Wang Ying, Yao Yamin, An Rui, You Lisha, Wang Xinhong
Department of Chemistry, College of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People's Republic of China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jul 1;877(20-21):1820-6. doi: 10.1016/j.jchromb.2009.05.035. Epub 2009 May 27.
A specific ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method was developed for the simultaneous determination of puerarin, daidzein, baicalin, wogonoside and liquiritin in rat plasma. Chromatographic separation was performed on a C(18) column packed with 1.7 microm particles by a linear gradient elution. The analytes and carbamazepine (internal standard, I.S.) were monitored in a selected-ion reaction (SIR) mode with a positive electrospray ionization (ESI) interface by the following ions: m/z 417.2 for puerarin, m/z 255.2 for daidzein, m/z 271.0 for baicalin, m/z 461.0 for wogonoside, m/z 441.0 for liquiritin and m/z 237.2 for carbamazepine (I.S.), respectively. The calibration curves of these analytes were linear over the concentration ranges from 0.00254-1.02 microg mL(-1) to 0.0102-10.2 microg mL(-1). Within-batch and between-batch precisions (RSD%) were all within 15% and accuracy (RE%) ranged from -10% to 10%. The extraction recoveries were on average 79.8% for puerarin, 90.8% for daidzein, 74.4% for baicalin, 70.2% for wogonoside and 84.7% for liquiritin. The validated method was successfully applied to investigate the pharmacokinetics of five bioactive compounds of GegenQinlian decoction (GQD) in rats.
建立了一种超高效液相色谱 - 质谱联用(UPLC - MS)法,用于同时测定大鼠血浆中葛根素、大豆苷元、黄芩苷、汉黄芩苷和甘草苷。采用线性梯度洗脱,在填充有1.7微米颗粒的C(18)柱上进行色谱分离。通过正电喷雾电离(ESI)接口,在选择离子反应(SIR)模式下监测分析物和卡马西平(内标,I.S.),对应的离子分别为:葛根素m/z 417.2、大豆苷元m/z 255.2、黄芩苷m/z 271.0、汉黄芩苷m/z 461.0、甘草苷m/z 441.0和卡马西平(内标)m/z 237.2。这些分析物的校准曲线在0.00254 - 1.02微克/毫升至0.0102 - 10.2微克/毫升的浓度范围内呈线性。批内和批间精密度(RSD%)均在15%以内,准确度(RE%)范围为 - 10%至10%。葛根素的提取回收率平均为79.8%,大豆苷元为90.8%,黄芩苷为74.4%,汉黄芩苷为70.2%,甘草苷为84.7%。该验证方法成功应用于研究葛根芩连汤(GQD)中五种生物活性成分在大鼠体内的药代动力学。