Simultaneous determination by UPLC-MS/MS of seven bioactive compounds in rat plasma after oral administration of Ginkgo biloba tablets: application to a pharmacokinetic study

UPLC-MS/MS法测定口服银杏叶片的大鼠血浆中七种活性成分及药代动力学研究(英文)

A rapid, reliable, and sensitive method was developed using ultra-performance liquid chromatography- tandem mass spectrometry （UPLC-MS/MS） with an electrospray ionization （ESI） source for determination of seven bioactive compounds in rat plasma after oral administration of Ginkgo biloba tablets （GBTs）. The method simultane- ously detects bilobalide （BB）, ginkgolide A （GA）, ginkgolide B （GB）, ginkgolide C （GC）, quercetin （QCT）, kaempferol （KMF）, and isorhamnetin （ISR） for pharmacokinetic study. The analytes and internal standard （IS） were extracted from rat plasma by acetidin. An MS/MS detection was conducted using multiple reaction monitoring （MRM） and operating in the negative ionization mode. The calibration curve ranges were 5-500, 5-500, 2.5-250, 1-100, 1-100, 1-100, and 1-100 ng/ml for BB, GA, GB, GC, QCT, KMF, and ISR, respectively. The mean recovery of the analytes ranged from 68.11% to 84.42%. The intra- and inter-day precisions were in the range of 2.33%-9.86% and the accuracies were between 87.67% and 108.37%. The method was used successfully in a pharmacokinetic study of GBTs. The phar- macokinetic parameters of seven compounds were analyzed using a non-compartment model. Plasma concentrations of the seven compounds were determined up to 48 h after administration, and their pharmacokinetic parameters were in agreement with previous studies.

研究目的:建立一个快速灵敏且具有专属性的体内分析方法用于银杏叶片的药代动力学研究。创新要点:运用超高压液相色谱-二级质谱连用法(UPLC-MS/MS)进行大鼠血浆中七种活性成分的测定,方法简单快速,适合于银杏叶制剂的药代动力研究。研究方法:采用液液萃取进行血浆样品处理,该方法简单且快速。采用超高压液相色谱(UPLC)进行七种活性成分分离,达到了多种成分快速分离的目的。采用二级质谱(MS/MS)进行检测,优化质谱条件,减小了内源性干扰并提高检测灵敏度。经过专属性、线性、回收率、基质效应、精密度、准确度和稳定性考察,验证了该方法的适用性。经验证,该方法适用于银杏叶片的药代动力学研究。重要结论:液液萃取处理血浆样品简单可靠,回收率稳定,无基质效应(表4)。UPLC-MS/MS能够快速、灵敏、专属性地分析大鼠血浆七种活性成分的血药浓度(表3和图3)。该方法适用于银杏叶片的七种活性成分的药代动力学研究(图4和5)。