX II

Toxicity of nonylphenol and its ethoxylates to marine aquatic life

through water column exposure
APPENDIX IIa. Toxicity of nonylphenol to marine algae through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Skeletonema costatum

(algae)

NR 96h S, M growth

inhibition

(cell count)

EC50

NOEC

27

10

7.9-9.6 NR 30 21-22 1 Ward and Boeri

1990d

NR 96h F, NR NR EC50

NOEC

LOEC

MATC

27

10

20

14.1

NR NR NR NR U Weeks et al. 1996

(also cited in Naylor

1995)

NR = not reported

* duration reported in hours (h)

Test type:S = static, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment

Conc. = concentration

D.O. = dissolved oxygen

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable
APPENDIX IIb. Toxicity of nonylphenol to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Crustaceans

Acartia tonsa

(calanoid copepod)

ADT

(10-12d)

48h S, N mortality LC50

LC50

LC50

190

280

360

NR NR 18 20 U Kusk and Wollenberger 1999

Americamysis bahia1

(mysid)

NEO 28d F, M mean total

length

NOEL

LOEL

MATC

3.9

6.7

5.1

7.4-8.3 5.0-8.5 20-21 23.2-26.4 1 Ward and Boeri 1991a

NR 28d F, NR length MATC

NOEC

LOEC

5.1

3.9

6.7

NR NR NR NR U Weeks et al. 1996 (also cited

in Naylor 1995)

NEO 28d F, M number of

young per

surviving

female

NOEL

LOEL

MATC

6.7

9.1

7.8

7.4-8.3 5.0-8.5 20-21 23.2-26.4 1 Ward and Boeri 1991a

NEO 28d F, M survival NOEL

LOEL

MATC

LC50

6.7

9.1

7.8

>21

7.4-8.3 5.0-8.5 20-21 23.2-26.4 1 Ward and Boeri 1991a

NR 28d F, NR reproduction MATC

NOEC

LOEC

7.8

6.7

9.1

NR NR NR NR U Weeks et al. 1996 (also cited

in Naylor 1995)

NR 28d F, NR survival MATC

NOEC

LOEC

7.8

6.7

9.1

NR NR NR NR U Weeks et al. 1996 (also cited

in Naylor 1995)

NEO

(2.8 mg)

96h F, M mortality LC50

NOEC

43

18

7.3-8.2 6.5-7.8 20 23.8-25.3 1 Ward and Boeri 1990b

NR 96h F, NR mortality LC50 43 NR NR NR NR U Weeks et al. 1996 (also cited

in Naylor 1995)

postlarvae

1-day

96h F, M mortality LC50 60.6 7.8-8.2 NR 30-31 24-25 1 Lussier et al. 2000

Balanus amphitrite

(barnacle)

LAR 1-6d R, M cypris major

protein

production

50%

increase

0.023 NR NR NR 28 U Billinghurst et al. 2000

LAR

(day-3

cypris)

48h R, M reduced

settlement

LOEC 0.059 NR NR NR 28 2 Billinghurst et al. 1998

LAR 1-6d R, M cypris major

protein

production

2-fold

increase

0.693 NR NR NR 28 U Billinghurst et al. 2000
APPENDIX IIb (continued). Toxicity of nonylphenol to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Balanus amphitrite

(barnacle)

LAR (day2 cypris)48h in

NP,

48h in

clean

water

R, M reduced

settlement

LOEC 0.693 NR NR NR 25 U Billinghurst et al. 1998

Corophium volutator

Pallas (amphipod)

<5d JUV

mean

1.02 mm

120d S , N density and

population

growth

LOEC 10 NR NR 25 15 U Brown et al. 1999

<5d JUV

mean

1.02 mm

120d S , N time to

maturity;

length;

sex ratio

NOEC 10 NR NR 25 15 U Brown et al. 1999

mean

4.38 mm

42d S , N mortality 96h LC50

10d LC50

30d LC50

1670

620

270

NR NR 25 15 U Brown et al. 1999

Crangon crangon

(brown shrimp)

NR 7d NR,

NR

mortality LC50 340 NR NR NR NR U Waldock & Thain

19912

NR 96h NR,

NR

mortality LC50 420 NR NR NR NR U Waldock & Thain

19912

NR 96h R,NR mortality LC50 600 NR NR 32 17 U Granmo et al. 1991a

Crangon septemspinosa

(sand shrimp)

2 g each 96h R, M mortality LC50 300 NR NR NR 10 U McLeese et al. 1981

1.3g 96h R, M mortality LC50 300 NR NR NR 10 U McLeese et al. 1980a

Crassostrea gigas

(Pacific oyster)

LAR 72h S, N delayed larval

development

LOEC 100 measure

d

NR 35 20 2 Nice et al. 2000

LAR 48h S, N deformity LOEC 100 measure

d

NR 35 20 2 Nice et al. 2000

Dyspanopeus sayii

(mudcrab)

zoea 4th &

5

th stage

96h F, M mortality LC50 >195 7.8-8.2 NR 30-31 24-25 2 Lussier et al. 2000

Homarus americanus

(American lobster)

zoea first

stage

96h R, N mortality LC50 71 7.8-8.2 NR 30-31 16-17 2 Lussier et al. 2000

NR (450g) 96h R, M mortality LC50 170 NR NR NR 10 U McLeese et al. 1980a

Leptocheirus plumulosus

(amphipod)

ADT 96h F, M mortality LC50 61.6 7.8-8.2 NR 30-31 24-25 1 Lussier et al. 2000

Nitocra spinipes

(harpacticoid)

NR 96h NR,

NR

mortality LC50 118, 139 NR NR NR NR U Bergström 19843

Paleomonetes vulgaris

(grass shrimp)

LAR (2-

day)

96h F, M mortality LC50 59.4 7.8-8.2 NR 30-31 24-25 1 Lussier et al. 2000
APPENDIX IIb (continued). Toxicity of nonylphenol to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Tisbe battagliai

(copepod)

nauplii <1

day old

53d R, M reduced survival of

offspring, intrinsic rate of

increase, egg product-ion

NOEC

LOEC

20

41

NR NR 25 10 1 Bechmann 1999

nauplii <1

day old

96h S, N mortality LC50 31-62 NR NR 25 10 2 Bechmann 1999

ADT 96h S, N mortality LC50 500-

1000

NR NR 25 10 2 Bechmann 1999

Molluscs

Mulinia lateralis

(coot clam)

EMB -

LAR

48h S, N inhibition of fertilization EC50 37.9 7.8-8.2 NR 32 18.3 2 Lussier et al. 2000

