SUMMARY REPORT

HAIR ANALYSIS PANEL DISCUSSION:

EXPLORING THE STATE OF THE SCIENCE

June 12–13, 2001

Prepared for:

The Agency for Toxic Substances and Disease Registry

Division of Health Assessment and Consultation

and

Division of Health Education and Promotion

Atlanta, Georgia

Prepared by:

Eastern Research Group

110 Hartwell Avenue

Lexington, MA 02421

December 2001
ATSDR Hair Analysis Panel Discussion

NOTE

This report was prepared by Eastern Research Group, Inc. (ERG), an ATSDR contractor, as a

general record of discussion for the “ATSDR Hair Analysis Panel Discussion: Exploring the

State of the Science.” As requested by ATSDR, this report captures the main points of scheduled

presentations and highlights discussions among the panelists. This report is not a verbatim

transcript of the meeting proceedings, nor does it embellish, interpret, or expand upon matters or

agenda topics that were incomplete, unclear, or not addressed. Statements are the individual

views of each panelist or meeting participant. Except as specifically noted, no statements in this

report represent analyses or positions of ATSDR or ERG. 

Any mention of trade names or commercial products does not constitute endorsement or

recommendation for use by ATSDR or the Department of Health and Human Services.
ATSDR Hair Analysis Panel Discussion

FOREWORD

The Agency for Toxic Substances and Disease Registry (ATSDR) has found the expert panel

process to be an effective tool for discussing and weighing scientific and public health issues.

ATSDR convened one such expert panel to discuss the state of the science related to analyzing

hair for environmental substances of concern found at hazardous waste sites. The panel consisted

of individuals who represented state and federal government agencies, academia, and private

practice and whose expertise, interests, and experience covered a wide range of technical

disciplines that were critical to the issues being discussed. ATSDR convened the expert panel as

part of an effort to begin formulating guidance on the use of hair analysis in exposure

assessments. The panel met to discuss their opinions regarding hair analysis for 11⁄2 days in June

2001 in Atlanta, Georgia. This document summarizes the panel discussions.

For ATSDR, the overarching objective of the panel discussion was to gain information on when

to consider using hair analysis for exposure assessments. Exposure assessments are a necessary

component of public health assessments and other related public health activities performed by

the Agency for communities near hazardous waste sites. The Agency sought information about

the overall utility, advantages, and limitations of hair analysis and how these factors would affect

informed decisions on a site-specific basis.

The panel was asked to address a series of general questions about the science of hair analysis.

These focused on exposure assessment and health interpretation of the results of hair analysis.

The panel was strongly encouraged to avoid discussing the merits of hair analysis for drug testing

or nutritional screening, unless such discussions involved a technical point that was directly

applicable to environmental exposure assessment at hazardous waste sites. ATSDR did not seek

consensus statements from the panel; rather, the panel was asked to discuss in detail specific

issues related to methodology, factors influencing the interpretation of results, toxicologic

considerations, data gaps, and research needs. The opinions expressed in the report are those of

the individual panelists and may or may not represent those of ATSDR.

ATSDR views the panel discussions as a first step to sorting through the scientific issues

regarding the advantages and disadvantages of hair analysis. ATSDR plans to weigh the

information and data presented at the panel meeting and, over the next few months, develop

interim guidance for its health assessors and other professionals who are asked by communities

about the virtues of hair analysis as it relates to exposure and health evaluations at hazardous

waste sites. 

RADM Robert C. Williams, P.E., DEE

Assistant Surgeon General

U.S. Public Health Service

and

Director, Division of Health Assessment and Consultation (DHAC)
ATSDR Hair Analysis Panel Discussion

i

Agency for Toxic Substances and Disease Registry

TABLE OF CONTENTS

LIST OF ABBREVIATIONS ................................................... iii

EXECUTIVE SUMMARY ...................................................... v

SECTION 1

INTRODUCTION ..................................................... 1-1

1.1 Background .................................................... 1-1

1.2 Selection of Panelists ............................................. 1-3

1.3 Charge to the Panelists ............................................ 1-4

1.4 The Meeting Format .............................................. 1-5

1.5 The Report Organization .......................................... 1-6

SECTION 2

OPENING REMARKS AND PRESENTATIONS ............................ 2-1

2.1 Welcome ...................................................... 2-1

2.2 Purpose of the Meeting and Charge to the Panelists ..................... 2-2

2.3 Impetus for Panel Discussions—A Case Example ...................... 2-6

2.4 General Physiology of Hair—An Overview ........................... 2-8

SECTION 3

SAMPLING AND ANALYTICAL METHODS .............................. 3-1

3.1 Sample Collection Methods ........................................ 3-2

3.2 Sample Preparation Methods ....................................... 3-3

3.3 Analytical Methods .............................................. 3-4

SECTION 4

FACTORS INFLUENCING THE INTERPRETATION OF ANALYTICAL

RESULTS ........................................................... 4-1

4.1 Distinguishing Between Endogenous and Exogenous Sources of Metals ..... 4-1

4.2 Temporal Considerations and Exposure Conditions ..................... 4-4

4.3 Reference/Background Ranges ..................................... 4-5

SECTION 5

TOXICOLOGIC CONSIDERATIONS ..................................... 5-1

5.1 Pharmacokinetic Issues ........................................... 5-2

5.2 Dose-Response and Clinical Relevance ............................... 5-3

5.3 Choosing the Best Biological Marker ................................ 5-5
ATSDR Hair Analysis Panel Discussion

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SECTION 6

CONCLUSIONS AND RECOMMENDATIONS ............................ 6-1

6.1 What is the State of the Science of Hair Analysis? ...................... 6-1

6.2 When Is It Appropriate to Consider Hair Analysis in Assessing Human 

Exposures to Environmental Contaminants? ........................... 6-1

6.3 What Are the Limitations of Hair Analysis? What Data Gaps and

Research Needs Exist? ............................................ 6-3

6.4 Recommendations ............................................... 6-5

6.5 Next Steps to be Taken by ATSDR .................................. 6-6

SECTION 7

OBSERVER COMMENTS .............................................. 7-1

SECTION 8

REFERENCES ....................................................... 8-1

Appendices

Appendix A List of Panelists and Panelist Biographical Sketches

Appendix B Charge to the Panel

Appendix C Pre-Meeting Comments

Appendix D Hair Analysis Bibliography

Appendix E Meeting Agenda

Appendix F List of Observers

Appendix G Post-Meeting Observer Comments
LIST OF ABBREVIATIONS

AA atomic absorption

ATSDR Agency for Toxic Substances and Disease Registry

CDC Centers for Disease Control and Prevention

CLIA Clinical Laboratory Improvement Act

DHAC Division of Health Assessment and Consultation

DHEP Division of Health Education and Promotion

EI exposure investigation

EPA U.S. Environmental Protection Agency

HIV human immunodeficiency virus

IARC International Agency for Research on Cancer

ICP-AES Inductively coupled argon plasma atomic emission spectrometry

ICP-MS Inductively coupled argon plasma mass spectrometry

ICP-OES Inductively coupled argon plasma optical emission spectrometry

:g/L micrograms per liter

NAA Neutron activation analysis

NHANES National Health and Nutrition Examination Survey

NRC National Research Council

NTP National Toxicology Program

PIXE Proton induced x-ray emission

ppm parts per million

PTH parathyroid hormone

QA/QC quality assurance/quality control

Abbreviations for Panelists’ Names

RB Dr. Robert Baratz

TC Dr. Thomas Clarkson

MG Dr. Michael Greenberg

MK Dr. Michael Kosnett

DP Dr. Dan Paschal

SS Dr. Sharon Seidel

LW Dr. LuAnn White
ATSDR Hair Analysis Panel Discussion

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EXECUTIVE SUMMARY

The Agency for Toxic Substances and Disease Registry (ATSDR) convened a seven-member

panel to review and discuss the current state of the science related to hair analysis, specifically its

use in assessing environmental exposures. ATSDR invited a cross section of scientific experts in

the fields of hair analysis, toxicology, and medicine to participate in 11⁄2 days of discussions on a

variety of topics, including analytical methods, factors affecting the interpretation of analytical

results, toxicologic considerations, and data gaps/research needs. The meeting was held June 12

and 13, 2001, in Atlanta, Georgia.

Background

ATSDR convened this panel in response to (1) a growing number of inquiries from community

members looking for assistance in interpreting hair analysis results and (2) agency interest in

learning more about the utility of hair analysis in evaluating exposures and health effects at

hazardous waste sites. The agency hopes to use the input received from this effort to develop

guidance for agency health assessors on the use and interpretation of hair analysis data.

The general questions that ATSDR seeks to answer include:

• For what substances do reliable hair analysis methods exist?

• When is it appropriate/inappropriate to consider hair analysis in assessing human

exposures to environmental contamination?

• What data gaps exist that limit the interpretation and use of hair analysis in the

assessment of environmental contaminants?

This summary report presents the findings of the panel discussions. Central discussion points are

highlighted below.
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Overview of Discussions

Panelists engaged in a series of discussions to address ATSDR’s questions, pointing to several

limitations—having to do with the current state of the knowledge—on the usefulness of hair

analysis in assessments of environmental exposures. Discussions focused primarily on metals

and trace elements in scalp hair. Panelists considered the distinct differences between using hair

analysis to identify exposures (Is the substance reaching people? Does a competed pathway

exist?) and using it to predict, diagnose, or treat disease (What do hair concentrations tell us

about the likelihood of harmful health effects?). Panelists noted that the latter is where the largest

data gaps exist.

Although they were not required to reach consensus, the panelists did agree on the following

summary statement related to the overall usefulness of hair analysis in evaluating environmental

exposures:

For most substances, insufficient data currently exist that would allow the prediction of a

health effect from the concentration of the substance in hair. The presence of a substance

in hair may indicate exposure (both internal and external), but does not necessarily

indicate the source of exposure.

For what substances do reliable hair analysis methods exist?

The group agreed that laboratory methods exist to measure the levels of some environmental

contaminants in hair, but procedures need to be standardized to help ensure more accurate and

reliable results (this includes ensuring that samples are collected by a trained person and

establishing consistent sampling protocols, washing protocols, quality control/quality assurance

procedures, etc.). Further, the panel agreed that testing should be targeted to the specific element

of interest.
ATSDR Hair Analysis Panel Discussion

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When is it appropriate/inappropriate to consider hair analysis in assessing human exposures to

environmental contamination?

In general, panelists agreed that, before determining the appropriateness of hair analysis as an

assessment tool, assessors should consider the following:

(1) The exposure type and period. Take exposure histories to understand the likelihood that a

particular substance will be in the hair at the time of testing and to identify other exposure

sources (e.g., hair treatments).

Because the growth rate of hair is on average 12 centimeters per year, the panel concluded

that hair analysis is not generally useful for evaluating very recent exposures or those

longer ago than 1 year. Segmental analysis of hair (i.e., looking at concentration trends

along the length of the hair) may have a role in documenting exposures over time (e.g.,

identification of a high-dose acute exposure). This would need to be considered on a

subject-, substance-, and situation-specific basis.

(2) The type of substance and its behavior in the body. Determine the biological plausibility

that a particular substance will be present in hair and whether it is a marker of external

contamination.

The group agreed that little is known about the transfer kinetics of substances into hair.

(3) The clinical relevance of a negative or positive finding. Determine whether any doseresponse relationship exists between chemical concentrations in hair and target organ

effects/illness. Without an understanding of a dose-response relationship, useful

interpretations will not be possible.

The panelists agreed that a relationship between contaminant concentrations in hair and

any kind of measurable outcome have only been established for methyl mercury (e.g., the

relation between maternal hair levels and observed developmental neurological

abnormalities in offspring) and to a limited extent for arsenic (e.g., segmental analysis for

forensic analysis), provided external contamination can be ruled out. There may be

unique forensic settings for other substances.

The group also indicated the need to evaluate, on a substance- and exposure-specific basis, the

extent to which hair analysis may be more advantageous than other biological sampling, such as

blood or urine analysis.
ATSDR Hair Analysis Panel Discussion

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What data gaps exist that limit the interpretation and use of hair analysis in the assessment of

environmental contaminants?

The group identified several factors that limit the interpretation of even the most accurate,

reliable, and reproducible laboratory results. These include:

• The lack of reference (or background) ranges in which to frame the interpretation of

results. Assessors need a greater understanding of what is expected to be in hair in the

absence of environmental exposures in order to determine whether detected levels are

elevated as a result of environmental releases, including possible geographical or regional

differences in background levels.

• Difficulties in distinguishing endogenous (internal) from exogenous (external)

contamination in hair. Being able to make this distinction is important in evaluating

internal doses of the substance of interest. The group voiced different views on the

effectiveness of washing hair prior to analysis to eliminate external contamination. Some

felt that the current literature suggests that there is no reliable washing method capable of

separating external contamination from internal deposition of elements. It was suggested

that identifying metabolites (or other unique markers of internal exposure) for substances

of interest, where possible, is most helpful in distinguishing internal from external

contamination.

• A lack of understanding of how and to what extent environmental contaminants are

incorporated into the hair. Little scientific information is available on the uptake or

incorporation of environmental contaminants into hair. Neither kinetic models nor

metabolite data are known or fully understood for metals or environmentally relevant

organic compounds.

• The lack of correlation between levels in hair and blood and other target tissues, as well

as the lack of epidemiologic data linking substance-specific hair levels with adverse

health effects. These correlations must be understood before hair analysis results can be

used as a diagnostic tool or to predict health endpoints. The panel noted that hair analysis

is not likely to play a role in evaluations of some of the more common health concerns

associated with hazardous waste sites (e.g., cancer, birth defects).

• Little information is available pertinent to the study of environmentally relevant organic

compounds in hair. The panel recommended taking advantage of what is known about

hair analysis for testing drugs of abuse.
ATSDR Hair Analysis Panel Discussion

viii

In moving forward, the panelists encouraged the standardization of sampling protocols and

identified possible research areas. Before hair analysis can be considered a valid tool for any

particular substance, research is needed to establish better reference ranges, gain a better

understanding of hair biology and pharmacokinetics, further explore possible dose-response

relationships, establish whether and when hair may serve as a better measure or predictor of

disease than other biological samples (e.g., blood or urine), and learn more about organic

compounds in hair.

Future ATSDR Activities

ATSDR plans to evaluate all the input received during the panel deliberations and generate a

report on lessons learned from the panel discussions. In addition, the agency anticipates that the

following activities will help all of ATSDR’s divisions as well as professionals in the

community.

• Providing education to physicians and other health professionals about hair analysis.

• Developing a generic fact sheet to help health assessors and communities communicate

and understand hair analysis issues.

• Continuing to develop substance-specific toxicological profiles. The profiles are an

excellent resource and contain information on biomarkers of exposure. In light of the

panel discussions, additional language may be added regarding hair analysis (e.g., in

terms of limitations, etc.).

• Developing guidance on hair analysis to support public health assessments and health

studies conducted by the agency. That is, developing criteria for determining when to

consider hair analysis as part of an ATSDR exposure investigation.
ATSDR Hair Analysis Panel Discussion

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SECTION 1

INTRODUCTION

ATSDR convened a panel of seven experts to discuss the state of the science related to hair

analysis, with specific focus on its utility in assessing environmental exposures. A 11⁄2-day

meeting held at the Radisson Executive Park in Atlanta, Georgia, on June 12 and 13, 2001,

served as a forum for the panelists to discuss scientific issues related to the analysis and

interpretation of hair data. The meeting, which was open to the public, also gave other interested

parties the opportunity to observe the discussions, ask questions, and provide input.

This section details ATSDR’s purpose for convening the panel (Section 1.1), how ATSDR

selected panel members (Section 1.2), the charge to the panel (see Section 1.3), the meeting

format (see Section 1.4), and the organization of this summary report (see Section 1.5).

1.1 Background

ATSDR conducts public health assessments to evaluate possible public health implications of

contamination associated with hazardous waste sites and other environmental releases. An

important step in ATSDR’s assessment process is examining exposures to contaminants under

site-specific conditions and determining whether people are being exposed to contaminants at

harmful levels. In most of the agency’s evaluations, the environmental concentration serves as a

surrogate for “exposure.”

Exposure concentrations, or estimated doses based on exposure concentrations, however,

represent only one factor in a continuum of events that ultimately determine whether exposures

will result in illness. Other factors include exposure conditions and various

pharmacokinetic/pharmacodynamic events (e.g., absorption, distribution, metabolism, excretion),
ATSDR Hair Analysis Panel Discussion

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as well as individual variability and susceptibility in the exposed population. To a large extent,

ATSDR evaluates these factors qualitatively in its public health assessments.

To refine its assessments and/or to fill data gaps, ATSDR seeks ways to more precisely quantify

exposures, such as measuring body burdens of a particular contaminant or its metabolites (e.g.,

lead in blood or arsenic and its metabolites in urine). On a site-by-site basis, ATSDR evaluates

what additional exposure data it might be practical and useful to obtain to further support public

health evaluations and ultimately to help determine the disease potential of a particular exposure. 

In convening this panel, ATSDR’s goal was to determine the overall utility of hair analysis as

one such exposure assessment tool. Hearing various points of view will help ATSDR draw

conclusions based on the best available science.

ATSDR plans to weigh the information and data presented at the panel meeting and, in the short

term (i.e., over the next several months), independently develop some interim guidance for its

health assessors and others at ATSDR who are asked by communities about the virtues of hair

analysis in understanding exposures to, or the disease potential of, particular chemicals. For the

purposes of the panel discussions, ATSDR was not seeking consensus of the panel on any

particular issue, but rather scientific input (consistent or varied) for consideration by the agency.

Also, the panel was not convened to discuss or evaluate the merits of hair analysis for other

purposes (e.g., testing for drugs of abuse or nutritional screening). Again, the focus was on

environmental exposures.

See the introductory remarks in Section 2 for additional background information.
ATSDR Hair Analysis Panel Discussion

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1.2 Selection of Panelists

ATSDR identified candidates for the expert panel by reviewing the scientific literature in the

field of hair analysis, researching professional organizations, and consulting with known experts

within research institutes and other academic centers. The agency sought individuals who were

experienced in the field of hair analysis and its interpretation for hazardous substances released

to the environment.

To help ensure that a broad range of views was brought to the table, the agency sought

individuals possessing a range of experience, interest, and expertise in the field of hair analysis.

Potential candidates were ranked based on their level of technical expertise (i.e., either high,

medium, or low) in each of the following categories:

• Hair analysis research

• Laboratory analysis 

• Pediatric medicine

• Occupational medicine

• Forensic medicine

• Exposure assessment

Based on these criteria, ATSDR selected seven panelists, each of whom had expertise in one or

more of the categories listed above. The collective expertise of the panel covered all categories,

and individuals on the panel represented state and federal government, academia, and private

practice.
ATSDR Hair Analysis Panel Discussion

1The papers provided to the panelists for their consideration included: Hopps 1977; Miekeley et

al. 1998; Sky-Peck 1990; Seidel et al. 2001; Steindel and Howanitz 2001 (editorial); Yoshinaga et al.

1990; Wennig 2000.

1-4

Appendix A lists the names and affiliations of the panelists who participated in the meeting as

well as a brief biographical sketch of each of the panelists.

1.3 Charge to the Panelists

ATSDR prepared a list of specific questions for the panel (referred to as the “charge”). Questions

included a wide variety of topics designed to prompt discussions at the meeting. The main topics

in the charge include:

• Analytical methodologies

• Factors influencing the interpretation of analytical results

• Toxicologic considerations

• Data gaps and research needs

• Identifying scenarios for which hair analysis may be appropriate

A copy of the charge to the panelists is included in this report as Appendix B.

Prior to the June 12 and 13, 2001, meeting, panelists were requested to review the charge and

prepare initial responses to the charge questions (in the form of pre-meeting comments). To

support their effort, panelists received six papers from the published literature and a bibliography

of additional literature pertaining to hair analysis.1 The purpose of this pre-meeting exercise was

to stimulate panelists’ thoughts in relation to the charge questions and to serve as a stepping-off

point for the 11⁄2 days of panel discussions. Appendix C contains the panelists’ pre-meeting

comments.
ATSDR Hair Analysis Panel Discussion

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Panelists provided additional relevant references with their pre-meeting comments and during

panel discussions. An expanded bibliography of hair analysis literature is provided in 

Appendix D.

1.4 The Meeting Format

After some introductory remarks by ATSDR and an overview of hair physiology by one of the

panelists, the panel engaged in open discussions related to individual charge questions. 

Discussions generally followed the meeting agenda, as shown in Appendix E. However, as might

be expected, some overlap occurred across topics due to the closely linked nature of the topics. 

Dr. LuAnn White led the panel discussions. At the beginning of the meeting, she clearly stated

the ground rules for the discussion:

• Focus on the scientific issues related to hair analysis.

• Focus on the specific charge topics. In the context of the charge questions, describe the

advantages and disadvantages of using hair analysis.

• Limit discussions to topics directly or indirectly related to environmental exposures.

Focus on the markers of environmental exposures or internal dose.

• Actively listen to one another and exchange ideas and different perspectives.

In addition to the panelists, approximately 50 observers attended one or both days of the meeting.

The observers included representatives from ATSDR, the Centers for Disease Control and

Prevention (CDC), other federal and state agencies, commercial laboratories, and professional

organizations. A list of the observers who attended the meeting is included in Appendix F.

Though the discussion at the meeting was largely among the panelists, observers were given

three separate opportunities during the meeting to comment or ask questions (see the agenda) and
ATSDR Hair Analysis Panel Discussion

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also were encouraged to provide written comments to ATSDR in response to the charge

questions and panel discussions. Written comments received from observers after the meeting are

included in Appendix G.

1.5 The Report Organization 

The organization of this report generally follows the list of topics outlined in the agenda and

charge to the panelists. Section 2 includes a summary of opening remarks. Sections 3, 4, and 5

summarize the panelists’ comments and discussions related to analytical methodologies, various

factors influencing the interpretation of results, and toxicological considerations. Section 6

reports overall conclusions drawn by the panel, including data gaps and research needs.

Comments provided by observers throughout the meeting are presented in Section 7. Section 8

lists references cited in this summary report. 

Note: In subsequent sections, the panelists’ initials are used to attribute comments. They are as

follows: Dr. Robert Baratz (RB), Dr. Thomas Clarkson (TC), Dr. Michael Greenberg

(MG), Dr. Michael Kosnett (MK), Dr. Dan Paschal (DP), Dr. Sharon Seidel (SS), Dr.

LuAnn White (LW).
ATSDR Hair Analysis Panel Discussion

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SECTION 2

OPENING REMARKS AND PRESENTATIONS

Dr. Robert Amler, ATSDR’s Chief Medical Officer, opened the meeting by welcoming panelists

and observers and describing how the hair analysis panel discussions would help support the

agency’s public health mission. Dr. Allan Susten and Dr. Deanna Harkins, technical coordinators

of the panel, reviewed the scientific issues related to hair analysis and the impetus for convening

the hair analysis panel. They briefly described how hair analysis fits into the agency’s public

health assessment process, the goals and objectives of the panel discussions, and how the agency

plans to use the scientific information obtained from panelists and observers.

To help ground subsequent discussions, panelist Dr. Robert Baratz provided an overview on the

anatomy and physiology of hair. ATSDR’s and Dr. Baratz’s presentations are summarized below.

2.1 Welcome

Robert Amler, M.D.

ATSDR Chief Medical Officer

After welcoming all in attendance, Dr. Amler stated that ATSDR’s overall mission is to protect

people’s health by identifying and preventing toxic exposures. Because recognizing problems

and knowing how to evaluate them are key to the agency’s ability to assess potential health

threats, discussions such as those anticipated during the hair analysis meeting are of key

importance. Dr. Amler noted that such discussions will help ATSDR sort through the advantages

and disadvantages of using hair analysis in its exposure and health assessments.

Dr. Amler explained that the panel process has been shown to be a very effective means for

discussing and weighing scientific issues. He further explained that these panel discussions will

serve as a first step in developing agency guidance on the appropriateness of using hair analysis.
ATSDR Hair Analysis Panel Discussion

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Because it is only a first step, additional areas of discussions may be necessary. Dr. Amler

acknowledged that it would not be possible to obtain all the answers in this forum.

Dr. Amler thanked the individuals who were instrumental in initiating and organizing the panel

discussions, including Mr. Robert Williams, Director of ATSDR’s Division of Health

Assessment and Consultation (DHAC); Dr. Gregory Christenson, Acting Director of ATSDR’s

Division of Health Education and Promotion (DHEP); Dr. Allan Susten, DHAC’s Assistant

Director for Science; and Dr. Deanna Harkins, Medical Officer, within DHEP. He also thanked

Dr. LuAnn White for moderating the meeting. Lastly, he thanked all participants for their

involvement in what promised to be fruitful discussions.

2.2 Purpose of the Meeting and Charge to the Panelists

Allan Susten, Ph.D., D.A.B.T.

Assistant Director for Science

ATSDR/DHAC

Dr. Susten described how the agency seeks to use the best available science in conducting its

public health assessments. He indicated that the overarching goal for the panel discussions is to

review the state of the science of the hair analysis field and help the agency evaluate the overall

utility of hair analysis in its public health assessments. Specifically, the agency seeks to

determine when it might be appropriate to use hair analysis in evaluating possible exposures

and/or possible adverse health effects associated with environmental toxicants. Dr. Susten

acknowledged that hair analysis is used for other purposes (e.g., drugs of abuse, forensics) but

said that the focus of this forum was on the relevance of hair analysis to hazardous waste site

evaluations.
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To help illustrate the nature of the scientific input that helps the agency evaluate exposures and

health effects, Dr. Susten displayed a “continuum” showing the components of ATSDR’s public

health assessment process (see Figure 2-1). In doing so, he described the following components

of the process:

• Exposure evaluation—Involves studying how hazardous substances can reach people,

studying the means by which people can come in contact with hazardous materials, and

determining the exposure concentration or dose at the point of contact.

• Target dose evaluation—Involves studying the distribution of a hazardous substance once

it enters the human body and determining the internal and biologically effective doses.

• Health effects evaluation—Takes a closer look at the dose-response relationships of the

substance(s) under evaluation and how the substance exerts its effect.

Dr. Susten explained that it is not enough to look at the estimated exposure concentration or

exposure dose when evaluating the potential that a particular exposure will lead to clinical

disease. To better understand the extent of exposures and the potential that a particular exposure

will lead to disease, one needs to study the biology and the toxicology of the substance involved.

Therefore, where possible, the agency seeks ways to estimate or measure internal dose and to

assess whether such exposures might be associated with adverse health effects.

Through its work over the past decade or so, ATSDR has recognized that knowledge of

environmental concentrations of hazardous substances alone is not enough to evaluate possible

health effects. In response, the agency established a special “exposure investigation” (EI) section

within DHAC to look specifically at biomonitoring and how it can be used to further inform the

public health assessment process. Dr. Susten presented the criteria developed by ATSDR to

determine whether biomonitoring should be considered to evaluate a site-specific exposure

situation:
ATSDR Hair Analysis Panel Discussion

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Figure 2-1. Continuum of events considered in the public health assessment process.
ATSDR Hair Analysis Panel Discussion

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• Can an exposed population be identified?

• Does a data gap exists that affects ATSDR’s ability to interpret whether a public health

hazard exists?

• Can the data gap be filled with an EI?

• How would the EI results impact public health decision-making?

Dr. Susten stated that the panel’s charge is to discuss scientific issues related to hair analysis that

will help the agency determine the criteria for determining when hair analysis might be a useful

tool in assessing public health exposures. He recognized that the science may not be available to

support all analyses and that research may be needed. 

In convening this panel, Dr. Susten emphasized, the agency’s goal was to receive panelist and

observer input on the following general questions:

• When is it appropriate to consider hair analysis in assessing human exposures to

environmental contaminants?

• When is it inappropriate to consider hair analysis in assessing human exposures to

environmental contaminants?

• What data gaps exist that limit the interpretation and use of hair analysis in the

assessment of environmental exposures? What research is needed to fill these gaps?

• For what substances do reliable hair analysis methods exist (e.g., trace elements, organic

compounds)?

ATSDR’s primary interest in hair analysis, as was reiterated throughout the meeting, is using the

best science when responding to an individual’s request to interpret hair analysis results and

determining when hair analysis at the population level may be helpful in demonstrating that an

environmental exposure has occurred.
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2.3 Impetus for Panel Discussions—A Case Example

Deanna Harkins, M.D., M.P.H.

Medical Officer

ATSDR/DHEP

Dr. Harkins described a recent site-specific scenario that served as a primary trigger for

organizing the hair analysis panel. Specifically, hair analysis issues raised at a plating facility

prompted ATSDR to look more closely at the criteria that should be considered when choosing

hair analysis as an exposure assessment tool and the best way to interpret hair analysis results. 

Dr. Harkins explained that the U.S. Environmental Protection Agency (EPA), ATSDR, and

relevant state and local agencies have been working together to address community health

concerns related to this particular facility. Dr. Harkins briefly reviewed how ATSDR evaluated

potential exposures associated with releases from the facility (i.e., by examining the nature and

extent of contamination and determining whether contaminants have moved from the source to a

point where people might contact them), noting that the agency studies both past and current

exposures. She re-emphasized Dr. Susten’s point that evaluating exposures is only one step in

evaluating possible public health hazards and that understanding the continuum of events

between exposure and resultant disease is critical to determining the likelihood that a given

exposure will have adverse effects.

Dr. Harkins provided the following summary of the issues reviewed during the assessment of the

facility:

• Investigations at and around the facility revealed the presence of Chromium VI (Cr6+) in

groundwater. In response, affected residences were supplied with bottled water since

1977 and municipal water since 1997. Therefore no recent exposures have occurred.

• Chromium is found naturally in rock/soils and can be found in three valence states (0, 3+,

and 6+). Also, Cr3+ is an essential nutrient: it is required for normal glucose metabolism

and in the potentiation of the action of insulin, and it aids in the metabolism of fat and
ATSDR Hair Analysis Panel Discussion

2-7

cholesterol (Anderson 1997; Schroeder 1968; Mertz 1969; Hunter 1974). The National

Academy of Sciences has established a safe and adequate daily intake for chromium in

adults of 50 to 200 micrograms per day (μg/day) (NRC 1989). It has been reported that

the daily dietary intake of chromium for a typical American is approximately half the

minimum safe and adequate daily intake of 50 μg/day (Anderson and Kozlovsky 1985)

Chromium deficiencies have been shown to result in glucose intolerance, peripheral

neuropathy, and decreased fertility (Anderson 1997). Because chromium is an essential

nutrient and part of normal diets, it is difficult to measure body burdens from

environmental sources.

• The primary health concerns expressed by site community members include birth defects,

miscarriages, and cancer. Neither birth defects nor miscarriages are known to be

associated with chromium exposures. Lung cancer and other respiratory effects have been

associated with chromium exposures, but only in occupational settings where high doses

of Cr6+ were received via inhalation. Cr3+ is not classified by EPA, the National

Toxicology Program (NTP), or the International Agency for Research on Cancer (IARC)

as a carcinogen.

• Site community members wanted to use chromium levels in hair as proof that they were

exposed to chromium and clinically ill. In response, ATSDR, in cooperation with EPA,

the state health department, and outside experts, held a series of meetings with the

community, including the local medical community, to communicate why hair analysis

was not appropriate for this site:

– ATSDR determined that estimated chromium doses based on detected levels of

chromium in groundwater were lower than those associated with any adverse

health effects. 

– Because of the stomach’s and gastric juices’ high capacity for the reduction of

Cr6+, ingested Cr6+ is reduced to Cr3+ within minutes (Kerger et al. 1996). As a

result, a person can tolerate ingestion of 50–100 milligrams of Cr6+ per day

without risk of systemic effects (Donaldson and Barreras 1966; DeFlora and

Wetterhan 1989).

– Measuring chromium in hair would not demonstrate past environmental

exposures.

– The health effects of concern to the community are not known to be linked with

chromium exposure.

Dr. Harkins stated that this case assessment led several ATSDR health assessors to inquire about
ATSDR Hair Analysis Panel Discussion

2-8

the overall utility of hair analysis. In turn, this has prompted the agency to look more closely at

the scientific issues associated with hair analysis and to work toward developing guidance on

when hair analysis might be useful in identifying environmental exposures and in evaluating

disease potential. 

2.4 General Physiology of Hair—An Overview

Robert Baratz, M.D., Ph.D., D.D.S.

