HPLC—MS/MS同时测定广金钱草中8种成分在大鼠胆汁中的浓度及其排泄动力学研究(1)
[摘要] 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 C18 column (4.6 mm×150 mm，5 μm)，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 mL·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。
, http://www.100md.com [关键词] 广金钱草； 黄酮； 酚酸； HPLC-MS/MS； 含量测定； 胆汁排泄
[Abstract] To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1， 1-2， 2-4， 4-6， 6-8， 8-12， 12-24 h. Diamonsil C18 column (4.6 mm×150 mm， 5 μm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL·min-1， and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity， with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%， and the accuracies (RE) ranged from -15% to 15%. The method had a good stability， and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%， and the RSD of IS-normalized matrix factors were no more than 15%， which met the requirements for analysis. According to the biliary excretion study， the bile excretion rates of the 8 analytes were relatively low， with great difference among the individuals. The results showed that the established method was simple， selective and specific， thus could be applied in the biliary excretion study of the 8 analytes in rat bile.
, 百拇医药
[Key words] Desmodii Styracifolii Herba； flavonoids； phenolic acid； HPLC-MS/MS； content determination； biliary excretion
廣金钱草为豆科植物广金钱草Desmodium styracifolium(Osb.)Merr.的干燥地上部分[1]，是我国常用的传统中药，具有清热排石、利胆利尿、抗炎镇痛的功效，可用于预防、治疗泌尿系统结石以及结石术后消炎止痛等 [2-3]。广金钱草中含有黄酮、酚酸、多糖、生物碱、鞣质、挥发性成分等[4]。其中，黄酮类成分能够显著减少高草酸尿大鼠肾结石的形成[5]，酚酸则具有抗炎的效果[6]。为更好的指导其临床应用，本课题组在对广金钱草在大鼠体内的药物代谢动力学研究的基础上[7]，进一步对其在大鼠体内的排泄进行研究。旨在建立一种HPLC-MS/MS同时测定广金钱草中7种黄酮和1种酚酸类化合物在大鼠胆汁中的浓度，从而考察其胆汁排泄的动力学过程，为全面研究广金钱草的体内过程奠定基础。, http://www.100md.com(樊雪艳 王鑫 刘晓晨 王春英)
, http://www.100md.com [关键词] 广金钱草； 黄酮； 酚酸； HPLC-MS/MS； 含量测定； 胆汁排泄
[Abstract] To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1， 1-2， 2-4， 4-6， 6-8， 8-12， 12-24 h. Diamonsil C18 column (4.6 mm×150 mm， 5 μm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL·min-1， and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity， with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%， and the accuracies (RE) ranged from -15% to 15%. The method had a good stability， and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%， and the RSD of IS-normalized matrix factors were no more than 15%， which met the requirements for analysis. According to the biliary excretion study， the bile excretion rates of the 8 analytes were relatively low， with great difference among the individuals. The results showed that the established method was simple， selective and specific， thus could be applied in the biliary excretion study of the 8 analytes in rat bile.
, 百拇医药
[Key words] Desmodii Styracifolii Herba； flavonoids； phenolic acid； HPLC-MS/MS； content determination； biliary excretion
廣金钱草为豆科植物广金钱草Desmodium styracifolium(Osb.)Merr.的干燥地上部分[1]，是我国常用的传统中药，具有清热排石、利胆利尿、抗炎镇痛的功效，可用于预防、治疗泌尿系统结石以及结石术后消炎止痛等 [2-3]。广金钱草中含有黄酮、酚酸、多糖、生物碱、鞣质、挥发性成分等[4]。其中，黄酮类成分能够显著减少高草酸尿大鼠肾结石的形成[5]，酚酸则具有抗炎的效果[6]。为更好的指导其临床应用，本课题组在对广金钱草在大鼠体内的药物代谢动力学研究的基础上[7]，进一步对其在大鼠体内的排泄进行研究。旨在建立一种HPLC-MS/MS同时测定广金钱草中7种黄酮和1种酚酸类化合物在大鼠胆汁中的浓度，从而考察其胆汁排泄的动力学过程，为全面研究广金钱草的体内过程奠定基础。, http://www.100md.com(樊雪艳 王鑫 刘晓晨 王春英)