Mya arenaria

(soft-shelled clam)

NR 144h R, M mortality NOEC >700 NR NR NR 10 U McLeese et al. 1980a

NR (20g) 360h R, M mortality NOEC >1000 NR NR NR 10 U McLeese et al. 1980a

Mytilus edulis

(common blue mussel)

ADT (40-

50mm)

30d F, N byssus strength NOEC

LOEC

32

56

NR NR 32 10 U Granmo et al. 1989

NR 32d NR,

NR

growth & strength LOEC 56 NR NR NR NR U Naylor 1995

NR 30d R,NR byssus strength LOEC 56 NR NR 32 17 U Granmo et al. 1991a

NR 850h R,NR mortality LC50 140 NR NR 32 17 U Granmo et al. 1991a

eggs to

veligers

72h NR,

N

fertilization, development NOEC 200 NR NR 32 17 U Granmo et al. 1989

NR 96h NR,

NR

mortality LC50 2600 NR NR NR NR U Naylor 1995

ADT (40-

50mm)

32d R, N mortality 96h LC50

360h

850h

3000

500

140

NR NR 32 17 U Granmo et al. 1989

NR 96h R,NR mortality LC50 3000 NR NR 32 17 U Granmo et al. 1991a

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent)

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration; D.O. = dissolved oxygen; Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

Critical study for marine Canadian Water Quality Guideline appears in bold

1

 This species was formerly named Mysidopsis bahia

2

 As cited in Servos et al. 2000

3

 As cited in Wahlberg et al. 1990
APPENDIX IIc. Toxicity of nonylphenol to marine fish through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Agonus cataphractus

(hook nose)

NR 7d NR, NR mortality LC50 360 NR NR NR NR U Waldock and Thain 19911

NR 96h NR, NR mortality LC50 510 NR NR NR NR U Waldock and Thain 19911

Cyprinodon bovinus

(Leon Springs pupfish)

NR 96h NR, NR NR LC50 480 NR NR NR NR U Brecken-Folse et al. 1995

Cyprinodon variegatus

(sheepshead minnow)

JUV 96h F, M mortality LC50 142 7.8-

8.2

NR 30-31 24-25 1 Lussier et al. 2000

JUV (mean

0.55g ww,

26.3 mm)

96h F, M mortality;

bloating,

lethargy,

etc.

LC50

NOEC

310

240

7.4-

8.1

7.0-8.8

(60-105%

saturation)

NR 22

(21.4-

23.0)

1 Ward and Boeri 1990c

NR 96h NR, NR NR LC50 460 NR NR NR NR U Brecken-Folse et al. 1995

Fundulus heteroclitus

(killifish)

LAR

(2 weeks

old)

96h R, N mortality LC50 209 NR NR 20 20 2 Kelly and Di Giulio 2000

EMB 10d R, N sublethal

abnormalitie

s

LOEC 2203 NR NR 20 20 2 Kelly and Di Giulio 2000

EMB 96h R, N mortality LC50 5441 NR NR 20 20 2 Kelly and Di Giulio 2000

Gadus morhua

(cod)

NR 21d F, NR mortality median

tolerance

limit

<100 NR NR NR 16 U Swedmark 1968

NR 96h F, NR mortality median

tolerance

limit

300 NR NR NR 16 U Swedmark 1968

NR 96h NR, NR mortality LC50 370 NR NR 32 NR U Granmo et al. 1991a

Gasterosteus aculeatus

(3-spined stickleback)

NR 96h R, NR mortality LC50 370 NR NR 32 17 U Granmo et al. 1991a

Menidia beryllina

(inland silversides)

JUV 96h F, M mortality LC50 70 7.8-

8.2

NR 30-31 24-25 1 Lussier et al. 2000

Platichthys flesus

(flounder)

ADT (male) 15d IPR, N dosedependent

increase in

vitellogenin

observed

NR 10 - 200

mg·kg-1·

week-1

NR NR NR NR U Bjerregaard et al. 1998
APPENDIX IIc (continued). Toxicity of nonylphenol to marine fish through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Platichthys flesus

(flounder)

ADT 21d F, M vitellogenin

induction

NOEC 24.5 NR >80% 34 10 2 Allen et al. 1999

ADT 21d F, M effect on

gonadosoma

-tic index

NOEC 24.5 NR >80% 34 10 2 Allen et al. 1999

ADT 21d F, M increased

hepatosomatic index

LOEC 24.5 NR >80% 34 10 2 Allen et al. 1999

Pleuronectes

americanus

(winter flounder)

LAR (2-day) 96h S, M mortality LC50 17 7.8-

8.2

NR 30-31 5 2 Lussier et al. 2000

Salmo salar

(Atlantic Salmon)

JUV (8 g

each)

96h F, M mortality LC50 130, 160 NR NR NR 10 U McLeese et al. 1981

JUV (8 g

each)

96h R, M mortality LC50 190 NR NR NR 10 U McLeese et al. 1981

hepatocytes

from males

or juveniles

48h R, N induction of

zona radiata

proteins

NOEC

LOEC

1 μM (220.3

μg·L-1)

5 μM (1100

μg·L-1)

NR NR NR 10 U Celius et al. 1999

JUV (4g) 96h R, NR mortality LC50 900 NR NR NR 10 U McLeese et al.

1980a

hepatocytes

from males

or juveniles

96h R, N vitellogenin

induction

NOEC

LOEC

5 μM (1100

μg·L-1)

10 μM (2203

μg·L-1)

NR NR NR 10 U Celius et al. 1999

JUV (ca. 1

year old)

167g

14d IPS, N increased

vitellogenin

and zona

radiata

protein levels

significant

effect

25 mg·kg-1 NR NR 34 10 U Arukwe et al. 2000a

JUV (ca. 1

year old)

75-120g

14d F

(water);

S (NP),

N

decreased

EROD

activity

LOEC

NOEC

125 mg·kg-1

fish

25 mg·kg-1

fish

NR NR NR 10 U Arukwe et al. 1997a
APPENDIX IIc (continued). Toxicity of nonylphenol to marine fish through water column exposure.

Species

(Common name)

Life

stage

Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Zoarces viviparus

(eelpout)

pregnant

females

(70g)

21d R, N increased

vitellogenin

conc.

LOEC

NOEC

100

50

NR NR 20 11 2 Korsgaard and

Pedersen 1998

EMB 21d R, N increased

vitellogenin

conc.