To provide a foundation for subsequent discussions, Dr. Baratz described the general

characteristics of hair and the underlying skin (e.g., structure, composition, growth patterns,

growth cycles). Understanding the characteristics of hair, the temporal and spatial patterns of hair

growth, and the factors that affect hair growth, for example, is important when collecting and

interpreting hair analysis data. Dr. Baratz’s presentation is summarized below.

• Anatomy of hair. Hair is encompassed in the follicle located below the skin surface in the

dermis, the fiber-rich layer that makes up the bulk of the skin. The follicle has a

connective tissue component (muscles) and glandular component (sebaceous glands). The

muscles elevate the hair and the glands lubricate the hair.

The primary components of the hair follicle are the dermal papilla and the follicle cells.

The dermal papilla is the “generative zone” of hair (it contains blood vessels, nerves, and

pigment-forming cells). The follicle cells generate the hair shaft; the hair shaft is

composed of essentially dead cells, which are the outermost layers of the epithelium and

form a solid cylinder in the dermis. Mitotic activity at the base of the hair follicle

generates different layers that will “keratinize” (see below).

• Keratinization of hair. Hair is composed of hard keratin (a family of proteins ranging in

size from 20,000 to 70,000 Daltons) and is chemically denser than other forms of keratin

(e.g., calluses, dander flakes). Keratinized cells contain more than 85% protein. Where

the hair shaft separates from the follicle it undergoes “disjunctive” keratizination, which

involves the splitting of layers and exposing surfaces not previously exposed.

Keratinized cells have a very distinctive appearance, and have tiny pores littering their

surfaces. The cells are flattened and tightly bound to their neighbors in a very complex

array. When they begin to split apart (by an unknown process), large “nooks and

crannies” are formed. These types of anatomical features allow external environmental
ATSDR Hair Analysis Panel Discussion

2-9

agents to be easily trapped in the outer surface of the hair.

• Elements found in hair. Because so many elements are ubiquitous in the environment and

therefore found in the human body, merely finding a particular element in the hair does

not prove that it got there via a specific route/source, or that finding it has clinical

significance.

• Growth rates. Hair growth varies depending on body region. For example, average

eyelash/brow growth rates have been reported at 0.16 millimeters (mm) per day, scalp

hair at 0.34 to 0.36 mm/day, and beard hair at 0.38 mm/day. Growth rates also are

affected by age, gender, hair color, and ethnicity. For example, scalp hair in a

prepubescent, adolescent, adult, and older adult have been reported at 0.41, 0.30, 0.34,

and 0.32 mm/day, respectively (Myers and Hamilton 1951).

Interindividual variability also occurs. Scalp hair grows at an average rate of 1 centimeter

(cm) per month, but can range from 0.6 to 3.36 cm/month (Harkey 1993). Thus, 12 cm

can represent 31⁄2 to 20 months of hair growth.

• Growth cycles. Hair grows in phases (see Figure 2-2). Usually, more than 90% of the hair

is in the growing (or anagen) phase. The length of anagen varies from 2 to 6 years. The

longer the hair, generally the longer the phases. For example, long hair tends to grow

more slowly. Through apoptosis, the hair will begin to enter the relatively short catagen

phase, during which the follicle will begin to regress and move toward the surface (the

papilli will essentially disappear). During the next phase, telogen, the hair will actually

fall out. If the cycle is complete, a resting phase will follow and then the follicle will

resume the anagen phase. However, hair can “exit” the cycle and cease being a terminal

hair. For example, it can become a vellus hair (non-pigmented “peach-fuzz” hair) or the

hair follicle may permanently disappear, as is the case with male-pattern baldness.

Events known to affect the hair follicle and its cycle include local signaling events (e.g,

cytokines, hormones, adhesion molecules). However, no firm theory of cycle control

exists. Hypotheses include the presence of (1) a morphogenesis clock, (2) a cycling

inducer, (3) a desynchronizer, and (4) an actual cycle clock, but none of these are

specifically known.

• Generation, cycling, and “patterning” of hair. The hair growth cycle changes throughout

life and varies based on species and body location. Patterning of hair is important to the

generation and cycling of hair, and to how it relates to its neighbors (e.g., signaling goes

on in various regions to space follicles in even arrays). Because of similarities in hair

growth patterns, studying sheep hair growth has been useful in understanding human hair

growth patterns. Rodent hair growth models, on the other hand, may not be applicable to
ATSDR Hair Analysis Panel Discussion

2-10

humans because rodents have

regional variation in hair

growth; the hair cycles, but in

waves across the body.

Figure 2-2. The hair growth cycle.

• Substances affecting hair growth. A great number of substances can affect hair growth.

For example, some drugs, such as alkalating agents, are cytotoxic and can make hair fall

out (e.g., cancer chemotherapeutic agents). Other agents drive hair into telogen (e.g.,

heparin, Vitamin A, $-blockers, L-dopa, lithium, and some of the non-steroidals). Drugs

that inhibit hair growth include parathyroid hormone (PTH) and PTH-related proteins.

Variable agents also exist, such as Vitamin D. At low concentrations, Vitamin D may

simulate hair growth, but at high concentrations hair growth is inhibited.

Substances such as testosterone, danazol, adrenocorticotropin hormone, metyrapone,

anabolic steroids, glucocorticoids, retinoids, and insulin can lead to hirsutism (growth of

hair where it does not normally occur). Cyclosporin, minoxidil, diazoxide, and
ATSDR Hair Analysis Panel Discussion

2-11

chromakalin increase the growth rate and size of hair (hypertrichosis). However, some

regional variation may occur. For example, steroids will decrease the rate of growth of

eyebrows, lashes, and hair on the extremities, but estrogen and testosterone will generally

stimulate the growth of pubic and axillary hair. 

Other factors can potentiate or inhibit hair growth by affecting the growth of the dermal

papillae, hair, and follicle (see Table 2-1).
ATSDR Hair Analysis Panel Discussion

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Table 2-1. Factor Effects

Factor Effect on Hair Growth

Beta-fibroblast growth factor

Platelet-derived growth factor

Potentiate growth of dermal papillae.

Transforming growth factor beta Inhibits follicle proliferation, if induced by

mitogens.

Interleukin-1 alpha Inhibits growth of hair and follicle.

Epidermal growth factor Stimulates growth.

Fibroblast growth factor-5 Inhibits growth.

Keratinocyte growth factor Stimulates growth; induces keratinization.

Insulin-like growth factor-1 Accelerates growth of hair and follicle.

Skin damage (e.g., cut, scrape, burn,

irritation)

Forces telogen to anagen (well-illustrated in

rodent models).

Allergens (e.g., food) Major changes in the skin, including hair loss.

Malnutrition Protein/calorie deficiencies inhibit hair

growth.

Fungal infection Inhibits growth; hair may fall out.

Hypothyroidism Diminution of eyebrows.

Viral agents (e.g, HIV virus) Hair loss in patches.

Source: Jankovic and Jankovic 2001.
ATSDR Hair Analysis Panel Discussion

3-1

SECTION 3

SAMPLING AND ANALYTICAL METHODS

Panel discussions related to the sampling, handling, and laboratory methodologies used in hair

analysis centered around the strengths and weaknesses of existing procedures and the lack of

standardized methods for collecting and analyzing hair samples and reporting the results.

The group generally agreed that the technology exists to measure substances in hair, but

variations in sample collection, preparation, and analytical methods can drive what will be

measured in the final analysis. Therefore, the panelists encouraged the development of standard

protocols for hair analysis to help ensure the generation of reliable and reproducible analytical

results. In the interim, panelists encouraged laboratories to clearly document procedures used in

their analyses, and encouraged users to be cognizant of these procedures when interpreting

results. The group acknowledged that even if standard protocols were in place, the greatest

challenge would still be interpreting the results from a practical and toxicologic perspective (see

Sections 4 and 5).

Panelist Dr. Dan Paschal, research chemist at CDC, opened discussions on this topic with a brief

overview of the advantages and limitations of existing analytical methods and approaches related

to hair analysis. He emphasized that hair has real advantages in that (1) it can contain relatively

high levels of hazardous substances of potential interest, including elements and some organic

compounds, (2) it is easy to collect by relatively non-invasive methods, and (3) it is a stable

specimen. He also commented on some of the limitations: lack of precision and accuracy of hair

analysis results, external contamination, interindividual variations, lack of correlation with health

effects, and lack of believable reference intervals. 

In setting the stage for discussions on analytical methods, Dr. Paschal commented on published

work related to reference intervals, detection limits, and hair concentrations of metals as a
ATSDR Hair Analysis Panel Discussion

3-2

function of age (DiPietro et al. 1989; Paschal et al. 1989). His specific comments are integrated

in the sections that follow.

3.1 Sample Collection Methods

The panelists offered some varying opinions regarding the best way to collect samples. Topics

discussed included preferred cutting tools, sampling location, and sample handling, as

summarized below.

• Selecting the appropriate cutting device. Panelists offered differing views on what type of

cutting tools should be used when collecting hair samples. One panelist noted that metals

can be released from scissors and therefore recommended using quartz instruments (RB).

One panelist pointed out that if a stainless steel device is used, chromium and nickel

results should be interpreted carefully, although he questioned whether use of stainless

steel would really make a significant difference in the analytical results. This panelist

questioned whether any data are available that document the extent to which chromium

and nickel in stainless steel contribute to sample levels compared to quartz tools (MK). 

In theory, said another panelist, labs that have used stainless steel scissors (for example)

should run a careful blank for chromium. It is, however, difficult to do so: the variable

concentration that is present in the specimen would be measured as well as the variable

amount being introduced by the scissors. A chromium-free hair sample would be needed,

which is not feasible (DP).

This same panelist stressed the importance of being sensitive to possible psychological

and cultural issues when choosing a cutting tool. For example, children may be

intimidated by certain types of shears or other cutting devices. Also, in certain cultures,

hair is considered sacred. Touching, never mind cutting, is prohibited (DP).

One panelist suggested that if interferences due to the cutting instrument used are proven

to be significant, a new instrument might need to be created that would be practical for

field use (e.g., a relatively small tool with a quartz blade) (LW).

• Collection location. Because of differences in growth rates in different regions of the

scalp, the location from which a sample is taken must be carefully considered to ensure

consistency in measurements. For example, the anterior and the parietal regions grow
ATSDR Hair Analysis Panel Discussion

3-3

differently than the vertex (top), occipital (back), and temporal (side) regions (RB). In

response to a question whether an optimal location exists, one panelist noted that defining

an optimal sampling protocol is difficult (DP). At a minimum, it is important to choose a

protocol that is practical in the field setting.

Another panelist noted the desire to identify a reproducible point on the skull. He

suggested taking a sample from the nape of the neck (using a caliper to take the midpoint

between the external auditory meatus), an area where hair is known to grow in a

particular way (RB). Another panelist recommended sampling from the occipital region

(SS).

In its 1989 study, CDC looked for a standard protocol but could not find one. Therefore,

CDC defined its protocol as follows: Approximately 500 to 1,000 milligrams of occipital

hair was collected using stainless steel scissors. Hair was pre-washed (using a non-ionic

detergent). Samples were stored in pre-cleaned plastic bags that were rigorously tested.

Therefore, within the context of the reference interval being generated, data were from

specimens collected in a like fashion (DP).

• Sample storage. One panelist stated that plastic bags or other plasticware should not be

used for storing hair samples unless the containers have been washed or cleaned. Zinc, he

said, is used in plastic molding processes. Because detection limits are precise and

relatively low, it is easy to record contamination from external sources; therefore,

whatever container is used needs to be looked at with great scrutiny (RB). 

• Who should collect the sample. One panelist stressed that people not be allowed to collect

their own samples, put them in plastic bags, and ship them off to the laboratory (MG).

Others agreed: only trained professionals should collect hair samples. 

3.2 Sample Preparation Methods

Panel discussions on sample preparation focused on washing protocols. The group agreed that

washing hair prior to analysis was an important consideration when external sources of the

substance(s) being studied exist. Panelist-specific comments follow.

• One panelist stated that no washing method can distinguish between external

contamination and internally deposited elements. She noted that a number of washing

procedure “camps” exist, including the “no wash hypothesis” (Chittleborough 1980), use

of a mild detergent, the washing procedure recommended by the International Atomic
ATSDR Hair Analysis Panel Discussion

3-4

Energy Agency that uses a solvent in water (adopted by many research groups), and more

radical procedures that use chelating agents. Wide differences in results have been

observed depending on the washing method (SS).

• External interferences can be especially significant with small children, so CDC uses a

standard washing protocol (DP).

• The extent to which washing is necessary depends on the substance being studied and

how the sample is being used. For example, washing is not necessary when one is testing

for a substance for which no external source exists (e.g., methyl mercury). Other key

questions to consider include: Are you looking at a spectrum or a specific agent/element

at a hazardous waste site? Are you sampling for exposure information? Are you sampling

to determine changes in exposures over time? (TC)

See Section 4 for additional discussions on hair washing, specifically as it relates to

distinguishing between endogenous and exogenous sources of metals.

3.3 Analytical Methods

The group noted that reliable analytical methodologies do exist to measure and verify the

presence of various substances in hair. Several panelists specified methods currently used for hair

analysis in their pre-meeting comments; throughout the meeting, they mentioned some of the

methods’ strengths and weaknesses, as well as their applicability. This section highlights the

points made during the meeting, but should not be considered an exhaustive discussion of

existing methodologies. The methods discussed include:

• Cold Vapor Atomic Absorption (AA). It was noted that this is the preferred methodology

for measuring methyl mercury. 

• Inductively Coupled Argon Plasma Mass Spectrometry (ICP-MS). ICP-MS has

widespread use in commercial laboratories. It can be used to measure methyl mercury, but

it is difficult to get reproducible calibrations (DP). With certain types of mass

spectrometry, stable isotope studies can help show the incorporation rates of certain

elements in hair, which may help to answer some of the toxicology questions. For
ATSDR Hair Analysis Panel Discussion

3-5

example, a 20-day delay has been shown between the appearance of lead in blood and its

appearance in hair (TC).

• Inductively Coupled Argon Plasma Optical Emission Spectrometry (ICP-OES) or

Inductively Coupled Argon Plasma Atomic Emission Spectrometry (ICP-AES). Of the

commonly used methods, ICP-OES/AES is used the most (DP). This method makes it

possible to generate a large amount of data on a large number of elements. It is a “quick

and dirty” way of getting a global picture of the elemental composition of a hair sample.

It was noted that, historically, CDC used Jarrell Ash Model 1160 AtomComp (e.g., to

generate the data cited in DiPietro et al. 1989). CDC presently uses a Jobin Yvon Ultima

C (DP). 

• Neutron Activation Analysis (NAA). NAA has been used in forensics to measure trace

elements in small quantities of hair. It can be used for segmental analysis of hair.

Segmental analysis can reveal isolated elevations of contamination along the hair and

provide information regarding the contamination of the length of the hair over time.

Identifying patterns over time can help distinguish whether exposure is endogenous or

exogenous (see also Section 4). These techniques are not widely commercially available,

however (MK).

• X-ray Fluorescence. This technique is amenable, nondestructive, and multi-element. It

also has the advantage of measuring the mass of hair as well as the amount of the element

present in that segment of hair (TC). Another panelist noted that the distribution of

mercury in segments along the length of a single strand of hair may be determined by xray fluorescence. 

• Proton Induced X-ray Emission (PIXE) Spectrometry. This method was brought into play

approximately 30 years ago. This method studies a cross section of hair, enabling

identification of external versus internal contamination. This method has not been used

very much because the instrument is expensive (TC). One panelist noted that singlestrand analysis can be problematic if hair is in the non-growing phase (RB), although it

was noted that this is not a problem if the sample is taken from the root (TC).

Another panelist commented on the variable success of the PIXE method. For example,

differentiating internal and external arsenic may not always be that straightforward. In

cases of internal uptake, peaks of arsenic on the external shaft of hair may be a

consequence of appreciable cysteine residues and sulphydral groups. In cases of external

contamination, washing procedures may lead to greater incorporation of external

contamination into the shaft (MK).
ATSDR Hair Analysis Panel Discussion

3-6

Given the various methodologies that might be used, several panelists pointed out the importance

of understanding the method and analytical equipment used when interpreting hair analysis

results, noting that it is the laboratory’s responsibility to clearly report the method used, quality

assurance measures taken, any possible interferences, etc. Further, the data user should carefully

consider this information when evaluating the results.

As stated by one panelist, it is easy to standardize measurements by using good standards and

good laboratory practice (use of blanks, use of external verification) (DP). While the group

recognized that valid methods exist, several panelists stressed that the challenge lies in the

interpretation (see Section 4 of this report) 

3.4 Other Methodological Considerations

The group discussed a number of other issues that influence the analytical results and should be

considered when choosing methods and evaluating analytical data.

• What amount of hair is needed for reliable analyses? CDC has used between 500 and

1,000 milligrams of hair in its studies (DP). Another panelist commented that the amount

selected depends on the analytical method used, but he is more accustomed to sample

sizes in the 50 milligram range (TC). Down the road, there may be an interplay between

the sensitivity of the method and the quantity of hair needed for analysis.

• To what extent should multi-element analytical approaches be used? The group agreed

that a targeted (single-chemical) approach is preferable when analyzing hair for a

particular environmental contaminant. The analytical method selected needs to be

considered in the specific context of the substance and exposure situation under

evaluation; both time and element need to be targeted (RB, MG, MK). 

Serious interference problems can exist with instruments that test for a spectrum of

metals (e.g., ICP instruments) (DP). According to one panelist’s observations,

laboratories do not always appear to account for these interferences: inconsistencies in

approaches are seen across laboratories using ICP-MS and ICP-OES (SS). When

performing OES, one must take a lot of care in choosing the emission wavelength used in
ATSDR Hair Analysis Panel Discussion

3-7

the measurement. Interferences from other elements can occur and must be considered.

This is particularly true when one uses ICP-MS for elements with masses less than 80.

Peaks can be the result of molecules made in the process of generating the ions. These

can interfere with the peaks you are trying to measure (e.g., argon chloride and arsenic,

both with nominal masses of 75 atomic mass units). A high-resolution MS, however, can

resolve two such peaks (DP). 

• Other interferences. Metals in acid solutions, as well as paint, dusts, gloves, etc. in the

laboratory setting can be detected by the instruments used for hair analysis. Looking at

low-level metals in a hair sample is therefore not a simple exercise (RB). These

interferences might potentially overwhelm the amount that you may be seeking to

measure in the hair sample (MK).

• What about organic compounds? A hair assay for benzene is being developed that is

evaluating metabolic products in hair (data are proprietary). Such an assay may have a

great impact on determining the feasibility of using hair analysis for organic chemicals

(MG).

• Quality assurance and quality control. It is the responsibility of the laboratory to

demonstrate its quality control procedures, such as standardizing procedures, running

blank measurements, calibrating equipment, and verifying measurements externally

through proficiency testing programs.
ATSDR Hair Analysis Panel Discussion

4-1

SECTION 4

FACTORS INFLUENCING THE INTERPRETATION OF ANALYTICAL RESULTS

Panelists identified several factors that influence, and more often than not can limit, the

interpretation of hair analysis results. In light of these factors, the panelists generally concluded

that hair analysis findings need to be used and interpreted very cautiously. Even if issues related

to the reliability and reproducibility of the data are resolved, panelists stressed repeatedly, several

factors limit the utility of hair analysis as an exposure and diagnostic tool. Scientists and

clinicians currently know little about what such measurements mean in terms of predicting or

treating clinical disease (see also Section 5, Toxicologic Considerations).

During the meeting, several factors influencing the interpretation of hair analysis data were

discussed:

• Inconsistent sample collection and preparation methods (e.g., sample location, cutting

method, sample storage). (Panelist discussions related to methodological issues are

detailed in Section 3.)

• Difficulties in distinguishing metals deposited externally from those incorporated

internally from the hair follicle. 

• Exposure chronology and conditions (e.g., exposure period of interest, hair growth cycle,

other exposures, etc.).

• The questionable reliability and variability of “reference” ranges. That is, what defines an

“elevated” level?

4.1 Distinguishing Between Endogenous and Exogenous Sources of Metals

All of the panelists agreed that using hair analysis as an exposure or diagnostic tool for metal

contamination is severely limited by difficulties in distinguishing between internal and external

sources of metals. It is further complicated by the natural occurrence of many of the trace
ATSDR Hair Analysis Panel Discussion

4-2

elements (several of which are essential nutrients) within the body. The group recognized,

however, that this distinction is not a limitation when a metabolite or a substance with no

external source is being measured (e.g., organic compounds such as methyl mercury or many

drugs of abuse).

Dr. Kosnett led discussions on the difficulties that exist in distinguishing endogenous and

exogenous substances in hair. Other panelists expanded upon these issues. Individual points are

summarized below.

• Hair analysis data do not necessarily enable you to determine where the measured

contaminant came from and how it got there. High hair levels may provide “markers of

potential exposure,” but that does not tell us how much is internally incorporated. If hair

analysis is used in ATSDR’s evaluations of exposures to contaminants in air (e.g., in the

form of particulates), water, or soil/dust, it must be realized that this distinction cannot

necessarily be made (MK).

• An Alaskan study of arsenic levels in tap water, urine, and nails (Harrington et al. 1978),

reveals some interesting trends. Individuals drinking bottled water, but bathing in tap

water with arsenic averaging 345 micrograms per liter (μg/L), had higher average levels

of arsenic in hair (5.7 parts per million, or ppm) compared to those drinking and bathing

in tap water with arsenic containing 30 μg/L (0.46 ppm arsenic in hair). Urine levels were

similar, however. This example helps illustrate the difficulties in using hair

concentrations alone to draw inferences regarding the magnitude of the internally

absorbed dose of a metal (MK).

• Though they are not applicable to the example above (based on arsenic toxicokinetics),

another reviewer noted that the following caveats could further confound the

interpretation of such a scenario: (1) other exposures could be occurring (e.g., cooking,

brushing teeth), (2) dermal absorption could be occurring, and/or (3) a pool of the

contaminant could be sequestered in and later released from the bone (e.g., this can be

true with tetracycline) (RB).

• Effect of washing hair. Dr. Kosnett described various studies that have looked at the role

and/or effectiveness of washing hair in order to distinguish between endogenous and

exogenous sources of arsenic. These studies suggest that no truly good washing method

exists to remove arsenic: If hair is not washed aggressively, exogenous arsenic will

remain. If hair is washed too aggressively, endogenous arsenic may be removed.
ATSDR Hair Analysis Panel Discussion

4-3

– Smith (1964) showed that detected concentrations of arsenic in hair will vary

depending on washing method, with no method shown to be capable of removing

all arsenic. The results of applying different washing methods (to hair purposely

externally contaminated with 12.08 ppm arsenic) are highlighted in Table 4-1. The

arsenic concentration in hair before contamination was measured at 0.14 ppm.

Table 4-1. Effect of washing method and time on arsenic levels in hair

Washing Method Washing Time (mins) Arsenic (ppm)

Water 5 9.16

15 5.78

30 5.05

60 5.03–6.21

Detergent (5%) 60 4.20–4.93

HCl (N) 60 4.92–6.26

NaOH (N) 60 0.40–0.70

 Source: Smith 1964.

– Van den Berg et al. (1968) showed similar findings. Depending on the washing

regime, this study revealed that even after 1,600 minutes of washing, externally

deposited arsenic was still detected (MK).

• Measuring total concentrations in hair. Depending on the purpose of your testing, it may

not be critical to distinguish between internal and external contamination. For example, in

an industrial hygiene exposure investigation, identifying elevated levels of an element

may be enough to suggest that the potential for exposure exists and protective measures

are needed. While urine data may reveal that existing protective measures have prevented

internal exposures, knowledge that employees have exposure potential may be important

(e.g., contamination could be carried home) (MK, TC, MG). Several panelists reiterated,

however, the limitations of using such data for clinical evaluation or interpretation.
ATSDR Hair Analysis Panel Discussion

4-4

4.2 Temporal Considerations and Exposure Conditions

Panelists agreed that, in determining whether to use hair analysis and in interpreting analytical

results, one must carefully consider exposure chronology and conditions. Because hair growth is

a factor in evaluating when a contaminant might become incorporated in the hair, one must

consider it when deciding whether sampling hair will identify exposures over the period of

interest. With regard to this, the panelists discussed these topics:

• Window of exposure that hair levels may represent. Growth rate is a key consideration.

Assuming growth at approximately 1 centimeter a month, the hair on the average person’s

head generally represents a year or less of time. Therefore, hair analysis is not the best

biological medium to serve as an indicator of very recent exposure or past exposures

(greater than 1 year) (RB).

• Using segmental analysis to study exposures over time. If hair is looked at in a micro or

segmental way, temporal patterns of exposure may be identified. Understanding when

exposure might have occurred may be useful in documenting some historic exposures. As

mentioned in Section 3, neutron activation analysis has been used to identify isolated

elevations along small segments of a hair (e.g, millimeter[s] in length) (MK).

Segmental analysis has been shown to find isolated arsenic peaks at distal points along

the hair shaft. For example, studies of past acute suicidal exposures to arsenic show

distinct peaks migrating away from the scalp (Leslie and Smith 1978). Such analysis can

reveal past exposures even when current urinary levels are normal. Curry and Pounds

(1977) demonstrated peak concentrations of arsenic in hair migrating away from the scalp

following the administration of medicinal arsenic (1 hour to 72 days after ingestion).

Segmental analysis may help ATSDR scientists identify past elevated exposures (e.g.,

acute high exposures from a spill event). Segmental analysis may also rule out exposures.

Houtman et al. (1978), for example, studied hair in a population exposed to an accidental

release of arsenic dust. Segmental hair analysis revealed that concentrations on the distal

parts of hairs were elevated. However, it was determined that the higher levels were

detected on portions of hair that would have been fully formed before the accident, thus

establishing that the arsenic in hair was the result of external contamination. In some

settings, a relatively uniform distribution of a metal such as arsenic along the length of

sampled hair can reflect relatively stable, chronic ingestion, but even in those settings the

contribution of external contamination cannot always be readily determined (MK).
ATSDR Hair Analysis Panel Discussion

4-5

The challenge of using segmental analysis to demonstrate exposure patterns is that it

requires techniques that will enable the analysis of small quantities of hair (e.g.,

subcentimeter sections). It also requires collection of hair in a careful way, to preserve the

orientation of the hair. Further, it has been shown that uptake of arsenic—even on

deliberate external contamination—was not uniform. It has been hypothesized that the

use of shampoos might account for the uneven distribution. This observation might limit

the interpretation of segmental analysis for measuring patterns of endogenous levels

(MK). 

It was also noted that concentration increases towards the tip of the hair because it is

exposed longer. This pattern is typical with external lead exposures. Increased

concentration toward the tip is a useful clue regarding the extent of external

contamination (MK, TC).

• Understanding exposure conditions/histories. Panelists suggested obtaining complete

exposure histories as part of any hair analysis evaluation. A clinician or health assessor

needs to understand the exposure situation and work within a framework of knowing

when data may have a valid use. Using an exposure questionnaire as part of any hair

analysis exercise will help the clinician/assessor identify sources of exposure, both siteand non-site related. Such information will ultimately help the assessor put available data

into perspective (DP, TC, SS).

• Age. The age of the individual or population tested can affect the results and

interpretation of hair analysis. Studies suggest, for example, that alkaline earths and zinc

are not excreted as much in early years of life. The opposite is true with aluminum, of

which children excrete higher levels than adults (Paschal 1989). When skeletal growth

stops, the excretion of these substances into hair is relatively constant. As part of its

National Health and Nutrition Examination Survey 99+ (NHANES), CDC studied

mercury levels in the hair of children and women of childbearing age. Data suggest that

children had lower mercury levels than adults (CDC 2001a; CDC 2001b) (DP).

4.3 Reference/Background Ranges

Discussions regarding reference ranges focused on uncertainties associated with levels of metals,

etc., in “healthy” or “unexposed” individuals and the variability of reference ranges used by

different laboratories. The panelists discussed how the uncertainties play out when one tries to

interpret results of hair analysis.
ATSDR Hair Analysis Panel Discussion

4-6

Individual comments regarding currently available reference range data and inherent limitations

are detailed below:

• The panelists discussed the importance of first clearly defining the term “reference

range.” Two panelists expressed concern about using the term synonymously with

“normal” because it implies that knowledge exists about associated health status, when in

fact such information is largely unavailable. Reference ranges do not represent

“background” or “controls,” nor do we know baseline levels for “normal” states of health

(SS). We do not know what should be present in “healthy” hair (LW). Others suggested

that it is background data that is ultimately being sought, noting that possible geographic

and demographic differences need to be considered.

• A “normal” range does not exist for many elements. Unlike drugs, the presence of which

would be considered “abnormal,” normal ranges need to be identified for metals.

Building the database of normal levels would help assessors better understand and

interpret hair analysis results. Considering hair data in the absence of reference data

against which to compare them is therefore of limited utility (RB). 

• The availability of a reference range does not mean we know the background or typical

levels of endogenous incorporation. It does not necessarily represent what occurs

naturally. It may represent external exposures to ubiquitous levels of contaminants (e.g.,

lead dust, etc.) (MK, TC). For ATSDR’s purposes, the key is distinguishing site

exposures from non-site exposures (e.g., What are background levels where no known

external exposure sources exist?). For example, will we be able to discern whether levels

of a contaminant of interest are elevated in a potentially exposed population (LW)?

• DiPietro et al. (1989) reported analytical results for 271 adults, ages 20 to 73 years, for

selected elements. In comparing the findings of this study with mean hair concentrations

of the same elements reported by others, investigators concluded that results compare

relatively well, given limitations and variability in hair analysis (DP).

• It might be useful to draw a distinction between essential trace elements and nonessential trace elements. One would expect a reference level of the essential trace

elements in hair. The presence of non-essential elements, on the other hand, would

suggest environmental exposure, deposited internally or externally (TC).

• According to two panelists, available reference ranges are often biased and based on

small numbers. Some reference ranges are based on one or two old case reports (RB,

MG).
ATSDR Hair Analysis Panel Discussion

4-7

• The validity of samples used to develop a reference range in the first place is unknown

(RB).

• Available reference values may not relate to the population under study (RB).

• Reference ranges with an approximate 100-fold difference have been used by different

commercial laboratories. What does this really mean from a biological perspective? (SS) 

• One panelist emphasized that more important than understanding reference ranges is

gaining an understanding of whether chemical-specific value have toxicologic or clinical

significance. The availability of reference range information alone is inadequate to assess

the clinical significance of a particular laboratory result; the fact that a reference range has

been exceeded does not establish that the individual sustained a toxicologically

significant dose (MK). Another panelist reminded the group that establishing reliable

reference levels will inform assessors about the possible extent of exposures (LW).

• One panelist questioned whether CDC might consider additional hair analysis as part

NHANES efforts—providing an opportunity to collect data on a cross section of the

population. It was speculated that if the science supported the need for such data

collection, it could be proposed (funding aside) (LW, DP). NHANES 99+ did measure

hair mercury of a selected subpopulation (children ages 1 to 5 and females 16 to 49 years)

(CDC 2001a).
ATSDR Hair Analysis Panel Discussion

5-1

SECTION 5

TOXICOLOGIC CONSIDERATIONS

The panelists agreed that, in order to interpret hair analysis data in any meaningful way, scientists

need a greater understanding of substance-specific relationships between levels in hair and other

body compartments, including target tissues, and how those levels relate to adverse health

outcomes. Much of the toxicology discussion, accordingly, centered around data gaps and

research needs.

The panel chair stressed the importance of understanding to what extent a particular substance

might enter the body, what could conceivably get into hair, and ultimately how such information

can be used as an indicator of exposure and/or of possible clinical effects. Specific questions to

consider included:

• What are the substance-specific pharmacokinetic factors (e.g., intake, absorption,

distribution, excretion) that can influence the biologic uptake of specific substances and

the concentration delivered and incorporated into the hair? How should half-life and

possible storage pools within the body be considered?

• What substances are transported to the hair, and by what mechanism are they transported

(e.g., how are environmental substances of interest incorporated into the hair)?

• What is the dose that causes effect at the target organ? If this is known, how does it relate

to the concentration in the hair matrix?

• How do different patterns of exposure over time (e.g., as may be revealed by segmental

analysis) help us understand possible acute versus long-term exposures, and how might

these patterns correlate with potential health effects?
ATSDR Hair Analysis Panel Discussion

5-2

5.1 Pharmacokinetic Issues

The group acknowledged that little is known about the transfer kinetics of substances into hair

(i.e., their “normal” percolation or rate of appearance in hair). Factors such as transit times, pools

in the body, permeability of basal membranes, and co-factors that may be involved in transit are

not known. Without this knowledge, interpretation of hair analysis results is greatly limited.

