UPLC-MS/MS 同时测定小鼠血中 6 种生物碱及其在药代动力学和生物利用度中的应用。
Simultaneous Determination of Six Alkaloids in Mouse Blood by UPLC-MS/MS and Its Application in Pharmacokinetics and Bioavailability.
机构信息
The Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou People's Hospital, Wenzhou 325000, China.
School of Basic Medicine, Wenzhou Medical University, Wenzhou 325035, China.
出版信息
Biomed Res Int. 2020 Aug 17;2020:1030269. doi: 10.1155/2020/1030269. eCollection 2020.
A specific ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the simultaneous determination of six alkaloids in mouse blood with midazolam as the internal standard (IS). Only 20 L blood was needed for sample preparation, and the protein was precipitated with acetonitrile. The UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 m) was used for chromatographic separation. The mobile phase consisted of 0.1% formic acid and acetonitrile with gradient elution within 5.5 min. Multiple reaction monitoring (MRM) and the positive electrospray ionization model were used for quantitative analysis. The accuracy of the UPLC-MS/MS method ranged from 86.5% to 110.4%. The precision for intraday and interday was ≤15% each. The mean recovery and the matrix effects were found to be 64.4-86.8% and 94.1-109.4%, respectively. The calibration curves in blood were linear in the range of 1-1000 ng/mL with a favorable correlation coefficient ( ) of 0.995. The pharmacokinetic results showed that six alkaloids metabolized rapidly in mice with a half-life between 0.6 h and 4.4 h. The bioavailability of corynoxeine, isocorynoxeine, rhynchophylline, isorhynchophylline, hirsutine, and hirsuteine was 27.3%, 32.7%, 49.4%, 29.5%, 68.9%, and 51.0%, respectively, which showed satisfactory oral absorption of each alkaloid.
建立了一种同时测定小鼠血中 6 种生物碱的超高效液相色谱-串联质谱（UPLC-MS/MS）方法，以咪达唑仑为内标（IS）。样品制备仅需 20 μL 血，用乙腈沉淀蛋白。采用 UPLC BEH C18 柱（2.1mm×100mm，1.7μm）进行色谱分离。流动相由 0.1%甲酸和乙腈组成，在 5.5min 内进行梯度洗脱。采用多反应监测（MRM）和正电喷雾离子化模式进行定量分析。UPLC-MS/MS 方法的准确度在 86.5%至 110.4%之间。日内和日间精密度均≤15%。平均回收率和基质效应分别为 64.4%-86.8%和 94.1%-109.4%。血中校准曲线在 1-1000ng/mL 范围内呈线性，相关系数（ ）良好，为 0.995。药代动力学结果表明，6 种 生物碱在小鼠体内迅速代谢，半衰期在 0.6-4.4h 之间。柯诺辛、异柯诺辛、钩藤碱、异钩藤碱、毛钩藤碱和毛钩藤碱的生物利用度分别为 27.3%、32.7%、49.4%、29.5%、68.9%和 51.0%，表明各生物碱具有良好的口服吸收。
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