Determination of thiabendazole and 5-hydroxythiabendazole in human serum by fluorescence-detected high-performance...

We have developed a rapid, sensitive and precise high-performance liquid chromatographic method using fluorescence detection for the simultaneous determination of thiabendazole and unconjugated 5-hydroxythiabendazole in serum. Sample pretreatment consists only of protein precipitation with acetonitrile containing the internal standard, 2-methylindole. Detection limits were found to be 0.1 microgram/ml serum for thiabendazole and 0.4 microgram/ml serum for 5-hydroxythiabendazole. Between-day analytical precision coefficients of variation for serum-based controls were 7% and 11% for thiabendazole levels of 1 and 5 micrograms/ml, respectively; and 43% and 8% for 5-hydroxythiabendazole levels of 6 and 60 micrograms/ml, respectively. We also devised a microenzymatic method for the conversion of the glucuronide and sulfate esters of 5-hydroxythiabendazole using beta-glucuronidase [EC 3.2.1.31] and sulfatase [EC 3.1.6.1]. Thus, quantitation of the separate metabolites was possible. We also utilized a special adaptation of the chromatographic procedure for the determination of the 5-hydroxythiabendazole metabolites in the sera of uremic patients, which can contain large amounts of interfering fluorescent substances. The method should be particularly useful for monitoring thiabendazole therapy in patients unable to eliminate the potentially toxic metabolites.