Individual panelist input focused on possible ways to fill data gaps. Specifically:

• Hair is a nonvascular tissue (separate from liquid phase transfer kinetics). Understanding

the rate of uptake in the hair, if any, for substances of interest is of critical importance.

Experimental models are needed (RB, MK).

• Implanting human hair on hairless mice, administering a radioactive isotope, and

following its movement to hair may be an effective method for determining the

incorporation of metals into hair (TC).

• Studying the uptake of arsenite used in the treatment of leukemia might be a possible

human model to use to increase our understanding of pharmacokinetics and doseresponse relationships, realizing that administered doses are much higher than they would

ever be expected in an environmental setting (MK).

• Identifying the “transportable” form or metabolite(s) of substances of interest may

provide the best biomarker. Methyl mercury may serve as a model. The key is

understanding the transport mechanism. It may be worthwhile to pursue organo metals

and their behavior (e.g, dimethyl arsenic acid, butyl tin); they may serve as more unique

markers of exposure (TC, MK, SS, LW).

• When interpreting data, studying nutritional status should be considered because it may

play a role in the uptake and distribution of metals. For example, iron and calcium can

increase the uptake of lead into the hair. Zinc levels in hair may be high in “failure to

thrive” cases because hair has stopped growing (LW, SS).

• Obtaining data to better correlate exposure, blood/urine, and hair levels would enable a

better understanding of the relationship of elements in the various body compartments. It

would help correlate external concentration with internal doses. Few such data exist, with

the exception of NHANES data, which evaluate lead levels across hair, blood, and urine
ATSDR Hair Analysis Panel Discussion

2EPA has established a methyl mercury benchmark dose (in maternal hair) of 11 ppm. This is

equivalent to 46 to 49 micrograms of methyl mercury per liter of maternal blood; the critical effect is

developmental neurological abnormalities in offspring (U.S. EPA 2001).

5-3

(which correlated poorly). It was speculated that such substance-specific relationships

could be studied further as part of the NHANES program (DP, LW).

5.2 Dose-Response and Clinical Relevance

The panelists concurred that relationships between hair and any kind of measurable outcome

have only been established for methyl mercury and arsenic. The relationship between maternal

hair and fetal brain levels of methyl mercury is the only well-documented hair/target tissue

relationship; one panelist pointed to the benchmark dose of methyl mercury of 11 ppm in hair

established by EPA2 (RB, SS, TC, MK). Data for arsenic relate largely to forensic examinations;

data do not exist for arsenic that offer disease-predictive value (e.g., long-term health outcomes).

The group could not identify any other environmental substances for which any hard and fast

clinical relationship has been established. Dose-response curves simply do not exist.

Panel discussions regarding current knowledge and the implications for using hair analysis are

highlighted below:

• Can hair analysis predict cancer and other common community health concerns?

Common community health concerns relate to health outcomes such as cancer and birth

defects (according to Dr. Harkins, ATSDR). Questions relate to what harm may have

been done or what future risk may exist as a result of environmental exposures.

One panelist stated that it is not likely for hair analysis to be used to any large extent to

address public health or individual concerns related to teratology or carcinogenicity

(MG). This panelist did note, however, that current efforts to measure benzene in hair

might in the future provide some predictive value for aplastic anemia, but only because of

the known association between benzene and aplastic anemia. Another panelist reemphasized that hair only provides an approximate 1-year time frame in terms of possible

exposures, further supporting the conclusion that hair analysis has little predictive value

in studies of the carcinogenic potential of environmental exposures (RB). Judging from
ATSDR Hair Analysis Panel Discussion

5-4

the current understanding of underlying science (particularly for carcinogens), another

panelist commented, he would rather have exposure history instead of hair analysis data

(DP).

• Importance of establishing a clinical basis prior to testing. A fair amount of discussion

occurred regarding the criticality of establishing a clinical basis before pursuing hair

analysis. Several panelists questioned the relevance of measured levels if they cannot be

used to predict health endpoints. As in other discussions, the dichotomy of using hair

analysis as an exposure tool versus a clinical tool was very evident. 

The physicians on the panel strongly stated that a clinical basis must be established before

hair analysis can be considered a useful tool. One panelist stressed that one should not

collect data that one is not prepared to use (RB). In response to an acknowledgment that a

community might press for hair analysis—for example, even in the absence of

supportable scientific data—two panelists were emphatic that science must be the focus:

politics, litigation, and any other underlying agendas must be put aside (RB, MG). In

general, one must consider what doses, under what circumstances, are relevant (RB). Part

of the challenge lies in communicating to the public what the current science enables us

to do. No absolutes exist in toxicology and medicine. The exposure, the form, the

presentation, and the distribution must be placed in the right context (RB). 

Another panelist strongly stated that the predictive value of the test result must be

weighed and communicated. He emphasized that should the science clearly show no

plausible correlation for a particular substance or exposure situation, then hair analysis

should not be considered (MK).

One panelist reiterated that in the absence of dose-response data, hair analysis may simply

give us a better sense of exposure; it “raises some suspicion” of possible exposure and

effects (TC). Measurements of particular substances in hair may be indicative of

exposure, but not the risk of disease (LW). 

• Understanding the function of various elements in hair. In order to ultimately understand

dose-response relationships and the clinical significance of exposures, scientists need a

better understanding of the role of various elements in the hair. Two panelists briefly

commented on the basic lack of understanding of the function, if any, of metals, cations,

etc., in hair. From a practical point of view, keratinized cells “are on their way out” with

the purpose of protecting the skin and providing warmth. It is therefore difficult to

determine the biological meaning of individual components in hair. Some elements

maintain homeostasis (e.g. potassium). Other elements are co-factors in synthesis (e.g.,

chromium in collagen synthesis). Some elements, on the other hand, are ubiquitous and

have no known purpose (e.g., lead, uranium) (RB, LW).
ATSDR Hair Analysis Panel Discussion

5-5

• Substances for which hair analysis might prove useful. Panelists provided a couple of

examples of other elements for which hair analysis may hold some promise. The panelists

agreed that if strong hypotheses exist, the scientific merit of these types of relationships

may be worth pursuing (RB, DP, TC, SS).

– Thallium might be useful in hair because it is an unusual toxicant. (Thallium was

suggested based on a “classic picture” of thallium intoxication studied by CDC in

Florida.) (DP)

– The possible correlation between excessive manganese levels (as measured in

hair) and violent and other antisocial behaviors has been studied in incarcerated

populations. While study findings suggest some correlation and have some merit

on the surface, many potentially confounding factors exist that need to be

examined more closely, such as hair color, race, and social context (DP). Panelists

questioned the overall scientific merit of the correlation, based on the possible

lack of biological plausibility—that is, symptoms are not necessarily consistent

with documented health effects associated with manganese (DP). One panelist

noted that manganese exposures would more likely be expected to cause

neurological effects that lead to more withdrawn or inactive behavior (e.g.,

Parkinson-like symptoms) (SS). Another panelist noted that, because manganese

is an essential trace element, it is reasonable that it will get into hair (TC).

Another study, by Bader et al. (1999), showed some correlation between axillary

hair and airborne manganese (attributed to contamination by dust and water), but

overall did not support the use of hair for manganese analysis (SS).

5.3 Choosing the Best Biological Marker

Panelists briefly discussed if and when hair may be more advantageous than other biological

samples, such as blood or urine. From both an exposure and clinical perspective, panelists

considered which approaches were most productive. Generally, based on current science, they

concluded that hair may be used to provide historical exposure perspective within a fairly small

window of time (i.e., 1 year). Panelists’ views are highlighted below:

• Two panelists emphasized that the following question needs to be answered in making

such a determination: When might a substance be detected in hair, but not in urine

(measure of excreted amount) or blood (measure of body compartment) (MG, LW)?

Another panelist encouraged consideration of the following question: For what
ATSDR Hair Analysis Panel Discussion

5-6

substances do we have knowledge of the toxicologic implication of the measurement of

the substance in hair compared to the measurement of the substance in other biological

specimens (e.g., urine, blood, bone) (MK)? 

• How do we move toward establishing the “gold standard?” Could hair samples be a better

way to non-invasively get a sample? Is it a valid measure and how does that relate back to

blood or target organ levels (LW)?

• Hair samples may be considered preferable or less invasive under certain situations (e.g.,

pediatric exposures) (SS). Others commented that collecting blood or urine samples did

not appear to be that much of an obstacle (MK, LW).

• Hair may be considered for retrospective purposes when blood and urine are no longer

expected to contain a particular contaminant. Again, the distinction between the use of

hair analysis as an exposure tool, rather than a diagnostic tool, was made (LW).

• From a clinical point of view, it is important to focus on what substances are of greatest

interest, then ask what is the best way to analyze them. Is hair analysis the best way to

measure body burden (instead of blood or urine)? For example, we may be able to

analyze/identify many elements in hair, but it still may be more useful to look at blood

levels. Blood may simply be the better body compartment to test from a scientific point of

view regardless of whether we can test for a particular substance in hair. That is, what can

potential levels in hair tell us that blood levels do not (RB)?

• An acute spike in hair might help document exposure, but generally will not help from a

diagnostic perspective (MG, LW). Acute exposures are best measured through blood or

urine (RB).

• Growth rate is a key consideration. Assuming growth at approximately 1 centimeter a

month, the hair on the average person’s head generally represents a year or less of time.

Hair analysis will therefore have limited usefulness in cases where exposures occurred

more than a year prior to an exposure assessment (RB). While hair analysis may provide a

snapshot of exposure conditions, it is not likely to predict long-term exposures (SS).
ATSDR Hair Analysis Panel Discussion

3This statement addresses only exposure to environmental contaminants and does not address

substances of abuse.

6-1

SECTION 6

CONCLUSIONS AND RECOMMENDATIONS

On the second day of the meeting, panelists reviewed earlier discussions and drew overall

conclusions. During these final deliberations, panelists commented on the overall state of the

science in hair analysis, the major limitations of hair analysis, topics for which a complete

scientific understanding is not available, and research that might permit a better understanding of

the science. The panel’s general conclusions and recommendations are summarized below.

6.1 What Is the State of the Science of Hair Analysis?

Although consensus was not required, the panelists did agree on the following overall conclusion

statement:

For most substances, insufficient data currently exist that would allow the prediction of a

health effect from the concentration of the substance in hair.3

 The presence of a substance

in hair may indicate exposure (both internal and external), but does not necessarily

indicate the source of exposure.

6.2 When Is It Appropriate To Consider Hair Analysis in Assessing Human Exposures

to Environmental Contaminants?

The panelists recognized that hair analysis can serve two distinct purposes: (1) as a tool in

identifying exposures (Is the substance reaching people? Does a competed pathway exist?) and

(2) as a clinical tool (What is the threshold for adverse health effects?) The latter is where the

largest data gaps exist. The panelists agreed that a body of literature describes specific conditions

and uses of hair analysis for methyl mercury and arsenic. There may be a unique forensic setting
ATSDR Hair Analysis Panel Discussion

6-2

for other metals. Segmental analysis with ultra-sensitive techniques may have a role in special

cases—that is, subject-, substance-, and situation-specific cases (e.g., identification of high-dose

acute exposure).

The group agreed on the general criteria that need to be fulfilled in order to consider hair analysis

a valid assessment tool. Panelists encourage assessors to ask: What is the predictive value of a

positive or negative test? Are data available to determine whether the measured level is of

sufficient magnitude to be of pathological or public health importance? The following factors are

key to that determination:

(1) Defining the type of exposure that may have occurred and over what time period. (What

do exposure histories tell us about the likelihood that a particular substance will be in hair

at the time of testing?)

(2) Understanding the type of substance and its behavior in the body. (Are data available that

relate exposure to proportional uptake in hair? Is uptake in hair biologically plausible? Is

it a marker of external exposure?)

(3) Identifying the clinical relevance of a positive or negative finding. (Are any dose-response

data available that will make useful interpretations possible?)

The panel provided this specific input on when hair analysis can be useful:

• From an exposure perspective, hair analysis can be useful for simply identifying or

confirming exposures. Issues raised or reiterated included (1) the difficulty in

distinguishing between internal and external contamination, (2) the qualitative nature of

any such finding, (3) the inability to confirm the source of the substance under study, (4)

the dilemma of not being able to “take it to the next step” (i.e., to use the results as a

clinical tool). To overcome issues 1 through 3, it was noted, it may be more feasible for

some substances to confirm the contamination source (e.g., based on the specific

signature of the substance[s] of interest). Also, more sophisticated studies (e.g., looking at

stable isotopes of certain metals) may now be possible (TC, MK, SS, LW).
ATSDR Hair Analysis Panel Discussion

6-3

• According to the current science, the primary utility of hair analysis is as a measure of

historical exposure. The research focus needs to be on seeking data that establish doseresponse relationships (SS). 

• From a clinical perspective, the following conditions must be satisfied before hair

analysis can be viewed as a reliable means to measure a particular substance: (1) hair

contains a substance concentration that correlates with body organs, tissues, or fluids; (2)

correlates exist and are predictive from a clinical and/or forensic perspective; and (3) hair

can be used reliably to sample individuals, groups, and/or populations to measure the

substance (RB).

• Theoretically, potential substances for which hair analysis may be useful include those for

which the route of exposure would limit external contamination and those for which a

metabolite might be measurable (MK).

• Because of general hair growth and cutting patterns, for exposures longer ago than a year

or quite recent, hair analysis is not useful (RB, LW).

• Depending on the test or element under study, a negative test can help to rule out an

exposure and any potential problem. Again, “negative” needs to be defined. That is, what

is elevated (RB, MK, TC, LW)?

• Before considering hair analysis, a practical consideration is questioning whether there

are any laboratories available that provide cost-effective services and reliable results

(DP).

6.3 What Are the Limitations of Hair Analysis? What Data Gaps and Research Needs

Exist?

Throughout the 11⁄2-day meeting, the group identified various factors that currently limit the use

of hair analysis in evaluations of environmental exposures. No specific research agenda was
ATSDR Hair Analysis Panel Discussion

4One panelist cited a pre-print of a paper by Jason Ditton, professor of criminology at Sheffield

University, England, as a good overview of the potential problems associated with interpreting hair

analysis results, which he felt were on par with panel discussions. The paper highlights uncertainties and

intra-individual variability in hair growth rates and substance-specific incorporation rates. It also

describes the challenges of external contamination issues, including variability in results depending on

wash procedures. The paper concludes that hair analysis is not an “absolute dosimeter,” but rather a

“chronometrically operating relativistic dosimeter” (RB).

6-4

proposed, but gaps in the scientific data were clearly identified.4

 The limitations and data gaps

were recapped by the panelists as follows:

• The lack of standard procedures for sample collection.

• The lack of standardization of methods and quality assurance/quality control (QA/QC)

among laboratories.

• The possible over-interpretation of results far beyond the current body of scientific data

and in light of limitations of techniques and procedures.

• External contamination from a variety of sources, which lowers sensitivity (e.g.,

environmental, hair treatments, personal hygiene, and others). 

• The lack of a body of evidence to demonstrate the effect of washing hair on analytical

results.

• The lack of reference ranges in which to frame the interpretation of results. Reliable

reference ranges are needed—specifically, background or expected ranges in different

geographical areas or regions. Reference ranges should be applicable to population of

interest. The DiPietro (1989) data are a good start, but more data characterizing regional

differences are needed.

• The lack of data related to uptake/incorporation of environmental contaminants into hair.

For both metals and organic compounds, neither kinetic models nor metabolite data are

known or fully understood. Identifying metabolites of substances of interest would be

helpful, because they could serve as markers of internal exposure.

• The lack of correlation between levels in hair and blood and other target tissues.

• The lack of an epidemiologic database linking substance-specific hair levels and health

end points. 
ATSDR Hair Analysis Panel Discussion

6-5

It was re-emphasized that identifying measurable levels of particular substance in hair

does not mean an adverse effect will occur or has occurred. From a medical perspective,

many panelists felt strongly that there is little point in performing hair analysis for a

substance if the findings cannot be used as a diagnostic aid. Justification needs to be

provided for choosing hair analysis over blood or urine analysis, and a connection to a

clinical endpoint is needed.

• A limited knowledge of the biological variations of hair growth with age, gender, race,

and ethnicity.

• Insufficient data on environmentally relevant organic compounds in hair. However,

information on testing for pharmaceuticals and drugs of abuse may have value for those

looking at organic compounds.

Panelists repeated, throughout the discussions, the risk communication challenges that exist with

any exposure or diagnostic tool. The limits of the state of knowledge need to be communicated as

clearly as possible by laboratories, practitioners, ATSDR, etc. (RB, MG).

6.4 Recommendations

Panelists’ recommendations focused on measures to standardize sampling protocols. The group

agreed that such efforts would improve the overall usability and reliability of testing data. The

group discussed sample collection, handling, and processing procedures. One panelist

recommended considering hair analysis results only if the laboratory documents good practice in

terms of handling and validation protocols (MK). It was also recommended that the

governmental, commercial, and research laboratories pool their experience and help develop

standard protocols (SS). Panelists offered the following specific recommendations:

• Standardize sample collection procedures. Samples should be ordered by a physician,

taken for a defined reason, properly collected, and dealt with according to proper chain of

custody procedures. A determination needs to be made regarding the best location on and

distance from the scalp to test. No consensus was reached on the preferred cutting device.

To avoid metal contamination, some panelists recommend using quartz or plastic or

teflon-coated shears. Others questioned whether it really made that much of a difference.
ATSDR Hair Analysis Panel Discussion

6-6

Most important, everyone agreed, is for the laboratory to demonstrate the extent of

contamination introduced, if any, during sample collection. Lastly, sample handling

(chain of custody) procedures should be the same as those applied to other environmental

samples.

• Collect exposure histories. Several panelists recommended obtaining exposure histories

concurrent with collecting hair samples. Information should be collected for the year prior

to the collection date, although one panelist pointed out that recall bias may likely be a

limiting factor. Histories should consider environmental and treatment exposures. It was

recommended that the questionnaire that has been used by CDC be used as a starting

point or model. Lastly, any such questionnaire should be substance-specific.

• Establish quality assurance protocols. Use quality assurance methods for laboratory

analyses recommended by the World Health Organization (1994). Specifically, (1)

reference samples of the same matrix (hair) with known concentrations of the metal

should be used as standards, (2) reference samples should contain the metal at

approximately the same concentration as the sample, (3) if such reference materials are

not available, analysis of quality-control samples at different laboratories by different

analytical methods must be used, and (4) because results may vary over time and for

different metals, results should be present for the corresponding time periods and metals

(SS).

• Require external validation. Require performance evaluations of hair testing laboratories

in the form of proficiency testing (e.g., running reference samples and evaluation of

materials of unknown content). The Center for Toxicology in Quebec occasionally offers

a hair analysis sample for ICP-MS (DP).

• Require documentation. Testing laboratories need to be challenged to make a deliberate

day-to-day effort to demonstrate internal and external validation. Calibration and quality

assurance methods need to be well-documented (DP, MK).

• Encourage targeted analyses. Target testing to the specific element of interest. Testing

for multiple analytes increases uncertainty. Overlapping peaks may lead to the

misinterpretation of results (MK).

• Develop washing protocols. Differing opinions were voiced regarding whether hair

samples should be washed, but the panelists generally agreed that the effects of washing,

when performed, need to be clearly documented by the laboratory. Individual panelist

input is summarized below. 
ATSDR Hair Analysis Panel Discussion

6-7

– The determination of whether or not to wash the sample is a substance-specific

decision (SS). 

– Insufficient data exist to measure the true effects of washing, so washing adds

another layer of uncertainty when data are interpreted (MK).

– One panelist recommended examining the wash solution when washing (RB), but

others questioned how to interpret the resulting data, fearing that it may add yet

another layer of uncertainty (DP, MK).

6.5 Next Steps To Be Taken by ATSDR

Dr. Susten described ATSDR’s anticipated next steps related to evaluating the utility of hair

analysis. First, a summary report of this panel meeting will be generated and released (the report

will be posted on ATSDR’s Web site). Second, ATSDR will generate a report related to lessons

learned from the panel discussions (and possibly publish it in the open literature). In addition,

internally, the agency plans to do the following:

• Continue to provide education to physicians and other health professionals.

• Develop a generic fact sheet to help health assessors and communities communicate and

understand hair analysis issues.

• Continue to develop substance-specific toxicological profiles. The profiles are an

excellent resource and contain information on biomarkers of exposure. In light of the

panel discussions, additional language may be added regarding hair analysis (e.g., in

terms of limitations, etc.) on a substance-specific basis.

• Develop guidance on hair analysis to support public health assessments and health studies

conducted by the agency. That is, develop criteria for determining when to consider hair

analysis as part of an ATSDR exposure investigation.

These activities will help all of ATSDR’s divisions as well as professionals in the community.
ATSDR Hair Analysis Panel Discussion

7-1

SECTION 7

OBSERVER COMMENTS

On both days of the panel discussions, observers were given the opportunity to provide input on

issues related to the charge questions and panel deliberations. Observer comments received

during the meeting are summarized below, alphabetized by observer’s name. A full list of

observers and their respective positions and affiliations is included in Appendix F. Observers

were asked to provide appropriate references and data to support their statements where possible.

Statements provided without reference are included, but have not been verified or validated by

ATSDR or the panel. In some cases panelists responded to a particular observer comment or

question; such responses are summarized in this section as well.

Observers were also encouraged to provide written comments after the June 12–13, 2001, panel

discussions. Appendix G includes written comments from two individuals.

Erik Auf der Heide

ATSDR

Dr. Auf der Heide commented that considering sensitivity, specificity, and predictive value is as

important as the reference range when interpreting laboratory data.

Sherlita Amler

ATSDR

Dr. Amler, a pediatrician, stressed her observations of over-interpretation and misinterpretation

of hair analysis results.

She noted that a lack of knowledge exists among health care providers in terms of how to use

hair analysis, citing two examples. She described a case of an autistic child with reportedly high
ATSDR Hair Analysis Panel Discussion

7-2

levels of mercury in his hair. The physician presumed that the elevations were due to his

immunizations and ordered chelation in hopes of improving the autism. In another case, the

interpretation of hair analysis results of a Down’s Syndrome child as a dietary insufficiency led

to the administration of high vitamin doses and an unusual diet. (Dr. Clarkson raised the point

that misuse or misinterpretation of laboratory tests is not unique to hair analysis.)

Gary Campbell

ATSDR

Dr. Campbell emphasized the need to clearly define “normal” and “reference” ranges and to

describe how these ranges are developed in the various laboratories. Understanding the meaning

and derivation of such ranges is very important to individuals who need to interpret site-specific

hair analysis results and understanding whether results may be elevated. Further, Dr. Campbell

questioned what is known about possible geographical or regional differences in background

concentrations of various substances in hair. 

Robert Jones

CDC

Dr. Jones requested that the panel and ATSDR consider the following:

• Evaluate substances on a species-specific basis, not just on an element basis. Looking at

the form in which elements such as arsenic, mercury, and selenium are present in hair

may help to distinguish exposures due to the form released from a Superfund site from

exposures to a form originating from another source.

• If ATSDR is considering hair analysis in its public health assessments, begin the process

of generating substance- and species-specific quality control reference materials as soon

as possible. Generation of such reference materials can take years.

• Include handling procedures and short- and long-term storage requirements (e.g.,

container and climatic conditions) in any standard protocol.
ATSDR Hair Analysis Panel Discussion

7-3

• Do not standardize hair analysis procedures too highly or you risk stifling innovation by

laboratories. Strict standardization will not guarantee good quality control. Specific

procedures or technologies should not be required as long as the laboratory can

demonstrate the quality of its results. Proficiency systems (daily and longer-term), as

recommended by the Clinical Laboratory Improvement Act (CLIA), are encouraged.

Melody Kawamoto

CDC/National Institute for Occupational Safety and Health

Dr. Kawamoto presented a schematic that integrated many of the concepts and issues being

discussed by the panel (see Figure 7-1). She explained the interface between the many

compartments within the body and how different testing methods help piece exposure

information together. Specifically, Dr. Kawamoto discussed how different methods help

assessors identify potential (environmental media sampling), external (wipes, breathing zone air

samples, hair), and internal (hair, blood, urine) exposures to a particular substance and how that

information may be integrated to evaluate potential health effects. She emphasized the

importance of establishing a framework under which to conduct exposure and health effects

evaluations, including clearly identifying the problem and the hypothesis under which you will

proceed, identifying study design issues, and understanding sampling and analytical issues.

David Mellard

ATSDR

In reference to the arsenic conference held in San Diego in 2000, Dr. Mellard commented on a

study in which a single volunteer showered in arsenic-contaminated water to help better

understand internal versus external contamination. The study revealed that up to a certain level,

no change in arsenic levels in hair were observed. Dr. Mellard suggested that perhaps further

study is worthwhile to see if, for relatively low levels of arsenic in water, hair could be used as a

measure of internal contamination, without worrying about external contamination.
ATSDR Hair Analysis Panel Discussion

7-4

Dr. Kosnett responded with a few words of caution: In vitro experiments have shown that

external absorption is dependent on time. Therefore a single showering episode may not reflect a

longer-term exposure or exposure through bathing. Having reviewed the literature, Dr. Kosnett 

indicated that he is not convinced yet that any cut-off point exists at which there is no element of

external uptake of arsenic in hair from bathing.
ATSDR Hair Analysis Panel Discussion

7-5

Figure 7-1. Evaluation and Solution of Environmental and Occupational Health Problems: Critical Analysis

in Practice1

Model: CauseEffect

<

Source <Exposure <Effect

Indicators of

exposure and

effects

Substances

known to

be present

at site

Potential

dose

<

External dose <

Internal

dose

“Abnor

mality”

<

Disease or

other

health

effect

Possible

detectable or

measurable

parameters

(examples)

Inventories

of raw

materials,

byproducts

, and final

products

Concentr

a-tions in

air, soil,

water, and

surface

wipe

samples

Personal air

samples,

wipe samples

of skin, hair

analysis;

dosimetry

Concentrations in

blood,

urine, and

other body

tissues and

fluids;

dosimetry

Biomarkers

or other

indicators

Biomarkers

or other

indicators

Type of

assessment

Document

review

Environmental Biological Biological and other

I

S

S

U

E

S

Model • Hypothesis or problem statement clearly defined

• Scientific plausibility (e.g.,toxicity, biokinetics, relationship between time of exposure and time of

assessment)

• Motives and desired results

• application of scientific methods to research theoretical questions

• application of available scientific knowledge to provide answers to questions from the

public

Public

health

criteria

and study

design

• Selected parameters to be measured are valid with respect to the model

• Interpretation possible (e.g., dose-response relationship, population norms, intra- and

interindividual variability)

• Relevance of interpretation to the problem statement (e.g., prevention possible) or to questions

from the public (e.g., predictive value, risk communication)

• Feasibility (technical feasibility and cost feasibility)

• Timeliness

• Ethics

Collection

and

handling

• Contamination

• Stability during transport or storage

• Preparation methods (e.g., cleaning, digestion)

Laboratory

methods

• Validity, reliability, accuracy, precision, sensitivity, specificity

• Quality control (proficiency tests, coefficient of variation)

1 Dr. Kawamoto provided this schematic following the meeting, as a work in progress, as a visual

display of the various concepts presented as part of the panel discussions. It is an expanded version

of a hand-drawn figure presented at the meeting.
ATSDR Hair Analysis Panel Discussion

7-6

David Quig

Doctor’s Data

Day #1

Dr. Quig, from Doctor’s Data (a commercial laboratory), expressed extreme gratitude for being

invited to this meeting and offered his opinion on a variety of topics related to analytical methods

and factors affecting the interpretation of laboratory results:

• As a screening tool, no one laboratory test exists that is absolutely definitive. It is critical

that hair analysis results be looked at in careful consideration of patient symptoms and

exposures. Hair analysis is not a test to end all tests.

• A targeted approach is necessary for certain elements. There is no question, for example,

that chromium is extremely difficult to measure. One laboratory using high-resolution

mass spectrometry is getting closer to being able to measure Cr6+ in blood. However,

interference problems do not exist for all the elements.

• Hair treatment is an important issue and clearly affects hair analysis results. Dr. Quig has

worked on a study of 150 hair products (pre-published status); the most common

contaminants identified include tin, aluminum, silicone, and phosphorous. Only two

products have been found to contain mercury and arsenic (Denorex and Aquanet), which

could confound hair analysis for these elements.

• Ethnicity/race needs to be factored in when evaluating hair analysis results. For example,

the reference ranges for Caucasians should not be used for African Americans. The basic

profile is very different between the two.

• With respect to growth rates, the difference between the very young and the very old is

significant.

• Distinguishing internal versus external levels is impossible. Some laboratories claim they

have an algorithm for making such distinctions. Any such claim should be seriously

questioned.

• In Dr. Quig’s experience, laboratories do take into account the type of container in which

samples are stored.
ATSDR Hair Analysis Panel Discussion

7-7

• Using hair analysis for an individual can be acceptable and useful—for example, when

tracking occupational exposures of a particular person over time (e.g., a worker exposed

to lead).

• Washing procedures are a critical part of the hair analysis protocol (with the possible

exception of methyl mercury testing). It would not be desirable, for example, to test

unwashed dreadlocks.

• The only time Dr. Quig has seen significantly elevated mercury in hair levels in non-fisheating individuals is with dentists exposed occupationally to mercury vapor. In

questioning whether this was internal or external contamination, a comparison of scalp

and pubic hair confirmed equally high levels; this suggested internal exposure. Again, it

is critical to look at hair analysis screening in context of other measurements (e.g., blood).

• As indicated by the panel, it is important to realize that the presence of organic toxins

(e.g., DDT) is not “normal.” It is equally important to recognize that we are all subjected

to exposure to a variety of organic compounds and toxic metals. It is therefore important

to consider multiple exposures.

• Standardization of laboratories is a necessity. The same methods and sensitivities should

be required. It is not surprising that Seidel et al. (2001) found different reference ranges

across the laboratories studied, because the laboratories used different analytical methods

(i.e., ICP-MS versus OES) that have a 1,000-fold difference in the detection limits. This

discrepancy should not be used as a reason for not using hair analysis, but as the impetus

for advocating standard protocols. 

Day #2

Dr. Quig provided more comments toward the end of the second day of the meeting. His stated

opinions are summarized below:

• If done correctly, hair analysis can be a useful tool.

• No question exists that gross ineptness has been observed at some commercial

laboratories. The issue of interlaboratory differences is not sufficient reason, however, to

conclude that hair analysis is not of value. It is simply a question of tightening up

sampling/analytical protocols and QA/QC procedures.
ATSDR Hair Analysis Panel Discussion

7-8

• Regarding quality control issues, Doctor’s Data has been pressing for the establishment of

standardized procedures for hair analysis under CLIA and the Health Care Financing

Administration. The fact that procedures are not yet in place is not a reason not to do hair

analysis; it is a matter of the organizations catching up with the needs of the time.

• Regarding washing protocols: A laboratory should produce a reasonable report describing

its washing protocol. The user of the data should look for this information before

interpreting the data.

• A standardized procedure can and should be set for sample collection.

• Statements by panelists regarding the over-interpretation and misuse of hair analysis were

not relevant to the specific charge of this meeting and should not be of concern to

ATSDR.

• Doctor’s Data only accepts hair samples from licensed physicians or for research

purposes. Dr. Quig agreed that hair samples should only be submitted by trained

practitioners.

• Dr. Quig suggested looking at research conducted by Needleman (University of

Pittsburgh) and Masters (Dartmouth) before dismissing the utility of hair analysis for

evaluating lead exposures.

• Sound literature does exist on manganese and aberrant behavior, although the literature is

criticized by the panel. Dr. Quig referenced a follow-up study comparing manganese

levels in prisoners committing violent versus nonviolent crimes. With regards to the

symptoms and the neurotoxicity of manganese, psychological effects range from apathy

progressing to violent reactions and loss of tolerance. The physiology of manganese

toxicity is well-established in the literature. Manganese has a high propensity to bind to

myelin pigmented dopaminergic neurons in the brain.

• Reference ranges are not based exclusively on small data pools (e.g., “n=2”), as suggested

during some of the panel discussions. Available reference ranges are based on 28 years of

doing hair analysis. As methods improve, so will reference ranges. Data sets are

expanding to include documentation of variations in levels of elements between

Caucasians and African Americans, as well as transcontinental differences.
ATSDR Hair Analysis Panel Discussion

7-9

Barry Sample

Quest Diagnostics

Dr. Sample speculated on the possible value of measuring wash solutions as well as washed hair

in attempts to further distinguish between internal and external exposures. Wash solution may

provide a better sense of external levels and the hair may provide a better indication of the total

internal burden. At a minimum, Dr. Sample suggested incorporating wash evaluation into any

standard protocol.