LOEC 1000 NR NR 20 NR U Korsgaard and

Pedersen 1998

males 21d R, N vitellogenin

increased

10000-fold

NR 1000 NR NR NR 12 U Christiansen et al.

1998b,c

ADT (male) 25d IPR, N vitellogenin

increased

10000-fold

NR 0.1 mg·g-1·

week-1

NR NR NR 12 U Christiansen et al.

1998b,c

males 25d IPR, N vitellogenin

increased

1000-fold

NR (70.1 mg·g-1·

week-1

NR NR NR 12 U Christiansen et al.

1998b,c

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent), IPS = single intraperitoneal injection, IPR = repeated intraperitoneal injections

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Servos et al. 2000
APPENDIX IId. Toxicity of nonylphenol ethoxylates to marine algae and macrophytes through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

12 species marine

phytoplankton

NR NP9EO 14d NR, NR growth

inhibition

EC50 10000 NR NR NR NR U Ukeles 1965

NR NP30EO 14d NR, NR growth

inhibition

EC50 1000000 NR NR NR NR U Ukeles 1965

NR = not reported

* duration reported in days (d)

Conc. = concentration

D.O. = dissolved oxygen

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable
APPENDIX IIe. Toxicity of nonylphenol ethoxylates to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Crustaceans

Americamysis bahia 1

(mysid)

3-8 days old NP1.5EO 48h R, M mortality LC50 110 7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

Americamysis bahia1

(mysid)

NR NP2EO 48h R, NR mortality LC50 ca. 150 NR NR NR NR U Patoczka and Pulliam

1990

Americamysis bahia1

(mysid)

3-8 days old NP9EO 48h R, N mortality LC50 900 - 2000

710 - 1560

7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

NR NP9EO 48h R, NR mortality LC50 1230 NR NR NR NR U Patoczka and Pulliam

1990

NR NP9EOPO3

-2 48h R, NR mortality LC50 4600 NR NR NR NR U Patoczka and Pulliam

1990

3-8 days old NP9EO

phosphated

48h R, N mortality LC50 5230 7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

3-8 days old NP9EO

sulphonated

48h R, N mortality LC50 29600

24300

7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

NR NP9EOSO3

-

48h R, NR mortality LC50 29600 NR NR NR NR U Patoczka and Pulliam

1990

Balanus balanoides

(common barnacle)

LAR (stage II

nauplius)

NP10EO 96h R, N mortality LC50 1500 NR NR NR 6-8 U Swedmark et al. 1971

ADT NP10EO 96h F, N mortality LC50 <25000 NR NR NR 6-8,

15-17

U Swedmark et al. 1971

Carcinus maenas.

(shore crab)

ADT NP10EO 96h F, N mortality LC50 >100000 NR NR NR 6-8 U Swedmark et al. 1971

Eupagurus bernhardus

(hermit crab)

ADT NP10EO 96h F, N mortality LC50 >100000 NR NR NR 6-8 U Swedmark et al. 1971

Hyas araneus

(spider crab)

LAR (stage 1

zoea)

NP10EO 96h S, N mortality LC50 10000 NR NR NR 6-8 U Swedmark et al. 1971

ADT NP10EO 96h F, N mortality LC50 >100000 NR NR NR 6-8 U Swedmark et al. 1971

Leander adspersus

(decapod)

postmoult

phase

NP10EO 96h F, N mortality LC50 10000 NR NR NR 15-17 U Swedmark et al. 1971

intermoult

phase

NP10EO 96h F, N mortality LC50 50000 NR NR NR 15-17 U Swedmark et al. 1971

ADT NP10EO 96h F, N mortality LC50 >100000 NR NR NR 6-8 U Swedmark et al. 1971

Leander squilla

(decapod)

ADT NP10EO 96h F, N mortality LC50 >100000 NR NR NR 6-8 U Swedmark et al. 1971
APPENDIX IIe (continued). Toxicity of nonylphenol ethoxylates to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Crangon crangon

(brown shrimp)

NR NP12EO 48h NR,

NR

mortality LC50 89500 NR NR NR NR U Portmann and Wilson

19712

Carcinus maenus

(shore crab)

NR NP12EO 48h NR,

NR

mortality LC50 >100000 NR NR NR NR U Portmann and Wilson

19712

NR NP12EO 48h NR,

NR

mortality LC50 >100000 NR NR NR NR U Waldock and Thain

19913

Pandalus montagui

(pink shrimp)

NR NP12EO 48h NR,

NR

mortality LC50 19300 NR NR NR NR U Portmann and Wilson

19712

Americamysis bahia1

(mysid)

NR NP15EO 48h R, NR mortality LC50 ca. 2000 NR NR NR NR U Patoczka and Pulliam

1990

3-8 days old tp-NP15EO 48h R, N mortality LC50 2570 7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

Americamysis bahia1

(mysid)

3-8 days old tp-NP40EO 48h R, N mortality LC50 >100000 7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

NR NP40EO 48h R, NR mortality LC50 ca.

100000

NR NR NR NR U Patoczka and Pulliam

1990

Americamysis bahia1

(mysid)

NR NP50EO 48h R, NR mortality LC50 ca.

 4000000

NR NR NR NR U Patoczka and Pulliam

1990

3-8 days old tp-NP50EO 48h R, N mortality LC50 >4110000 7.7-8.0 >90%

saturation

NR 25 2 Hall et al. 1989

Americamysis bahia1

(mysid)

NR NP1EC 96h NR,

NR

mortality LC50

NOEC

9400

4100

NR NR NR NR U Naylor et al. 1997

Mollusca

Monodonta turbiformis

(snail)

NR NP9.5EO 14d NR,

NR

differential

survival of

allozyme

genotypes

NR 7000-

117000

8.3 NR NR 22 U Lavie et al. 1984

Monodonta turbinata

(snail)

NR NP9.5EO 14d NR,

NR

differential

survival of

allozyme

genotypes

NR 6740-

67400

8.3 NR NR 22 U Lavie et al. 1984
APPENDIX IIe (continued). Toxicity of nonylphenol ethoxylates to marine invertebrates through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Cardium edule

(cockle)

ADT NP10EO 96h F, N mortality LC50 5000 NR NR NR 6-8 U Swedmark et al.

1971

JUV NP10EO 96h F, N mortality LC50 <<10000 NR NR NR 15-17 U Swedmark et al.

1971

Mya arenaria

(clam)

ADT NP10EO 96h F, N mortality LC50 <10000 NR NR NR 15-17 U Swedmark et al.

1971

ADT NP10EO 96h F, N mortality LC50 18000 NR NR NR 6-8 U Swedmark et al.

