Watts M T, Raisys V A, Bauer L A
J Chromatogr. 1982 Jun 11;230(1):79-86. doi: 10.1016/s0378-4347(00)81432-2.
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.
我们开发了一种快速、灵敏且精确的高效液相色谱法，采用荧光检测同时测定血清中的噻苯达唑和未结合的5-羟基噻苯达唑。样品预处理仅包括用含内标物2-甲基吲哚的乙腈进行蛋白沉淀。噻苯达唑的检测限为0.1微克/毫升血清，5-羟基噻苯达唑的检测限为0.4微克/毫升血清。基于血清的对照品，噻苯达唑水平为1微克/毫升和5微克/毫升时，日间分析精密度变异系数分别为7%和11%；5-羟基噻苯达唑水平为6微克/毫升和60微克/毫升时，日间分析精密度变异系数分别为43%和8%。我们还设计了一种微酶法，使用β-葡萄糖醛酸酶[EC 3.2.1.31]和硫酸酯酶[EC 3.1.6.1]将5-羟基噻苯达唑的葡萄糖醛酸酯和硫酸酯进行转化。因此，可以对各个代谢物进行定量。我们还对色谱程序进行了特殊调整，用于测定尿毒症患者血清中的5-羟基噻苯达唑代谢物，这些患者的血清可能含有大量干扰荧光物质。该方法对于监测无法消除潜在有毒代谢物的患者的噻苯达唑治疗应该特别有用。