Based on his experience looking at drugs, Dr. Sample acknowledged that data may not exist to

set the “normal range.” In order to do so, one needs to understand the different rates and methods

of incorporation into the hair. He suggested that there may be some value, in an occupational

setting, in developing an individual reference range.

In response, Dr. Kosnett commented that workers may not be the best population to study for

normal ranges because of the potential for external exposures in various work places. Dr. Seidel

noted that further research is needed into the utility of studying wash water. Studies suggesting

that easily removed fractions represent exogenous sources and the not so easily removed fraction

represents endogenous sources have been disproved.

Michael Schaffer

Pyschemedics Corporation

Day #1

Dr. Schaffer, a trained industrial toxicologist with an interest in criminal justice and forensics,

explained that Pyschemedics performs hair analysis as part of workplace drug testing. He asked

participants to keep an open mind and consider the science of hair analysis very carefully.

Knowledge gained from the last 10 years of testing hair for drugs of abuse can, he said, be used
ATSDR Hair Analysis Panel Discussion

5Dr. Baratz clarified that his purpose in presenting the Ditton paper was to summarize some of

the key aspects and possible pitfalls of hair analysis. Dr. Baratz noted that the author, a chemist, has done

studies on drugs of abuse and has shown the validity of hair analysis for testing drugs of abuse.

7-10

to enhance the knowledge base for using hair analysis for environmental/public health

evaluations. He stressed that his experience in the drug testing arena has revealed that hair

analysis is not totally unreliable. Good science and good analyses have supported legal cases. If

the proper analytical tools and washing procedures are used, valid interpretations can be made.

Dr. Schaffer recognizes that drugs of abuse are different than trace metals. Working with mass

spectrometry, metabolite profiling has helped identify uniquely internal measures of the

substance of concern. It has taken 10 years, but such tools are now available.

Dr. Schaffer stressed that hair offers a unique matrix, recognizing that there is much that is not

known or understood. In time, he feels, hair analysis will likely provide a lot of useful

information.

Day #2

Dr. Schaffer expressed concern that some of the statements made during the panel discussions

could be misinterpreted or used inappropriately. Specifically, he wanted to make certain that

caveats were provided with panel conclusion statements so that it is clear that hair analysis for

substances of abuse is appropriate and based on good science; the conclusions drawn by the

panel should apply to environmental contaminants only.

Dr. Schaffer also responded directly to Dr. Baratz’s overview of the Ditton paper.5

 He took

exception to the implication that hair analysis may not be suitable for testing drugs of abuse. He

stated that conducting hair testing with the proper safeguards is defensible and has been upheld

by the courts. He noted that no hair color or ethnicity bias exists. In vitro studies have shown
ATSDR Hair Analysis Panel Discussion

7-11

incorporation of drugs in different types of hair, but those drugs can be removed by washing as

quickly as they are bound to hair. The Department of Health and Human Services (Substance

Abuse/Mental Health Services Administration) is currently writing draft guidelines for the

incorporation of hair analysis into the federal workplace drug testing program. A pilot

proficiency survey is also available to help address quality control issues; the model is urine drug

testing.

Subsequent to the June 12–13, 2001, panel discussions, Dr. Schaffer submitted additional

comments and supporting literature. He provided (1) a partial listing of those cases

demonstrating judicial acceptance of the Psychemedics hair analysis method, (2) information on

hair testing and racial or color bias, and (3) information on the effectiveness of Psychemedics’

washing procedures for ruling out external contamination. (See Appendix G.)

Margaret Schonbeck

Colorado Department of Public Health and Environment

Ms. Schonbeck questioned whether hair analysis would be a valid consideration at an arsenic

exposure site (soil pica/soil ingestion) where urine sampling is already planned.

Dr. Kosnett commented that a hair assay could reveal the potential for exposure, but that

environmental and urine data will have already provided that information. It is not likely that hair

analysis would provide additional insight. Dr. Baratz re-emphasized that one must examine the

clinical utility before considering hair analysis. Does it have any predictive value? Without

symptom or disease history, or unless you have a quantifiable dose-response relationship, hair

analysis data will not help. Dr. Baratz expressed concern that collecting hair samples as another

means of documenting exposure will only muddy the waters. Dr. Seidel suggested collecting,

analyzing, and archiving the data, but being clear with the community up front what the data can

and cannot be used for. Dr. White emphasized the distinction between medicine and public
ATSDR Hair Analysis Panel Discussion

7-12

health, which can sometimes cause confusion and tension in the community. That is, medicine is

looking at the individual and treatment options, while public health is looking at populations and

possible risk factors.
ATSDR Hair Analysis Panel Discussion

7-13

Anthony Suruda

Association of Occupational Environmental Clinics

Rocky Mountain Center for Occupational and Environmental Health 

Dr. Suruda questioned whether nails are more susceptible to external contamination by metals

than hair. In response, Dr. Kosnett noted that, in some forensic investigations, the distal portions

of nails have shown correlation with poisoning. Some studies have investigated whether the inner

surface of the nail may be less likely to contain elevated levels of arsenic as a result of external

contamination. Study findings suggest that external contamination of nails is an issue as it is in

hair. For example, a study that measured arsenic in nails over time following arsenic ingestion

revealed the following: (1) elevated levels of arsenic were measured in distal segments of

unscraped nails (believed to be deposited by sweat); (2) scraped nails during the same period did

not reveal elevated levels; and (3) samples of scraped nails taken later in time showed elevated

arsenic levels (as a result of the ingestion episode). As with hair, it is questionable whether

methods exist to clearly distinguish between externally and internally deposited contamination.

Dr. Suruda indicated that he was requested to evaluate an individual with peripheral neuropathy 9

months after possible exposures to lead and arsenic. Total arsenic urinalysis had been performed

closer to the time of exposure, but not a fractionated analysis. To evaluate past exposures, a

toenail sample was taken down to the growth plate, which was negative. These results were used

to conclude that the individual had not been exposed to arsenic within the past year.

Dr. Suruda noted that the charge to the panel was to examine aspects of hair analysis related to

public health assessments. Dr. Suruda commented that he is more often faced with questions

from individuals (practitioners, community members) looking for assistance in interpreting hair

analysis results. He expressed hope that the panel and ATSDR will consider the utility of hair

analysis in the assessment of public health as well as for individual assessment. Dr. Suruda noted

that ATSDR’s toxicological profiles and other agency documents have great credibility within

the scientific community and that he looks forward to further guidance (e.g., biological
ATSDR Hair Analysis Panel Discussion

7-14

monitoring guidelines) to assist in his evaluations. Even if all the answers are not available, Dr.

Suruda said, hair analysis should be ranked with other methods of monitoring (e.g, blood, urine).

Regarding research needs, Dr. Suruda indicated the need for a population-based study on how

hair analysis is used and what impact it has had. Questions to consider include: Can it be used to

identify poisoned individuals? How many people are unnecessarily alarmed or mistreated on the

basis of hair analysis? What type of reports do practitioners receive on hair analysis? Dr. Suruda

expressed concern regarding what he referred to as “junk science.” For example, he pointed to a

laboratory report that indicated “lead is slightly above detection limit” and that the “zinc to

mercury ratio is extremely high.” The report indicated that these ratios do not indicate disease;

however, it also indicated that research has shown that this “will eventually lead to other

disturbances in metabolic function.” Physicians and other practitioners need to recognize that

they do not often know what results mean and should be cautious in what they report.
ATSDR Hair Analysis Panel Discussion

8-1

SECTION 8

REFERENCES

The following papers were reviewed by the panelists prior to the meeting for their consideration

when reviewing the charge questions.

Hopps H. The biologic bases for using hair and nail analyses for trace elements. Sci Total

Environ. 1977; 7:71-89.

Miekeley N, Dias Carneiro MTW, Porto da Silveira CL. How reliable are human hair

reference intervals for trace elements? Sci Total Environ. 1998; 218:9-17.

Seidel S, Kreutzer R, Smith D, McNeel S, Gilliss D. Assessment of commercial

laboratories performing hair mineral analysis. JAMA. 2001 Jan 3; 285(1):67-72.

Sky-Peck H. Distribution of trace elements in human hair. Clin Physiol Biochem. 1990;

8:70-80.

Steindel S, Howanitz P. The uncertainty of hair analysis for trace metals. JAMA. 2001

Jan 3; 285(1):83-5. Editorial. 

Wennig R. Potential problems with the interpretation of hair analysis results. Forensic Sci

Int. 2000 Jan 10; 107(1-3):5-12. Review.

Yoshinaga J, Imai H, Nakazawa M, Suzuki T. Lack of significantly positive correlations

between elemental concentrations in hair and in organs. Sci Total Environ. 1990; 99:125-

35.

The next set of references represents those cited in the main text of this report through direct

reference during the meeting or provided after the meeting to support statements made during the

meeting.

Anderson RA. 1997. Chromium as an essential nutrient for humans. Regulatory

Toxicology and Pharmacology. 1997; 26:S35-S41.
ATSDR Hair Analysis Panel Discussion

8-2

Anderson RA, Kozlovsky AS. 1985. Chromium intake, absorption, and excretion of

sumbects consuming self-selected diets. Am J Clin Nutr. 1985; 41:1177-83.

Bader M, Dietz MC, Ihrig A, Triebeg G. 1999. Biomonitoring of manganese in blood,

urine and axillary hair following low-dose exposure during the manufacture of dry cell

batteries. Int. Arch. Occup. Environ. Health. 1999; 72:521-27.

CDC. 2001a. Blood and hair mercury levels in young children and women of childbearing

age-United States, 1999. MMWR Weekly. 2001 Mar 2; 50(8):140-3.

CDC. 2001b. National Report on Human Exposure to Environmental Chemicals. NCEH

Publication 01-0379. http://www.cdc.gov/nceh/dls/report

Chittleborough G. 1980. A chemist’s view of the analysis of human hair for trace

elements. Sci Total Environ. 1980; 14:53-75.

Curry AS, Pounds CA. 1977. Arsenic in hair. J. For. Sci. Soc. 1977; 17:37-44.

DeFlora S, Wetterhahn RF. 1989. Mechanisms of chromium metabolism and

genotoxicity. Life Chem Reports. 1989; 7:169-244.

DiPietro ES, Phillips DL, Paschal DC, Neese JW. 1989. Determination of trace elements

in human hair. Reference intervals for 28 elements in nonoccupationally exposed adults

in the US and effects of hair treatments. Biol Trace Elem Res. 1989 Oct; 22(1):83-100.

Donaldson RM, Jr., Barreras RF. 1966. Intestinal absorption of trace quantities of

chromium. J Lab Clin Med. 1966; 68:484-93.

Harkey MR. 1993. Anatomy and physiology of hair. Forensic Sci Int. 1993 Dec;

63(1-3):9-18.

Harrington, JM et al. 1978. A survey of a population exposed to high concentrations of

arsenic in well water in Fairbanks, Alaska. Am. J. Epid. 1978; 108(5):377-85.

Hunter D. 1974. The Diseases of Occupations. 5th Edition. Boston: Little, Brown. 1974

Jankovic SM, Jankovic SV. The control of hair growth. Dermatology Online Journal.

4(1):2. http://dermatology.cdlib.org/DOJvol4num1/original/jankovi.html

Kerger BD, Richter RO, Chute SM, Dodge DG, Overman SK, Liang L, Finley BL,

Paustenbach DJ. 1996. Refined exposure assessment for ingestion of tapwater
ATSDR Hair Analysis Panel Discussion

8-3

contaminated with hexavalent chromium: consideration of exogenous and endogenous

reducing agents. Journal of Exposure Anal. and Environ. Epid. 1996; 6(2):163-79.

Leslie ACD, Smith H. 1978. Napolean Bonaparte’s exposure to arsenic during 1816.

Arch Toxicol. 1978; 41:163-7.

Mertz W. 1969. Chromium occurrence and function in biological systems. Physiol Rev.

1969; 49(2):165-239.

Myers RJ, Hamilton. 1951. Regeneration and rate of growth of hairs in man. Ann NY

Acad Sci. 1951; 27(53):562-8.

National Research Council (NRC). 1989. Recommended Dietary Allowances.

Washington, DC: National Academy of Sciences. 1989.

Paschal DC, DiPietro ES, Phillips DL, Gunter EW. 1989. Age dependence of metals in

hair in a selected U.S. population. Environ Res. 1989 Feb; 48(1):17-28.

Schroeder HA. 1968. The role of chromium in mammalian nutrition. Am J Clin Nutr.

1968; 21(3):230-44.

Smith H. 1964. The interpretation of the arsenic content in human hair. J. For. Sci. Soc.

1964; 4:192-99.

U.S. EPA. 2001. Integrated Risk Information System. Methyl mercury (last significant

revision: 7/21/01). http://www.epa.gov/iris/subst/0073.htm

Van den Berg AJ et al. 1968. Arsenic content of human hair after washing as determined

by activation analysis, in DeVoe JR [ed.] Modern Trends in Activation Analysis, Vol. I.

Washington, DC: NBS. 1968; pp. 272-82.

World Health Organization (WHO). 1994. Biological Monitoring of Metals. Geneva:

WHO. 1994.

See also the bibliography of hair analysis references provided in Appendix D.
ATSDR Hair Analysis Panel Discussion

APPENDIX A

List of Panelists 

Panelist Biographical Sketches
 

Agency for Toxic Substances and Disease Registry

Division of Health Assessment and Consultation

 

Hair Analysis: Exploring the State of the Science

Panel Participants

LuAnn White, Chair

Associate Professor

Director, Center of Environmental Public Health

Tulane School of Public Health

1430 Tulane Avenue

New Orleans, LA 70112

Phone: 770-488-8474

Fax: 770-488-8200

E-mail: lawhite@tulane.edu

Robert Baratz

President

Internal Medical Consultation Services, Inc.

159 Bellevue Street

Newton, MA 02458

Phone: 617-594-7776

Fax: 617-332-3063

E-mail: imcsi@rcn.com

Thomas Clarkson

Professor

University of Rochester

P.O. Box EHSC

Rochester, NY 14642

Phone: 716-275-3911

Fax: 716-256-2591

E-mail: twc30@aol.com

Michael Greenberg

Professor of Emergency Medicine

MCP Hahnemann University

3300 Henry Avenue

Philadelphia, PA 19129

Phone: 215-842-6545

Fax: 610-964-9003

E-mail: mg43@drexel.edu

Michael Kosnett

Associate Clinical Professor

University of Colorado Health Sciences Center

Division of Clinical Pharmacology & Toxicology

1630 Welton Street - Suite 300

Denver, CO 80202

Phone: 303-571-5778

Fax: 303-892-5628

E-mail: michael.kosnett@uchsc.edu

Dan Paschal

Research Chemist

Centers for Disease Control

4770 Buford Highway, NE (MS F18)

Atlanta, GA 30341

Phone: 770-488-7985

Fax: 770-488-4097

E-mail: dcp2@cdc.gov

Sharon Seidel

Toxicologist

California Department of Health Services

Environmental Health Investigations Branch

1515 Clay Street - Suite 1700

Oakland, CA 94612

Phone: 510-622-4500

Fax: 510-622-4505

E-mail: sseidel@dhs.ca.gov

Technical Coordinators:

Deanna Harkins

Medical Officer

Division of Health Education and Promotion

Agency for Toxic Substances and Disease Registry

1600 Clifton Rd., NE

Atlanta, GA 30333

Phone: 404-639-4669

Fax: 404-639-6207

E-mail: dharkins@cdc.gov

Allan Susten

Assistant Director for Science

Division of Health Assessment and Consultation

Agency for Toxic Substances and Disease Registry

1600 Clifton Rd., NE

Atlanta, GA 30333

Phone: 404-639-0625

Fax: 404-639-0654

E-mail: asusten@cdc.gov
ATSDR Hair Analysis Panel Discussion

A-3

Hair Analysis: Exploring the State of the Science

The Panel

Robert Baratz

Dr. Baratz is the founder of two corporations in the medical device area. He was also the

associate medical director for Harbor Health Services in Boston. He currently provides

consulting services for clients such as the U.S. Food and Drug Administration, ABC News, CBS

News, NBC News, American Dental Association, and the states of Iowa, California, Colorado,

and Minnesota. His area of research interest is the development and formation of the skin and its

appendages, including hair. He is currently working with the state of Wisconsin regarding

chelation therapy and use of hair analyses. Dr. Baratz is the national spokesperson for the

American Dental Association regarding alleged mercury toxicity. He received his A.B. in biology

and his M.D. from Boston University. Dr. Baratz received his Ph.D. in cell biology and anatomy

and his D.D.S. from Northwestern University. He has published more than 150 papers.

Thomas Clarkson

Dr. Clarkson has been a professor at the University of Rochester for over 20 years, where much

of his research has involved conducting epidemiologic-toxicologic studies on populations

exposed to mercury and its compounds and studying the pharmacokinetics of mercury in animals

and humans. He has studied the toxicology of hair analysis, including coauthoring two studies on

the subject, entitled “The biological monitoring of mercury in the Seychelles study” and

“Monitoring methyl mercury during pregnancy: maternal hair predicts fetal brain exposure.” Dr.

Clarkson served as a panelist at an NIEHS Workshop on Assessment of Mercury Exposure. He

has also co-edited various books, including Biological Monitoring of Toxic Metals and Advances

in Mercury Toxicology. Dr. Clarkson is a member of the Society of Toxicology and the Society

for Trace Element Research in Humans. He serves as a reviewer for numerous national and

international scientific journals, such as Journal of Applied Toxicology, British Journal of

Industrial Medicine, and New England Journal of Medicine. Dr. Clarkson received his B.S. in

chemistry and his Ph.D. in biochemistry from the University of Manchester. 
ATSDR Hair Analysis Panel Discussion

A-4

Michael Greenberg

Dr. Greenberg is a professor and toxicology fellowship program director at the Medical College

of Pennsylvania–Hahnemann School of Medicine. He also serves as chief of the Division of

Occupational, Environmental and Hyperbaric Emergency Medicine at the Allegheny University

of the Health Services School of Medicine (Department of Emergency Medicine). In addition,

Dr. Greenberg is a senior consultant for the Philadelphia Poison Control Center. He has

performed more than 1,000 exposure assessments and has extensive experience both interpreting

and clinically utilizing hair testing in forensic, occupational, and environmental toxicology

venues. Dr. Greenberg has published more than 100 articles and abstracts in occupational and

environmental toxicology. He also is the editor in chief of the book Occupational, Industrial and

Environmental Toxicology, published by Mosby, St Louis, Missouri. Dr. Greenberg received his

B.A. and M.A. in biology from Hofstra University, his M.D. from Temple University School of

Medicine, and his M.P.H. in occupational medicine from the Medical College of Wisconsin. He

is board certified in Occupational/Environmental Medicine, Medical Toxicology, and Emergency

Medicine. 

Michael Kosnett

Dr. Kosnett is an associate clinical professor at the University of Colorado Health Sciences

Center. His main specialty area is occupational and environmental toxicology. One of his

research areas is the application of laboratory data to clinical epidemiology and clinical

assessment in humans exposed to heavy metals, particularly arsenic. He has been regularly

involved in reviewing and utilizing quantitative information on human exposures to toxic

substances and has designed and implemented biological monitoring programs. He has served as

a consultant to ATSDR at the Bunker Hill Superfund site in Idaho, and acts as a toxicology

consultant to the California Department of Health Services, Occupational Lead Poisoning

Prevention Program. He also has experience in assessing the impact of toxic substances on

children. Dr. Kosnett was a member of the National Research Council’s (NRC’s) Subcommittee

on Arsenic in Drinking Water and testified on the subcommittee’s behalf to a U.S. Senate

Committee on Environment and Public Health. He contributed to sections of the 1999 NRC

report Arsenic in Drinking Water which discussed the use of hair analysis in assessing exposures

to arsenic in drinking water. He also has served on the Committee on Toxicology of the NRC,

and as vice president of the American College of Medical Toxicology. Dr. Kosnett recently gave

the keynote address at the 2001 ATSDR Partners in Public Health Meeting, entitled “Elemental

Mercury Exposure and Human Health: Controversial Issues Regarding Low Level Exposure.”

Dr. Kosnett received his B.S. in molecular biophysics and biochemistry from Yale University, his

M.D. from the University of California San Francisco, and his M.P.H. in environmental health

sciences from the University of California at Berkeley.
ATSDR Hair Analysis Panel Discussion

A-5

Dan Paschal

Dr. Paschal has more than 20 years experience as a research chemist in CDC’s Nutritional

Biochemistry Branch, Division of Environmental Health Laboratory Sciences, within the

National Center for Environmental Health. He is responsible for establishing and maintaining

“state of the art” analytical methods for toxic elements in biological samples, participating in

transfer of appropriate technology to state or other environmental health laboratories, and

supervision of technical research involving method development and specimen collection

protocols. Dr. Paschal has studied various aspects of hair analysis and published papers related to

the “age-dependence of metals in hair” and “reference intervals for 28 elements in

nonoccupationally exposed adults in the United States and effects of hair treatment.” Dr. Paschal

also consults with a wide variety of state and local health professionals to evaluate analytical

methods, quality control procedures, and exposure assessment approaches. Dr. Paschal received

his B.S. in chemistry and his Ph.D. in analytical chemistry from the Georgia Institute of

Technology.

Sharon Seidel

Dr. Seidel is a toxicologist for Impact Assessment, Inc., a contractor for the California

Department of Health Services. Prior to that she served as a toxicologist for the U.S.

Environmental Protection Agency Superfund Program in Region 9. She has studied the health

implications of a number of environmental exposures, including children’s exposures to

pesticides. Her primary involvement with hair analysis is a recent comprehensive assessment of

practices of commercial laboratories performing a broad suite of mineral analyses being used as a

clinical assessment tool and to identify toxic exposures. Her findings were published in the

January 2001 Journal of the American Medical Association. Dr. Seidel received her B.A. in

chemistry from the University of California, San Diego, and her Ph.D. in pharmacology from the

University of Iowa.

LuAnn White

Dr. White is an associate professor and director of the Center for Applied Environmental Public

Health at the Tulane School of Public Health and Tropical Medicine. She has designed and

implemented the key elements of the Tulane/HAMMER project for the U.S. Department of

Energy to improve environmental restoration and waste management worker health and safety

through education and training. She is an innovator in the use of distant learning technologies.

Dr. White is a diplomate, American Board of Toxicology, and has promoted the use of

Geographic Information Systems for toxicologic research. She developed a model for curriculum

development in environmental health which provides a skills-based approach to environmental

education. Dr. White also serves on ATSDR’s Board of Scientific Counselors. Dr. White

received a B.S. in chemistry from St. Mary’s Dominican College and a Ph.D. in

pharmacology/toxicology from Tulane University.
ATSDR Hair Analysis Panel Discussion

APPENDIX B

Charge to the Panel
ATSDR Hair Analysis Panel Discussion

B-1

Charge to the Panel

Hair Analysis: Exploring the State of the Science

The Agency for Toxic Substances and Disease Registry (ATSDR) is holding a panel discussion

to review and discuss the current state of the science related to hair analysis. ATSDR has invited

a cross section of scientific experts in the fields of hair analysis, toxicology, and medicine to

participate in 11⁄2 days of discussions on a variety of topics, including analytical methods, factors

affecting the interpretation of analytical results, toxicologic considerations, and data

gaps/research needs. The panel will discuss whether hair analysis is a useful tool in evaluating

exposures to hazardous substances present in the environment. The agency will use the input

received during the discussions to develop a framework for determining when measuring

contaminant levels in hair can help support scientifically defensible public health evaluations.

Background 

ATSDR conducts public health assessments to evaluate possible public health implications of

contaminants associated with hazardous waste sites and other environmental releases. An

important step in ATSDR’s assessment process is examining exposures to contaminants under

site-specific conditions and determining whether people are being exposed to contaminants at

harmful levels. In most of the agency’s evaluations, the environmental concentration serves as a

surrogate for “exposure.”

Exposure concentrations, or estimated doses based on exposure concentrations, however,

represent only one factor in a continuum of events that ultimately determine whether exposures

will result in illness. Other factors include exposure conditions and various

pharmacokinetic/pharmacodynamic events (e.g., absorption, metabolism, excretion), as well as

individual variability and susceptibility in the exposed population. To a large extent, ATSDR

evaluates these factors qualitatively in its public health assessments.

To refine its assessments and/or to fill data gaps, ATSDR sometimes identifies ways to more

precisely quantify exposures, such as measuring body burdens of a particular contaminant or its

metabolites (e.g., lead in blood). On a site-by-site basis, ATSDR evaluates what additional

exposure data might be practical and useful to obtain to further support public health evaluations

and ultimately to help determine the disease potential of a particular exposure. ATSDR seeks to

determine the overall utility of hair analysis as one such exposure assessment tool.
ATSDR Hair Analysis Panel Discussion

B-2

Charge to Panel Members

General Questions

ATSDR’s overall goal is to receive expert opinion on the following four general questions

related to hair analysis. Panelists should keep these questions in mind when answering the

specific charge questions that follow.

# When is it appropriate to consider hair analysis in assessing human exposures to

environmental contaminants?

# When is it inappropriate to consider hair analysis in assessing human exposures to

environmental contaminants?

# What data gaps exist that limit the interpretation and use of hair analysis in the

assessment of environmental exposures? What research is needed to fill these data gaps?

# For what substances do reliable hair analysis methods exist (e.g., trace elements, organic

compounds)?

Specific Charge Questions

Discussions on the first day of the meeting will focus on answering questions that pertain to

Topics #1, #2, and #3 below. In asking these questions, ATSDR seeks a critical review and

assessment of the state of the science pertaining to hair analysis. The second day of the meeting

will be devoted to identifying critical data gaps and research needs, and also identifying scenarios

for which hair analysis should/should not be considered in light of limitations in analyzing and

interpreting hair data (Topics #4 and #5).

Topic #1: Analytical Methods. Discuss/review basic sampling and laboratory methodology

used in hair analysis.

# What analytical methods currently exist?

# For what substances do reliable analytical methods exist? 

# For what purposes are these methods typically used (e.g., diagnostics, forensics, industrial

hygiene)?

# What amount of hair is needed? Is it dependent on the substance being tested? If so,

specify substance-specific requirements.
ATSDR Hair Analysis Panel Discussion

B-3

# To what extent are multi-element analytical approaches used? Concern: Accuracy and/or

sensitivity for a specific element may be sacrificed.

# Intralaboratory variability: How variable are results and interpretations?

# Interlaboratory variability: How variable are reference ranges, results, and interpretations?

Topic #2: Factors Influencing the Interpretation of Analytical Results. Discuss to what

extent (qualitatively and/or quantitatively) the following factors influence the interpretation of

hair analysis data. Provide substance-specific examples. Note any additional factors not

highlighted below that are critical to or limit the interpretation of hair analysis data. 

# Variations in sample collection and preparation methods, including:

< Hair sample scalp location and homogenization.

< Laboratory sample preparation and washing methods.

< Laboratory calibration standards and proficiency testing programs (QA/QC

procedures).

< “Normal” reference ranges.

< “Abnormal” or “toxic” concentration ranges.

# Exposure of hair sample to the external environment (e.g., shampoos, bleaches, dyes,

permanent waving, relaxers, styling products, hair sprays, hot dryers and curlers, tobacco

smoke).

# Distinguishing between endogenous and exogenous sources of metals in hair. 

# Distinguishing between exposures associated with site contamination versus exposures

from typical background and other sources.

# Hair color, location of hair on the scalp, and hair diameter.

# Gender, ethnicity/race, diet, age, geographical location, and season.

# Rate of hair growth.
ATSDR Hair Analysis Panel Discussion

B-4

Topic #3: Toxicologic Considerations. Discuss to what extent hair analysis data can be used to

predict adverse health outcomes. Cite specific examples/substances for which information is

available.

# What is known about biological uptake of specific substances and the concentration

delivered to and incorporated into hair?

# What is the relationship between chemical concentrations in the hair and blood

compartment and target organs? For what chemicals does a correlation exist between

specific chemical concentrations in other body tissues, organs, fluids, subcellular

fractions, or metabolic pools?

# What is the dose-response relationship between chemical concentration in hair and target

organ effects?

# Ultimately, what is the relationship between chemical concentrations in hair and disease?

What is the disease-predictive value?

# Is information available defining “normal” ranges of chemical concentrations in hair that

have physiological and health-related significance?

Topic #4: Data Gaps and Research Needs. In light of Topics #1–#3, provide recommendations

to fill data gaps and overcome/clarify limitations in hair analysis and interpretation.

# Specify data gaps and limitations that most significantly limit the use of hair analysis in

public health evaluations, both in terms of analytical methodologies and toxicologic

interpretations.

# Identify hair analysis research needs for ATSDR’s research agenda; identify specific

recommendations for future studies.
ATSDR Hair Analysis Panel Discussion

B-5

Topic #5: Identifying Scenarios for Which Hair Analysis May Be Appropriate. Considering

the factors identified in the matrix below, and in light of issues identified under Topics #1–#4,

discuss when hair analysis may or may not be appropriate for evaluating exposures to

environmental contaminants. This matrix will assist ATSDR in developing a framework or

decision logic for determining when to conduct hair analysis.

Exposure Scenario Chemical/

Exposure

Pathway

Exposure

Chronology

Exposure

Duration

Measurable

Health Effects

(Y/N)

- Individual

- Community 

- Population

(List both

appropriate and

inappropriate

scenarios)

E.g., ingesting

lead in soils

- Past

- Present

- Acute

- Chronic

- Specify

Reference Materials

When addressing the charge questions, please provide citations for references that ATSDR

should consider when evaluating hair analysis issues. ATSDR has developed a bibliography

for your review (primarily post-1985). Please identify additional key studies or papers.

The following journal articles and papers have been provided for panelist review and

consideration. ATSDR selected these papers to represent the breadth of the issues to be discussed

by the panel. Their selection does not indicate ATSDR’s position on any particular issue.

ATSDR recognizes that these only represent a sampling of the many peer-reviewed papers on the

subject of hair analysis. The purpose of reviewing these papers is to help stimulate thought and

discussion related to the charge questions.

1) Hopps H. The biologic bases for using hair and nail analyses for trace elements. Sci Total

Environ. 1977; 7:71-89.

2) Miekeley N, Dias Carneiro MTW, and Porto da Silveira CL. How reliable are human hair

reference intervals for trace elements? Sci Total Environ. 1998; 218:9-17.

3) Seidel S, Kreutzer R, Smith D, McNeel S, Gilliss D. Assessment of commercial

laboratories performing hair mineral analysis. JAMA. 2001 Jan 3; 285(1):67-72.
ATSDR Hair Analysis Panel Discussion

B-6

4) Sky-Peck H. Distribution of trace elements in human hair. Clin Physiol Biochem. 1990;

8:70-80.

5) Steindel S, Howanitz P. The uncertainty of hair analysis for trace metals. JAMA. 2001

Jan 3; 285(1):83-85. Editorial. 

6) Wennig R. Potential problems with the interpretation of hair analysis results. Forensic Sci

Int. 2000 Jan 10; 107(1-3):5-12. Review.

7) Yoshinaga J, Imai H, Nakazawa M, and Suzuki T. Lack of significantly positive

correlations between elemental concentrations in hair and in organs. Sci Total Environ.

1990; 99:125-135.
ATSDR Hair Analysis Panel Discussion

APPENDIX C

Pre-Meeting Comments
Hair Analysis Panel Discussion:

Exploring the State of the Science

Pre-Meeting Comments
June 6, 2001

Notice

This booklet includes the panelists’ pre-meeting responses to the charge questions. It should

be noted that the pre-meeting comments are preliminary in nature. The purpose of these

comments is to stimulate meeting discussions. Some panelists’ technical findings might change

based on discussions during the meeting; therefore, pre-meeting comments should not

necessarily be considered the panelists' final opinions.

