Abstract
Urinary dialkylphosphate (DAP) metabolites have been used to estimate human exposure to organophosphorus pesticides. We developed a method for quantifying the six DAP urinary metabolites of at least 28 organophosphorus pesticides using lyophilization and chemical derivatization followed by analysis using isotope-dilution gas chromatography–tandem mass spectrometry (GC–MS/MS). Urine samples were spiked with stable isotope analogues of the DAPs and the water was removed from the samples using a lyophilizer. The dried residue was dissolved in acetonitrile and diethyl ether, and the DAPs were chemically derivatized to their respective chloropropyl phosphate esters. The chloropropyl phosphate esters were concentrated, and analyzed using GC–MS/MS. The limits of detection of the method were in the low μg/l (parts per billion) to mid pg/ml range (parts per trillion) with coefficients of variation of 7–14%. The use of stable isotope analogues as internal standards for each of these metabolites allows for sample-specific adjustment for recovery and thus permits a high degree of accuracy and precision. Use of this method with approximately 1100 urine samples collected from pregnant women and children indicate that the low limits of detection allow this method to be used in general population studies.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 6 print issues and online access
$259.00 per year
only $43.17 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Anwar W.A. Biomarkers of human exposure to pesticides. Environ Health Perspect 1997: 105(Suppl. 4): 801–806.
Aprea C., Sciarra G., and Lunghini L. Analytical method for the determination of urinary alkylphosphates in subjects occupationally exposed to organophosphorus pesticides and in the general population. J Anal Toxicol 1996a: 20: 559–563.
Aprea C., Sciarra G., Orsi D., Boccalon P., Sartorelli P., and Sartorelli E. Urinary excretion of alkylphosphates in the general population (Italy). Sci Total Environ 1996b: 177: 37–41.
Aprea C., Sciarra G., Sartorelli P., Ceccarelli F., and Centi L. Multiroute exposure assessment and excretion of urinary metabolites of fenitrothion during manual operations on treated ornamental plants in greenhouses. Arch Environ Contamin Toxicol 1999: 36: 490–497.
Aprea C., Sciarra G., Sartorelli P., Desideri E., Amati R., and Sartorelli E. Biological monitoring of exposure to organophosphorus insecticides by assay of urinary alkylphosphates: influence of protective measures during manual operations with treated plants. Int Arch Occup Environ Health 1994: 66: 333–338.
Aprea C., Strambi M., Novelli M.T., Lunghini L., and Bozzi N. Biologic monitoring of exposure to organophosphorus pesticides in 195 Italian children. Environ Health Perspect 2000: 108: 521–525.
Azaroff L.S. Biomarkers of exposure to organophosphorous insecticides among farmers' families in rural El Salvador: factors associated with exposure. Environ Res 1999: 80: 138–147.
Bardarov V., and Mitewa M. High-performance liquid and gas chromatography of dialkylphosphates, dialkylthiophosphates and dialkyldithiophosphates as their pentafluorobenzyl derivatives. J Chromatogr 1989: 462: 233–241.
Bradway D.E., and Shafik T.M. Malathion exposure studies. Determination of mono- and dicarboxylic acids and alkyl phosphates in urine. J Agric Food Chem 1977: 25: 1342–1344.
Bravo R., Driskell W.J., Whitehead R.D., Needham L.L., and Barr D.B. Quantification of dialkyl phosphate metabolites of organophosphate pesticides in human urine using GC-MS/MS with isotope dilution method. J Anal Toxicol 2002: 26: 245–252.
CDC. National Report on Human Exposure to Environmental Chemicals. 2002, Available: www.cdc.gov/nceh/dls/. (Accessed January 2002.).
Cocker J., Mason H.J., Garfitt S.J., and Jones K. Biological monitoring of exposure to organophosphate pesticides. Toxicol Lett 2002: 134: 97–103.
Davies J.E., and Peterson J.C. Surveillance of occupational, accidental, and incidental exposure to organophosphate pesticides using urine alkyl phosphate and phenolic metabolite measurements. Ann NY Acad Sci 1997: 837: 257–268.
