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建立同时测定灌胃给予大鼠广金钱草提取物后胆汁中7种黄酮和1种酚酸类化合物浓度的HPLC-MS/MS,并用于胆汁排泄研究。大鼠灌胃给予广金钱草提取物后,分别在0~1,1~2,2~4,4~6,6~8,8~12,12~24 h时间段收集胆汁。色谱柱采用Diamonsil C18column(4.6 mm×150 mm,5μm),柱温40℃。以甲醇-0.01%乙酸水溶液为流动相,梯度洗脱,流速0.8 m L·min~(-1)。质谱采用电喷雾离子源(ESI),负离子模式下检测,多反应离子监测(MRM)模式进行定量。结果表明8种成分在测定浓度范围内线性关系良好(r≥0.991 5),日内、日间精密度RSD<15%,准确度RE-15%~15%,稳定性良好。提取回收率>63.2%,内标归一化的基质因子RSD均未超过15%,符合要求。大鼠胆汁排泄研究结果显示,8种成分在胆汁中的原型排泄率均比较小,并且不同个体之间差别较大。该研究方法灵敏度高,选择性好,样品处理简单,可用于灌胃给予大鼠广金钱草提取物后胆汁中8种成分的排泄动力学研究。
To establish a HPLC-MS / MS method for the simultaneous determination of 7 flavonoids and 1 phenolic acid in bile by intragastric administration of extract of Desmodium pullulans, and to study the biliary excretion. After gavage, the rats were given ginsenoside extract, and the bile was collected at 0 ~ 1, 1 ~ 2, 2 ~ 4, 4 ~ 6, 6 ~ 8, 8 ~ 12 and 12 ~ 24 h respectively. The column was Diamonsil C18column (4.6 mm × 150 mm, 5 μm) with a column temperature of 40 ° C. The mobile phase consisted of methanol - 0.01% acetic acid in water with a gradient of 0.8 m L · min ~ (-1). Mass spectra were quantified using electrospray ionization (ESI), negative ion mode detection and multiple reaction ion monitoring (MRM) mode. The results showed that there was a good linearity (r ≥ 0.9951) for the eight components in the range of determination concentration, RSD <15%, accuracy RE-15% -15%, and good stability. Extraction recovery> 63.2%, internal standard normalized matrix factor RSD did not exceed 15%, in line with the requirements. The results of bile excretion in rats showed that the eight components in the bile in the prototype excretion rate is relatively small, and the difference between different individuals larger. The method has the advantages of high sensitivity, good selectivity and simple sample processing, and can be used for the excretion kinetics study of eight components in the bile after Gavagenil extract.