Any mention of trade names or commercial products does not constitute endorsement or

recommendation for use.
Table of Contents

Panelist Comments

LuAnn White, Chair .............. .............. .............. ....... C-1

Robert Baratz .............. .............. .............. ............ C-5

Thomas Clarkson .............. .............. .............. ....... C-19

Michael Greenberg .............. .............. .............. ....... C-29

Michael Kosnett .............. .............. .............. ......... C-39

Dan Paschal .............. .............. .............. ............ C-47

Sharon Seidel .............. .............. .............. .......... C-57

Note: Comments have been printed as received
C-1

LuAnn White, Chair
LuAnn White

Overview/Perspective from the Panel Chair

The overriding question for the use of hair analysis in environmental public health is the need to

find reliable methods for assessing chemical exposure of people living in communities near

hazardous waste sites. Hair sampling is tantalizing because it is a biological material that is

readily available, noninvasive, and easy to collect. However, much controversy exists regarding

the use of hair samples as an indicator for environmental exposure, health status, or disease state.

The use and misuse of results from hair sampling has stimulated debate and at times, cast a

shadow over the issue.

Complex questions linger regarding three overarching issues: 1) accuracy and reliability because

of laboratory methods; 2) toxicokinetics of compounds and the biological variability among

individuals; and 3) the relationship of the results to exposure and/or potential disease. Within

each of these issues, multiple questions arise that include, but are not limited to, the reliability

and reproducibility of the analytical methods; interlaboratory variability; types of compounds

suitable for hair analysis; baseline of elements and compounds found in hair—for an individual

and/or populations; influence of distribution, metabolism, storage and excretion on incorporation

of compounds and elements into hair; and duration and level of exposure. Even if all of the

methodological and toxicological questions can be answered, there are still great gaps in our

knowledge as to the relationship between the concentration of a compound/element in hair and

environmental exposure, and then between exposure and disease or reduced health status. Indeed,

a lack of knowledge of these complex interrelationships exists with any biological sample and

prevents full answers to many questions.

While there is much we do not know, there is a body of knowledge on hair analysis. The

challenge is to define the parameters whereby hair analysis can be a valuable tool to assist in

exposure investigations, but to guard against overinterpretations beyond our knowledge and

experiences. Perhaps, identifying the issues will open the door to stimulate research to answer

questions and fill our knowledge gaps.
Robert Baratz
Robert S. Baratz

C-4

Hair Analysis

By Robert S. Baratz, MD, PhD, DDS

Introduction

When physicians study a disease or process, they look for ways to evaluate that process in the

body. Blood and urine are taken for testing because they are easily obtained and can be readily

standardized. Normal values for populations can be set easily with such testing. Blood generally

represents what is inside the body, and urine represents what is excreted from the body.

Hair testing has very limited usefulness in medical practice, because it does not represent either

the tissues inside the body or what is excreted. Hair analysis is only useful for detecting exotic

compounds that are not normally found in the body. Thus, for example, a medicine that someone

is taking, might be detected in the hair. Poisons, such as arsenic, also show up in the hair.

Elements normally found in the body -- such as copper, chromium, zinc, and even lead, mercury,

and uranium-- will show up in the hair, but the levels are quite variable and have little or no

practical or clinical significance.

Analysis of hair won’t tell you about the source of an element found in the hair. Most minerals

obtained by the body come from food or water. Foods are grown all over the country and thus,

their constituents, more likely than not, have come from another region, in some cases, from

another country. People are more commonly drinking bottled water and juices which also are

coming from other regions. Thus, finding something in the hair or body in no way indicates the

source of the material. This is especially problematic when dealing with elements that are

somewhat ubiquitous in the environment. The most common source of lead, for example, can be

from the solder joints of household plumbing. However, lead could also be introduced through

any number of foods, and/or beverages. In some cases, even unglazed pottery used for serving

food can be a source of lead contamination. 

When hair analyses have been done rigorously in quantitative laboratory settings, it has been

pointed out that great care must be taken to avoid possible sources of contamination. First, the

hair itself must be processed in a uniform fashion to avoid introducing any exogenous material.

Metal cutting instruments in sampling hair should be avoided. The hair sample must be

standardized as to region of the scalp, length from the scalp and any washing done of the hair

during processing. Even so, because hair grows at different rates in different people, there is still

a great deal of uncertainty regarding even hair obtained close to the scalp. Many contend that

such hair is of more recent vintage and thus more “representative” of the “body composition”.

There are no data, however, that confirm this idea. Hair seems to grow at an average rate of about

1 cm per month. However, a considerable portion of the hair shaft lies within the skin and thus

hair that has been sampled that has already grown out represents hair that may be as many as

several months old. 
Robert S. Baratz

C-5

Even if hair analysis was a highly reproducible and valid test, done properly, there are virtually

no data for correlation of findings with levels of elemental minerals found in other tissues or

organs. Given the element of interest in the Colorado plateau region, it is important to point out

that radioactive compounds from tailings are unlikely to go into hair. The agent of most interest

is radon which ends up in the body as lead. Because radon exists primarily as a gas, the major

organ that is affected is the lung. This reviewer is unaware of any studies that correlate amounts

of material found in lungs with amounts found in hair. 

As hair is handled in the laboratory, a number of possible contaminants can enter the hair from

solutions used in processing. Rigorous care must be taken to check each and every reagent used

in the laboratory. Even acid reagents can have significant amounts of trace elements within them

when parts per billion are at issue. Water used in the laboratory for washing, hand washing or

even wiping down counters may contaminate samples. Use of vaporization techniques such as

atomic absorption spectroscopy can release agents into the laboratory air which would then end

up contaminating other samples or solutions or both. Laboratory dust must be excluded since it

too can act as a source of contamination. Even powder used to cover gloves of laboratory

workers can result in significant contamination of the laboratory environment. Analyses are often

done at the level of parts per billion and it takes very little contaminating material to change

findings dramatically. 

Many laboratories that handle hair fail to take into account that exogenous contaminants such as

hair shampoos, swimming pools, shower water and the like can all add exogenous agents to hair.

These include: selenium, bromine, zinc, copper and even arsenic. Some elements are removed by

pre-washing before hair analysis. Acetone, a common washing agent, has been shown to remove

sodium, bromine, and calcium. The same solution is known to add copper, iron, manganese, zinc,

and mercury. Even the pH of washing solutions can effect the amounts of lead, mercury and

cadmium found in hair samples.

Topic I – Analytic Methods

Types of Analytical Methods

The principal means for analysis of hair depends on the object of the analysis(es). Simple small

molecules such as trace minerals, can be analyzed either using atomic absorption spectroscopy or

mass spectroscopy. Analysis for organic compounds would depend on the specific compound

being tested. 

Some of these methods are exquisitely sensitive and small quantities of contaminants found in

laboratory air from vaporization, dust, or coatings on lab-ware and/or contamination of test

solutions can significantly affect results. 
Robert S. Baratz

C-6

For trace minerals, results are commonly in the range of parts per billion, or smaller. Thus,

exquisite attention to detail and lab cleanliness must be followed. Reliable analytical methods

exist for detection of most trace minerals, however, quantification becomes an issue particularly

when multiple overlapping peaks with spectroscopy occur. Moreover, sampling errors, and the

nature of the starting material, often inhibit precise quantification. More commonly, qualitative

results can be observed reliably.

For reasons to be discussed in later sections numeric quantitative evaluation of trace mineral

substances is not clinically, forensically, or for matters of industrial hygiene, useful. This is

largely due to the fact that normal ranges have not been established, cannot be established, or are

irrelevant. A finding of an exotic substance that is never normally present is significant.

Similarly, a change in order of magnitude of a trace substance that is normally present, and may,

at high doses, be poisonous, often has clinical and other relevancies. 

Hair analysis has been shown to be quantitatively useful for the detection of arsenic, and methyl

mercury. Other validated uses of hair analysis are for the finding the presence of drugs of abuse

or the presence of certain pharmacological agents. 

In this reviewer’s experience, commercial laboratories, as opposed to research laboratories, have

been observed to have considerable variation in their performance. This variability is the result of

inconsistent specimen preparation, source, and handling, inconsistent use of standards, and lack

of multiple runs of the same material. Typically, only single samples are run and thus any

variability within the laboratory and/or method are often unknown. Where multiple samples have

been observed from the same laboratory on the same material, wide variations have been shown

to exist. In the case of commercial laboratories, interpretation of results suggests that results are

often misleading, inappropriate, and lack sufficient information to make them useful. Two

reports by Barrett ( 1985) and Seidel and colleagues (2001) show that, at least in the case of

commercial laboratories, reference ranges, results and interpretations vary considerably from

laboratory to laboratory. This is not surprising considering the milieu in which this work is done,

and the factors described above.

Topic II- Factors Influencing the Interpretation of Analytical Results

Regionally, there can be marked differences in elemental composition of hair even for the same

element. For example, in 16 different regions of the scalp, antimony content was shown to vary

considerably. Even with a person with a standardized diet and living conditions, the composition

of hair at different distances out from the scalp itself can vary. This has been shown, in particular,

for copper and zinc. 

Additional problems in doing hair analysis show that there are difficulties in trying to measure

more than two or three elements at the same time. In atomic absorption spectroscopy, one of the
Robert S. Baratz

C-7

more common methods for hair mineral content analysis, many elements give off multiple peaks

which overlap. These absorption peaks obscure each other, negating the ability to do quantitative

analyses accurately.

There is often a lack of precision and standardization in the amount of hair taken from any

particular subject. Unless a uniform sample was taken from which all analyses were done, the

validity of the analysis can be called to question. 

Racial differences among subjects have also been found. There is considerable variability in

calcium, iron, nickel, chromium, manganese, arsenic and lead levels between Caucasian subjects

and blacks. 

Similarly, age is a significant factor in metal composition of hair. Paschal and co-workers (1989)

found marked differences in concentrations of 28 different metals in hair samples of 199 children

compared with 322 adults. Age-dependent increases in calcium, barium, magnesium, zinc and

strontium all occur up to about 12-14 years of age. Aluminum is shown to decrease with age. It

has been hypothesized that metal composition of hair is related to skeletal and bony growth.

Thus, adults undergoing osteoporosis would have differences in their hair composition related to

those who did not have this problem. Similarly, anyone with any kind of bone abnormality would

have findings that are non-standard. 

Most analyses on hair do not correlate positively with concentrations found in organs (Yoshinaga

and co-workers 1990). It is intuitively obvious why this is likely so, since tissue concentrations

involve both uptake and release which vary over time. Hair is essentially a one-way path out of

the body. Likewise, some elements have significant diurnal variation. A good example is

chromium (Sheard and co-workers 1980).

Many elements, when analyzed in the presence of other elements, can give false readings. The

interaction of chromium with other anions and cations in hair may affect analytical results

(Sheard and coworkers 1980). Merely painting the laboratory with particular types of paint,

failure to use HEPA filters on the air intake and the presence of dust can easily affect sensitive

analytical measurements. 

A variety of hair treatments have been shown to alter hair trace element concentrations.

(McKenzie, 1978). Other common issues are dyeing and permanent waving, shampooing, hair

color, sex, seasonal variations, age, and growth rates. It is generally assumed that hair grows

approximately 1 cm per month, however this must be verified in each individual tested. Hair

growth is a function of individual factors as well as protein in the diet.

Many commercial laboratories claim to be able to detect and measure more than 20 elements in a

single sample of hair, however, this is often accomplished without any specific knowledge of the

patient’s medical history. What is more troubling, is that there is no definition of a normal range
Robert S. Baratz

C-8

(Hambridge 1982, Rivlin 1983; Manson and Zlotkin 1985, Barrett, 1985, Druyan and others

1998 and Seidel and others 2001;). Quality control in the laboratory is essential towards having

useful data. Rigorous attention to detail, methodology, and sampling techniques must be

followed. Even when known standards were used, because of the sensitivity of instrumentation,

data varied commonly by up to 10%. (Nowak and Kozkowski 1998). 

Variations in Sample Collection and Preparation Methods

A number of compounding variables limit interpretation of results from hair analysis. It is well

known from the literature that the rate of hair growth varies from person to person, with

nutritional and disease states, with the presence of particular drugs, with gender, age,

ethnicity/race, with site on the scalp and/or other body parts. While some of the factors may be

known in an individual case, others are unknown, or cannot be known. Thus, hair analysis from a

particular individual is fraught with a series of uncontrolled variables and unknown data. It

should be obvious that these belie making any precise quantitative diagnostic or forensic

analysis. This becomes even more of a problem when dealing with trace minerals that are

normally found ubiquitously in the environment and characteristically in foods, water, and air.

Many trace minerals occur in human hair normally. Thus, finding them there is expected. Making

interpretations based upon the quantitative analysis of these is fraught with uncertainty due to the

unreliability of the data regarding exposure, timing, hair growth, treatment of the hair, diet,

nutrition, and a host of other factors mentioned above. 

Similarly, considerable variation exists from laboratory to laboratory in terms of sample

preparation, whether a sample is washed, how it is digested, how long it is digested, and how it is

handled after digestion. 

Even more problematic is the development of “normal ranges” or “reference standards”

(“reference ranges”). In most cases, population standards have not been developed. Thus, each

laboratory has developed its own “reference range”. The major problem with this is that the

source of the specimens used to create the “reference range” in a particular lab may be biased.

Many commercial laboratories accept samples from a variety of practitioners, patients, and other

sources. From reports this reviewer has seen, the precision of medical knowledge and facts

regarding the source material is often poorly documented. Careful attention to uniform sample

collection techniques is often also a problem.

So-called “normal” reference ranges do not exist for most trace minerals found in hair. The

reasons for this are obvious. Considerable variation exists from person to person and the variety

of unknown variables enter into the equation. Thus, there are no standardized “reference ranges”

for most normal trace minerals. This has to do, in part, with the composition of hair. In essence,

hair consists of keratinized or cornified cells packed into tight arrays in the hair shaft. These cells

are fundamentally similar to the epidermis however contain proportionally more keratin fibrils

and somewhat different materials in the thickened cell membrane that is left when the cells
Robert S. Baratz

C-9

keratinize. Hair, nails, horn, and some portions of the filiform papillae of some animal tongues

are a so-called “hard” keratin compared with so-called “soft” keratins found in epidermis and

oral and other, mucosae. All keratinized cells contain virtually no aqueous phase after

keratinization. It is unclear if minerals are removed from these cells when they mature, or merely

remain in the intracellular matrix. Along these lines, even if some minerals were found to be left

“inside” such cells, each and every individual trace mineral would have to be studied in prekeratinized cells and keratinized cells to see how it was handled. 

Moreover, epithelium and its derivatives (hair) is an a vascular tissue with little intercellular

space or material. What little extracellular material exists is primarily a lipid that forms a barrier

to diffusion. The epithelium is neither a gland nor excretory organ but merely forms a protective

layer. Thus, substances that would normally be excreted into various body fluids are normally not

present in epidermal epithelium. Regionally, there is variability in the thickness of the epidermal

epithelium and indeed there is some variability in the consistency of thickness of hair in different

regions of the scalp. Hair in other body locations, axillary, pubic, limb, peri-anal, eyebrows, and

eyelashes, all vary considerably in their structure, function, and growth rates.

Since hair is principally protein in nature, there is little need for trace minerals in the hair cells

themselves. Trace minerals in the body are usually present as co-factors for enzymes. Keratinized

cells are generally non-metabollic. After the filaments of keratine aggregate and are coated by

other proteinatious material, the cell contents become essentially inert. Nuclear materials,

enzymes, carbohydrates, and even lipids are essentially not present in the internal milieu of

keratinized cells. Consequently, there would be no need for a regular array of minerals present

from a functional point of view.

Some heavy metals may distribute into hair and become complexed with hair proteins. This

would be due largely to interaction with free side chains on amino acids and/or forming

crosslinks among protein chains as they may be denatured by heavy metals. Some heavy metals

are well known as protein denaturants, e.g. mercuric chloride. They may become trapped in hair

cells before they become completely keratinized. Whether or not this happens is largely

unknown.

Finding trace minerals in hair is neither surprising nor a consistent finding. Because hair shafts

consist of essentially of two portions, intra-epithelial and extra-epithelial, the possible absorption

of extraneous material is possible in the extra-epithelial portion. The extra-epithelial portion is

essentially free in the environment. Thus it is subject to washing, drying, chemical alteration,

cosmetics, environmental pollutants present in the water or air, and a host of other chemical and

physical insults. Not only may things be adsorbed and absorbed by the hair, but, substances may

also be leached from the hair as well. Prolonged immersion and wetting of hair can cause some

swelling of the cells of which hair is composed. This can diminish the barriers to diffusion of

things both from the outside in, and from the inside out. Moreover, hair is being constantly

exposed to scalp oils, and other glandular products excreted into the hair shaft space by

sebaceous and other glands present in skin. These provide an additional source of extraneous

material to be adsorbed or absorbed onto or into the hair.
Robert S. Baratz

C-10

A common and highly variable factor in hair is hair growth. Hair growth can occur in several

different ways. First, is the fact that hair undergoes a cycle in its normal growth. That is, hair is

regularly shed from the scalp and other locations, and replaced by “new hairs”. The stages of the

growth (catagen, anagen, telogen) each have unknown times associated with them in particular

subjects. Further complicating an understanding of growth is the fact that hair in humans is

known to grow in a mosaic across the scalp. That is, any particular hair may be in a different state

than its neighbors. A long list of drugs, hormones, and other factors can either accelerate or

prolong the time a hair stays in a particular part of its growth cycle. Moreover, a number of other

factors such as diet, nutrition, age, sex, hair color, and other factors are known to influence

growth rates. 

Growth can occur both longitudinally and in diameter. Hair in general varies from individual to

individual in shape and not all individuals have a circular cross section of their hair. In particular,

individuals with highly curled or “kinky” hair have hair that is somewhat flattened to a ribbonlike shape. 

In general, hair growth in length is often described at approximately 1 cm per month. However,

there is considerable variation in this from individual to individual and results can vary by a

factor of 2 either in increase or decrease in rate of growth.

Topic #3: Toxicologic Considerations

As previously mentioned, among the mineral toxic agents studied only arsenic and methyl

mercury have been shown to have reliable information on their presence and distribution in hair

when viewed in comparison to their distribution in other organs.

To have predictive value, the values obtained from analysis of hair of a particular subject must be

capable of yielding data that would be predictive for disease in general. This may prove to be

considerably problematic in the case of heavy metals as the agents themselves may affect hair

growth directly.

This reviewer is less well versed in arsenic and methyl mercury studies than others on the panel

and wishes to defer to their knowledge and experience.

Topic #4: Data Gaps and Research Needs

Many of the data gaps in our knowledge of hair physiology and growth have been discussed

earlier.

In one sense each trace mineral must be independently studied with regard to the best source for

analytical material. In most cases it is likely that hair will prove to be a problematic source.

While hair theoretically gives a longitudinal history of prior events, the speed of that history is

largely unknown, and may even change over time. Whether this theory meets practice is also
Robert S. Baratz

C-11

unknown. A better understanding of the physiology of hair growth is obviously an important area

of research. This, of course, begs the question that hair may be useful at all for mineral analyses.

This later point is yet unproven. A variety of data would suggest that hair is not useful for

mineral analyses for most minerals, and that other body sources would be better—e.g. bone,

teeth. 

Knowledge of the dynamics of incorporation of a variety of environmental toxins, principally

organic compounds, into hair would be desirable. Attendant to such a study would be studies of

absorption, adsorption and leaching of such compounds. 

Studies of the nature of differences in incorporation of materials into hair at different ages, by

different sexes, different ethnic groups, and different hair colors would also be useful.

Topic #5: Identifying Scenarios for Which Hair Analysis May Be Appropriate

Hair analysis appears useful only for population studies where much of the individual variability

can be eliminated. If a number of factors were known—duration of exposure, rates of

incorporation into hair, effects on growth, amounts of leaching, sources of material that were

found in hair, etc.—then useful data on exposure could be extracted. Correlating these with

clinical findings is more problematic, since such are best done on the individual level, where hair

analyses are likely more useful only for population studies.

Particularly for small molecules such as trace minerals, hair is unlikely to prove a reliable source

of material for meaningful study. 

Organic compounds that can be shown to incorporate into hair may be an area where hair

analysis could be appropriate for following exposures to environmental toxins.
Robert S. Baratz

C-12

Selected References

Anderson RA et al. “Designing a Biological Monitoring Program to Assess Community

Exposure to Chromium: Conclusions of an Expert Panel” Journal of Toxicology and

Environmental Heath, 40:555-583, 1993.

Barrett S. “Commercial Hair Analysis: Science or Scam?”; JAMA; 254(8): 1041-1045; 1985.

Barrett, S. “Commercial Hair Analysis: A Cardinal Sign of Quackery”, www.quackwatch.com,

2000.

Centers for Disease Control, “Blood and Hair Mercury Levels in Young Children and Women of

Childbearing Age—United States, 1999”, MMWR, 50 (08); 140-3; March 2, 2001.

Chittleborough G. “A Chemist’s View of the Analysis of Human Hair for Trace Elements”, The

Science of the Total Environment, 14: 53-75, 1980.

Clarkson TW. “The Toxicology of Mercury”, Crit Rev Clin Lab Sci, 34(4):369-403; 1997.

Cornelis R. “Neutron activation analysis of hair failure of a mission” J Radioanal Chem; 15:305-

316; 1973.

Deening SB and Wever CW; “Hair Analysis of trace minerals in human subjects as influenced by

age, sex and contraceptive drug” Am J Clin Nutr ; 31:1175-1180; 1978.

Druyan ME et al. “Determination of Reference Ranges for Elements in Human Scalp Hair”,

Biological Trace Element Research, Vol 62, pp 183-197, 1998.

Fletcher DJ. “Hair Analysis: Proven and Problematic Applications” Postgraduate Medicine, Vol

72, No 5, pp 79-81,84,87-88, 1982.

Gaillard Y and Pepin G. “Testing Hair for Pharmaceuticals”; J Chromatogr B Biomed Sci Appl,

733(1-2):231-46; 1999.

Hambidge KM. “Hair Analyses: worthless for vitamins, limited for minerals”, Am J Clin Nutr;

36(5): 943-9; 1982

Hopps HC. “The Biologic Bases for Using Hair and Nail for Analysis of Trace Elements”, The

Science of the Toxic Environment, 7: 71-89; 1977.

Kidwell DA, Lee EH, and DeLauder SF. “Evidence for bias in hair testing and procedures to

correct bias”; Forensic Sci Int; 107(1-3):39-61; 2000.
Robert S. Baratz

C-13

Klevay LM et al. “Hair Analysis in clinical and experimental medicine” Am J Clin Nutr; 46:233-

6; 1987.

Lamand M., Faviert A., and Pineau A. “La détermination des oligoéléments dans les poils et dans

les cheveux: intérêt et limites”, Annales de Biologie Clinique, 48, 433-442, 1990.

Manson P and Zlotkin S. “Hair Analysis- a critical review”; Can Med Assoc J; 133:186-188;

1985.

McKenzie JM.. “Alteration of the zinc and copper concentration of hair”; Am J Clin Nutr,

31:470-476; 1978.

Mertz W. “Confirmation: Chromium levels in serum, hair and sweat decline with age”; Nutr

Rev; 55(10): 373-5; 1997.

Nowak B and Chmielnicka J. “Relationship of lead and cadmium to Essential Elements in Hair,

Teeth, and Nails of Environmentally Exposed People”; Ecotoxicol Environ Saf; 46(3):265-274;

2000.

Nowak B and Kozlowski H. “Heavy Metals in Human Hair and Teeth: The Correlation with

Metal Concentration in the Environment”, Biological Trace Element Research, Vol 62, 1998.

Paschal DC et al. “Age Dependence of Metals in Hair in a Selected U.S. Population”,

Environmental Research, 48, 17-28, 1989.

Ponce RA et al. “Uncertainty analysis methods for comparing predictive models and biomarkers:

A Case study of dietary methyl mercury exposure”; Regul Toxicol Pharmacol; 28(2): 96-105;

1998.

Rivlin RS. “Misuse of Hair Analysis for Nutritional Assessment”; Amer Jour of Med; Vol 75:

489-493; 1983.

Seidel S. personal communication to John L. Watson, March 6, 2001.

Sheard EA, Johnson MK, and Carter RJ. “The Determination of Chromium in Hair and other

Biological Materials” Hair, Trace Elements and Human Illness, Praeger, NY, 1980.

Sky-Peck HH. “Distribution of Trace Elements in Human Hair”, Clin Physiol Biochem:8:70-80,

1990.

Teresa M, Vasconcelos SD, and Tavares HMF. “Trace Element Concentration in Blood and Hair

of Young Apprentices of a Technical-professional School”, The Science of the Total

Environment, 205, 189-199, 1997.
Robert S. Baratz

C-14

Vir SC and Love AHG. “Zinc and copper nutriture of women taking oral contraceptive agents”

Am J Clin Nutr; 34:1479-1483; 1981

Wennig R. “Potential problems with the interpretation of hair analysis results”; Forensic Sci Int

107(1-3); 5-12; 2000.

Willhelm M and Idel H. “Hair Analysis in Environmental Medicine”, Zeutralblatt for Hygiene

and Unweltmedizin; 198:485-501; 1996.

Yoshinaga J et al. “Lack of Significantly Positive Correlations Between Elemental

Concentrations in Hair and in Organs”; The Science of the Total Environment, 99: 125-135;

1990.

Zlotkin SH, “Hair Analysis: A Useful Tool or a Waste of Money?” International Journal of

Dermatology, Vol 24, 161-164; 1985.
Thomas Clarkson
C-16

Hopps (1977)

He provides background physiology and histology of human hair formation and growth

General questions

He gives no information

Topic 1 Analytical methods

No information

Topic 2 Factors influencing the interpretation of analytical results

He notes various pathways of metal into hair:

1) via the follicle into the hair matrix

2) secretion of metals in the sebum on to the hair surface

3) secretion of metal in exocrine sweat on to the surface of the hair

4) secretion of metals in apocrine sweat on to the surface of the hair.

He notes that apocrine sweat may not be important for scalp hair.

He discusses the relative merits of head versus pubic hair and concludes that scalp hair is to be

preferred

He discusses some reports where lead and arsenic have been measured in scalp hair. The metal

level depend on the distance from the scalp/ Lead tends to increase towards the tip of the hair

strand.

Arsenic appears to be accumulated in hair and may present a historical record of tissue levels.

However hair can accumulate external arsenic in the form of arsenite. Animal experiments

indicate arsenic is excreted in sweat.

Variable data have been obtained with cadmium

He gives a table of normal levels of metals in hair.

He notes that attempts to distinguish external versus internal uptake of metals have usually been

unsuccessful

Topic 4 Toxicological consideration

No information
C-17

Topic 5 Data gaps and research needs

No information

Miekelay et al (1998)

Compared two methods of measuring metal in samples of scalp hair taken from 1,091 adults

living in Rio de Janeiro. They also sent a test sample to commercial laboratories for comparison.

General questions

The article indicates the need to revise reference interval for normal levels of metals in hair

Topic 1 Analytical methods

The article claims that ICP-AES (inductively coupled plasma atomic emission spectrometry is

out of date with poor detection limits but is still used by most commercial laboratories. The

article claims that ICP-MS (inductively coupled mass spectrometry) is the method of choice.

Tables are presented comparing reference limits published by five commercial laboratories

indicating wide differences between laboratories for certain metals. Tables are also presented

indicating wide differences in results for certain metals on two hair samples circulated blind to

the same five commercial laboratories. However, results for some metals yielded reasonable

agreement. These metals included Na, Ca, Mg, Mn, Cu, Fe, Zn. The following metals gave

reasonable agreement if results from one of the laboratories were excluded: Pb, Cd, Ba, Ni, Li, P,

B, Cr, Mo.

Topic 2 Factors influencing the interpretation of analytical results

No information

Topic 3 Toxicological consideration

No information

Topic 4 Data gaps and research needs

The study indicates the need to revise reference limits for some metals.

Sky-Peck (1990)

He performed X-ray fluorescence analysis in six carefully aligned samples of hair from 987

employees and their families at a major medical center in Cook County, Illinois. The purpose

was to elucidate factors that might affect concentrations of trace elements in human scalp hair
C-18

General questions

He concludes that hair analysis should only be used as a screening method along with other

measures of the nutritional status of the patient.

More data are needed on factors affecting trace elements in hair before hair can be used as a

quantitative tool to assess the nutritional status of any trace element.

Topic 1 Analytical methods

He used only X-ray fluorescence analysis. He did not describe how the weight of hair was

obtained. Usually Compton scattering is used to measure the hair mass. This does not appear to

be the method used in this report

Topic 2 Factors influencing the interpretation of analytical results

The method of washing the hair sample can influence the levels of certain trace elements. The

mild washing procedure used in the report did not affect levels of 14 selected trace elements.

Treatment with peroxide produced a statistically significant reduction in S, Ca, Fe, and Zn. The

reduction in Ca was almost complete and Zn was reduced substantially. Hg levels were not

affected.

Permanent waving produced a statistically significant increase in levels of 6 trace elements.

Levels of Ca, Ni and As were more than doubled. Mercury was unaffected.

Brunettes and blondes differed significantly in only three trace elements, F (slightly lower in

blondes); Mn (slightly lower in blondes); and Pb (almost double in blondes). Compared to

brunettes and redheads differed statistically in 5 trace elements. Iron was almost doubled in red

heads. Mercury was slightly reduced.

Blacks differed from Caucasians in 10 trace elements. Ni, As, and Pb in blacks were more than

twice as high as in Caucasians. Orientals differed from Caucasians in 9 trace elements. Ca and Pb

in Orientals were a factor of 2 below corresponding levels in Caucasians. Mercury was the same.

Note: Elements differed according to age. Ca in the older group was less than 50% of the

younger group. Br was five times higher. Hg was unaffected.

The longitudinal profiles differ according to the trace element. The levels of As, Hg, Cu, Fe, Zn,

S and Se were steady and unaffected by distance from the root end. On the other hand, the levels

of Pb, Ni and Mn rose sharply towards the tip of the hair strands suggestion external

contamination. Ca and Sr showed less pronounced changes.

The results indicate that the levels of certain trace elements are influenced by a number of

factors. It would appear that Pb, Ni and Mn are affected by external contamination.
C-19

On the other hand, levels of Hg appear to be robust and unaffected by all but one of the factors

tested in this report. For one factor, natural hair color of redheads versus brunettes, there was a

statistically significant difference in mercury levels, but this difference was quantitatively small.

The most stable trace elements were S, Cu, Zn, Se, Cr, and Rb because these were not changed

by more than a factor of 2 by any of the factors tested in this study. 

The most unstable elements were Ca, which was affected by more than a factor of 2 by five of

the six factors tested. Pb was affected by four factors, and Ni. Br, and Sr by three factors.

Topic 3 Toxicological consideration

No information

Topic 4 Data gaps and research needs

The paper stresses the need for more data on factors affecting levels of trace elements in hair

Seidel et al. (2001)

The authors sent a common hair sample to six commercial laboratories for trace element analysis

Different levels were obtained However, it is difficult to evaluate the data without knowing the

correct level. These levels can be compared to the normal ranges for each laboratory. 

The authors also checked on the accreditation of the labs and on the dietary advice given on the

basis on the findings

General questions

The authors argue that there are few if any trace elements that have been validated as indicators

of dietary sufficiency or of toxicity. Methyl mercury may be the only substance for which toxic

dose response relationships have been established.

Topic 1 Analytical methods

The labs tests used atomic fluorescence or mass spectrometry detection methods. The authors

note that the mass spec. method is much lower detection limits

Topic 2 Factors influencing the interpretation of analytical results

The labs can be compared in terms of identifying with elements are outside their normal range.

All six labs agreed that the following elements were with their normal range: Ba, Be, B, Cd, S

and Ti. All labs agreed that Mn and Mo were outside their normal range. For the following

elements all labs except one agreed on classifying according to their normal range: Al, As, Pb,

Mg, Hg, Ni, and Zn.
C-20

Thus, for approximately half the elements tested, there was reasonable agreement between the

commercial labs.

Topic 3 Toxicological consideration

There is lack of toxicological information of the value of hair element concentration as a

biomarker for tissue levels, especially levels in the target tissue. This information is available

only for methyl mercury.

Topic 4 Data gaps and research needs

As mentioned above, the data gaps are in dose response information and in relating hair levels to

levels in the target tissue

Steindel & Howanitz (2001)

The authors provide editorial comment on the paper by Seidel and provide a discussion of

proficiency testing in clinical chemistry laboratories.

General questions

They point out that the current lack of normal ranges for trace elements in hair make

interpretation of results impossible. They comment of the difficulty of making nutritional

conclusions from hair data

Topic 1 Analytical methods

No information

Topic 2 Factors influencing the interpretation of analytical results

The authors listed many problems in interpretation of hair data including external contamination

and the absence of reliable reference standards and uniform methods for processing the hair

samples.