1971

Mytilus edulis

(common blue mussel)

ADT NP10EO 96h F, N mortality LC50 <10000 NR NR NR 15-17 U Swedmark et al.

1971

ADT NP10EO 96h F, N mortality LC50 12000 NR NR NR 6-8 U Swedmark et al.

1971

Pecten maximus

(scallop)

ADT NP10EO 96h F, N mortality LC50 <<5000 NR NR NR 15-17 U Swedmark et al.

1971

Pecten opercularis ADT NP10EO 96h F, N mortality LC50 <<10000 NR NR NR 15-17 U Swedmark et al.

1971

Cardium edule

(cockle)

NR NP12EO 48h NR,

NR

mortality LC50 92500 NR NR NR NR U Portmann and

Wilson 19712

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h) or days (d)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent), IPS = single intraperitoneal injection, IPR = repeated intraperitoneal injections

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration

D.O. = dissolved oxygen

Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 This species was formerly named Mysidopsis bahia

2

 As cited in Talmage 1994

3

 As cited in Servos et al. 2000
APPENDIX IIf. Toxicity of nonylphenol ethoxylates to marine fish through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

 (70.7 mm)

NP5EO 24h exposure, 24h

recovery

S, M mortality LC50 1200 7.92 95-98%

saturation

NR 12 2 Miossec and Bocquéné

1986

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP8EO 24h exposure, 24h

recovery

S, M mortality LC50 3000 7.78 94-104%

saturation

NR 12 2 Miossec and Bocquéné

1986

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP10EO 24h exposure, 24h

recovery

S, M mortality LC50 3300 7.89 94-95%

saturation

NR 12 2 Miossec and Bocquéné

1986

Gadus morhua

(cod)

NR NP10EO NR NR, NR avoidance

behaviour

>20 NR NR NR NR U Hoglund 19761

ADT (30 cm) NP10EO 96h F, N mortality LC50 2500 NR NR NR 15-17 U Swedmark et al. 1971

ADT (30 cm) NP10EO 96h F, N mortality LC50 6000 NR NR NR 6-8 U Swedmark et al. 1971

Pleuronectes flesus

(flounder)

ADT NP10EO 96h F, N mortality LC50 3000 NR NR NR 15-17 U Swedmark et al. 1971

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP12EO 24h exposure, 24h

recovery

S, M mortality LC50 7500 7.95 93-94%

saturation

NR 12 2 Miossec and Bocquéné

1986

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP17EO 24h exposure, 24h

recovery

S, M mortality LC50 95000 7.84 93-96%

saturation

NR 12 2 Miossec and Bocquéné

1986

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP20EO 24h exposure, 24h

recovery

S, M mortality LC50 970000 7.9 96-98%

saturation

NR 12 2 Miossec and Bocquéné

1986
APPENDIX IIf (continued). Toxicity of nonylphenol ethoxylates to marine fish through water column exposure.

Species

(Common name)

Life

stage

Chemical Dur.* Test

type

Effect Endpoint Conc.

(μg·L-1)

pH D.O.

(mg·L-1)

Salinity

(‰)

Temp.

(°C)

Evaluation

Rank

Reference

Anguilla anguilla

(European eel)

- spawn in saltwater

post-LAR

(elvers)

(70.7 mm)

NP30EO 24h exposure, 24h

recovery

S, M mortality LC50 >1 x 107 NR NR NR 12 2 Miossec and Bocquéné

1986

NR = not reported

Life Stage (at start of assay): ADT = adult, JUV = juvenile, NEO = neonates, EMB = embryo

* duration reported in hours (h)

Test type:S = static, R = static-renewal, F = flow-through (continuous or intermittent), IPS = single intraperitoneal injection, IPR = repeated intraperitoneal injections

M = measured concentrations throughout experiment, N = nominal concentration

Conc. = concentration; D.O. = dissolved oxygen; Temp. = temperature

Evaluation Rank: 1 = primary, 2 = secondary, U = unacceptable

1

 As cited in Servos et al. 2000
APPENDIX III

Toxicity of nonylphenol to sediment-dwelling life
Appendix IIIa. Toxicity of nonylphenol to freshwater life through spiked-sediment exposure.

Organism Life

Stage

Endpoint NP

(mg·kg-1 dw)

Temperature (°C)

pH

% TOC

Study Conditions Reference

LETHAL TESTS

Chironomus tentans

(midge)

larvae 14-d exposure, 10%

mortality, not toxic

(NOEC)

20.1 19 to 21

5.5 (sediment)

1.27

- based on standard ASTM and USEPA test

methods

- clean, flow-through water

- system equilibrated for 1 day

England and Bussard

1993

larvae 14-d exposure, 31%

mortality, toxic (LOEC)

34.2 19 to 21

5.5 (sediment)

1.27

- based on standard ASTM and USEPA test

methods

- clean, flow-through water

- system equilibrated for 1 day

England and Bussard

1993

Rana catesbiana

(bullfrog)

tadpole 30-d exposure, 10%

mortality, not toxic

(NOEC)

155.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d LC50, (toxic) 260.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d exposure, 85%

mortality, toxic (LOEC)

390.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

SUB-LETHAL TESTS

Chironomus tentans

(midge)

larvae 14-d exposure, 6%

growth reduction, not

toxic (NOEC)

20.1 19 to 21

5.5 (sediment)

- based on standard ASTM and USEPA test

methods

- clean, flow-through water

- system equilibrated for 1 day

England and Bussard

1993

larvae 14-d exposure, 27%

growth reduction, toxic

(LOEC)

34.2 19 to 21

5.5 (sediment)

- based on standard ASTM and USEPA test

methods

- clean, flow-through water

- system equilibrated for 1 day

England and Bussard

1993
Appendix IIIa (continued). Toxicity of nonylphenol to freshwater life through spiked-sediment exposure.

Organism Life

Stage

Endpoint NP

(mg·kg-1 dw)

Temperature (°C)

pH

% TOC

Study Conditions Reference

Rana catesbiana

(bullfrog)

tadpole 30-d exposure, 10%

sublethal effects

(lethargy, hemmorhaging,

tail lesions), not toxic

(NOEC)

155.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d exposure, 25%

growth reduction, not

toxic (NOEC)

155.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d EC50 for growth and

other sublethal effects

220.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d exposure, 29%

growth reduction, toxic

(LOEC)

390.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

tadpole 30-d exposure, 100%

sublethal effects

(lethargy, hemmorhaging,

tail lesions), toxic (LOEC)

390.0 17.1 to 18.8

6.9 to 8.3 (water)

0.05

- based on USEPA test methods

- clean, flow-through water

- system equilibrated for 2.5 hrs

Ward and Boeri 1992

NOEC = no observed effect concentration

LOEC = lowest observed effect concentration

LC50 = median lethal concentration

EC50 = median effective concentration

dw = dry weight
Appendix IIIb. Toxicity of nonylphenol to marine life through spiked-sediment exposure.

