Analysis of 2,4-Dichlorophenoxyacetic Acid and 2-Methyl-4-Chloro-Phenoxyacetic Acid in Human Urine

  • Cristina Aprea
  • Gianfranco Sciarra
  • Nanda Bozzi
  • Liana Lunghini
Protocol
Part of the Methods in Biotechnology book series (MIBT, volume 19)

Abstract

Two methods for the quantitative analysis of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-methyl-4-chlorophenoxyacetic acid (MCPA) in urine are reported. Hydrochloric acid, sodium chloride, and internal standards (2,3-dichlorophenoxyacetic acid [2,3-D] or 4-chlorophenoxyacetic acid [4-CPA]) are added to the urine. The mixture is extracted with dichloromethane. For the high-performance liquid chromatographic (HPLC) method, the concentrated extract is drawn into a previously conditioned silica cartridge in which 2,4-D and MCPA are enriched. For the gas chromatographic (GC) method, after extraction the two compounds are converted to their pentafluorobenzyl esters and purified in a previously conditioned silica cartridge. Calibration is carried out using standard solutions in urine, which are processed in the same way as the urine samples and are determined by HPLC with diode array detector (DAD) or GC with electron capture detection (ECD). The peak areas of the chlorophenoxycarboxylic acids obtained are divided by the peak areas of the respective internal standard. The resulting quotients are plotted as a function of the concentration of the chlorophenoxycarboxylic acids to obtain a calibration curve. The two methods have detection limits of about 15 µg/L and 1 µg/L for 2,4-D and MCPA, respectively.

Key Words

Chlorophenoxycarboxylic acids general population GC analysis herbicides HPLC analysis occupational exposure pentafluorobenzylbromide derivatization 

References

  1. 1.
    Fjellstad, P. and Wannag, A. (1977) Human urinary excretion of the herbicide 2-methylchlorophenoxyacetic acid. Scand. J. Work Environ. Health 3, 100–103.Google Scholar
  2. 2.
    Sauerhoff, M. W., Braun, W. H., Blau, G. E., and Gehering, P. J. (1977) The fate of 2,4-dichlorophenoxyacetic acid (2,4 D) following oral administration to man. Toxicology 8, 8–11.CrossRefGoogle Scholar
  3. 3.
    Kolmodin-Hedman, B., Sverker, H., Ake, S., and Malin, A. (1983) Studies on phenoxy acid herbicides. II. Oral and dermal uptake and elimination in urine of MCPA in humans. Arch. Toxicol. 54, 267–273.CrossRefPubMedGoogle Scholar
  4. 4.
    Kanna, S. and Fang, S. C. (1966) Metabolism of C14-labeled 2,4-dichlorophenoxyacetic acid in rats. J. Agric. Food Chem. 14, 500–503.CrossRefGoogle Scholar
  5. 5.
    World Health Organization. (1984) Environmental Health Criteria 29. 2,4-Dichlorophenoxyacetic Acid (2,4-D). WHO, Geneva.Google Scholar
  6. 6.
    Nidasio, G. F., Burzi, F., and Sozzé, T. (1984) Determination of 2-methyl-4-chlorophenoxyacetic acid (MCPA) in urine using combined TLC and HPLC methods. G. Ital. Med. Lav. 6, 261–264.PubMedGoogle Scholar
  7. 7.
    Hutta, M., Kaniansky, D., Kovalcikova, E., et al. (1995) Preparative capillary isotachophoresis as a sample pretreatment technique for complex ionic matrices in high performance liquid chromatography. J. Chromatogr. A 689, 123–133.CrossRefGoogle Scholar
  8. 8.
    Vural, N. and Burgaz, S. (1984) A gas chromatographic method for determination of 2,4-D residues in urine after occupational exposure. Bull. Environ. Contam. Toxicol. 35, 51872–524.Google Scholar
  9. 9.
    Smith, A. E. and Hayden, B. J. (1979) Method for the determination of 2,4-dichlorophenoxyacetic acid residues in urine. J. Chromatogr. 171, 482–485.CrossRefPubMedGoogle Scholar
  10. 10.
    Hughes, D. L., Ritter, D. J., and Wilson, R. D. (2001) Determination of 2,4-dichlorophenoxyacetic acid (2,4-D) in human urine with mass selective detection. J. Environ. Sci. Hetalth B36, 755–764.CrossRefGoogle Scholar
  11. 11.
    Holler, J. S., Fast, D. F., Hill, R. H., et al. (1989) Quantification of selected herbicides and chlorinated phenols in urine by using gas chromatography/mass spectrometry/mass spectrometry. J. Anal. Toxicol. 13, 152–157.PubMedGoogle Scholar
  12. 12.
    Manninen, A., Kangas, J., Klen, T., and Savolainen H. (1986) Exposure of Finnish farm workers to phenoxy acid herbicides. Arch. Environ. Contam. Toxicol. 15, 107–111.CrossRefPubMedGoogle Scholar
  13. 13.
    Knopp, D. and Glass, S. (1991) Biological monitoring of 2,4-dichlorophenoxyacetic acid-exposed workers in agriculture and forestry. Int. Arch. Occup. Environ. Health 63, 329–333.CrossRefPubMedGoogle Scholar
  14. 14.
    Knopp, D. (1994) Assessment of exposure to 2,4-dichlorophenoxyacetic acid in the chemical industry: results of a 5 year biological monitoring study. Occup. Environ. Med. 51, 152–159.CrossRefPubMedGoogle Scholar
  15. 15.
    Aprea, C., Sciarra, G., and Bozzi, N. (1997) Analytical methods for the determination of urinary 2,4-dichlorophenoxyacetic acid and 2-methyl-4-chlorophenoxyacetic acid in occupationally exposed subjects and in the general population. J. Anal. Toxicol. 21, 262–267.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Cristina Aprea
    • 1
  • Gianfranco Sciarra
    • 1
  • Nanda Bozzi
    • 1
  • Liana Lunghini
    • 1
  1. 1.Unitàa Funzionale di Igiene Industriale e Tossicologia Occupazionale (Department of Industrial Hygiene and Occupational Toxicology)Laboratorio di Sanitàa Pubblica, Azienda USL 7 (Public Health Laboratory, National Health Service Local Unit 7)SienaItaly

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