Topic 3 Toxicological considerations

No information

Topic 4 Data gaps and research needs

More data are needed on inter-laboratory comparisons
C-21

Wennig (2000)

This is a review article on the incorporation of drugs into hair. It presents a useful review of hair

physiology and biochemistry. It gives recommendations for collection and storage of hair

samples.

It gives no information on trace elements in hair.

Yoshinaga et al (1990)

The paper compares the concentration of a number of trace elements in hair with corresponding

concentrations in several organs and tissues obtained at autopsy.

Unfortunately, little detail was given on how the hair samples were collected or on the length of

the hair samples.

General questions

Topic 1 Analytical methods

A commonly used analytical method was used (ICP-AES). Quality control tests were made.

Topic 2 Factors influencing the interpretation of analytical results

The varying length of the hair samples may have influenced the result and accounted for the poor

correlations.

Topic 3 Toxicological consideration

The main finding was that levels of Ca, Mg, P, and Zn in hair did not correlate with tissue levels

or body burden

They were not able to draw any conclusions about Fe, Cu or Se as the appropriate tissues were

not available for analysis

Topic 4 Data gaps and research needs

More information is needed on hair versus levels in autopsy tissues. The hair length should be

restricted to a short segment close to the scalp.
C-22

Hopps HC. (1977). The biologic bases for using hair and nail for analyses of trace elements. The

Science of the Total Environment, 7:71-89.

Miekeley N, Dias Carneiro MW, Porto da Silveira CL. (1998). How reliable are human hair

reference intervals for trace elements? The Science of the Total Environment, 218:9-17.

Sky-Peck HH (1990). Distribution of trace elements in human hair. Clin. Physiol. Biochem. 8:70-

80.

Seidel S, Kreutzer R, Smith D, McNeel S, Gilliss D. (2001). Assessment of commercial

laboratories performing hair mineral analysis. JAMA 285(1):67-72.

Steindel SJ, Howanitz PJ. (2001). The uncertainty of hair analysis for trace metals. JAMA

285(2):83-85.

Wennig R. (2000). Potential problems with the interpretation of hair analysis results. Forensic

Science International 107:5-12.

Yoshinaga J, et al.(1990). The Science of the Total Environment 99:125-135.
Michael Greenberg
Michael I. Greenberg, M.D., MPH

Topic #1: Analytical Methods

Comments:

The laboratory analytical methods available are capable of defining the qualitative existence of a

variety of pharmaceuticals, drugs of abuse, and occupational/environmental toxicants. The

operative word here is “qualitative”. Quantitation of specific levels are not, in my opinion and

experience, either generally reliably reproducible and/or clinically useful. Specific analyte levels

are essentially of little or no value in the determination of so-called cut-off levels (e.g., PELs,

TLVs, “safe levels”), “normal levels,” or other designators which rely on reference levels. In

addition, the analytical techniques currently in use are capable of providing “segmental analysis”

of hair, which in turn can provide a historical picture of various qualitative (not quantitative)

exposures over time. In addition, hair analysis may help to derive an essential time frame which

may indicate, based on the average rate of hair growth, the time of inception for various

exposures.

The amount of hair needed for analysis may be dependent on the specific analyte sought as well

as the temporal relationship between exposure and hair harvest.

One of the most important shortcomings for hair analysis, as it currently exists, is the fact that

reference ranges may often be unreliable. Laboratories frequently base their reference ranges for

specific analytes on limited case reports in the medical literature or exclusively on data derived

from animals, which has limited applicability to humans. These facts contribute to substantial

limitations with regard to interpretation of results. Variability undoubtedly exists from one

laboratory to another. Certainly these facts limit the clinician’s ability to interpret and utilize

hair-derived values beyond the potential qualitative information that might come from hair

testing of any individual.

Thus, at this time, it may be prudent to recommend that hair testing for all substances (drugs of

abuse, occupational toxicants, environmental toxicants) be limited to qualitative determinations
Michael I. Greenberg, M.D., MPH

C-25

as opposed to quantitative determinations. The goal of quantitation for any laboratory analyte is

to derive clinical algorithms that translate into levels that indicate disease, dysfunction, or

specific risks for disease or dysfunction. With regard to hair testing in its current state, there is

little evidence that there is sufficient reliability to use quantitation for these purposes. 

Laboratory washing procedures prior to digestion may significantly alter the hair content of

various analytes. For example, when hair is tested for THC, if it is washed with methanol, THC

concentrations may be reduced by as much as 85% (Forensic Drug Abuse Advisor, 1996) by

virtue of this process. It is reasonable to expect that similar degradations in analyte concentration

occur when other analytes are involved.

Hair pigmentation is a critical factor in the interpretation of the concentration of certain

compounds and their metabolites incorporated into hair. Melanin is responsible for the

pigmentation. The color and the melanin content of human hair samples differs over a wide

range. Once deposited into hair, chemicals may remain detectable for a period of months to

years. However, if disposition into hair is influenced by those properties attributed to hair color,

then certain persons may test positive more frequently than other persons. Removal of the

melanin from hair digests prior to hair analysis may reduce the effect of melanin on the total

chemical concentration by excluding the drug bound to the pigment. In one study (Hold KM, et

al), the effect of melanin removal by centrifugation of hair digests on cocaine concentrations was

investigated. Two sets of hair samples from five cocaine users were analyzed for cocaine and

metabolites. A solution consisting of 10 mL of 0.5M Tris buffer (pH 6.4) to which is added 60

mg D,L-dithiothreitol, 200 mg SDS, and 200 U Proteinase K, was used to digest the hair. Two

milliliters of this solution was added to 20 mg of hair and incubated at 37 degrees in a shaking

water bath (90 oscillations/min) overnight. The samples were removed from the water bath and

mixed. One set was centrifuged at 2000 rpm and divided into supernatant and melanin pellet. The

other set was not centrifuged. Internal standards were added to all tubes. The samples were

further extracted, derivatized, and analyzed by gas chromatography-mass spectrometry. A mean

of 8.8% (standard deviation [SD] 7.0%) of the total cocaine concentration (supernatant and
Michael I. Greenberg, M.D., MPH

C-26

pellet) was left behind in the pellet. The same experiment was repeated—except that the melanin

pellet was redigested with 0.1 N HCl. After redigestion of the melanin pellet, the mean cocaine

concentration in the pellet was 3.8% +/- 4.0% (mean +/- SD) of the total cocaine concentration in

hair. These investigators felt that their data demonstrate that removal of melanin from hair

digests by centrifugation does not eliminate hair color bias when interpreting cocaine

concentrations. 

TOPIC #2: FACTORS INFLUENCING THE INTERPRETATION OF ANALYTICAL 

RESULTS

Comments:

Exposure of hair sample to the external environment could be an important factor 

in confounding results on both a quantitative and qualitative basis. By way of example, 

many over the counter hair coloring preparations contain lead acetate (e.g., Grecian formula).

This may persist for long periods on hair shafts and thus confound hair testing results for lead. It

is also unclear if the use of coloring agents containing lead acetate alters or enhances the hairs

ability to bind other analytes or potential toxicants. 

Based on the medical literature that describes the use of hair testing for substances of abuse there

are differences in hair uptake of various substances based on ethnicity. For example, negroid hair

has been suggested to bind cocaine residues with greater affinity than caucasoid hair.

There are also reports in the literature that the ability to bind various chemicals and drugs may

depend on endogenous hair color as well as if hair has undergone bleaching. For example,

bleached hair radically lowers the drugs [of abuse] content of hair. This may explain the

observation that many competitors on the professional biking circuit sport bleach blond hair

(Kintz). Blond hair has been shown to not bind cocaine or its metabolites as well as pigmented

hair (Hubbard). In addition, there was no evidence of a dose-related incorporation of these drugs
Michael I. Greenberg, M.D., MPH

C-27

and metabolites into non-pigmented hair. The concern is that similar circumstances may occur

with regard to specific occupational or environmental toxicants and chemicals. 

Based on a study presented by Reid et al. indicating that gray hair takes up less cocaine than nongray hair, it is possible that gray hair may also alter the utility of hair analysis in other settings.

The Reid study evaluated cocaine levels in the same individuals by comparing the levels in gray

and non gray hairs from the same person. In a similar study (Rothe et al.), hair samples from 15

patients receiving medical treatment with amitriptyline, carbamazepine, chlorprothixene,

diclofenac, doxepine, indomethacine, maprotiline, or metoclopramide, or with a chronic heroin

and cocaine abuse, were separated into white and pigmented fibers and both fractions were

independently investigated by GC-MS. The drugs were found in pigmented fibers as well as in

white fibers, but the concentrations in the white fibers were smaller than in the pigmented ones

for most of the samples investigated. The concentration ratio of the drugs or their metabolites in

both hair fractions (white/pigmented) was found to be between 0.09 and 1.57 (mean 0.70, 30

concentration pairs). There are large differences in this ratio between different subjects with the

same drug; whereas for different drugs in the same subject—in many cases—similar ratios were

measured. As reasons, a different grade of pigmentation of the hair and the influence of the drug

structure are discussed. From these results it follows that the natural hair color is an important

parameter in the evaluation of drug concentration in hair. Again, similar effects may be seen

when dealing with occupational and environmental toxicants. 

The rate of hair growth may be an important factor in the ability to identify the presence of

various materials based on time of exposure. Sources usually indicate that head hair grows at the

rate of 1-2 cm per month. That in itself represents a range encompassing up to a 100% difference

in hair growth rate. Obviously, comparisons of individuals whose hair growth rates differ by a

factor of 100% is problematic.
Michael I. Greenberg, M.D., MPH

C-28

TOPIC #3: TOXICOLOGIC CONSIDERATIONS

Relatively little is known about the biological uptake of specific substances with regard to the

concentration delivered to and incorporated into hair. There is essentially no data that reliably

establishes the relationship between chemical concentrations in the hair and blood or other target

organs for most chemcials. More specifically, and more importantly, no dose-response data

currently exists with regard to chemical concentrations in the hair and blood or other target

organs. In addition, no disease predictive value exists for any quantatative data that has been

derived to date with regard to the hair concentration of drugs or chemicals. 

Rollins et al have suggested that the ionization state of any given chemical is what determines

whether or not it will bind with hair melanin. These investigators reported that catiomnic drugs

are more likely to bind with melanin when compared with anionic drugs. This study may provide

some guidance with regard to the binding ability of other toxicants of concern.

TOPIC # 4: DATA GAPS AND RESEARCH NEEDS

Comments:

The data gaps that most significantly limit the use of hair testing in public health evaluations are

1) the lack of accurate and reliable reference range data and 2) the lack of specific information

about dose response relationships with regard to the relationship between chemical

concentrations in the hair and blood or other target organs. In my estimation, these two items

constitute the most pressing research needs with regard to hair testing. 

Future studies must address these basic data gaps in order to even begin to decide if hair testing

has clinical screening or other clinical usefulness.
Michael I. Greenberg, M.D., MPH

C-29

TOPIC # 5: IDENTIFYING SCENARIOS FOR WHICH HAIR ANALYSIS MAY BE

APPROPRIATE

Comments:

Hair testing for acute exposures is clearly not the best alternative for determination of either dose

or exposure with regard to any potential toxicant. If acute exposure is defined as the predistribution time frame, then blood or urine testing would be far superior to hair testing in any

scenario. However, in the event of a single exposure (as opposed to an ongoing exposure) the use

of hair testing after the completion of the pre-distribution phase of kinetics may be helpful in

qualitatively identifying the fact that exposure has indeed occurred and/or generally timing that

exposure. The use of hair analysis in this setting may have forensic as well as public health value.

In the setting of chronic exposures, hair analysis may have value in identifying and documenting

a given exposure. This, again, may have forensic, civil-legal, and risk assessment value for

individuals as well as communities and populations. Obviously, the length of any given

individual’s hair may limit the use of hair analysis, as well as how frequently the hair is cut. 

In any scenario, however, the state of the art is such that specific and measurable health effects

will generally not be uncovered by hair analysis. In addition, public health and/or individual risk

assessment determinations will be limited by whatever conclusions may be drawn by what is

essentially a qualitative and not quantitative toxicological evaluation.
Michael I. Greenberg, M.D., MPH

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ADDITIONAL CONSIDERATIONS:

Comments:

One interesting study (Al-Delaimy, et al) used hair analysis to measure the relation between

workplace smoking policies and exposures to environmental tobacco smoke (ETS) of workers in

bars and restaurants. In this study, 114 workers were questioned about sources of exposure to

ETS and smoking habits, and details of the smoke-free policy in their work place were recorded.

A hair sample was collected from each participant and tested for nicotine. Among non-smoking

workers, hair nicotine levels varied strongly according to the smoke-free policy at their place of

work. Those working in 100% smoke-free restaurants had much lower levels than staff working

in bars with no restrictions on smoking, and levels were intermediate for staff working in places

with a partial smoking ban. Hair nicotine levels among nonsmokers working in places with no

restriction on smoking were similar to hair nicotine levels of active smokers. The findings from

this study highlight the substantial levels of exposure of bar and restaurant staff from patrons'

smoking. 

The potential sources for confounding variables in the hair testing arena are truly legion. This

fact is demonstrated in one instance by a paper from Japan wherein investigators sought to draw

a relationship between head hair mercury and health. However, in the end, these investigators

discovered that “some subjects who showed a high total mercury level made habitual use of toilet

soap containing much mercury.” Thus, the confounding effect of an unusual source for a heavy

metal can interfere with effective hair analysis. 

Additional References:

Kintz P et al. Abstract presented at American Academy of Forensic Sciences meeting, Reno,

Nevada, February 2000.

Hubbard D et al. Society of Forensic Toxicologists meeting, Snowbird, Utah, 2000.
Michael I. Greenberg, M.D., MPH

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Reid R. et al. Society of Forensic Toxicologists meeting, Snowbird, Utah, 2000.

Rollins D et al. Society of Forensic Toxicologists meeting, Snowbird, Utah, 2000.

Rothe M et al. Effect of pigmentation on the drug deposition in hair of grey-haired subjects.

Forensic Sci Int 1997 Jan 17;84(1-3):53-60.

Harada M et al. Monitoring of mercury pollution in Tanzania: relation between head hair

mercury and health. Sci Total Environ 1999 Mar 9;227(2-3):249-56.

Holde et al. Quantitation of cocaine in human hair: the effect of centrifugation of hair digests. J

Anal Toxicol 1998 Oct;22(6):414-7.

Al-Delaimy W et al. Nicotine in hair of bar and restaurant workers. N Z Med J 2001 Mar

9;114(1127):80-3.
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Michael Kosnett
Michael J. Kosnett, MD, MPH

June 4, 2001

The following are preliminary comments regarding some topics that constitute the charge to the

panel. However, I am still in the process of reviewing some relevant studies and therefore may

revise or amend this material in a subsequent submission. 

Topic #1: Analytical methods

The key analytical methods currently used by clinical laboratories to measure trace elements in

hair appear to be inductively coupled plasma atomic emission spectrometry, and inductively

coupled plasma mass spectrometry (Miekeley et al, 1998; Seidel et al, 2001). Graphite furnace

atomic absorption spectrometry has been used to measure arsenic in hair, with reported limits of

detection of 0.005 to 0.01 :g/g (Rebel et al, 1998; Hewlett et al, 1995). Total and inorganic

mercury in hair has been determined by cold vapor atomic absorption (Boischio and Cernichiari,

1998; NRC, 2000), and the difference between total and inorganic Hg yielded by this method has

been used as a surrogate for the methyl mercury hair content. Methyl mercury in hair has also

been determined directly by gas chromatography using a tritium foil electron capture detector

(Smith et al, 1997). Selenium in hair has been measured fluorometrically after complexation with

2,3-diaminonaphthalene (Yoshinaga et al, 1990). The preceding methods appear to have

generally required a hair specimen size on the order of 50 mg or more. Although commercial

laboratories commonly measure the submitted hair sample in bulk, the methodology is

sufficiently sensitive to allow investigators to yield segmental analysis (3 1 cm) on bundles of

hair for which information on the alignment and distance from the root has been preserved.

Segmental analysis may potentially offer information on the temporal pattern of exposure to the

element in question that is of value in epidemiological and forensic investigations. 

Neutron activation analysis (NAA) has been used in forensic investigations and occasionally in

epidemiological or clinical studies for the sensitive determination of certain trace elements in

minute quantities of hair. For example, neutron activation analysis has been used to measure

arsenic in 2 mm segments of an individual hair, each segment weighing approximately 3 :g

(Smith, 1964; Curry and Pounds, 1977). NAA has also been used to measure the hair content of

Zn, Au, Cu, Mn, Hg, Sb, and Th (Jervis, 1968; Cornelius, 1973). The distribution of mercury in 2

mm segments along the length of a single strand of hair may be determined by nondestructive xray fluorescence (Cox et al, 1989, cited by NRC, 2000). Proton induced x-ray emission has been

used to measure the spatial distribution of multiple elements in 10 micron increments across

axial cross section of a single shaft of hair (Cookson and Pilling, 1975; Hindmarsh et al, 1999). 

A multitude of factors influence the quality control of laboratory hair analysis. These include the

finite limitations of the assay method (ideal method recovery and precision), and the variability

associated with within-run and day to day operation of the assay (actual method recovery and

precision). Although not necessarily reflective of a systematic review of the literature, a few

references may be cited as offering examples of operational precision in research investigations.

Using NAA to measure 7 elements in a single specimen of hair, the coefficient of variation

ranged from 5.92% in the case of Mn (mean concentration 1.65 ppm) to 15.7% in the case of Sb
Michael J. Kosnett, MD, MPH

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(mean concentration 0.18 ppm) (Cornelius, 1973). Wilhelm et al (1989) reported a day to day

coefficient of variation of approximately 6% for atomic absorption measurement of Zn, Pb, Cu

and Cd in hair. The issue of inter-laboratory variability of multi-element hair analysis for trace

elements provided by commercial laboratories using ICP-AES and ICP-MS has recently been

addressed by Miekeley et al (1998) and Seidel et al (2001), both of whom obtained widely

discrepant results from split samples sent to 4 to 6 different commercial laboratories. 

Topic #2: Factors Influencing the Interpretation of Analytical Results

One of the most fundamental factors impacting the potential utility of hair analysis as an

exposure assessment tool in public health evaluations is the limited capacity of such

measurements to distinguish external contamination from internal incorporation. In particular,

multiple studies have noted that toxic metals may become incorporated into hair following

external contact with metal containing dust, soil, water or hair care products. There is no reliable

analytical approach that can distinguish this external contamination from elevations in hair metal

content that result from metal ingestion or inhalation (Chittleborough, 1980). Although preanalysis washing or rinsing methods are often used in an attempt to selectively remove external

contamination, there is no standardized approach that has been shown to achieve the desired

result. 

The experience with arsenic, a toxic metalloid that is often encountered through environmental

exposures, is a case in point. In vitro studies have demonstrated that hair incorporates appreciable

amounts of arsenate and arsenite from aqueous solutions, and that the extent of absorption

increases with duration of contact time and moderate decrements in pH (e.g. pH 3 to 5) (Atalla et

al, 1965; Bate, 1966; Van den Berg et al, 1967; Fergusson et al, 1983). Adsorption of arsenic to

hair may also be substantial following contact with arsenic containing dust (Atalla et al, 1965).

The extent of adsorption may vary significantly along the length of a single hair (Maes and Pate,

1977). Adsorption-desorption experiments demonstrate that externally deposited arsenic cannot

be completely removed from hair by a variety of washing and rinsing techniques (Smith, 1964;

Atalla et al, 1965; Van den Berg et al, 1968). Moreover, washing may complicate interpretation

further by partially removing arsenic present in hair as a result of internal incorporation (Atalla et

al, 1965; Van den Berg et al, 1968; Young and Rice, 1944). Studies with other metals have

reported similar findings with respect to adsorption onto hair from external contamination, and

variable removal of both internal and externally derived traces by washing regimens

(Chittleborough, 1980; Fergusson et al, 1983; Wilhelm et al, 1989). 

The problems posed by this inability to distinguish external adsorption from internal

incorporation places substantial constraints on what can be learned from the results of hair

analysis for an environmental toxin where the suspected route of human exposure is via contact

with contaminated dust, soil, airborne particulate, or tap water. Although these routes of

exposure might result in ingestion or inhalation of an environmental toxin and its subsequent

appearance in hair through incorporation at the hair follicle, they also create ample opportunity
Michael J. Kosnett, MD, MPH

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for the agent to become externally adsorbed onto hair via airborne deposition, hand to hair

contact, or bathing. In such settings (which are probably characteristic of the majority of sites

subject to ATSDR health assessments), the finding of elevated levels of a environmental toxin in

the hair of a given subject or a study population is limited at best to establishing the potential for

that subject or population to have come into contact with the agent in a manner that may have

resulted in ingestion or inhalation. In addition to being a test of low specificity, the information

on potential exposure gleaned from an elevated hair level in such settings is likely to be

qualitative in nature. That is because with the notable exception of methyl mercury, quantitative

information on the relationship between ingestion or inhalation of a environmental toxin and its

concentration in hair is limited, and appears to be subject to considerable inter-subject and interpopulation variability. 

Again, an example derived from the measurement of arsenic in hair is instructive. Although

several epidemiological studies have noted a correlation between levels of arsenic in hair and

arsenic in dust, soil, or water, (e.g. Bencko and Symon, 1977; Hartwell et al, 1983; Valentine et

al, 1979), the hair arsenic levels may not correlate with levels of arsenic in urine (Harrington et

al, 1978; Hewlett et al, 1995). For example, Harrington et al (1978) studied hair and urine arsenic

levels in a community near Fairbanks, Alaska, where the arsenic concentration of water obtained

from domestic wells averaged 224 :g/L (range 1.0 to 2450 :g/L). A subset of subjects whose

wells contained arsenic averaging 345 :g/L consumed only bottled water. Although they had

relatively low arsenic levels in urine (average 43 :g/L), the arsenic concentration of their hair

was high, averaging 5.74 ppm. Subjects consuming water from domestic wells with the lowest

levels of arsenic (less than 50 :g/L in water) had hair arsenic concentrations averaging 0.46 ppm,

and urine arsenic levels averaging 38 :g/L. Thus, the arsenic level in hair varied by 14-fold,

despite similar levels of arsenic in urine. The authors noted the likely implication that the

elevated hair arsenic levels were probably due to external contamination derived from bathing in,

but not drinking, the high arsenic well water. 

Topic #3 To what extent may hair analysis be used to predict adverse health outcomes? and

Topic #5, Under what scenarios may hair analysis be appropriate for evaluating exposures to

environmental contaminants?

From a medical standpoint, there appears to be no disease or illness caused by an environmental

toxin for which there is a general medical consensus that the results of hair analysis would form

the basis for specific medical treatment. 

In the case of methyl mercury, segmental maternal hair analysis may have diagnostic value as a

biomarker of fetal exposure to levels of this neurotoxin that are associated with a postnatal risk

of adverse neurobehavioral development (NRC, 2000). Some data suggest that the level of hair

methylmercury in children and adults may also be a biomarker of exposure associated with

adverse effects on neurological function and other health endpoints (NRC, 2000). Because most

contemporary exposure to methylmercury is confined to ingestion via seafood, there is little
Michael J. Kosnett, MD, MPH

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potential for high hair levels of methylmercury to be a result of external contamination. In most

populations whose level of seafood ingestion is of a sufficient magnitude to pose a potential

health risk from methylmercury, measurement of total mercury in hair may be an acceptable

surrogate for measurement of methylmercury in hair.

In certain settings, segmental hair measurement of arsenic (and potentially other toxins such as

thallium) may be of diagnostic and/or forensic value in identifying or confirming a high dose

toxic exposure or poisoning that terminated months (but not years) in the past. For example,

segmental analysis of a sufficiently long hair might help to confirm a suspicion that an episode or

outbreak of severe gastroenteritis followed by peripheral neuropathy that occurred 8 to 10 months

in the past was likely to have been the consequence of acute arsenic or thallium poisoning.

Months after the exposure ended, levels of arsenic or thallium in the urine may have fallen to

normal values, and high peak levels in the hair (or nails) may offer the only remaining

confirmatory forensic evidence. It should be noted that although the hair measurements in such

scenarios might conceivably be of value in confirming past poisoning, the epidemiological

database on hair analysis is insufficient to use these measurements to predict the risk of latent

diseases such as cancer.

Supplemental comments from Michael J. Kosnett, MD, MPH (submitted June 21, 2001)

1. A key factor to be addressed prior to ATSDR’s use or interpretation of hair testing is the

predictive value of a positive or negative test with respect to detecting an exposure and/or

internally absorbed dose of a toxic substance of sufficient magnitude to be of pathological

or public health significance.

2. One of the inherent limitations of hair analysis arises from the fact that hair represents a

matrix that is in direct contact with the external environment and as such may be subject

to greater contamination than other analytes traditionally used in biological monitoring,

such as blood, urine, or even expired air.

Supplemental references submitted by Michael J. Kosnett, MD, MPH (June 21, 2001)

Atalla L, Silva CM, Lima FW. Activation Analysis of Arsenic in Human Hair—Some

Observations on the Problems of External Contamination. Ann. Acad. Bras. Cien., 1965, 37:432-

441.

Bate LC. Adsorption and Elution of Trace Elements on Human Hair. Int. J. Appl. Rad. Isot.,

1966, 17:417-423.
Michael J. Kosnett, MD, MPH

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Bencko V, Symon K. Health Aspects of Burning Coal with a High Arsenic Content. I. Arsenic in

Hair, Urine, and Blood in Children Residing in a Polluted Area. Environ. Res., 1977, 13:378-385.

Chittleborough G. A Chemist’s View of the Analysis of Human Hair for Trace Elements. Sci.

Tot. Envir. 1980, 14:-75.

Cornelis R. Neutron Activation Analysis of Hair: Failure of a Mission. J. Radioanal. Chem.,

1973, 15:305-316.

Cox C, Clarkson TW, Marsh DO, Amin-Zaki L, Tikriti S, and Myers GG. 1989. Dose-response

analysis of infants prenatally exposed to methyl mercury: An application of a single compartment

model to single-strand hair analysis. Environ. Res. 49(2):318-322.

Curry AS, Pounds CA. Arsenic in Hair. J. For. Sci. Soc., 1977, 17:37-44.

Ferguson JE, Holzbecher J, Ryan DE. The Sorption of Copper [II], Manganese [II], Zinc [II], and

Arsenic [III] into Human Hair and Their Desorption. Sci. Tot. Envir., 1983, 26:121-135.

Gebel TW, Suchenwirth RHR, Bolten C et al. Human biomonitoring of arsenic and antimony in

case of an elevated geogenic exposure. Environ Health Persp 1998; 106:33-39

Harrington JM, Middaugh JP, Morse DL et al. A Survey of a Population Exposed to High

Concentrations of Arsenic in Well Water in Fairbanks, Alaska. Am. J. Epid., 1978, 108(5):377-

385.

Hartwell TD, Handy RW, Harris BS et al. Heavy Metal Exposure in Populations Living Around

Zinc and Copper Smelters. Arch. Environ. Health, 1983, 38(5):284-295.

Henke G, Nucci A, Queiroz LS. Detection of Repeated Arsenical Poisoning by Neutron

Activation Analysis of Foot Nail Segments. Arch. Toxicol, 1982, 50:125-131.

Hewitt DJ, Millner GC, Nye AC, et al. Investigation of arsenic exposure from soil at a Superfund

site. Environ Research 1995; 68:73-81

Houtman, JPW, de Bruin M., de Goeij JIM: Arsenic Levels of Human Hair as an Indicator for

Environmental Exposure, in Nuclear Activation Techniques in the Life Sciences, Vienna, IAEA,

1978, pp. 599-614.

Jervis RE, Present Status of Activation Analysis Applications in Criminalistics. Isot. Rad. Tech.,

1968, 6(1):57-70.
Michael J. Kosnett, MD, MPH

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Leslie ACD, Smith H. Napoleon Bonaparte’s Exposure to Arsenic During 1816. Arch. Toxicol.,

1978, 41:163-167.

Maes D, Pate BD.: The Absorption of Arsenic into Single Human Head Hairs. J. For. Sci., 1977.

22:89-99.

National Research Council. Toxicological Effects of Methyl Mercury. National Academy

Press: Washington, DC, 2000

Pounds CA, Pearson EF, Turner TD. Arsenic in Fingernails. J. For. Sci. Soc., 1979, 19:165-173.

Smith H. The Interpretation of the Arsenic Content of Human Hair. J. For. Sci. Soc., 1964,

4:192-199.

Valentine JL, King HK, Spivey G. Arsenic Levels in Human Blood, Urine, and Hair in Response

to Exposure via Drinking Water. Environ. Res., 1979, 20:24-32.

Van den Berg AJ, de Bruin M, Hortman JPW. Sorption Behavior of Trace Elements in Human

Hair, in Nuclear Activation Techniques in the Life Sciences, Vienna, IAEA, 1967, pp. 661-674.

Van den Berg AJ, de Geoij JJM, Houtman JPW et al. Arsenic Content of Human Hair After

Washing as Determined by Activation Analysis, in DeVoe JR [ed.] Modern Trends in Activation

Analysis, Vol. I. Washington, DC: NBS, 1968, pp. 272-282.

Wilhelm M, Ohnesorge FK, Lombeck I et al. Uptake of aluminum, cadmium, copper, lead, and

zinc by human scalp hair and elution of the absorbed dose. J Anal Toxicol 1989; 13:17-21

Young EG, Rice FAH. On the Occurrence of Arsenic in Human Hair and Its Medicological

Significance. J. Lab. Clin. Med., 1944, 29:439-446.
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Dan Paschal
Daniel C. Paschal–CDC

ATSDR Hair Analysis Workshop

June 12–13, 2001 

Atlanta, GA

Charge Questions for Panelists:

Analytical Methods

1) What analytical methods currently exist?

Analytical methods for hair analysis include cold vapor atomic absorption analysis (1); graphite

furnace atomic absorption (2); inductively coupled argon plasma optical emission spectrometry

(3,4); inductively coupled argon plasma mass spectrometry (5); proton induced X-Ray emission

(PIXE) spectrometry (6) ; X-Ray analysis (7); and neutron activation analysis (8).

2) Substances/elements for which reliable analyses exist include:

a) mercury- methyl and inorganic (1);

b) arsenic (2,8);

c) aluminum (3,4);

d) gold (3,4);

e) boron (3,4);

f) barium (3,4);

g) beryllium (2,3,4);

h) calcium (3,4);

i) cadmium (2,3,4);

j) cobalt (3,4);

k) chromium (2,3,4);

l) copper (2,3,4);

m) iron (3,4);

n) lithium (2,3,4);

o) magnesium (2,3,4);

p) manganese (2,3,4);
Daniel C. Paschal–CDC

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q) molybdenum (3,4,5 );

r) sodium (3,4);

s) nickel (2,3,4);

t) phosphorous (3,4);

u) lead (2,3,4,5);

v) antimony (3,4);

w) selenium (2,3,4,5);

x) strontium (3,4);

y) titanium (3,4);

z) thallium (2,3,4,5);

aa) vanadium (2,3,4);

bb) zinc (2,3,4);

cc) drugs of abuse -cocaine, PCP, opiates (9,10)

3) For what purposes are these methods typically used?

Forensics- As

Exposure evaluation- As, Cd, Cr, Hg, Mn, Pb, Se, Al

Diet/Nutrition Status- Ca, Mg, Na, Se, Sr, V, Zn, Cu, Co

4) What amount (g) of hair is needed?

0.1-0.5g (4,5)- Amount depends on type (occipital or other) and detection limit (4,5,9,10).

5) Intralaboratory variability (within-lab/run precision and accuracy)- MUST be evaluated

with a stable, homogeneous, well-characterized pooled material.
Daniel C. Paschal–CDC

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6) Interlaboratory variability-(among laboratories accuracy and precision)- 

evaluation can be by regulation (CLIA or state/county/city licenses) or voluntary participation in

Quality Assurance/Quality Control programs- e.g. Center for Toxicology of Quebec

(http://www.ctq.qc.ca/ctqintre.html http://www.ctq.qc.ca/icpms.html).

Factors Influencing the Interpretation of Analytical Results

Variations in sample collection 

A variety of sample preparations have been suggested to sort exogenous (presumable

contamination from exposure to the external environment) and endogenous metals and drugs

from collected hair specimens. These vary from no treatment, washing with

deionized/distilled/ultrapure water only to washing with ionic or non-ionic detergents, either

alone or in concert with organic solvent washes. For details and references, see (2).

