基于UPLC-Q-TOF-MS的α-别隐品碱在大鼠体内的代谢产物分析

Analysis of Metabolites ofα-allocryptopine in Rats Based on UPLC-Q-TOF-MS

目的分析α-别隐品碱经灌胃给药后在大鼠体内的代谢产物,阐明其在大鼠体内的代谢途径。方法大鼠灌胃给药α-别隐品碱20 mg·kg-1后,分别收集血浆、尿液及粪便样品,采用超高效液相色谱-四级杆飞行时间质谱联用(UPLC-Q-TOF-MS)技术进行分析,获得各样品中代谢产物的保留时间、精确分子量、二级碎片信息。利用MetabolitePilotTM代谢软件进行预测,预测结果结合原型化合物的质谱裂解规律进一步进行裂解行为的分析,鉴定α-别隐品碱在大鼠体内的代谢产物。结果共鉴定出25个代谢产物,Ⅰ相代谢产物主要为α-别隐品碱甲氧基去甲基化、亚甲二氧基开环及开环后去甲基、氧化产物,Ⅱ相代谢产物主要为Ⅰ相代谢产物的葡萄糖醛酸、硫酸结合产物及少量葡萄糖基化产物。结论采用UPLC-Q-TOF-MS结合MetabolitePilotTM软件及化合物的质谱裂解规律可实现α-别隐品碱代谢产物的快速分析,为进一步研究其药效物质基础提供参考。

Objective To identify the metabolites of α-allocryptopine in rats.Methods The plasma,urine and feces samples were collected processed and determined by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) after oral administration of α-alloc ryp topi ne.The metabolites were predicted using MetabolitePilotTM software.Theses predicted metabolites were further analyzed by MS2 spectra,and compared with the detailed fragmentation pathway of α-allocryptopine standard.Results A total of25 metabolites were identified in rat plasma,urine and feces samples after oral administration of α-allocryptopine.The α-allocryptopine underwent extensive phase Ⅰ and phase Ⅱ metabolism in rats.The phase Ⅰ biotransformation of α-allocryptopine consist of αdemethylation,ring opening of 2,3-thylenedioxy group and followed by 0-demethylation,oxidation.The phase Ⅱ biotransformations include glucuronide conjugation,sulfate conjugation and glucosylation.Conclusion UPLC-Q-TOF-MS combined with MetabolitePilotTM software and fragmentation pathway was efficient to analyze the metabolites of α-allocryptopine.The results would provide key information for understanding the material basis and function mechanism of α-allocryptopine.