Analysis of ethylenethiourea as a biomarker in human urine using liquid chromatography/triple quadrupole mass spectrometry

Ethylenebisdithiocarbamates (EBDCs) are widely used fungicides. Ethylenethiourea (ETU), the main metabolite and also a contaminant in the commercially available products, is of major toxicological concern. In this study, a method using liquid chromatography/triple quadrupole mass spectrometry (LC/MS/MS) is described for the analysis of ETU in human urine after a single‐step extractive derivatization using pentafluorobenzyl bromide (PFBBr). Analysis was carried out using selected reaction monitoring (SRM) in the positive ion mode. Quantification of ETU was performed using [ 2 H 4 ]‐labeled ETU as internal standard (IS). The limit of detection (LOD) was determined to 0.05 ng/mL. The method was linear in the range 0.1–54 ng/mL urine and had a within‐run precision of 3–5%. The between‐run precision was determined at an average urine level of 2 and 10 ng/mL urine and found to be 9%. The inter‐batch precision was 6%. To validate ETU as a biomarker of exposure, the method was applied in a human experimental oral exposure to the commercial fungicide Ridomil Gold, containing 64% mancozeb and 4.5% ETU. Two healthy volunteers received 8.9 µg/kg body weight (b.w.) Ridomil Gold in a single oral dose followed by urine sampling for 104 h post‐exposure. The elimination half‐life of ETU was estimated to 17–23 h. Copyright © 2008 John Wiley & Sons, Ltd.