Analytical method for the quantitative determination of urinary ethylenethiourea by liquid chromatography/electrospray ionization tandem mass spectrometry.
A direct, rapid and selective method for the quantitative determination of the ethylenethiourea (ETU) in human urine has been validated and is reported in the present study. It allows the accurate quantification of ETU in this complex matrix without the use of any internal standard as the sample cleanup is effective enough for the removal of interferences that could lead to ion suppression in the electrospray ionization (ESI) source. This simple and rapid purification system, based on the use of a Fluorosil phase of a BondElut column followed by a liquid-liquid extraction procedure, achieves mean extracted recoveries, assessed at three different concentrations (2.5, 10.0, and 25.0 microg/L), always more than 85%. High-performance liquid chromatography (HPLC) with positive ion tandem mass spectrometry, operating in selected multiple reaction monitoring (MRM) mode, is used to quantify ETU in human urine. The assay is linear over the range 0-50 microg/L, with a lower limit of quantification (LOQ) of 1.5 microg/L and a coefficient of variation (CV) of 8.9%. The lower limit of detection (LOD) is assessed at 0.5 microg/L. The overall precision and accuracy were determined on three different days. The values for within- and between-day precision are < or = 8.3 and 10.1%, respectively, and the accuracy is in the range 97-118%. The relative uncertainties for the LOQ and QC concentrations have been estimated to be 18 and 8%, respectively. The assay was applied to quantify ETU in human urine from growers that regularly handle ethylenebisdithiocarbamate pesticides in large crop plantations. The biological samples were collected at the start and end of the working day, and the ETU urine levels were found to vary between 1.9 and 8.2 microg/L.
Sottani
Cristina
C
Laboratory of Environmental Hygiene and Industrial Toxicology, S. Maugeri Foundation, via Ferrata 8, Pavia, Italy. csottani@fsm.it
Bettinelli
Maurizio
M
Lorena Fiorentino
Maria
M
Minoia
Claudio
C
D002853:Chromatography, Liquid / Q000379:methods*; D005031:Ethylenethiourea / Q000032:analysis*; D005659:Fungicides, Industrial / Q000032:analysis; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D005659:Fungicides, Industrial; D005031:Ethylenethiourea