Organism Life

Stage

Endpoint NP

(mg·kg-1 dw)

Temperature

(°C)

pH

% TOC

Study Conditions Reference

LETHAL TESTS

Ampelisca abdita

(amphipod)

juvenile

0.7-1.2

mm

10-d LC50 (toxic) 98.7 20

NR

NR

- based on USEPA test methods

- 29 to 32 g·L-1 seawater

- aeration of water column

Fay et al. 2000

juvenile

0.7-1.2

mm

10-d LC100 (toxic) 243 20

NR

NR

- based on USEPA test methods

- 29 to 32 g·L-1 seawater

- aeration of water column

Fay et al. 2000

SUB-LETHAL TESTS

Capitella sp. I

(polychaete)

larvae

to adult

78-d exposure, 17% delay in time to

first reproduction, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 0% difference in time to

first reproduction, not toxic (NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 46% increase in time

between reproductive events, toxic

(LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 3% increase in time

between reproductive events, not toxic

(NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 44% fewer broods per

reproductive individual, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 2% more broods per

reproductive individual, not toxic

(NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 78% fewer eggs per

reproductive individual, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 3% fewer eggs per

reproductive individual, not toxic

(NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 76% fewer eggs per

individual, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999
Appendix IIIb (continued). Toxicity of nonylphenol to marine life through spiked-sediment exposure.

Organism Life

Stage

Endpoint NP

(mg·kg-1 dw)

Temperature

(°C)

pH

% TOC

Study Conditions Reference

Capitella sp. I

(polychaete)

larvae

to adult

78-d exposure, 3% fewer eggs per

individual, not toxic (NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 33% decrease in size at

maturity, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 16% decrease in size at

maturity, not toxic (NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 24% decrease in

population growth rate, toxic (LOEC)

174 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

larvae

to adult

78-d exposure, 0% decrease in

population growth rate, not toxic

(NOEC)

52 20

NR

7.4

- 26 to 27 g·L-1 seawater

- renewal of water and sediment weekly

Hansen et al. 1999

NR = not reported

NOEC = no observed effect concentration

LOEC = lowest observed effect concentration

dw = dry weight
APPENDIX IV

Toxicity of nonylphenol and its ethoxylates to terrestrial life
APPENDIX IVa. Toxicity of nonylphenol to soil microbial processes.

Organisms / Effect Endpoint Concentration

(mg·kg-1)

Duration* Soil pH Test Substrate Reference

Selected

CO2 evolution

CO2 evolution (-64%)

NOEC

LOEC

100

1000

40d 6.8 mixture of sewage sludge compost (digested sludge and sawdust

matured for 9 months) & Permian sandstone (75% sand, 13%

clay, 12% silt) in ratio 1:2 (dw); adjusted to 60% field moisture

capacity with demineralized water

Trocmé et al. 1988

CO2 evolution

CO2 evolution (+26%)

NOEC

LOEC

10

500

3 mo. 7.3 silt loam, Udic Haploboroll, Calcaric Phaeozem, 13% clay, 42%

silt, 55% sand; dried, sieved, stored for 2 years; 3.5% CaCO3

Kirchmann et al. 1991

N-mineralization NOEC 500 3 mo. 7.3 silt loam, Udic Haploboroll, Calcaric Phaeozem, 13% clay, 42%

silt, 55% sand; dried, sieved, stored for 2 years; 3.5% CaCO3

Kirchmann et al. 1991

(nitrite accumulation)

Nitrification

Nitrification (-35%)

NOEC

LOEC

10

500

7d 7.3 silt loam, Udic Haploboroll, Calcaric Phaeozem, 13% clay, 42%

silt, 55% sand; dried, sieved, stored for 2 years; 3.5% CaCO3

Kirchmann et al. 1991

Consulted

Azotobacter spp. (bacteria)

growth

growth (-95%)

NOEC

LOEC

18.8 mg·L-1

37.6 mg·L-1

72h NR medium: CaCl2, FeSO4 · 7 H20, K2HP04,

MgSO4 · 7 H20, MnCl2, Na2MoO4, mannitol, deionized water

Mårtensson and

Torstensson 1996

Azotobacter spp. (bacteria)

N2-fixation

N2-fixation

NOEC

LOEC

18.8 mg·L-1

37.6 mg·L-1

72h NR medium: CaCl2, FeSO4 · 7 H20, K2HP04,

MgSO4 · 7 H20, MnCl2, Na2MoO4, mannitol, deionized water

Mårtensson and

Torstensson 1996

Biomass ATP evolution NOEC 1000 40d 6.8 mixture of sewage sludge compost (digested sludge and sawdust

matured for 9 months) & Permian sandstone (75% sand, 13%

clay, 12% silt) in ratio 1:2 (dw); adjusted to 60% field moisture

capacity with demineralized water

Trocmé et al. 1988

Candida albicans (yeast),

decrease in growth rate

EC50

EC98

28 μM (6.2 mg·L-1)

100 μM (22 mg·L-1)

8h NR yeast nitrogen base medium; incubated on orbital shaker Karley et al. 1997

Neurospora crassa

growth inhibition

growth inhibition

EC50

EC100

6 μM (1.3 mg·L-1)

100 μM (22 mg·L-1)

168h

(7d)

NR Vogel’s minimal media plus 2% (wt/vol) sucrose; incubated on

orbital shaker

Karley et al. 1997

Neurospora crassa

oxygen uptake rate inhibition

oxygen uptake rate inhibition

EC50

EC60

23 μM (5.1 mg·L-1)

100 μM (22 mg·L-1)

10 min. 5.8 buffer: 3,3-dimethylglutarate, CaCl2, glucose, KOH Karley et al. 1997
APPENDIX IVa (continued). Toxicity of nonylphenol to soil microbial processes.