Sampling methods

CDC has standardized the specimen collection and washing for hair, based on studies conducted

internally and reported (4,5) in the literature. We obtain about 0.5 grams of occipital hair, and

wash with a non-ionic detergent. Quality control is preformed by analysis of reference materials

from NIST (SRM 1643d-Trace Elements in Water; SRM 1641d Mercury in Water), and a

digested hair sample characterized by our operational method(s). Normal or “reference” ranges

for 28 elements were published (4). “Abnormal” ranges would be those outside (generally higher

than) the 95% upper limits for these analytes- toxic levels vary considerable depending on the

adverse health outcome for each individual toxicant.

Exposure of hair to external environment

includes copper from certain chlorinated swimming pools, lead from lead acetate “Grecian

Formula”, selenium from dandruff shampoo (“Selsun”); zinc from “herbal” shampoos (Herbal
Daniel C. Paschal–CDC

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Essence; Head and Shoulders), lead , cadmium, mercury and arsenic from dust, dirt, smoke, etc

(4,5,11).

Exogenous and endogenous

hair levels are difficult to distinguish, due to the high porosity of hair, and ineffective and nonstandard “washing” procedures. The ideal washing/cleaning procedure would remove ONLY

exogenous metals or other analytes- unfortunately, none have been reported (4,5,12,13,14).

Hair color

pigmentation (melanin?) (15) and location (4,5,11) have been demonstrated to affect hair

concentrations of several analytes.

Gender, ethnicity

affect hair metals concentrations due to presence or absence of gender-linked hair treatment

activities (e.g. coloring, permanent) and pigmentation (4,5,11).

Rate of Growth

of hair has been assumed by many investigators to be relatively “constant” at about 1 cm/month

(4,5,11) but is known to vary somewhat with age/gender/season (4,5,11).

Toxicologic Considerations

Biological uptake of metals (4,5,11,16,17) and drugs of abuse have been extensively studied and

described.

Relationship between hair and other tissue concentration levels, including urine (18) , whole

blood (1,19) and serum (20) as well as other tissues (21) has been studied and described to some

degree. The most complete and compelling evidence exist for hair mercury/blood mercury
Daniel C. Paschal–CDC

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(methylmercury) and for arsenic in hair/urine/fingernail/tissue (1,21,23,24). Other metals and

drugs of abuse are less well characterized (17).

Dose response relationships have been demonstrated in very few recognized studies—only hair

mercury and arsenic have been clearly associated with body burden and health (adverse) effects

(25,26). Other evidence, e.g. correlation between the concentration of manganese in hair and

behavioral disorder or violence, is less compelling (27). 

Data Gaps

Methodological- Quality control/quality assurance- although some laboratories are licensed for

trace metals determinations, there are very few (28) proficiency testing programs or reference

materials available (29,30) for evaluation and documentation of precision and accuracy of

laboratory analytical systems.

Toxicological- Serious disagreement exists as to “reference” (normal or expected) values for a

large number of elements. Drugs of abuse can often be detected at low concentrations; there is

some disagreement as to the correlation between results of hair testing for abused drugs and more

conventional determinations of drugs in urine, exhaled breath, or other (29).

Research Needs- Simply stated, carefully designed studies of exposure, body burden, and hair

concentrations are needed to move beyond “anecdotal” levels of documentation. These studies,

will, unfortunately, be limited by available funds and other resources.

Scenarios Where Hair Analysis May Be Appropriate

Exposure Pathway Chronology Exposure Measurable Health

Duration Effects (Y/N)

Individual Ingestion Past Chronic Yes (if high)

(MeHg)
Daniel C. Paschal–CDC

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Individual Ingestion Past Chronic Yes (if high)

Inhalation

(As 3/5)

Individual Ingestion Past Chronic Yes (if elevated)

Lead

REFERENCES

1) Greenwood MR, Dhahir P, Clarkson TW, Farant JP, Chatrand A, and Khayat AJ. Analyt.

Toxicol. 1, 265 (1977).

2) Tsalev DL. Atomic Absorption Spectrometry in Occupational and Environmental Health

Practice. Boca Raton FL: CRC,1995. 

3) DiPietro DS et al. Biological Trace Element Research, 22, pp. 83-100 (1989).

4) Paschal DC et al. Environmental Research 48, pp. 17-28 (1989). 

5) Miekeley N, Dias Carniero MTW, and Porto de Silviera CL. Science of the Total

Environment, 218, pp. 9-17 (1998).

6) Du Y, Mangelson NF, Rees LB, and Matheny RT, "PIXE Elemental Analysis of South

American Mummy Hair," Nucl. Instru. Meth. Phys. Res. B, (1996).

7) Stephenson,J. JAMA Vol. 281 No. 17,May 5, 1999.

8) Cornelis R., Speecke A. Neutron activation analysis of human hair collected at regular

intervals for 25 years J. Forensic Sci. Soc., 11/1 (1971), 29-46.

9) Tomoaki Sakamoto, Akira Tanaka, and Yuji Nakahara Journal of Analytical Toxicology, 20,

pp. 124-130 (1996).

10) Baumgartner WA, Hill VA, Blahd WH. Journal of Forensic Sciences , 34 , pp. 1433-1453

(1989).

11) Esteban E, Rubin C, Jones R, and Noonan G. Archives of Environmental Health, 54, pp.

436-440 (1999).

12) Assarian GS and Oberleas D. Clin. Chem. Vol. 23, pp. 1771-2 (1977).

13) Buckley RA and Dreosoti IE. Am. J. Clin. Nutr. Vol. 50., pp. 840-6 (1984).

14) Wilhelm M., Ohnesorge FK, Lombeck I, and Hafner,D. J. Anal. Toxicol. Vol. 13, pp. 17-21

(1989). 
Daniel C. Paschal–CDC

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15) Sky-Peck HH. Clin. Phys. Biochem., Vol. 8, pp. 70-80 (1990).

16) Koren G. Forensic Sci Int. Vol 70 (1-3) pp 77-82 (1995).

17) Sachs H. Forensic Sci Int. Vol 70 (1-3) pp 53-61 (1995).

18) Taneja SK, Mohajan M, Gupta S, Singh. KP, Biol Trace Elem Res , 62, pp. 255-264 (1998).

19) Pihl RO, Drake H, Vrana F. Department of Psychology, McGill University, Montreal,

Quebec, Canada.: Hair Analysis in Learning and Behavior Problems. Hair, Trace Elements, and

Human Illness. Brown, A. C.; Crounse, R. G., eds. Praeger Publications, 1980.

20) Yoshinaga J, Imai H, Nakazawa M,. and Suzuki T. Sci Total Environ. Vol. 99, pp. 125-135

(1990).

21) Sherlock JD, Lindsay DG, Hislop JE, Evans WH, and Collier TR. Archives of

Environmental Health, 37, pp. 271-8 (1982).

22) Wibowo AE, Herber RM, Das HA, Roeleveld N, and Zielhuis RL. Environmental Research,

40, pp. 346-56 (1986).

23) MMWR, March 02, 2001 / 50(08);140-3.

24) "High hair manganese in children with attention deficit-hyperactivity disorder," EJ Cordova,

FM Crinella, and JE Ericson, unpublished study. Address: F. M. Crinella, UC Irvine, Child

Development Center, 19722 MacArthur Blvd., Irvine, CA 92612.

25) Kirschmann G & J. Nutrition Almanac, 4th ed. New York: McGraw Hill, 1996.

26) http://www.doctorsdata.com/RESPONSE.HTM (PT)

27) http://www.ctq.qc.ca/icpms.html (CTQ ICP-MS hair specimens)

28) Steindel SJ, Howanitz, PJ. JAMA, 285(1), (2001).

29) Wennig R. Forensic Science International, 107, pp. 5-12 (2000).

30) http://www.iaea.or.at/programmes/nahunet/e4/nmrm/material/ (IAEA reference materials)
Sharon Seidel
ATSDR Hair Analysis Workshop - Charge questions:

Topic #1: Analytical Methods.

Atomic absorption spectroscopy (AAS) is commonly used for individual elements, and

can now do more than one element at a time. Lead, for example, is commonly measured by

graphite furnace AAS. A well-established conventional laboratory with forensic services

typically measures individual elements or a small panel of elements in hair for chronic exposure

(e.g., first panel - mercury by cold vapor AAS; lead, arsenic, chromium and cadmium by graphite

furnace (GF)-AAS; second panel - cadmium, manganese, nickel and thallium, all by GF-AAS).

The AAS methods are considered well-established methods. The amount of hair required for

either AAS panel (above) is 0.5 gram. Other analytical methods have the potential to measure a

number of elements simultaneously, including inductively coupled plasma-atomic emission

spectroscopy (ICP-AES) and mass spectroscopy (ICP-MS). Newer ICP-AES instruments can

attain a sensitivity equivalent to single element AAS. ICP-MS is a more sensitive method than

AES. 

In a carefully conducted study, a major research laboratory at the Centers for Disease

Control and Prevention (CDC) reported the determination of 28 elements in hair from nonoccupationally exposed U.S. populations.1

 These investigators used ICP-AES for all elements

except mercury, which was measured with an LDC mercury monitor. The required minimum hair

sample weight was 0.5 gram. Miekeley et al. more recently reported results for ICP-MS analysis

of a suite of elements from hair in a Brazilian population, with improved sensitivity compared to

ICP-AES.2 The amount of hair required was approximately 0.3 gram.

Of the 9 commercial “nutritional hair analysis” laboratories currently operating in the

United States, 3 indicate that they primarily use ICP-MS, 4 primarily use ICP-AES, and 1 reports

use of directly coupled plasma (DCP)-AES. DCP-AES is an older technique that is potentially

less stable than ICP-AES. On average, these laboratories measure 26 elements per hair sample.

Nutritional hair analysis laboratories require between 0.3 and 1 gram for the AES methods, and

0.25-1 gram for ICP-MS. Puchyr et al. also discuss preparation of hair for elemental analysis by

ICP-MS from a nutritional hair analysis laboratory perspective. 3
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Other investigative techniques for measuring elements in hair are reported in the

scientific literature. A general discussion of common methods is provided by Jacobs and by

Haraguchi et al. 4,5 Various other methods and example references, e.g: Differential pulse

voltametric (DPV); 6 Instrumental Neutron Activation Analysis (INAA); 7,8 Microwave-Induced

Plasma Mass Spectroscopy (MIP-MS); 9 Capillary electrophoresis (CE) and High Performance

Liquid Chromatography (HPLC); 10 and Particle Induced X-ray Emission (PIXE).11

Laboratory variability has been investigated for the commercial “nutritional hair analysis”

laboratories on several occasions.2,12-14 Inter-laboratory variability was high for reference ranges,

results, interpretations and health advice. For example, for one hair sample that was split and sent

to six of the laboratories, there was a difference of an order of magnitude or more between

laboratories in reported results for over 10 elements, including arsenic, lead, and mercury.13 In

the same split hair sample, no two laboratories flagged the same element as high, and laboratories

had conflicting health interpretations and dietary recommendations based on their analysis of the

sample. When intra-laboratory variability was investigated for nutritional hair analysis

laboratories, results were similarly discrepant.12

Topic #2: Factors Influencing the Interpretation of Analytical Results.

A.) Sample collection and analysis:

Sample collection and preparation methods can have a significant impact on the data

collected. Hopps notes that scalp hair has about 90% of follicles in the growth phase at any given

time, growing at about 0.45 mm/day.15 Scalp hair grows in a mosaic pattern over the scalp, with

similar growth activity in the various regions of the scalp. However, sampling near the face is

usually avoided due to increased likelihood of contamination from sebaceous secretions and

facial hygiene products/cosmetics. Miekeley et al. note that larger samples of scalp hair (50 g.),

cut into <1cm pieces and manually homogenized, showed homogeneity in repeated analyses of

aliquots of the samples.2
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Commercial nutritional hair analysis laboratories frequently offer the option of collecting

samples of axillary or pubic hair. Hair from these regions of the body grows more slowly, with a

much greater proportion in the resting phase, and is likely to be subject to external contamination

from apocrine gland secretions, in addition to use of personal hygiene products, clothing, etc.

There are no published reference ranges for elements from non-scalp hair. A lack of correlation

has been shown between scalp and pubic hair for Ca, Cu, Fe, Mg and Zn.16

Homogenization can be a concern, particularly if long lengths of hair are collected.

Concentrations in hair of a number of environmentally-important elements have been shown to

increase from the proximal to distal end of hair, e.g. Pb, Cu, Fe, Mn, and Zn.17,18 Contamination

is also a concern if the laboratory uses a grinding tool that introduces contaminants, as occurred

in the preparation of one hair reference material, where Al, Fe, Ti, Mn, and Mg contamination

were introduced through use of an agate ball grinding mill.19

Sample preparation and washing methods vary greatly and can cause different analytical

results. Chittleborough provides a detailed review of these issues.20 Various washing

recommendations include: no-wash;20 use of a standardized washing procedure recommended by

the International Atomic Energy Agency (IAEA) which uses a nonpolar solvent-acetone and

deionized water;21 a mild ionic detergent-sodium lauryl sulfate emulating a detergent shampoo;1

and more extreme methods including a chelating agent, EDTA;22 and others (see review by

Chittleborough).20 There is no washing method presently available which is capable of reliably

removing external contaminants without also affecting endogenously-deposited elements.20,23-25

While a no-wash approach offers the least disturbance to endogenous elements, the

demonstration by scanning electron microscopy of dust, dead skin, etc., adhering to much of the

length of unwashed hair samples discourages use of this approach.26 Other aspects of laboratory

sample preparation that may be critical include procedures which minimize loss of more volatile

elements, such as mercury, during sample dissolution.
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A major stumbling block in interpreting metals data for hair is laboratory analytical error.

The World Health Organization recommends the following quality assurance methods for

laboratory analyses. 1) Reference samples of the same matrix (hair) with known concentrations

of the metal (element) should be used as standards. 2) Reference samples should contain the

metal (element) at about the same concentration as the samples. 3) If such reference materials are

not available, analysis of quality-control samples at different laboratories by different analytical

methods must be used (i.e., split samples). 4) Since results may vary over time and for different

metals (elements), results should be presented for the corresponding time periods and elements.27

There are various certified reference materials (CRM), for one (mercury) or multiple

elements in hair, which meet certification requirements including certified values with a stated

level of confidence in each value.19,28-30 There is no certified hair reference material for all

elements currently analyzed by commercial “nutritional hair analysis” laboratories. The Chinese

hair CRM, reportedly used by four of these laboratories, certifies 17 elements: Al, As, Ca, Cd,

Co, Cr, Cu, Fe, Hg, Mg, Mn, Na, Ni, Pb, Se, Sr, and Zn - about half the elements tested by these

laboratories. A common practice among these and other laboratories is to use aqueous element

standards, or other non-hair standards such as bovine liver. The difficulty with this is the

possibility of complex matrix interferences in the hair sample that are not accounted for by the

calibration standard. Reference ranges cited by commercial U.S. nutritional hair analysis

laboratories show some rather broad inter-laboratory variations, e.g. arsenic (<0.06 vs. <5 ppm),

lead (<0.8 vs. 2-20 ppm), and lithium (0.0035-0.025 vs. 1.25-3 ppm).13

Investigations of “nutritional hair analysis” laboratory practices using split samples have

shown wide discrepancies.2,12,13 An approved proficiency testing program for hair element

analysis is not available under the Clinical Laboratory Improvement Act (CLIA). This type of

analysis is classified as a high-complexity test, with method and accuracy verification left up to

the individual laboratory.
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“Normal” reference ranges are largely undefined, due to the wide variation in elemental

hair concentrations in presumed healthy populations. Contributing factors include geography,

age, sex, ethnicity, hair type, hair treatments and other exogenous exposures. Examples of U.S.

studies follow. DiPietro et al. published reference intervals for 28 elements in a nonoccupationally exposed U.S. adult population.1

 These investigators used extensive questionnaire

data to control for many of these factors. A partial list of geometric means for healthy adults in

this study includes: arsenic (0.15 ppm); cadmium (<0.15 ppm); nickel (0.39 ppm); and lead (2.43

ppm). A number of population studies have been conducted for mercury in hair. For

methylmercury, the geometric mean hair concentration for U.S. women reporting some seafood

consumption was 0.36 ppm, and 0.24 ppm for no seafood consumption.31 Published clinical

references for biomonitoring for metals/elements in hair are sparse. These include arsenic (<1

ppm) and thallium ( 5-10 ppb)32 and mercury (<1ppm) and nickel (0.01-1.8 ppm).33 These are

secondary to the established blood and/or urine reference levels, and the problem of external

contamination is noted as a major stumbling block which limits the use of the hair references. 

Generally speaking the use of the term “normal” is misleading. What is being estimated is

a background or baseline level for a population, typically by geographic region, rather than a state

of health. Methylmercury data are an exception. Methylmercury exposures commonly occur

through consumption of fish and seafood. Clear dose-response relationships have been

demonstrated between dietary consumption of mercury-contaminated fish and concentrations in

human hair. Methylmercury is the only metal (compound) which has a health benchmark based

on hair concentrations. The U.S. EPA has a reference dose (RfD) for methylmercury of 0.0001

mg/kg body wt/day. This is based on a benchmark dose of 11 ppm in maternal hair, equivalent to

a maternal blood level of 44 micrograms/L, for developmental neurological abnormalities in

infants.34 Several reference range studies for methylmercury are available.31,35,36

B.) Other factors influencing analytical results:
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Hopps notes the sources of elements in hair as: 1) papilla (contacted by blood and lymph)

during hair formation; 2) sebaceous glands, sweat glands, desquamating skin cells (endogenous

exposures not necessarily related to blood/organ concentrations); 3) and exogenous materials.2

Salts of sodium, potassium, and calcium predominate in sweat, but minor amounts of other

elements are also found, e.g., zinc.15 There is evidence for an extra input route from the root

sheaths into the hair shaft, other than longitudinal growth, complicating the picture of a simple

blood compartment:hair relationship.11 Finally, the lipids and waxes in sebum and skin may

contribute to sealing exogenous contaminants into the hair shaft.

Exogenous contaminants can range from personal care products to elements present in

air, water, soil, occupational environments, etc. As mentioned above, there is currently no

washing method capable of removing exogenous elemental contaminants while leaving

endogenous elements undisturbed. Chemicals such as methylmercury, which are generally from

dietary sources, suffer less from this drawback, provided unusual sources of inorganic mercury

do not complicate the picture, e.g., mercury vapor in occupational settings. Practically speaking,

public health concerns are often related to exposure, and hair can serve as an index of overall

exposure, if not of biological uptake.

Examples of external contaminants of hair include both personal care products and

environmental sources. Hair is a porous material (witness the rapid uptake of water and increase

in weight during washing) and may bind through weak ion-exchange sites (e.g., Na, K, Ca, Mg),

and through stronger bonds, particularly with sulfur, (e.g, arsenic). Arsenic binds avidly to hair,

due to the sulfur content of keratin. Exogenous arsenic is readily taken up by hair and cannot be

differentiated from endogenous arsenic.15 It has been shown that adsorption of other metals such

as Al, Cd, Cu, Pb and Zn into scalp hair from aqueous solutions cannot be reversed even by

extreme washing methods.25 Hair treatments such as permanents can alter such binding.37

Dandruff shampoos containing selenium can contaminate hair.38 DiPietro et al. noted significant

difference between dandruff shampoo vs. regular shampoo for Na, Se, and Ti for men, and
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between permanents/color and any shampoo for Ba, Ca, Cu, Mg, Na, and Sr for women.1 Hair

dyes may contain metals, e.g., lead in “Grecian Formula.”39 Sky-Peck also found that peroxide

bleaches and permanents altered S, Ca, Fe, and Ni in hair, peroxide altered Zn, and permanents

increased Cu and As concentrations.

39

Soil, house dust, and water may contribute contaminants.40 Air serves as a contamination

source.41 This is a major concern in occupational settings. Cadmium is an example of a metal

where environmental sources contribute to concentrations in hair, e.g., drinking water and dust

levels and seasonal influences.42

As noted, gender, ethnicity, diet, age, geographic location, and season are capable of

influencing hair reference ranges in populations. Sky-Peck found the following for a healthy

midwestern U.S. population: 1) gender – females had higher Ca and Ni and lower Pb, Br and Se

compared to males; 2) hair color - blondes had less Fe than brunettes, red-heads had more Fe and

Cu; 3) ethnicity/race - Blacks had increased Ca, Fe, Ni, Cr, Mn, As, and Pb, and decreased Hg,

compared to Caucasians; Orientals had decreased Ca, Fe, Cu, Mn and Pb; 4) age – a decrease in

S, Ca and Sr, and an increase in Pb with age; 5) geography – increased hair strontium in areas

with elevated strontium in drinking water, and increased hair lead in industrial/older residential

areas.39 Sky-Peck notes that some of the differences in gender and ethnicity may be due to

differences in hair treatment and/or environmental exposure. While Sky-Peck found no

differences between gray hair and natural hair, other investigators have noted pigmentation

effects,43 and it is known that various chemicals, including metals, will bind to melanin.44

Other investigators have studied age-related differences in hair elements. Paschal et al.

observed age-dependent increases in Ca, Ba, Mg and Sr (Group 2A alkali elements) and Zn up to

12-14 yrs in U.S. residents.

45 In comparison, an Italian study showed increases in Cu, Zn, Cr and

Br, and decreases in Fe, Mn and Sr up to 8 yrs.46 In Japanese children, Zn decreased up to 12-14
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yrs, and Cu showed a similar trend.47 The reason for differences between laboratories and/or

populations is not presently known.

Baseline reference values for elements in clinical specimens, including hair, have been

referenced by international location.48,49 International differences are identified for hair Zn, Cd,

Cu, Mn and Pb. Some of this geographical difference may be due to differences in environmental

metal concentrations, industrialization, etc. Seasonal differences in hair element concentrations,

e.g. cadmium, may be due to time spent outdoors and contact with soil, dust, etc.

Topic #3: Toxicological Considerations.

As discussed above, methylmercury is the only element (compound) for which sufficient

data exist to define the relationship between concentrations in blood, concentrations in hair, and

effects on the target (the developing fetus). It is also the only element (compound) with a health

benchmark, the U.S. EPA reference dose, based on a threshold concentration in human hair. It

should be noted that this threshold was identified based on massive poisoning incidents in human

populations and not on typical (dietary) exposures.34,50 Forensic medicine has used hair to assess

poisoning by other elements, e.g, arsenic and lead. However, these document overwhelming

poisoning exposures, rather than a threshold for earliest/most subtle adverse health effects. Nor is

there a need in these instances to differentiate between a “normal” background and subtle

increases in exposures. Such a distinction is difficult due to the wide variations in background

reference ranges. This has caused a number of investigators to conclude that results for an

individual are not likely to be meaningful with respect to less drastic environmental/dietary

exposures, and that statistical analyses of group data must be employed.13,42,51,52 Finally, if the

goal is also to provide an index of body burden, rather than simply document exposure to

environmental contaminants, the lack of a washing technique capable of reliably separating

exogenous contaminants from biologically-deposited elements is a substantial concern and must

be addressed. 
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Of the trace elements that have been tested in hair, only a few have research data relating

hair concentrations to blood levels and/or tissue concentrations. Aside from mercury, the focus

has largely been on aluminum, arsenic, cadmium, chromium, copper, lead, selenium, and zinc.

Data highlights are summarized below.

Aluminum (Al) – Aluminum is elevated in hair only in extreme exposures (and even then

is inconsistent), and is unrelated to serum or bone aluminum.53-56 Aluminum dietary intake is

unrelated to aluminum in hair, even with controlled dietary intake.57 Aluminum in hair is not a

useful biological indicator of exposure.

Arsenic (As) – Arsenic is well taken up in hair. Animals show a dose-related increase in

hair arsenic.42 Forensic hair tests can determine the time-course of chronic arsenic poisoning.58

Increased arsenic in soil (<20 to 370 ppm soil As) show a slight correlation with slightly elevated

hair arsenic using group statistics (0.02 ppm to 0.06 ppm hair As).59 Consumption of drinking

water with elevated arsenic concentrations showed a correlation with hair arsenic, using group

comparisons.60-62 This correlation was not seen in a study where drinking water exposure was

only modestly above a legal threshold.63 Group statistics show elevated hair As in patients with

Blackfoot disease.64

Cadmium (Cd) – Animal studies show conflicting results with respect to any correlation

between cadmium in hair and the target organ, the kidney.42,52 The most significant nonoccupational exposure to cadmium occurs through tobacco smoke. Smokers have elevated blood

cadmium levels compared to nonsmokers. Studies show conflicting results with respect to hair

cadmium concentrations in smokers versus non-smokers.65-67 A nationwide German

environmental survey found little correlation with cadmium in hair and active cigarette smoking,

although it was the major predictor for blood and urine cadmium concentrations. In contrast,

outdoor activities, seasonality, and cadmium in tap water were more important predictors in hair

cadmium concentrations, emphasizing the role of exogenous deposition of cadmium into hair.67

Copper (Cu) – Taylor’s review notes that animal studies showed a proportional

relationship between copper in hair and liver.52 Yoshinaga et al. found no significant correlation

between hair copper and various internal organs, including the liver, in autopsy samples.68
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Literature studies of human populations show conflicting results with respect to hair versus

serum copper.52,69,70 Serum copper is generally higher in women than in men.33,49 However, hair

copper is inconsistent with respect to sex. Contiera and Folin found no effect of sex on hair

copper.71 Sky-Peck found a modest correlation (p<0.025) for higher hair copper in women

compared to men (24 vs. 20 ppm).39 In human patients with biliary cirrhosis, or Wilson’s disease

(systemic copper intoxication), with increased liver copper, hair copper was typically not

increased.52 Further studies of Wilson’s disease confirmed these findings, with no increase in hair

copper in patients with this disease.72 In copper deficiencies (malnutrition or Menkes syndrome),

hair copper was not significantly reduced.52

Chromium (Cr) – Studies of hair chromium are somewhat limited. A large study (40,872

patients) in England found age-related decreases in hair chromium for males and females [0.98

ppm (mean at age 1-4 yrs) to 0.5 ppm (mean at age 70 plus yrs)], slightly lower hair chromium in

males ages 25-49 years, and a correlation between hair and serum chromium, all statistically

significant.73 In comparison, a U.S. study found no difference in hair chromium by sex or age in

987 individuals.39 Hair chromium has been hypothesized to increase in gestational diabetes (in

early pregnancy), compared to non-diabetic pregnant women.74 Hair chromium measurements

have been used in monitoring occupational exposures, although blood and urine chromium are

the standard biological indices.75

Lead (Pb) – There are a number of studies relating lead exposure to tissue concentrations,

including hair. Animal studies show a dose-dependent correlated increase in lead in bone and

hair during the exposure period.76 Isotopic tracer studies have shown the deposition of lead into

human facial hair, interpreted as the integral of a blood lead pool over approximately 3 months.77

In humans, hair analysis can be used to demonstrate lead poisoning.72 Occupational exposures

show a correlation between blood and hair lead.52,78 Lower-level exposures have more variable

results,52 but larger studies appear to support a relationship between hair and blood lead.42

Exogenous deposition of lead onto scalp hair may be influential, e.g., season, dust exposure, and

hair treatment.42 Centers for Disease Control (CDC) investigators compared hair and blood

samples from 189 children to gauge the accuracy of using hair to screen for lead poisoning (mean
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blood lead 9.8 ug/dl; mean hair lead 7.2 ppm).79 Hair lead as a screening method had a 57%

sensitivity and 18% false-negative rate. The investigators concluded that hair lead measurements

are NOT an adequate method of screening for childhood lead poisoning. The reliable measure of

individual lead exposure is a blood lead test.

Selenium (Se) - Animal studies show that: 1) hair selenium is strongly influenced by the

chemical form of selenium and the level in the diet, with a greater increase for Lselenomethionine than sodium selenate 2) sodium selenate increases hair selenium but not

muscle selenium (the largest body Se pool); and 3) dietary methionine deficiency increases

selenium deposition in hair.80 These observations suggest caution when evaluating environmental

selenium exposures. Population measurements have shown a correlation between low hair

selenium and selenium-deficient soils.81 The hair-to-blood selenium ratio is calculated to be 3 in

dietary selenium deficiency, increasing to 10 as toxic levels are approached. Hair selenium will

continue to rise far beyond the plasma saturation concentration, indicating contribution from

another body pool.82 A hair concentration of >5 ppm Se is reported to be associated with elevated

exposure, while a concentration <0.12 ppm Se is reported to be associated with chronic selenium

deficiency.83 However, most population studies have preferred blood or urine to indicate

selenium exposure.84 Exogenous contamination with selenium-containing dandruff shampoos is a

serious confounding factor in developed countries.1

 Yoshinaga et al. found no significant

correlation between selenium concentrations in hair and in internal organs.68

Zinc (Zn) – Zinc in hair has been reviewed by several authors.

52,85-88 These reviewers note

that hair is a difficult medium for interpretation of zinc status. The interpretation of zinc

concentrations in hair can be obscured by confounders such as sex, body composition, and hair

treatment.89 In severe zinc deficiency, hair growth slows, producing normal or even elevated hair

zinc concentrations.87 Yoshinaga et al. found no significant correlation between concentrations of

zinc in hair and in various internal organs.68 Administration of zinc in the diet did not increase

zinc in beard hair.90 Serum zinc is typically decreased in dialysis patients. Hair zinc in these

patients is not consistent with serum findings.91
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In conclusion, with the exception of methylmercury, there is no good indication that hair

analysis offers any improvement over currently available clinical tests to determine individual

biological exposure to metals/metalloids of concern.92 Occupational texts note that hair analysis

is unproven to detect toxic chemicals in the body to account for symptoms and inappropriate in

the diagnosis of “environmental” illness.93 Group statistics on hair data, preferably geometric

means, may be useful in population screening for exposure to some of these metals (e.g.,

arsenic). Confounding factors, such as hair treatments, must be controlled for in these studies.

Analysis of hair minerals to predict nutritional status is a practice not supported by the state of

the science.

Topic #4. Data Gaps and Research Needs.

Generally speaking, further information is needed on concentrations of elements in the

hair of individuals with known exposures to trace elements, particularly where environmental

exposures are of concern. Laboratory studies of elemental concentrations in blood and target

tissues compared to hair concentrations are needed. Such data are important if one is to

hypothesize that there is a relationship between hair element concentrations and critical/target

organ effects. Clinical studies correlating hair concentrations with clinical conditions

(deficiencies or elevations) may also be helpful. Further work is needed on sample washing

methods. Standardization on one washing method is important for comparison of studies.

Specific recommendations:

• Do not use hair analysis for individual nutritional assessment. The state of the science does not

support this application.

• If hair analysis is undertaken for comparison of groups, choose element(s) for which the

literature supports such an approach, e.g., methylmercury, e.g., NOT aluminum.

• When studying control versus exposed groups, chose a group size of sufficient statistical power

to determine differences between group means, based on current literature findings.

• Use geometric means in analyzing group data.
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• Collect blood and/or urine samples for comparison with the hair results in the analysis of group

data. If this is not feasible for the entire study population, choose a subset of sufficient size to

provide statistically meaningful comparison data.

• A questionnaire should be administered to each individual in the study, determining: age, sex,

ethnicity, hair wash and hair treatment history including products used on hair, swimming habits,

time spent outdoors, occupation, smoking history, etc. (e.g, DiPietro et al., 1989).
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Topic #5: Identifying scenarios for which hair analysis may be appropriate.