Drevenkar V., Radic Z., Vasilic Z., and Reiner E. Dialkylphosphorus metabolites in the urine and activities of esterases in the serum as biochemical indices for human absorption of organophosphorus pesticides. Arch Environ Contamin Toxicol 1991: 20: 417–422.
Driskell W.J ., Barr D.B., Beeson M.D., and Harmon I.R. GC/MS/MS analysis of dialkyl phosphate metabolites of organophosphate pesticides traces in urine. Presented at The American Society for Mass Spectrometry Conference, Dallas, TX, June, 1999, Poster No. 167.
EPA. Organophosphate Pesticides 1999. Available: www.epa.gov/pesticides/op. (Accessed December 2001).
Eskenazi B., Bradman A., and Castorina R. Exposures of children to organophosphate pesticides and their potential adverse health effects. Environ Health Perspect 1999: 107(Suppl. 3): 409–419.
Eskenazi B., Bradman A., Gladstone E.A., Jaramillo S., Birch K., and Holland N. CHAMACOS, a longitudinal birth cohort study: lessons from the field. Child Health 2003: 1(1): 3–27.
Fenske R.A., and Leffingwell J.T. Method for the determination of dialkyl phosphate metabolites in urine for studies of human exposure to malathion. J Agric Food Chem 1989: 37: 995–998.
Fenske R.A., Lu C., Simcox N.J., Loewenherz C., Touchstone J., Moate T.F., Allen E.H., and Kissel J.C. Strategies for assessing children's organophosphorus pesticide exposures in agricultural communities. J Expo's Anal Environ Epidemiol 2000: 10: 662–671.
Garfitt S.J., Jones K., Mason H.J., and Cocker J. Development of a urinary biomarker for exposure to the organophosphate propetamphos: data from an oral and dermal human volunteer study. Biomarkers 2002a: 7: 113–122.
Garfitt S.J., Jones K., Mason H.J., and Cocker J. Exposure to the organophosphate diazinon: data from a human volunteer study with oral and dermal doses. Toxicol Lett 2002b: 134: 105–113.
Gustafsson J.E., and Uzqueda H.R. The influence of citrate and phosphate on the Mancini single radial immunodiffusion technique and suggested improvements for the determination of urinary albumin. Clin Chim Acta 1978: 90: 249–257.
Hardt J., and Angerer J. Determination of dialkyl phosphates in human urine using gas chromatography–mass spectrometry. J Anal Toxicol 2000: 24: 678–684.
Hernández F., Sancho J.V., and Pozo O.J. Direct determination of alkyl phosphates in human urine by liquid chromatography/electrospray tandem mass spectrometry. Rapid Commun Mass Spectrom 2002: 16: 1766–1773.
Heudorf U., and Angerer J. Metabolites of organophosphorous insecticides in urine specimens from inhabitants of a residential area. Environ Res 2001: 86: 80–87.
Loewenherz C., Fenske R.A., Simcox N.J., Bellamy G., and Kalman D. Biological monitoring of organophosphorus pesticide exposure among children of agricultural workers in central Washington State. Environ Health Perspect 1997: 105: 1344–1353.
Lores E.M., and Bradway D.E. Extraction and recovery of organophosphorus metabolites from urine using an anion exchange resin. J Agric Food Chem 1976: 25: 75–79.
Lu C., Knutson D.E., Fisker-Andersen J., and Fenske R.A. Biological monitoring survey of organophosphorus pesticide exposure among pre-school children in the Seattle metropolitan area. Environ Health Perspect 2001: 109: 299–303.
Moate T.F., Lu C., Fenske R.A., Hahne R.M., and Kalman D.A. Improved cleanup and determination of dialkyl phosphates in the urine of children exposed to organophosphorus insecticides. J Anal Toxicol 1999: 23: 230–236.
Needham L.L., Ashley D.L., and Patterson Jr. D.G. Case studies of the use of biomarkers to assess exposures. Toxicol Lett 1995: 82-83: 373–378.