Organisms / Effect Endpoint Concentration

(mg·kg-1)

Duration

*

Soil pH Test Substrate Reference

Nitrification NOEC 10 g·kg-1 sludge dw

(concentration of

sludge/soil not

reported)

6h NR sandy loam from Flakkebjerg, Denmark; sieved (2 mm); stored at -

20C; activated sludge aerobically stabilized to remove organic

contaminants, spiked with NP, mixed with soil

Madsen et al. 1998

Photobacterium

phosphoreum (bacteria),

bioluminescence

EC50 5.6 μmol·L-1

(1.2 mg·L-1)

30 min. NR NR (Microtox test) Carlson-Ekvall and

Morrison 1995

NR = not reported

dw = dry weight

* duration reported in minutes (min.), hours (h), days (d), or months (mo.)
APPENDIX IVb. Toxicity of nonylphenol to soil invertebrates.

Organism Life stage Duration* Effect

(% decrease)

Endpoint Conc.

(mg·kg-1)

Soil

pH

Test substrate Reference

Selected

Apporectodea calignosa

(earthworm)

ADT 21d reproduction

(cocoon

production)

EC10 

EC50

3.44

13.7

NR sandy soil from Lundgård site, 89.7% sand,

3.8% silt, 4.3% clay, 2.2% humus, moisture

set to 16% of dw

Krogh et al. 1996

JUV 21d growth EC50 23.9 NR sandy soil from Lundgård site, 89.7% sand,

3.8% silt, 4.3% clay, 2.2% humus, moisture

set to 16% of dw

Krogh et al. 1996

ADT 21d mortality EC10 

EC50

>40

>40

NR sandy soil from Lundgård site, 89.7% sand,

3.8% silt, 4.3% clay, 2.2% humus, moisture

set to 16% of dw

Krogh et al. 1996

Folsomia candida

(springtail)

JUV 28d reproduction EC50 16 - 30 NR loamy sand from Jyndevad, Denmark; sieved

(2 mm); defaunated; mixed with spiked

sludge

Madsen et al. 1998

JUV 28d adult survival LC50 >30 NR loamy sand from Jyndevad, Denmark; sieved

(2 mm); defaunated; mixed with spiked

sludge

Madsen et al. 1998

Folsomia fimetaria

(springtail)

23-26 day 21d reproduction EC10

EC50

23.6

65.5

NR LUFA sand (German standard soil), 89.4%

sand, 5.6% silt, 5.1% clay, 3.9% humus

Krogh et al. 1996

Folsomia fimetaria

(springtail)

NR 21d reproduction EC10

EC50

27

39

NR sandy soil Holm as cited in

Jensen et al. 1997

Folsomia fimetaria

(springtail)

NR 21d reproduction EC10

EC50

48

59

NR sandy soil, mixed with sludge Holm as cited in

Jensen et al. 1997

Folsomia fimetaria

(springtail)

23-26 day 21d adult survival LC10

LC50

75.1

151.3

NR LUFA sand (German standard soil), 89.4%

sand, 5.6% silt, 5.1% clay, 3.9% humus

Krogh et al. 1996

Consulted

Folsomia fimetaria

(springtail)

23-26 day 21d reproduction EC50

EC10

44.2

27.2

NR Lundgård soil, 89-92% sand, 3-4% silt, 3-4%

clay, 1-2% humus

Krogh et al. 1997

Folsomia fimetaria

(springtail)

23-26 day 21d reproduction EC50

EC10

58.4

48.7

NR Lundgård soil, 89-92% sand, 3-4% silt, 3-4%

clay, 1-2% humus; mixed with spiked sludge

Krogh et al. 1997

Folsomia fimetaria

(springtail)

23-26 day 21d adult survival LC50

LC10

85.9

42.7

NR Lundgård soil, 89-92% sand, 3-4% silt, 3-4%

clay, 1-2% humus

Krogh et al. 1997

Folsomia fimetaria

(springtail)

23-26 day 21d adult survival LC50 >100 NR Lundgård soil, 89-92% sand, 3-4% silt, 3-4%

clay, 1-2% humus; mixed with spiked sludge

Krogh et al. 1997

NR = not reported

Life stage (at start of assay): ADT = adult, JUV = juvenile

* duration reported in days (d)

dw = dry weight
APPENDIX IVc. Toxicity of nonylphenol to terrestrial plants.

Organism Life stage Duration* Effect

(% decrease)

Endpoint Conc.

(mg·kg-1)

Soil

pH

Test substrate Reference

Selected

Brassica rapa ssp. rapa

(turnip)

seedling 14d growth inhibition EC5 10 6.1 sandy loam soil, 54.3% sand, 35.7%

silt, 9.9% clay; max water-holding

capacity 23.2g·100g-1 soil)

Günther and Pestemer 1992

Glycine max 

(soybean)

seedling 21d growth NOEC

EC50

100

1000

5.8 grit/loam soil Windeatt and Tapp 1986

Helianthrus rodeo

(sunflower)

seedling 21d growth NOEC

EC50

100

1000

5.8 grit/loam soil Windeatt and Tapp 1986

Lactuca sativa

(lettuce)

seed to

seedling

7d

14d

growth

growth

EC50

EC50

559

625

7.5 agricultural loam, 24% clay, 1.8%

organic matter, sieved (4 mm), 25-

30% moisture content (i.e. 80% waterholding capacity)

Hulzebos et al. 1993

Sorghum bicolor

(sorghum)

seedling 21d growth NOEC

EC50

100

1000

5.8 grit/loam soil Windeatt and Tapp 1986

Consulted

Atriplex hortensis

(garden orache)

intact plant 4d reduced dry

weight yield

EC50 0.01-0.05 NR grown aseptically in nutrient medium Bokern et al. 1998

cell culture 7-14d reduction of

plant cell growth

EC50 22-110 mg·L-1

(100-500 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Beta vulgaris var.

altissima L. cv. Bush

Mono Leaves

(sugarbeet)

cell culture 7-14d reduction of

plant cell growth

EC50 2.2-11 mg·L-1

(10-50 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Beta vulgaris var.

esculenta L. cv.

Monogold (beet)

cell culture 7-14d reduction of

plant cell growth

EC50 22-110 mg·L-1

(100-500 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Betula papyrifera Marsh

(white birch)

seedling

roots

9-11d electrical

conductivity (ion

leakage)

EC50 5 mg·L-1 NR media bathing roots Weinberger and Greenhalgh 1984

seeds 21d reduction in

sprout length

EC85 5-10 mg·L-1 NR no soil – seeds in petri dishes with NP Weinberger and Greenhalgh 1984

seeds 6d germination NOEC 10 mg·L-1 NR no soil – seeds in petri dishes with NP Weinberger and Greenhalgh 1984

seedlings 21d reduced sprout

growth

EC80 20 mg·L-1 NR no soil -- seeds in petri dishes with NP Weinberger and Vladut 1981

seeds 21d enhanced

germination

EC26 20 mg·L-1 NR no soil -- seeds in petri dishes with NP Weinberger and Vladut 1981

seeds 21d reduction in

percent

germination

EC26 100 mg·L-1 NR no soil -- seeds in petri dishes with NP Weinberger and Vladut 1981
APPENDIX IVc (continued). Toxicity of nonylphenol to terrestrial plants.

