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

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


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 


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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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