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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。
OBJECTIVE: To study the metabolism of intestinal flora of Pulsatilla in rats, and to provide references for the study of the metabolic characteristics of saponins in intestinal flora. Methods: Forsythin as internal standard was used to establish the UPLC-ESI-Q-TOF-MS / MS. The mobile phase consisted of gradient elution of acetonitrile-water (B) % A; 8-10 min, 95% A; 10-10.10 min, 95% -10% A) with a flow rate of 0.3 m L · min -1. The scanning mode was electrospray ionization Voltage of -3 500 V, collision voltage of 130 V, dry gas flow rate of 12 L · min -1, temperature of 350 ℃ and sheath gas flow rate of 12 L · min -1 at 350 ℃. The fragmentation voltages were 20,35,50 V. Total ion current full scan range m / z 100 ~ 1 100. Negative ion and multi-reactive ion monitoring mode was used to analyze the metabolites of Pulsanion saponin B_3 co-incubated with intestinal flora of rats. Results: The 3-position side chain desugar metabolites of Pulsatilla saponin B3 and the hydroxylation, carboxylation and decarboxylation, methylation and demethylation of metabolites in the aglycones were identified from the drug- 8 species. CONCLUSION: The intestinal flora of rats can effectively metabolize Pulsatilla saponin B3 under the experimental conditions, suggesting that the above metabolites are bioactive components.