Nordholm L, Dalgaard L
J Chromatogr. 1984 Feb 10;305(2):391-9. doi: 10.1016/s0378-4347(00)83353-8.
A high-performance liquid chromatographic method for the determination of trimethoprim metabolites in pig urine was developed. The metabolites-glucuronic acid and sulphuric acid conjugates of phenolic metabolites formed by demethylation of trimethoprim-were quantitated after treatment of urine with beta-glucuronidase (Escherichia coli). The sulphuric acid conjugate was not susceptible to enzymatic hydrolysis and was therefore assayed as the conjugate by use of ion-pair chromatography on the reversed-phase column. In order to find suitable conditions for enzymatic hydrolysis of the glucuronides, the conjugates were obtained by gel chromatography of urine from a [14C] trimethoprim-treated pig.
建立了一种测定猪尿液中甲氧苄啶代谢物的高效液相色谱法。甲氧苄啶去甲基化形成的酚类代谢物的代谢物——葡糖醛酸和硫酸结合物，在用β-葡糖醛酸酶（大肠杆菌）处理尿液后进行定量。硫酸结合物不易被酶水解，因此通过反相柱上的离子对色谱法作为结合物进行测定。为了找到葡糖醛酸苷酶促水解的合适条件，通过对用[14C]甲氧苄啶处理的猪的尿液进行凝胶色谱法获得结合物。
