UPLC-Q-trap-MS鉴定白头翁皂苷D在大鼠离体肠道菌群中的代谢产物

Identification of metabolites of Pulsatilla saponin D in intestinal microflora of rats in vitro by UPLC-Q-trap-MS

目的鉴定白头翁皂苷D在大鼠离体肠道菌群体外代谢产物。方法运用超高效液相色谱-三重四级杆线性离子肼复合质谱(UPLC-Q-trap-MS)分析白头翁皂苷D体外肠道菌群孵育样品,采用Analyst 1.5软件进行数据处理,根据保留时间和各色谱峰质谱裂解规律,比较确定白头翁皂苷D在体外肠道菌群中代谢产物,同时根据质谱二级裂解规律对代谢产物结构进行推测。结果在大鼠离体肠道菌群孵育样品中鉴定了白头翁皂苷D(M0)原型及常春藤皂苷元3-O-α-L-吡喃鼠李糖-(1→2)-α-L-吡喃阿拉伯糖苷(M1)、常春藤皂苷元3-O-β-D-吡喃葡萄糖-(1→4)-α-L-吡喃阿拉伯糖苷(M2)、常春藤皂苷元(M3)、白头翁皂苷D羟基化(M4)、白头翁皂苷D甲基化产物(M5、M6)、白头翁皂苷D脱氢产物(M7)等7个代谢产物。结论白头翁皂苷D能够在大鼠离体肠道菌群中发生广泛的代谢。

Objective To identify the metabolites of Pulsatilla saponin D （PSD） in the intestinal microflora of rats in vitro. Methods The sample of PSD incubated in intestinal microflora of rats in vitro was analyzed by a ultraperformance liquid chromatography/hybrid triple quadrupole linear ion trap mass spectrometry （UPLC-Q-trap-MS）. The data were processed with Analyst 1.5 software, and the metabolites of PSD were confirmed by comparing the retention time and MS/MS spectra. The possible structures of metabolites were proposed by analyzing the proposed fragmentation pathway. Results PSD （M0, parent） and seven metabolites were identified, including hederagenin 3-O-α-L-rhamnopyranosyl-（1→2）-α-L-arabinosyl （M 1）, hederagenin 3-O-α-L-glucopyranosyl-（1→4）-α-L-arabinosyl （M2）, hederagenin （M3）, hydroxylation of PSD （M4）, methylation of PSD （M5 and M6）, and dehydrogenation of PSD （M7）. Conclusion PSD has a wide range of metabolic in the intestinal microflora of rats in vitro.