UPLC-ESI-Q-TOF-MS/MS分析白头翁皂苷B3在大鼠肠道菌群中的代谢产物

目的:研究大鼠肠道菌群对白头翁皂苷B_3的代谢作用,为皂苷类物质在肠道菌群的代谢特征研究提供参考。方法:以连翘苷为内标物,建立UPLC-ESI-Q-TOF-MS/MS检测条件,流动相乙腈(A)-水(B)梯度洗脱(0~8 min,10%~95%A;8~10 min,95%A;10~10.10 min,95%~10%A),流速0.3 m L·min~(-1),扫描方式为电喷雾(ESI)负离子模式,离子喷雾电压-3 500 V,碰撞电压130 V,干燥气流量12 L·min~(-1),温度350℃,鞘气流量12 L·min~(-1),温度350℃,碎裂电压分别为20,35,50 V。总离子流全扫描的范围m/z 100~1 100。采用负离子和多反应离子监测模式分析白头翁皂苷B_3与大鼠肠道菌群共孵育后的代谢产物。结果:从药物-菌群孵育液中鉴定出白头翁皂苷B3的3-位侧链脱糖代谢产物及苷元母核上羟化、羧化和脱羧、甲基化和去甲基化代谢产物共8种。结论:大鼠肠道菌群在实验条件下能有效代谢白头翁皂苷B3,提示上述代谢产物是其生物活性成分。

Metabolites Analysis of Pulchinenoside B3 in Rat Intestinal Flora by UPLC-ESI-Q-TOF-MS/MS

Objective: To investigate metabolism of rat intestinal flora on pulchinenoside B₃,so as to provide scientific basis for metabolism characteristic analysis of saponins in intestinal flora. Method: With phillyrin as internal standard,UPLC-ESI-Q-TOF-MS/MS with negative ion and multiple reaction monitoring (MRM) mode was applied to analyze metabolites of pulchinenoside B₃ in rat intestinal flora. Result: Pulchinenoside B₃ prototype and other eight metabolites were detected in rat intestinal flora.Various types of metabolitic reactions happen to pulchinenoside B₃ in intestinal flora,including 3-side chain sugar-remove,hydroxylation,methylation, and methoxylation on the mother nucleus of aglycone. Conclusion: Pulchinenoside B₃ can be effective metabolism by rat intestinal flora under these experimental conditions,which shows these eight metabolites are active ingredients of pulchinenoside B₃.