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超高效液相色谱-质谱联用法同时测定大鼠血浆中葛根芩连汤的葛根素、大豆苷元、黄芩苷、汉黄芩苷和甘草苷

Simultaneous determination of puerarin, daidzein, baicalin, wogonoside and liquiritin of GegenQinlian decoction in rat plasma by ultra-performance liquid chromatography-mass spectrometry.

作者信息

Wang Ying, Yao Yamin, An Rui, You Lisha, Wang Xinhong

机构信息

Department of Chemistry, College of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People's Republic of China.

出版信息

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.

Abstract

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.

摘要

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。

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