UPLC-MS/MS法同时测定家兔血浆中6个黄酮类成分及其药代动力学研究

目的: 建立UPLC-MS/MS同时测定家兔血浆中注射用复方荭草冻干粉针的6个黄酮类化合物(异荭草素,荭草素,牡荆素,灯盏乙素,木犀草苷,槲皮苷)的分析方法,并计算其在家兔体内的药动学参数。方法: 血浆样品酸化后以甲醇沉淀蛋白;色谱采用Waters BEH C₁₈(2.1 mm×50 mm,1.7 μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液,梯度洗脱,质谱采用多反应监测(MRM)进行正离子检测,样品分析时间为3.5 min。结果: 6个黄酮类成分在家兔血浆中呈良好线性关系,提取回收率在79.4%～105.9%之间,日内、日间精密度和准确度良好,各检测成分的方法回收率分别为异荭草素96.8%～110.1%,荭草素85.8%～98.8%,牡荆素86.0%～110.6%,灯盏乙素86.7%～102.5%,木犀草苷84.1%～93.8%,槲皮苷96.5%～112.2%。6个黄酮类成分在家兔体内的平均滞留时间均较短,在16 min以内。结论: 该方法特异、快速、灵敏,可用于注射用复方荭草冻干粉针的药代动力学研究。

Objective: To develop an UPLC-MS/MS method for simultaneous determination of six flavonoids(isoorientin,orientin,vitexin,scutellarin,cynaroside,quercitrin) after intravenous administration of compound Hongcao freeze dried powder for injection and acqire the pharmacokinetic parameters in rabbit plasma. Methods: Following a protein precipitation with acidified methanol for plasma samples. The whole analysis was achieved within 3.5 min by gradient elution on an Waters BEH C₁₈(2.1 mm×50 mm, 1.7 μm) column and the mobile phase consisting of acetonitrile and water(containing 0.1% formic acid). Detection was carried out by the multiple reaction monitoring(MRM) positive ion mode. Results: Good linearity was achieved for the six compounds,the extraction recovery was between 79.4% to 105.9%, the intra-and inter-day precisions and the accuracy were good,the method recoveries of the six flavonoids were 96.8%-110.1% for isoorientin, 85.8%-98.8% for orientin, 86.0%-110.6% for vitexin, 86.7%-102.5% for scutellarin, 84.1%-93.8% for cynaroside, 96.5%-112.2% for quercitrin, respectively. The MRT of the six flavonoids in rabbit was all blow 16 min. Conclusion: The above mentioned method is specific, sensitive and suitable for the pharmacokinetic studies of compound Hongcao freeze dried powder for injection.

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