School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenhe District, Shenyang 110016, China.
School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenhe District, Shenyang 110016, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jan 1;1104:73-80. doi: 10.1016/j.jchromb.2018.11.012. Epub 2018 Nov 9.
A selective, sensitive and fully validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the determination of dihydromyricetin, dihydroquercetin, myricetin and quercetin in rat plasma after intragastric administration of Hoveniae Semen total flavonoids. Baicalin was selected as the internal standard. Analytes were extracted from the rat plasma by protein precipitation with acetonitrile and separated on a C18 chromatographic column (Agilent ZORBAX Eclipse Plus, 4.6 mm × 100 mm, 3.5 μm) using the mobile phase containing acetonitrile (A) and 0.1% formic acid-water (B) by gradient elution at 0.5 mL/min flow rate. A tandem mass spectrometer equipped with an electrospray ionization (ESI) source was used to detect analytes. The analytes were measured by multiple reaction monitoring (MRM) in the negative ionization mode. The lower limit of quantification of dihydromyricetin, dihydroquercetin, myricetin and quercetin were 0.70, 8.16, 1.62 and 0.56 ng/mL, respectively. The accuracy, intra-day and inter-day precision and recovery were all satisfactory and the compounds were stable in rat plasma under all tested conditions. The approach was successfully applied to study pharmacokinetic characteristics of the four bioactive flavonoids in plasma after administering Hoveniae Semen total flavonoids intragastrically to rat. Further investigation was carried out to assess pharmacokinetic comparability of the four bioactive flavonoids after intragastric administration of Hoveniae Semen total flavonoids to mixture of flavonoids.
建立了一种选择性、灵敏且完全验证的液相色谱-串联质谱（LC-MS/MS）方法，用于测定灌胃给予枳椇子总黄酮后大鼠血浆中二氢杨梅素、二氢槲皮素、杨梅素和槲皮素的浓度。以黄芩苷为内标。用乙腈沉淀蛋白从大鼠血浆中提取分析物，在 C18 色谱柱（Agilent ZORBAX Eclipse Plus，4.6mm×100mm，3.5μm）上，以含有乙腈（A）和 0.1%甲酸-水（B）的流动相进行梯度洗脱，流速为 0.5ml/min。采用电喷雾电离（ESI）源的串联质谱仪以负离子模式进行检测。分析物通过多反应监测（MRM）进行定量分析。二氢杨梅素、二氢槲皮素、杨梅素和槲皮素的定量下限分别为 0.70、8.16、1.62 和 0.56ng/ml。准确度、日内和日间精密度以及回收率均令人满意，在所有测试条件下，化合物在大鼠血浆中均稳定。该方法成功应用于研究灌胃给予枳椇子总黄酮后大鼠血浆中四种生物活性黄酮类化合物的药代动力学特征。进一步研究了灌胃给予枳椇子总黄酮后混合物中四种生物活性黄酮类化合物的药代动力学可比性。
