The aim of this study was to develop and validate a new, rapid, sensitive, selective and reliable liquid chromatography–tandem mass spectrometry method for simultaneous determination of 3-O-Acetyl-11-keto-β-boswellic acid (AKBA) and its active metabolite 3-O-Acetyl-11-hydroxy-β-boswellic acid (Ac-11-hydroxy-BA) in rat plasma. Both analytes (AKBA and Ac-11-hydroxy-BA) and the internal standard (IS, ursolic acid) were extracted from 100 μL of rat plasma by protein precipitation. Chromatographic separation was achieved on PRP-H1 RP-C18 column (75 mm × 2 mm, 1.6 μm) using acetonitrile–water (95.5 v/v) as the mobile phase. Mass detection was conducted by electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. A linear dynamic range of 1–1,000 ng/mL for both AKBA and Ac-11-hydroxy-BA was established with mean correlation coefficient (r (1)) of 0.999. Intra- and inter-day precision (% CV) of analysis were found in the range of 1.9–7.4%. The accuracy determined for these analytes ranged from 92.4 to 107.2%. The extraction recoveries for both analytes ranged from 92.6 to 97.3% for spiked plasma samples and were consistent. The % change in stability samples compared to nominal concentration ranged from 0.4 to 4.2%. This method was successfully tested to a pharmacokinetic (PK) study for estimation of AKBA and acetyl-11-hydroxy-BA in rat plasma following oral administration of AKBA. This method has been validated with the advantage of shorter run time that can be used for high-throughput analysis and has been successfully applied to the pharmacokinetic study of AKBA in rats.

中文翻译：

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同时测定3-O-乙酰基-11-酮基-β-乳香酸（AKBA）及其活性代谢物乙酰基-11-羟基-β-乳香酸（Ac-11-OH-）的有效LC-MS / MS方法BA）在大鼠血浆中的应用：在药代动力学研究中的应用。

这项研究的目的是开发和验证一种新的，快速，灵敏，选择性和可靠的液相色谱-串联质谱方法，用于同时测定3-O-乙酰基-11-酮-β-乳香酸（AKBA）及其大鼠血浆中的活性代谢物3-O-乙酰基-11-羟基-β-乳香酸（Ac-11-羟基-BA）。通过蛋白质沉淀从100μL大鼠血浆中提取分析物（AKBA和Ac-11-hydroxy-BA）和内标物（IS，熊果酸）。使用乙腈-水（95.5 v / v）在PRP-H1 RP-C18色谱柱（75 mm×2 mm，1.6μm）上进行色谱分离 ）作为流动相。通过电喷雾电离以正离子多反应监测（MRM）模式进行质量检测。AKBA和Ac-11-hydroxy-BA的线性动态范围均为1–1,000 ng / mL，平均相关系数（r（1））为0.999。日内和日间分析的精确度（％CV）在1.9–7.4％的范围内。这些分析物的准确度范围为92.4％至107.2％。对于加标血浆样品，两种分析物的萃取回收率在92.6％至97.3％之间，并且是一致的。与标称浓度相比，稳定性样品的变化百分比为0.4到4.2％。该方法已成功通过药代动力学（PK）研究，用于估计口服AKBA后大鼠血浆中的AKBA和乙酰基11-羟基-BA。