HPLC-MS/MS法测定银杏7种有效成分在犬血浆中浓度以及银杏叶酊剂的相对生物利用度

Simultaneous determination of seven bioactive components of Ginkgo biloba L. in dog plasma by HPLC-MS/MS and its application in relative bioavailability of the tincture extract of Ginkgo biloba leaves

目的:采用同时测定犬血浆中4种银杏内酯(银杏内酯A,B,C和白果内酯)和3种黄酮(槲皮素、山柰酚、异鼠李素)的高效液相色谱-串联质谱(HPLC-MS/MS)分析方法,研究了银杏叶酊剂的相对生物利用度。方法:相对生物利用度试验采用随机分组,两周期交叉设计。犬血浆样品经过酸化处理后,采用乙酸乙酯萃取。运用所建立的HPLC-MS/MS方法测定其血药浓度。结果:银杏叶酊剂和参比制剂(银杏叶提取物滴剂和银杏叶片剂)相比较,7个有效成分的吸收差异较大,其中白果内酯受试制剂吸收差于参比制剂;其他成分受试制剂吸收好于或者与参比制剂相当。结论:所采用的HPLC-MS/MS方法快速、灵敏、准确,受试制剂和参比制剂中有效成分的相对生物利用度存在一定差异。

Objective： To establish a high-performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method for determination of terpenoids（ginkgolides A,B and C;bilobalide） and flavonoids（quercetin,kaemferol and isorhamnetin） in dog plasma,and investigate the relative bioavailability of the tincture extract of Ginkgo biloba leaves.Methods： Three preparations were administrated to male beagle dogs according to a randomized cross design.Dog plasma samples were extracted with EtOAc after adding HCl（6 mol·L-1）.Then,the concentrations of 7 compounds in dog plasma were determined by HPLC-MS/MS.Results： Compare with reference preparations（extract drops and extract tablets of Ginkgo biloba leaves）,the 7 active compounds in tincture extract of Ginkgo biloba leaves were absorbed better or equal except biloblide.Conclusion： The HPLC-MS/MS method is rapid,sensitive and reliable.The relative bioavailability of the test preparations shows the difference from the reference preparations.