Determination of methyl 5-hydroxy-2-benzimidazole carbamate in urine by high-performance liquid chromatography with...

A high-performance liquid chromatographic (HPLC) assay for methyl 5-hydroxy-2-benzimidazole carbamate (5-HBC) in urine was developed in order to assess the exposure of workers to the pesticide carbendazim. 5-HBC is measured in urine after hydrolysis, sample clean-up through a strong cation-exchange (SCX) column and extraction with ethyl acetate. HPLC with electrochemical detection provides selective and sensitive determination of 5-HBC with a detection limit of 5 micrograms/l. A C18 reversed-phase column was used with 0.06 M ammonium acetate solution (pH 8)-methanol (73:27) as mobile phase. The method was validated with respect to hydrolysis of urine samples, analytical recovery of spiked 5-HBC, stability of 5-HBC conjugates, limit of detection, background and precision. The overall analytical recovery from urine was better than 60%. 5-HBC, excreted in urine as a conjugate, was stable for at least one year when stored at -20 degrees C. A background of ca. 5 micrograms/l was detected in urine from some non-occupationally exposed persons. Between-day coefficients of variation as calculated from the results of the stability test were 7, 4 and 4% for concentrations of 61, 244 and 295 micrograms/l 5-HBC, respectively (n = 16).