Organism Life stage Duration* Effect

(% decrease)

Endpoint Conc.

(mg·kg-1)

Soil

pH

Test substrate Reference

Chenopodium quinoa

(quinoa)

cell culture 7-14d reduction of

plant cell

growth

EC50 2.2-11 mg·L-1

(10-50 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Chenopodium rubrum

(red goosefoot)

cell culture 7-14d reduction of

plant cell

growth

EC50 11-22 mg·L-1

(50-100 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Daucus carota

(carrot)

cell culture 48h growth (dry

weight)

NOEC 1 mM NR cell culture medium Harms 1992

cell culture 7-14d reduction of

plant cell

growth

EC50 >220 mg·L-1

(>1000 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Glycine max

(soybean)

cell culture 7-14d reduction of

plant cell

growth

EC50 >220 mg·L-1

(>1000 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Hordeum vulgare

(barley)

cell culture 48h growth (dry

weight)

LOEC

NOEC

0.5 mM

0.1 mM

NR cell culture medium Harms 1992

cell culture 7-14d reduction of

plant cell

growth

EC50 22-110 mg·L-1

(100-500 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

Lactuca sativa

(lettuce)

seed to

seedling

21d growth EC50 >0.1, <0.32 NR for germination: perlite saturated with

nutrient solution; after germination 5

seedlings with roots >3cm long

transferred to nutrient solution only for

growth test

Hulzebos et al. 1993

cell culture 10d reduction of

plant cell

growth

EC50 110-220

mg·L-1 (500-

1000 μM)

NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mg·L-1 6-

benzylaminopurine; sucrose 20 g·L-1

Bokern and Harms 1997

Lepidium sativum

(garden cress)

seeds 72h inhibition of

radicle

elongation

EC50 6400 mg·L-1 NR on filter paper in petri dishes Günther and Pestemer 1992

Lupinus hartwegii

(Hartweg’s bluebonnet)

cell culture 7d reduction of

plant cell

growth

EC50 11-22 mg·L-1

(50-100 μM)

NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mM KH2PO4

Bokern and Harms 1997

root culture 7d growth

inhibition

EC50 0.1 mM NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mM KH2PO4;

cultured aseptically

Bokern et al. 1998
APPENDIX IVc (continued). Toxicity of nonylphenol to terrestrial plants.

Organism Life stage Duration* Effect

(% decrease)

Endpoint Conc.

(mg·kg-1)

Soil

pH

Test substrate Reference

Lupinus polyphyllus

(bigleaf lupine)

root culture 7d growth

inhibition

EC50 >1 mM NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mM KH2PO4;

cultured with microorganisms

Bokern et al. 1998

cell culture 7d reduction of

plant cell

growth

EC50 >220 mg·L-1

(>1000 μM)

NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mM KH2PO4

Bokern and Harms 1997

root culture 7d growth

inhibition

EC50 >1 mM NR MS medium + 2 mg·L-1 2,4-D + 0.25

mg·L-1 kinetine + 1 mM KH2PO4;

cultured aseptically

Bokern et al. 1998

Lycopersicon

esculentum

(tomato)

cell culture 7-14d reduction of

plant cell

growth

EC50 11-22 mg·L-1

(50-100 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

cell culture 48h growth (dry

weight)

LOEC

NOEC

0.1 mM

0.05 mM

NR cell culture medium Harms 1992

Pennisetum americanum

Rich.

(pearl millet)

cell culture 9d reduction of

plant cell

growth

EC50 11-22 mg·L-1

(50-100 μM)

NR MS medium + 5 mg·L-1 2,4-

dichlorophenoxyacetic acid

Bokern and Harms 1997

Pinus banksiana 

(jack pine)

seeds 14d reduction in

sprout length

EC60 5-10 mg·L-1 NR no soil – seeds in petri dishes with NP Weinberger and Greenhalgh

1984

seeds NR germination NOEC 10 mg·L-1 NR no soil – seeds in petri dishes with NP Weinberger and Greenhalgh

1984

seedling

roots

9-11d electrical

conductivity

(ion leakage)

EC50 10 mg·L-1 NR media bathing roots Weinberger and Greenhalgh

1984

seeds 14d reduction in

percent

germination

EC60 100 mg·L-1 NR no soil -- seeds in petri dishes with NP Weinberger and Vladut 1981

seedlings 14d reduced

sprout growth

EC20 100 mg·L-1 NR no soil -- seeds in petri dishes with NP Weinberger and Vladut 1981

Triticum aestivum

(winter wheat)

cell culture 7-14d reduction of

plant cell

growth

EC50 22-110 mg·L-1

(100-500 μM)

NR B-5 or MS medium; cultivated on

shaker at 120 rpm

Bokern and Harms 1997

NR = not reported

* duration reported in hours (h) or days (d)

Conc. = concentration
APPENDIX IVd. Toxicity of nonylphenol to terrestrial vertebrates.

Organism Effect

(% effect relative to control)

Endpoint Diet Concentration

(mg·kg-1)

Average Dose

(mg·kg-1 BW·d-1)

Exposure

Period

Reference

Selected

Rattus norvegicus

Sprague Dawley

(Norway rat)

F0 generation:

decreased litter size (9%)

increased male kidney weight (7%)

decreased female body weight (9%)

F1 generation:

decreased female body weight (7%)

earlier vaginal opening (47%)

increased pup vagina/uterus relative weight (14%)

increased estrous cycle length (14%)

increased male relative kidney weight (20%)

increased female relative kidney weight (11%)

decreased adult ovary weight (19%)

decreased adult testis weight (6%)

decreased adult male body weight (9%)

F2 generation:

increased gestation length (2%)

sooner to vaginal opening

decreased sperm density (8%)

increased male relative kidney weight (9%)

decreased ovary weight (12%)

decreased adult male body weight (8%)

increased estrous cycle length (18%)

increased male pup weight adjusted for litter size

decreased number of spermatids (13%)

increased relative kidney weight in female pups (9%)

decreased adult female body weight (10%)

F3 generation:

decreased adult female body weight (10%)

sooner to vaginal opening

decreased adult epididymis weight (13%)

decreased adult male body weight (7%)

decreased adult ovary weight

reproductive effects

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

EC

NOAEL

2000

650

2000

650

650

650

2000

2000

2000

2000

2000

2000

200

650

650

650

650

650

2000

2000

2000

2000

2000

650

650

650

2000

2000

200

88-185

28-63

88-185

30-108

30-108

30-108

100-353

100-353

100-353

100-353

100-353

100-353

10-35

31-115

31-115

31-115

31-115

31-115

109-360

109-360

109-360

109-360

109-360

NR

NR

NR

NR

NR

NR

8 weeks

11weeks

11 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks

21 weeks/

generation

Chapin et al. 1999
APPENDIX IVd (continued). Toxicity of nonylphenol to terrestrial vertebrates.