Exposure Scenario Chemical/

Exposure Pathway

Exposure

Chronology

Exposure Duration Measurable

Health Effects

(Y/N)

Individual –

severe

poisoning/forensic

Group/population:

Mercury, Arsenic,

Lead

Methylmercurydiet (fish, seafood)

Arsenic,

Cadmium, Lead

Past / present

Past / present

Past / present

Acute (1-2 months

min.); chronic

Acute (1-2 months

min.); chronic

Acute (1-2 months

min.); chronic

Possible with very

high exposure

Unlikely unless

very high

exposure

Unlikely unless

very high

exposure

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ATSDR Hair Analysis Panel Discussion

APPENDIX D

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D-5

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D-6

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D-7

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ATSDR Hair Analysis Panel Discussion

D-19

Zlotkin SH. Hair analysis: a useful tool or a waste of money? Int J Dermatol. 1985 Apr;

24(3):161-4. Commentary.

http://www.doctorsdata.com/RESPONSE.HTM (Initial response to the recent JAMA article on

hair analysis, dated January 8, 2001) 

http://www.ctq.qc.ca/icpms.html (Centre de Toxicologie du Quebec’s Interlaboratory ICP-MS

Comparison Program)
ATSDR Hair Analysis Panel Discussion

APPENDIX E

Meeting Agenda
(over)

Agency for Toxic Substances and Disease Registry

Division of Health Assessment and Consultation

Hair Analysis: Exploring the State of the Science

Agenda

Day 1 Tuesday, June 12, 2001

 Discussing the State of the Science in Hair Analysis 

8:00 AM Registration

8:30 AM Introductory remarks .......... .......... .......... ..... Robert Amler, MD

ATSDR Chief Medical Officer

8:45 AM Purpose of meeting and review of the charge ......... Allan Susten, PhD, DABT

Assistant Director for Science, DHAC

9:00 AM Impetus for panel discussions—a case example .... Deanna K. Harkins, MD, MPH

Medical Officer, DHEP

9:10 AM Introduction of panelists .......... .......... ....... LuAnn White, PhD, DABT

Panel Chair

9:20 AM General physiology of hair—an overview .......... Robert Baratz, MD, PhD, DDS

9:40 AM Topic #1: Analytical methods .......... .......... .......... ...... Panelists

10:30 AM Break 

10:45 AM Topic #1: Analytical methods (continued) .......... .......... ...... Panelists

11:30 AM Observer comments

12:00 PM Lunch

1:00 PM Topic #2: Factors influencing the interpretation of analytical results ...... Panelists

3:15 PM Break

3:30 PM Topic #3: Toxicologic considerations .......... .......... ......... Panelists 

4:45 PM Observer comments

5:15 PM Adjourn
Day 2 Wednesday, June 13, 2001

Developing Recommendations

8:00 AM Review of day 1 issues .......... .......... .......... ........ Panel chair

8:15 AM Topic #4: Data gaps and research needs .......... .......... ...... Panelists

9:00 AM Topic #5: Identifying scenarios for which hair analysis may be appropriate . Panelists

10:15 AM Break

10:30 AM Observer comments

11:00 AM Conclusions/recommendations .......... .......... .......... .... Panelists

12:30 PM Adjourn
ATSDR Hair Analysis Panel Discussion

APPENDIX F

List of Observers
Agency for Toxic Substances and Disease Registry

Division of Health Assessment and Consultation

Hair Analysis: Exploring the State of the Science

Observers

Sherlita Amler

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-29)

Atlanta, GA 30333

404-498-0694

Fax: 404-498-0094

E-mail: sgr6@cdc.gov

J. Thomas Bernert

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS F-19)

Atlanta, GA 30341

770-488-7911 

Fax: 770-488-4609

E-mail: jtb2@cdc.gov

Mel Bishop

President-elect

Am erican Association of Bioanalysts

917 Locust Street - Suite 1100 

St. Louis, MO 63101-1419

314-241-1445 

Fax: 314-241-1449

E-mail: aab@aab.org

Kathleen Caldwell

Assistant Section Chief Elem ental Analysis

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS F-18)

Atlanta, GA 30341-3724

770-488-7990 

Fax: 770-488-4097

E-mail: klc7@cdc.gov

Gary Campbell

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-56)

Atlanta, GA 30333

404-498-0384

Fax: 404-498-0063

E-mail: ghc1@cdc.gov

Pat Carpenter

Director of Toxicology Operations

American Medical Laboratories

4230 Burnham Avenue 

Las Vegas, NV 89119

702-733-7866 Ext: 3443

Fax: 702-733-0318

E-mail: patrickc@apllabs.com

Selene Chou

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-29)

Atlanta, GA 30333

404-498-0705

Fax: 404-498-0092

E-mail: cjc3@cdc.gov

Susan Crampton

Technical Assistant

Research Triangle Institute

3040 Cornwallis Road - Building 3 

Research Triangle Park, NC 27709

919-541-6176 

Fax: 919-541-7042

E-mail: sdc@rti.org

Pam Floener

North American Representative for DMPS

Heyltex Corporation

5885 Cumm ing Highway (108-291)

Sugar Hill, GA 30518

770-831-8606 

Fax: 770-831-8610

E-mail: pfloener@aol.com

Laura Frazier

Geologist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-56)

Atlanta, GA 30333

404-498-0386

E-mail: lyb5@cdc.gov
Buck Grissom

Toxicologist

National Institute of Environmental Health Sciences

P.O. Box 12233 

111 T.W. Alexander Drive

Research Triangle Park, NC 27709

919-541-3365 

Fax: 919-541-4737

E-mail: grissom1@niehs.nih.gov

M. Olivia Harris

Senior Environmental Health Scientist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-28)

Atlanta, GA 30333

404-498-0620 

Fax: 404-498-0081

E-mail: oharris@cdc.gov

Rubina Imtiaz

Chief, Program Development Branch

Epidemiology Program Office

Division of International Health

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS K-72)

Atlanta, GA 30342-3724

770-488-4322

Fax: 770-488-8455

E-mail: rxi0@cdc.gov

Steve Inserra

Epidemiologist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-31)

Atlanta, GA 30333

404-498-0564 

Fax: 404-498-0079

E-mail: sai0@cdc.gov

Robert H. Johnson

Medical Officer

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0498

E-mail: rdj2@cdc.gov

Robert Jones

Chief, Elemental Analysis Laboratory

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS F-18)

Atlanta, GA 30341-3724

770-488-7991 

Fax: 770-488-4097

E-mail: rljones@cdc.gov

Laurie Kamimoto

Medical Officer

Centers for Disease Control & Prevention

1600 Clifton Road, NE (MS E-61)

Atlanta, GA 30333

404-639-8946

Fax: 404-639-8616

E-mail: lek0@cdc.gov

Melody Kawamoto

Centers for Disease Control & Prevention

4676 Columbia Parkway (MS R-10)

Cincinnati, OH 45226

513-841-4526 

Fax: 513-458-7105

E-mail: mm k2@cdc.gov

David Mellard

Toxicologist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0005

E-mail: dam7@cdc.gov

Susan Metcalf

Section Chief, Exposure Investigation Section

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0493 

Fax: 404-498-0655

E-mail: swm1@cdc.gov

Ketna Mistry

Medical Officer

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0451

E-mail: kwm5@cdc.gov

Jane Mitchell

Research Scientist

Colorado Department of 

Public Health & Environment

4300 Cherry Creek Drive South (DCEED-EE-A3)

Denver, CO 80246

303-692-2644 

Fax: 303-782-0904

E-mail: jane.mitchell@state.co.us
Daphne Moffett

Environmental Health Scientist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0456

Fax: 404-498-0073

E-mail: dmoffett@cdc.gov

Ken Orloff

Senior Toxicologist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0506

E-mail: keo1@cdc.gov

Chris Poulet

ATSDR Regional Representative

Agency for Toxic Substances & Disease Registry

Region 8 (MS 8ATSDR)

999 18th Street - Suite 300

Denver, CO 80202

303-312-7013 

Fax: 303-312-7018

E-m ail: cgp8@cdc.gov

 poulet.chris@epa.gov

David Quig

Director of Research & Clinical Counseling

Doctor's Data, Inc.

3755 Illinois Avenue 

St. Charles, IL 60174

630-377-8139, Ext: 149

Fax: 630-587-7861

E-mail: dquig@doctorsdata.com

John Risher

Senior Toxicologist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-29)

Atlanta, GA 30333

404-498-0737

Fax: 404-498-0092

E-mail: jzr8@cdc.gov

Nickolette Roney

Environmental Health Scientist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-29)

Atlanta, GA 30333

404-498-0708

Fax: 404-498-0092

E-mail: nroney@cdc.gov

R.H. Barry Sample

Director, Science & Technology

Quest Diagnostics

3175 Presidential Drive 

Atlanta, GA 30340

770-936-5024 

Fax: 770-936-5035

E-mail: barry.x.sample@questdiagnostics.com

Michael Schaeffer

Vice President, Lab Operations

Psychemedics Corporation

5832 Uplander W ay 

Culver City, CA 90230

310-216-7776 

E-mail: michaels@psychemedics.com

Margaret Schonbeck

Program Manager, Rocky Flats Health Studies

Colorado Department of 

Public Health & Environment

4300 Cherry Creek Drive South (RFHS A-3)

Denver, CO 80246

303-692-2636 

E-mail: mschonb@smtpgate.dphe.state.co.us

Connie Sosnoff

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS F-19)

Atlanta, GA 30341

770-488-7910 

E-mail: css3@cdc.gov

Steven Steindel

Supervisory Health Scientist

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS G-23)

Atlanta, GA 30341

770-488-8126

Fax: 770-488-8275

E-mail: sns6@cdc.gov

Anthony Suruda

Board Treasurer

Association of Occupational Environmental Clinics

Rocky Mountain Center for

Occupational & Environmental Health 

University of Utah 

75 South 2000 East

Salt Lake City, UT 84112-0512

801-581-3850 

Fax: 801-585-3759

E-mail: asuruda@dfpm.utah.edu
Francisco Tomei-Torres

Program Manager for Environmental Justice

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-28)

Atlanta, GA 30333

404-498-0664 

Fax: 404-498-0087

E-mail: fbt3@cdc.gov

Ben Wells

Manager, Research & Development

Quest Diagnostics

3175 Presidential Drive 

Atlanta, GA 30340

770-936-5020 

Fax: 770-936-5012

E-mail: ben.r.wells@questdiagnostics.com

Clement Welsh

Environmental Scientist

Agency for Toxic Substances & Disease Registry

1600 Clifton Road, NE (MS E-32)

Atlanta, GA 30333

404-498-0448

E-mail: cqw9@cdc.gov

Suzanne Wolfe

Centers for Disease Control & Prevention

4770 Buford Highway, NE (MS F-19)

Chamblee, GA 30341

770-488-7158 

Fax: 770-488-4609

E-mail: zer6@cdc.gov
APPENDIX G

Post-Meeting Observer Comments

Buck Grissom

Michael Schaffer
Buck Grissom, Ph.D.

Health Science Administrator (Biomedical Sciences)

National Institute of Environmental Health Sciences

Comments Concerning the Interpretation of Hair Analysis

I have used only data from hair analysis to help determine when an exposure occurred (see Item 1). There

are too many variables to use hair data for any other purpose (see Item 2). I have commented on several

instances in which hair data had been misused/misinterpreted by citizens and health care providers.

Example: I received a call from a concerned parent. His son’s symptoms were as follows: dizziness, poor

skin color, poor mental acuity, and blackouts (petit mal seizure like symptoms). A physician had

analyzed his son’s hair for metals and recommended chelation therapy at a cost of $6,000. His son’s hair

levels were within levels typically reported for control groups in hair analysis studies. Neither a source

nor a pathway had been identified. No one else was in the area was having similar health problems. Their

water was not contaminated. I told the parent to get a second opinion. Additional testing was needed to

determine his son’s health problems; hair analysis was inadequate. Moreover, chelation therapy is not

risk free. I suggested the closest Association of Occupational Environmental Clinics clinic or a

pediatrician trained to diagnose neurological symptoms before proceeding with chelation therapy.

General Questions (page 2)

1. When is it appropriate to consider hair analysis in assessing human exposures to environmental

contaminants?

COMMENT: If a source and pathway have been identified, hair analysis may provide information

concerning episodic exposures (i.e., frequency and duration of exposure).

2. When is it inappropriate to consider hair analysis in assessing human exposures to environmental

contaminants?

COMMENT: Information concerning sources, pathways, etc. (i.e., is exposure plausible), of interest to

ATSDR is required before attempting to interpret hair data. Interpretation of data from hair analysis in

the absence of environmental data is conjectural.

3. What data gaps exist that limit the interpretation and use of hair analysis in the assessment of

environmental exposures? What research is needed to fill these data gaps?

COMMENT: Internal sources of metals detected in hair need to be distinguished from external sources.

COMMENT: External sources of arsenic need to be distinguished (e.g., air, food, water, medicinals, and

hair dyes). Is there a hazardous waste site involved?

COMMENT: Analytical methods that result in elimination of intra- and inter-laboratory data variability

are needed.
ATSDR Hair Analysis Panel Discussion

G-2

Specific Questions

4. Topic #2 (page 3)

# Factors Influencing the Interpretation of Analytical Results.

COMMENT: This section lists many of the factors that confound interpretation of hair data. I agree with

Dr. Baratz’s comment: I do not want information that I cannot interpret. Even under the best

circumstances, hair data are exceedingly difficult to interpret. In cases where hair levels exceed levels

expected in a population, additional information is needed. 

5. Topic #3, Toxicologic Considerations (page 4) 

# Is information available defining “normal” ranges of chemical concentrations in hair that have

physiological and health significance?

COMMENT: Terms such as standard or normal hair levels of metals or reference ranges for metal levels

in hair need to be carefully defined—i.e., what constitutes normal? All reference values for hair need to

be representative of the population being evaluated. For example, groundwater levels of arsenic (e.g., 100

ppb) have been reported to be elevated in some areas of Utah, Michigan, and Maine. Hair levels of

arsenic in these areas are likely to be greater than hair levels in areas with low levels of arsenic in

groundwater (e.g., 2 ppb).

COMMENT: What do hair levels above a reference value mean? How will hair data be interpreted?

Reference values are frequently used for purposes for which they were not intended. A law firm sent a

letter to the U.S. EPA citing the CDC lead guidance as a basis for not conducting an environmental

investigation requested by EPA. The letter stated that the blood lead levels in the community were not

above 20 microgams per deciliter (μg/dL) and did not consistently exceed 15 μg/dL; therefore, an

environmental investigation was not needed. 
ATSDR Hair Analysis Panel Discussion

G-3

Michael Schaffer

Psychemedics Corporation

• Partial listing of cases demonstrating judicial acceptance of the Psychemedics

hair analysis method.

• Information on hair testing and racial or color bias.

• Information on the effectiveness of Psychemedics washing procedures for ruling

out external contamination
PARTIAL LISTING OF THOSE CASES DEMONSTRATING JUDICIAL

 ACCEPTANCE OF THE PSYCHEMEDICS HAIR ANALYSIS METHOD

A. Employment Cases

Scott v. The City of New York. et al., Civil Action No. 98-C V-1902 (ERK), (U.S.D.C., Eastern

Dist. NY, March 21, 2001), a case involving a claim of constructive discharge based on race and

gender, was dismissed via summary judgment. In making its decision, the court relied on the

plaintiff’s hair test result, which was positive for marijuana, as well as plaintiff’s prior

admission of use.

In Jones et al. v. City of Chicago, Civil Action No. 99 C 8201, (U.S.D.C., Northern Dist. IL,

November 28, 2000), a case involving claims of race bias in hair testing, the United States

District Court granted summary judgment in favor of the City of Chicago and dismissed the case.

The Court found that not only was some of the evidence inadmissible, but also that “the

remaining admissible evidence would be insufficient for a trier of fact to find that the

[Psychemedics] hair test is more likely to result in false positive results for African-American

applicants that for white applicants...”

In Cruse v. Whirlpool Corp., Civil Action No. 99-2129, (U.S.D.C., Dist. AR, June 23, 2000), the

United States District Court found no merit to plaintiff’s allegations that the Psychemedics hair

test (“RIAH”) was racially biased against African Americans and, as such, granted the

defendant’s motion for summary judgment. “Summary judgment is not appropriate unless all the

evidence points toward one conclusion . . .” (citing Hardin v. Hussman Corp., 45 F. 3d 262 (8th

Cir. 1995)). The defendant’s expert offered through written testimony that “there is absolutely

no scientific support for the notion that plaintiff’s test result could be positive because of her

race.” The court considered the plaintiff’s failure to offer any statistical evidence in support of

her claim of racial bias in granting the defendant’s motion.

In Gregory Hicks et al. v. City of New York et al., Index No. 119154 (1999), the Supreme Court

of the State of New York upheld the termination of three officers through the use of

Psychemedics’ hair analysis drug testing.

In Brinson v. Howard Safir, et al, 680 N.Y.S. 2d 500, 255 A.D. 2d 247, (N.Y.A.D. 1 Dept.

1998), the New York Supreme Court Appellate Division upheld the lower court’s determination

of the accuracy of Psychemedics hair testing performed on an NYPD officer. Subsequent to this

decision, the plaintiff filed suit in federal court, (E.D.N.Y. Civil Action No. 98-CV-2784

(ERK)(JMA)), claiming, in part, that he had not been afforded procedural due process before he

was terminated from his position. The court, in granting the defendants’ motion for summary

judgment, found that the plaintiff was afforded, and took advantage of, every opportunity to

appeal his dismissal. The court also referenced the Appellate Division’s holding that “there was

reasonable suspicion to order the testing...and there was no reason to doubt the accuracy of the

test results.”

In Matter of Brown v. City of New York, 250 AD2d 546, 673 NYS2d 643, (1998), the New York

Supreme Court Appellate Division affirmed the New York Police Department’s discharge of a

New York City police officer for failure to pass a Psychemedics hair analysis drug test. Claims

of contamination and inadequacies of testing were determined to be devoid of merit.
ATSDR Hair Analysis Panel Discussion

G-5

In Nevada Employment Security Department et al. v. Cynthia Holmes, 914 P.2d 611 (Nev.1996),

the Nevada Supreme Court held the following with regard to a stand alone Psychemedics hair

test utilized to deny unemployment benefits:

We acknowledge that there are, arguably, no certainties in science. See Daubert v.

Merrell Dow Pharmaceuticals, Inc., U.S. , 113 S.Ct. 2786, 2795 (1993). Nonetheless,

we conclude that RIA [hair] testing especially when coupled with a confirmatory

GC/MS test, is now an accepted and reliable scientific methodology for detecting illicit

drug use.

...we conclude that Holmes’ ingestion of cocaine, subsequently proven by the RIA

screening and confirmatory GC/MS test constitutes misconduct within the definition of

NRS. 6 12.385.

In Bass v. Florida Department of Law Enforcement, Criminal Justice Standards and Training

Commission. 627 So.2d 1321 (Fla. Dist. Ct. of Appeals, 1993), the plaintiff, a corrections

officer, appealed from the decision of a hearing officer that her criminal justice certification

should be revoked based on a positive urinalysis. The Court in Bass held that evidence of a

negative Psychemedics hair analysis was erroneously excluded and that “the radioimmunoassay

analysis of human hair to determine cocaine use is generally accepted in the scientific

community.” On remand, the hearing officer disregarded the hair analysis results as well as a

subsequent negative urinalysis result and again recommended the revocation of the plaintiff’s

certification. The plaintiff appealed a second time in Bass v. Fla. Dept. of Law Enforcement, 712

So. 2d 1171 (Ct. App. Fla 1998), in which case the Court affirmed the ruling of the lower court

holding that hair analysis should be admitted as it is “precisely the tool which is used when there

is a claim of error in a urinalysis for cocaine.”

B. Parole Revocation

In United States v. Medina, 749 F. Supp. 59 (E.D. N.Y. 1990), the court ordered a hair test to

determine if a probationer, in a parole revocation hearing, had violated his parole by utilizing

drugs in the preceding months. In revoking parole, after a positive Psychemedics hair test, the

court found that:

Extensive scientific writings on RIAH hair analysis establishes both its reliability and its

acceptance in the field of forensic toxicology when used to determine cocaine use.

In his decision, Judge Weinstein, the author of a treatise on evidence, analyzed the admissibility

of hair analysis in the Medina case under the Federal Rules of Evidence as well as the older Frye

evidence standard and concluded hair analysis was admissible under both. In addition, Judge

Weinstein took judicial notice of extensive writings which support the acceptance of the

reliability of RIAH.
ATSDR Hair Analysis Panel Discussion

G-6

C. Unemployment Insurance Appeal Board/Administrative Law Judge Decisions

The decisions of the Department of Labor to deny benefits to claimants who are terminated after

receiving positive hair test results for drugs of abuse are routinely upheld by Administrative Law

Judges and the State’s respective appeal or review boards. The decisions are upheld based on the

established reliability of Psychemedics’ hair analysis, which is demonstrated in numerous peer

reviewed scientific publications.

In In the Matter of Patrick Forte, New York Appeal Board No. 477610 (4/7/00), the

Unemployment Insurance Appeal Board upheld the determination of the Administrative Law

Judge, (A.L.J. Case No. 097-0852 1), in affirming the decision of the Department of Labor to

disqualify a probationary police officer, (“claimant”), from receiving benefits. The claimant was

disqualified after his termination due to willful misconduct. The claimant submitted to a hair test,

which results were positive for cocaine use. The claimant argued that either the hair sample was

contaminated due to his exposure to crack cocaine vapors, or that he “passively ingested” small

amounts of cocaine. The Appeal Board found that due to the fact that the claimant’s results

showed a cocaine level 4-8 times the cutoff level and that benzoylecgonine, a cocaine metabolite,

was also detected, it was unlikely that the claimant “passively ingested” cocaine. The Appeals

Board recognized that it had previously been demonstrated to the Board successfully that

Psychemedics’ laboratory’s washing techniques eliminated the issue of external contamination.

In In re Claim of Delbert Otto, B 95-02542-000 (1996), the State of Ohio Unemployment

Compensation Board of Review, (“Board of Review”), overturned the Hearing Officer’s ruling

that the claimant was discharged without just cause and was entitled to benefits. The Board of

Review found that expert testimony demonstrated the reliability of the Psychemedics hair test

which detected quantities of marijuana in the claimant’s hair.

See also In the Matter of Otis K. McBride, State of New York, A.L.J. Case No. 099-17766

(1999); In the Matter of James Rawls, State of New York, S.S.A. No.120-42-0562 (1998); In the

Matter of Claimant, State of Indiana, Case No. 93-1BA-l lOB (1994); and In the Matter of Brian

J. Berrigan, State of New York, Index No. 121899 (1998).

D. Arbitrations

Hair analysis has been upheld in arbitrations between Anheuser-Busch, Inc. and its unions:

• In an October 1999 decision, the collection of body hair for analysis was upheld.

• In a July 1999 decision, union claims of improper specimen collection, and age, race and gender

bias related to slow hair growth were found to have no merit and the issues were resolved in favor of

the Company.

• In an August 2000 decision, it was determined that random hair testing of employees in safety

sensitive positions did not violate their state constitutional rights to privacy. The Psychemedics hair
ATSDR Hair Analysis Panel Discussion

G-7

test was deemed “a reliable method for detecting employee drug use, [which] therefore served to

further the Employer’s legitimate safety interest.”

In United States Steel, A Division of USX Corp. and United Steelworkers of America, Local 1557, Case

No. USS-38, 287 (1999), the Arbitrator ruled:

We find that hair testing for drugs is legitimate under the LCA and scientifically valid.

Psychemedics’ wash procedures are effective in removing environmental contamination.

The 5.0ng/10mg cutoff level for cocaine is appropriate in light of field studies. There was no

bias here on the basis of race or hair color. The chain of custody was unbroken. The

Company has satisfied us that Grievant ingested cocaine during the period covered by the

Last Chance Agreement. That material violation of the LCA was proper cause for discharge.

Hair analysis was also upheld in US Steelworkers Local 4134 & Lone Star Steel Co., Case No. D22-96

(1997); Battle Mountain Gold Co. & Operating Engineers Local 3 (1998); Cooper Tools and United

Automobile, Aerospace, Agricultural Implement Workers of America, AFL-CIO, Local 1774, Grievance

No. 005 (2000); and United States Steel, A Division of USX Corp. and United States Steelworkers of

America, Local 1014, Case No. USS-41, 820 (2001).
ATSDR Hair Analysis Panel Discussion

G-8

Hair Testing and Racial or Color Bias

Every large scale population study dealing with race and or color bias has concluded that hair color or

race as factors do not lead to any statistically significant variations that would create a “bias.” Several

studies utilizing Psychemedics’ methodology, (extensive washing of the sample, complete digestion and

removal of melanin, the color component of hair), have established that there is no systematic bias

occurring with this specific technology.

A large study on the issue of possible racial bias and drug testing was originally reported in Forensic

Science International in 1993. The study involving 1200 real world cases showed that with all three

methods of reporting utilized, (self-reports, urine testing and hair analysis) the same positive percentage

ratio between Caucasians and African-Americans was achieved.

An even larger study, published in the July 1999 Journal of Occupational and Environmental Medicine

by Dr. Benjamin Hoffman, compared the 1997 results of hair and urine tests on over 1800 black and

white candidates for a large municipal police force. Again, no racial bias was found comparing hair

testing to urine testing.

In a 1999 study, published in Drug Testing Technology – An Assessment of Field Applications, “An

Analysis of the Racial Bias Controversy in the Use of Hair Assays” concluded from the analysis of

numerous data sets that any effect of hair color or race would be negligible as a factor in the outcome of a

hair test. The authors of the study reported that in side-by-side comparison with hair, urine and selfreports, the racial differential in positive rates compared to self-reports was actually greater in urine than

in hair analysis.

In January 2000, Dr. Mieczkowski’s meta-analysis of all available published studies that included data on

drug test results matched to race or hair color was published in Forensic Science International. These

studies included European research where participants were dosed with known quantities of drugs. In no

instance, in any study, was a statistical bias shown to exist.

Most recently, in the Bulletin of the International Association of Forensic Toxicologists, an analysis of

over 56,000 cases showed no significant relationship between hair color and a likelihood to test positive

for cocaine.

The “potential” to create bias issues exists with any specimen, including urine– as any element that

affects the matrix could arguably lead to a “biased” result.

A) Diet has a significant impact on urine excretion. Some ethnic diets may greatly influence a urine

result.

B) Patterns of water retention/urine excretion in women are influenced by menstrual cycles that may

create longer detection times in women.
ATSDR Hair Analysis Panel Discussion

G-9

C) Body weight and size influence the amount of drugs that would be found in urine.

D) Certain medications influence urine output and drug excretion rates.

E) The ability of the body to effectively process drugs is influenced by age which increases retention

times.

F) Water intake and activity dramatically influence drug excretion rates in urine. A sedentary person in

a wheelchair could retain drugs in urine significantly longer than an athletically active person who

hydrates his or her system.

None of these “potential bias” issues have presented much of a problem in workplace testing. This is

largely due to the fact that normal biovariability between individuals overwhelms any single element and,

of course, a person claiming any sort of bias would first have to admit drug use.

[Note: Dr. Schaffer provided copies of the following supporting journal articles]

Hoffman B. Analysis of race effects on drug-test results. Journal of Occupational and Environmental

Medicine. 41(1999) 612-614.

Mieczkowski T and Newel R. An evaluation of patterns of racial bias in hair assays for cocaine: black

and white arrestees compared. Forensic Science International. 63 (1993) 85-98.

Mieczkowski T and Newel R. An analysis of the racial bias controversy in the use of hair assays. In:

Drug Testing Technology: Assessment of Field Applications (ed: Mieczkowski, T.), CRC Press (1999),

Boca Raton, pp. 313-348.

Mieczkowski, T and Kruger, Michael. Assessing the effect of hair color on cocaine positive outcomes in

a large sample: a logistic regression on 56,445 cases using hair analysis. Bulletin of the International

Association of Forensic Toxicologists. (2001), 9-11.
ATSDR Hair Analysis Panel Discussion

1

Passive Contamination of Undercover Narcotics Officers by Cocaine: An Assessment of Their

Exposure Using Hair Analysis. Microgram, 1995.

Distinguishing Passive Contamination from Active Cocaine Consumption: Assessing the

Occupational Exposure of Narcotics Officers to Cocaine. Forensic Science International (84) 1997.

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Environmental Contamination

Psychemedics employs several independent approaches which in combination, rule out the

possibility of a positive result from external sources.

a) The rigorous chemical washing of hair for extended periods of time.

b) The analysis of the contents of these washes followed by a comparison of the drugs

remaining in the hair.

c) Measurement of metabolites, the unique compounds created by the body’s processing of the

drugs. These metabolites are normally not present in the environment or in smoke. For

example, marijuana smoke does not contain carboxy THC - the metabolite that Psychemedics

identifies in marijuana positives.

d) Use of cut-off levels with hair, as with urine, to prevent any passive internal exposure from

producing a positive result. Because of the constancy of drug concentrations in hair, these

cut-off levels more accurately reflect use, and are therefore safer than those used by

urinalysis.

Several studies by Dr. Thomas Mieczkowski of the University of South Florida1 dealt with the real world

issue of external contamination and its removal by appropriate wash procedures. The studies concerned

the passive contamination of undercover narcotic officers who, in the course of their duties, had

continuing and extensive contact with cocaine, operated in cocaine rich environments and interacted

frequently with cocaine users and cocaine dealers. The officers handled cocaine in the process of buying

and selling and when they made arrests or seized contraband.

These undercover officers effectively mimicked drug users in all respects, except usage. In his studies,

Dr. Mieczkowski found that the officers had some amount of detectable cocaine on the outside of their

hair as a contaminant. However, even in this extreme contamination scenario the hair was easily

cleansed. Dr. Mieczkowski concluded that the commercial wash procedures utilized (Psychemedics)

were effective methods for removing external contamination from hair and that external contamination

did not present a difficult problem with properly performed hair analysis.

In a contamination study utilizing an early Psychemedics wash procedure researchers exposed volunteers

to crack smoke in a small, unventilated room (2.5 x 3 x 2.5 m) and exposed cut hair to the equivalent of

smoke vapors from 5000 lines of cocaine in closed beakers. In all cases, after washing, the exposed
ATSDR Hair Analysis Panel Discussion

2Hair Analysis of Cocaine: Differentiation Between Systemic Exposure and External

Contamination. Journal of Clinical Pharmacology, 1992.

G-11

contaminated hair tested negative. The authors concluded that deposition of cocaine from even these

extreme contamination scenarios was washable.2 Also in the study, hair from admitted cocaine users

tested positive, hair from non-users tested negative and hair from non-users who admitted being present

in crack environments also tested negative. It is not likely that any employee would claim an exposure

scenario greater than being in an enclosed room while 5000 lines of cocaine were vaporized or handling

cocaine more frequently than an undercover narcotics officer or evidence technician.

Most recently, in a contamination study presented at the Society for Forensic Toxicologists this past

October, Psychemedics’ extensive wash procedures were compared to the short wash results obtained in

an earlier cocaine contamination study and were shown to be effective at distinguishing contaminated

hair from user hair.

Due to the hyper-sensitivity of urine tests, it is well recognized by the scientific community that false

positives due to passive internal exposure to drugs are far more likely for urinalysis than for hair analysis

(e.g., the opiate false positive problem of urinalysis due to poppy seed ingestion). The Department of

Health & Human Services found that over 87% of urine opiate confirmed positives were overturned by

medical review officers because ingestion of poppy seeds as well as some medications could cause urine

opiate cutoff levels to be exceeded. The studies of Dr. Hans Sachs and those of others have shown that

even the massive ingestion of poppy seeds is incapable of producing interpretive false positive hair

analysis results. Additionally, the hair of heroin users contains stable amounts of the heroin metabolite, 6

MAM (an absolute marker of heroin). Testing for the 6 MAM metabolite in urine is required under the

amended NIDA urine guidelines, (the amended guidelines also increase the cut off levels from 300

ng/mL to 2000 ng/mL). Unfortunately, while 6 MAM is identifiable in hair for months, it has an

extremely short half-life in urine and for all practical purposes will be detectable at best only in persons

who use heroin on the day oftheir urine test. This makes the confirmation of heroin use extremely

problematic for urine testing creating false negatives.

NIDA scientist, Dr. Cone, experimentally demonstrated that as little as one-hundredth of a line of cocaine

(i.e., 1 or 2 mg) can produce interpretive false positive urinalysis results.3 These small quantities can be

inadvertently ingested by a non-drug user (e.g., a spouse) who may be in the constant presence of a drug

abuser. In contrast to the resistance of hair to drug penetration, the lungs and gastrointestinal tract have

absolutely zero resistance. In actual fact, drugs are transported by active transport mechanisms into the

interior milieu, i.e., by breathing or by active membrane processes. Such active internalization can cause

interpretive false positive urine results by minute amounts of cocaine if the timing of the test is in close

proximity to the passive ingestion.

Unlike hair, there is no method to remove this contamination from urine or to differentiate between

active drug use and unknowing exposure to a drug that may rise above cut off levels, e.g., spiked food or

drink. Unlike urine, hair can be segmented to substantiate or refute these claims. Additionally, a

completely new hair sample can be obtained that will replicate the same time frame of the original

sample eliminating concerns or claims of sample mix-up. New samples replicating the same time frame

cannot be obtained with urine as most drugs are completely flushed from the system in a couple of days.