Measurement of dialkyl phosphate metabolites of organophosphorus pesticides in human urine using lyophilization with gas chromatography-tandem mass spectrometry and isotope dilution quantification

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 microg/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.