LC/DAD/MSD技术研究大鼠胆汁中盐酸非洛普的II相代谢产物

目的研究大鼠服药后胆汁中盐酸非洛普(DDPH)II相代谢产物.方法收集大鼠空白胆汁及给药后12h内的胆汁,以LC/DAD/MSD技术判断II相代谢产物峰位.以HPLC法制备II相代谢产物馏分并以β-葡糖醛酸酶水解,再进行分析;同时将与II相代谢产物相应的I相代谢产物对照品按相同条件进行分析对照.结果大鼠给药后胆汁色谱图中峰M₇,M₈和M₉为DDPH的II相代谢产物,它们的β-葡糖醛酸酶水解产物分别为M₃,M₂和M₁.结论β-1-O-{3,5-二甲基-4--2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₇),β-1-O-{2,4-二甲基-3-2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₈)和β-1-O-{2-甲氧基-4-1-甲基-2-(2,6-二甲基苯氧基)-乙氨基-乙基]-苯基}-葡糖醛酸(M₉)为大鼠ipDDPH后产生的II相代谢产物.

STUDY ON THE PHASE II METABOLITES OF PHENOPROLAMINE HYDROCHLORIDE IN RAT BILE BY LC/DAD/MSD

AIM: To study the phase II metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered by i.p. to bile duct-cannulated rats. Bile samples were collected before drug administration and up to 12 h after drug administration. After being purified and enriched with C-18 SPE columns the rat bile samples were analyzed by LC/DAD/MSD to identify the peaks of phase II metabolites. The fractions of phase II metabolites were prepared by HPLC and treated with beta-glucuronidase, and then were purified and enriched with C-18 SPE columns and analyzed by LC/DAD/MSD. The corresponding reference standards of DDPH phase I metabolites were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The peaks M7, M8 and M9 in the chromatograms of rat bile samples were the phase II metabolites of DDPH and the enzymatic hydrolysates of M7, M8 and M9 were 1-(2, 6-dimethyl-4-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M2) and 1-(2,6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethyl-amino)-propane (M1) respectively. CONCLUSION: beta-1-O-3,5-dimethyl-4--2-methyl-2-(3,4-dimethoxy-phenylethylamino)- ethoxy]-phenyl]-glucuronic acid (M7, glucuronide of M3), beta-1-O-2, 4-dimethyl-3-2-methyl-2-(3, 4-dimethoxy-phenylethylamino)-ethoxy]-phenyl]-glucuronic acid (M8, glucuronide of M2) and beta-1-O-2-methoxy-4-1-methyl-2-(2, 6-dimethylphenoxy)-ethylamino-ethyl]-phenyl]-glucuronic acid (M9, glucuronide of M1) were the phase II metabolites of DDPH in rat bile.