Hair Analysis Panel Discussion:
Section: Appendix C, Dan Paschal

Appendix C
Pre-Meeting Comments

Hair Analysis: Exploring the State of the Science

Dan Paschal

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);
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.

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 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 non-standard "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 (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 Duration	Measurable Health Effects (Y/N)
Individual	Ingestion (MeHg)	Past	Chronic	Yes (if high)
Individual	Individual Ingestion (As 3/5)	Past	Chronic	Yes (if high)
Individual	Ingestion Lead	Past	Chronic	Yes (if evaluated)

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).
    
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, 4^th 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)

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