Organism Effect

(% effect relative to control)

Endpoint Diet Concentration

(mg·kg-1)

Average Dose

(mg·kg-1 BW·d-1)

Exposure

Period

Reference

Long Evans rat increased uterine weight in prepubertal females (10%)

increased uterine weight in prepubertal females (65%)

increased uterine weight in ovariectomized females (10%)

increased uterine weight in ovariectomized females (50%)

sooner to vaginal opening (5%)

sooner to vaginal opening (16%)

disruption of estrous cyclicity (29%)

disruption of estrous cyclicity (35%)

NOEC

LOEC

NOEC

LOEC

NOEC

LOEC

NOEC

LOEC

25

50

10

50

25

50

50

100

NR

NR

NR

NR

NR

NR

NR

NR

3 days

3 days

3 days

3 days

15 days

15 days

25 days

25 days

Laws et al. 2000

Noble rat increased uterine weight in ovariectomized females LOEC NR 75 3 days Odum et al. 1999

Consulted

Rattus norvegicus

Sprague Dawley

(Norway rat)

reduced body weight (7%)

reduced body weight gain (10-16%)

mortality

increased male kidney weight

all adverse subchronic effects

EC

EC

NOEC

LOEC

NOAEL

2000

2000

2000

2000

650

129-149

129-149

129-149

129-149

45-50

90 days Cunny et al. 1997

decreased seminiferous tubule diameter

decreased epididymal mass and ratio

decreased testicular mass, testis ratio, and sperm count

LOEC

LOEC

LOEC

100

250

400

NR

NR

NR

10 weeks

10 weeks

10 weeks

De Jager et al.

1999a

decreased body mass, testicular mass, seminiferous

 tubule diameter of offspring

increased testis ratio, decreased epididymal mass,

 decreased sperm count in offspring

LOEC

LOEC

100

250

NR

NR

prenatal to 10

weeks after

weaning

De Jager et al.

1999b

NR = not reported
APPENDIX IVe. Toxicity of nonylphenol ethoxylates to terrestrial organisms.

Organism Life Stage Chemical

(trade name)

Duration

*

Effect (%

decrease)

Endpoint ConcentrationSoil pH Test substrate Reference

Bacteria/Fungi

Candida albicans

(unicellular fungus)

yeast form NP1EO 8h decrease in

growth rate

NOEC

LOEC

100 μM

200μM

NR yeast nitrogen base

medium; incubated on

orbital shaker

Karley et al. 1997

Nectria galligena NR t-NP9.5EO

(Triton N101)

NR growth

inhibition

EC42 500 μg·L-1 NR nutrient media Hislop et al. 1977

Trichoderma viride NR t-NP9.5EO

(Triton N101)

NR growth

inhibition

EC80 500 μg·L-1 NR nutrient media Hislop et al. 1977

Rhizobium trifolii

(N-fixing bacteria)

NR 4-t-NPnEO

(PP 222)

35d growth

(reduced

colony

diameter)

EC50 1000 mg·L-1 6.8 agar medium Fisher et al. 1978

Plants

Phaseolus vulgaris

(green beans)

started from

seed

NPnEO

(DME)

56d reduced dry

weight yield

NOEC 12.3 g·1.8

kg-1 soil

(=0.00683

mg·kg-1)

6.2 Dagor silt loam; 10% clay;

fine-loamy, mixed, mesic

Cumulic Haploxerolls

formed from calcareous

sandstones; top 50 cm;

air-dried, sieved (0.5 cm)

Miller et al. 1980

Zea mays var. saccharata

(sweet corn)

started from

seed

NPnEO

(DME)

56d reduced dry

weight yield

NOEC 12.3 g·1.8

kg-1 soil

(=0.00683

mg·kg-1)

6.2 Dagor silt loam; 10% clay;

fine-loamy, mixed, mesic

Cumulic Haploxerolls

formed from calcareous

sandstones; top 50 cm;

air-dried, sieved (0.5 cm)

Miller et al. 1980

Glycine max

(soya)

seedling NP2EO 21d growth NOEC

EC50

10

1000

5.8 grit/loam soil Windeatt and Tapp 1986

Helianthus rodeo

(sunflower)

seedling NP2EO 21d growth NOEC

EC50

100

1000

5.8 grit/loam soil Windeatt and Tapp 1986

Sorghum bicolor

(sorghum)

seedling NP2EO 21d growth NOEC

EC50

10

1000

5.8 grit/loam soil Windeatt and Tapp 1986
APPENDIX IVe (continued). Toxicity of nonylphenol ethoxylates to terrestrial organisms.

Organism Life Stage Chemical

(trade name)

Duration

*

Effect (%

decrease)

Endpoint ConcentrationSoil pH Test substrate Reference

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP4EO

(Synperonic)

24h potassium

leakage

ca. 22% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP6EO

(Synperonic)

24h potassium

leakage

ca. 58% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP8EO

(Synperonic)

24h potassium

leakage

ca. 82% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP10EO

(Synperonic)

24h potassium

leakage

ca. 80% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP12EO

(Synperonic)

24h potassium

leakage

ca. 78% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Vicia faba cv. Maris Bead

(bean)

leaf discs

(3rd & 4th

leaf pair), 21-

28d plants

NP20EO

(Synperonic)

24h potassium

leakage

ca. 47% of

total

0.1% w/v NR leaf discs exposed to

toxicant solution only

Silcox and Holloway

1986

Trifolium repens

(white clover)

seedling 4-t-NPnEO

(PP 222)

84d growth (plant

weight)

LOEC

NOEC

5000

1000

7 sterile John Innes

compost, innoculated with

Rhizobium trifolii

Fisher et al. 1978

NR = not reported

* duration reported in hours (d) or days (d)

w/v = weight per volume