Oglobline A.N., Elimelakh H., Tattam B., Geyer R., O'Donnell G.E., and Holder G. Negative ion chemical ionization GC/MS-MS analysis of dialkylphosphate metabolites of organophosphate pesticides in urine of non-occupationally exposed subjects. Analyst 2001a: 126: 1037–1041.
Oglobline A.N., O'Donnell G.E., Geyer R., Holder G.M., and Tattam B. Routine gas chromatographic determination of dialkylphosphate metabolites in the urine of workers occupationally exposed to organophosphorus insecticides. J Anal Toxicol 2001b: 25: 153–157.
Persson B.A., and Vessman McDowall R.D. Is your method specific or just selective? LC-GC North Am 1998: 16(6): 556–560.
Reid S.J., and Watts R.R. A method for the determination of dialkyl phosphate residues in urine. J Anal Toxicol 1981: 15: 126–132.
Simcox N.J., Camp J., Kalman D., Stebbins A., Bellamy G., Lee I.C., and Fenske R. Farmworker exposure to organophosphorus pesticide residues during apple thinning in central Washington State. Am Ind Hyg Assoc J 1999: 60: 752–761.
Taylor J.K. Principles of measurements. In: Quality Assurance of Chemical Measurements. Lewis Publisher, Inc., Chelsea, MI, 1987, pp. 75–93.
Weisskopf C.P ., and Seiber J . New approaches to analysis of organophosphate metabolites in the urine of field workers. In: Wang R.G. Franklin C.A. Honeycutt R.C. Reinert C. (Eds.). Biological Monitoring for Pesticide Exposure. Measurement, Estimation, and Risk Reduction. Washington DC, American Chemical Society, 1989, pp. 206–214.
Wessels D., Barr D.B., and Mendola P. Use of biomarkers to indicate exposure of children to organophosphate pesticides: implications for a longitudinal study of children's environmental health. Environ Health Perspect 2003, doi: 10.1289/ehp.6316 (online 15 Sept 2003).
Westgard J.O . Basic QC Practices, Training in Statistical Quality Control for Health Care Laboratorie, 2nd edn.. Westgard QC, Madison, Available: http://www.westgard.com/basicqcbook.htm [accessed 10 June 2002].
Whyatt R ., Barr D.B . Measurement of organophosphate metabolites in postpartum meconium as a potential biomarker of prenatal exposure: a validation study. Environ Health Perspect 2001: 109: 417–420.
Acknowledgements
We recognize and thank the efforts of Kimberly Denise Smith in preparing samples for this study. We thank Pam Olive and Rosemary Schleicher for urinary creatinine measurements. Also, we appreciate the contributions of Brenda Eskenazi, director of CHAMACOS. Partial support for this research (for Asa Bradman) was provided by the US. Environmental Protection Agency (EPA) Grant R82679-01-0, and National Institute of Environmental Health Sciences (NIEHS) Grant PO1ES09605-02.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bravo, R., Caltabiano, L., Weerasekera, G. et al. Measurement of dialkyl phosphate metabolites of organophosphorus pesticides in human urine using lyophilization with gas chromatography-tandem mass spectrometry and isotope dilution quantification. J Expo Sci Environ Epidemiol 14, 249–259 (2004). https://doi.org/10.1038/sj.jea.7500322
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.jea.7500322
Keywords
This article is cited by
-
Urinary dialkylphosphate metabolites in the assessment of exposure to organophosphate pesticides: from 2000 to 2022
Environmental Monitoring and Assessment (2024)
-
Biomarkers of organophosphate insecticides exposure and asthma in general US adults: findings from NHANES 1999–2018 data
Environmental Science and Pollution Research (2023)
-
The association between urinary organophosphate insecticide metabolites and erectile dysfunction in the United States
International Journal of Impotence Research (2022)
-
Validated single urinary assay designed for exposomic multi-class biomarkers of common environmental exposures
Analytical and Bioanalytical Chemistry (2022)
-
Derivatization gas chromatography negative chemical ionization mass spectrometry for the analysis of trace organic pollutants and their metabolites in human biological samples
Analytical and Bioanalytical Chemistry (2020)