基于UHPLC-Q-TOF/MS的地榆化学成分分析及大鼠体内代谢研究

目的  采用超高效液相色谱和-四级杆串联飞行时间质谱(UHPLC-Q-TOF/MS)法分析地榆水提物在大鼠血清、尿液及粪便中的原型成分及代谢产物, 探讨地榆中活性成分在大鼠体内的代谢途径。方法  采用Thermo AcclaimTM RSLC 120 C₁₈色谱柱(3.0 mm×100 mm, 2.2 μm), 以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱, 流速0.4 mL·min-1, 进样量3 μL, 柱温40 ℃。运用电喷雾离子源(ESI), 扫描范围m/z 50~1 500, 正、负离子模式采集数据。通过比较灌胃地榆水提物后大鼠血清、尿液和粪便样品和空白组样品的差异, 对原型成分及其代谢产物进行鉴定。结果  灌胃给予地榆水提物后, 从大鼠血清、尿液及粪便中鉴定出69个外源性成分, 包括24个原型成分和45个代谢产物。其中24个原型成分包括16个酚类成分、7个三萜类成分及1个单萜烯醇苷。地榆水提物在大鼠体内的代谢途径主要包括还原、甲基化、脱水、氧化、硫酸化及葡萄糖醛酸结合等。结论  所鉴定出的原型成分和代谢产物可为进一步解析地榆体内代谢过程, 阐明其药效物质基础提供实验依据。 

Identification of Chemical Composition of Sanguisorba officinalis L. and Its Major Metabolites in Rats by UHPLC-Q-TOF/MS

OBJECTIVE  To analyze the prototype components and metabolites of aqueous extract of Sanguisorba officinalis L. by UHPLC-Q-TOF/MS in feces, urine and plasma of rats and explore its metabolic pathways.METHODS  Chromatographic separation was performed on the Thermo AcclaimTM RSLC 120 C₁₈ column (3.0 mm×100 mm, 2.2 μm). The mobile phase was consisted of 0.1% formic acid in water (solvent A) and acetonitrile (solvent B). The sample injection volume was 3 μL and the flow rate was set to 0.4 mL·min-1. The column temperature was maintained at 40 ℃. The detection was carried out over a mass range of m/z 50-1 500 and equipped with an electrospray ionization (ESI) source operating in positive mode and negative mode. Chemical composition on Sanguisorba officinalis L. and its major metabolites were identified by comparing the differences in feces, urine and plasma between adminstration samples and blank samples in rats.RESULTS  After intragastric administration of the aqueous extract of Sanguisorba officinalis L., 69 exogenous components were identified from rat plasma, urine and feces, including 24 prototype components and 45 metabolites. 24 prototype components included 16 phenolics, 7 triterpenes, and a monoterpene glycoside. The metabolic pathways of the aqueous extract of Sanguisorba officinalis L. in rats mainly included reduction, methylation, dehydration, oxidation, sulfation, and glucuronidation.CONCLUSION  The identified prototype drugs and the metabolites can supply a clear material basis for the further study of efficacy of Sanguisorba officinalis L..