Fischer D, Breyer-Pfaff U
Department of Toxicology, University of Tübingen, Germany.
J Pharm Pharmacol. 1995 Jun;47(6):534-8. doi: 10.1111/j.2042-7158.1995.tb05845.x.
The activities of beta-glucuronidases from Helix pomatia, Escherichia coli and rat towards the N-glucuronides of amitriptyline and diphenhydramine were considerably lower than those towards standard substrates. The two N-glucuronides were analysed in urine samples by the following procedures: HPLC of the intact conjugate after solid-phase extraction on a cation exchanger cartridge or after direct injection of urine; HPLC of the aglycone after hydrolysis with beta-glucuronidase from H. pomatia or E. coli or after alkaline hydrolysis. Solid-phase extraction led to the highest recovery and precision, and sensitivity can be improved by extracting a larger volume of urine. On application to samples from patients under treatment with amitriptyline, the results of all procedures except alkaline hydrolysis were in good agreement. When diphenhydramine N-glucuronide was analysed in urine samples of volunteers, solid-phase extraction, hydrolysis by E. coli glucuronidase and alkaline hydrolysis resulted in similar values.
来自苹果螺、大肠杆菌和大鼠的β-葡萄糖醛酸酶对阿米替林和苯海拉明的N-葡萄糖醛酸苷的活性显著低于对标准底物的活性。通过以下方法分析尿液样本中的两种N-葡萄糖醛酸苷：在阳离子交换柱上进行固相萃取后或直接进样尿液后对完整缀合物进行高效液相色谱分析；用苹果螺或大肠杆菌的β-葡萄糖醛酸酶水解或碱性水解后对苷元进行高效液相色谱分析。固相萃取回收率最高、精密度最好，通过萃取更大体积的尿液可提高灵敏度。将这些方法应用于接受阿米替林治疗患者的样本时，除碱性水解外的所有方法结果均吻合良好。在对志愿者尿液样本中的苯海拉明N-葡萄糖醛酸苷进行分析时，固相萃取、大肠杆菌葡萄糖醛酸酶水解和碱性水解得到的值相近。
