Determination of 8,2′-diprenylquercetin 3-methyl ether in plasma by UPLC-MS-MS and its pharmacokinetic application in rats

UPLC-MS-MS法测定大鼠血浆中8,2′-二异戊烯基槲皮素-3-甲醚的浓度及其药物动力学研究（英文）

8,2＇-Diprenylquercetin 3-methyl ether with significant anti-breast cancer activity is the main constituent of Tibetan medicine Sinopodophylli Fructus. In the present study, we developed and validated a rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method for the determination of 8,2＇-diprenylquercetin 3-methyl ether in rat plasma. 8-Prenylkaempferol was used as the internal standard. The separation was carried out using Waters ACQUITY UPLC BEH C18 column （2.1 mm×100 ram, 1.7 μm） with a mobile phase consisting of acetonitrile and 0.1% formic acid in water on a gradient program at a flow rate of 0.4 mL＇min-1 and temperature of 30 ℃. Triple quadrupole mass spectrometric detection in negative ion mode was used for multiple-reaction monitoring of the transitions at m/z 451.30→177.25 and m/z 353.25→298.15 for 8,2＇-diprenylquercetin 3-methyl ether and 8-prenylkaempferol, respectively. The calibration curves were linear within the concentration range 0.1-2000 ng/mL （r = 0.9954）. The recoveries were 103%-115%, and the results were consistent across low, middle and high concentration levels. The intra- and inter-day precisions were within 15%, and the bias was between --6%-15%. This method was simple, rapid and sensitive, which could be applied to the determination of 8,2＇-diprenylquercetin 3-methyl ether in plasma and pharmacokinetic study in rats. Pharmacokinetic test indicated that the peak plasma concentration occurred in 2 h after the female rats were intragastrically administered with 8,2＇-diprenylquercetin 3-methyl ether at the dose of 100 mg/kg, and the biological half-life was 6.79 h. The blood drug concentration maintained equal amount for 20 h, which was conducive to the in vivo effects of drugs.

8,2′-二异戊烯基槲皮素-3-甲醚是藏药小叶莲中的主要成分,具有较好的抗乳腺癌活性。本文首次建立了快速、灵敏的测定大鼠血浆中8,2′-二异戊烯基槲皮素-3-甲醚浓度的UPLC-MS/MS方法。以8-异戊烯基山柰酚为内标,采用waters ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸水–乙腈为流动相梯度洗脱,流速为0.4 mL·min–1,柱温为30oC。通过电喷雾离子化四极杆串联质谱,负离子多反应监测(MRM)模式,用于定量分析的离子反应分别为m/z451.30→177.25(8,2′-二异戊烯基槲皮素-3-甲醚)、m/z 353.25→298.15(8-异戊烯基山柰酚)。8,2′-二异戊烯基槲皮素-3-甲醚浓度为0.1–2000 ng/m L时线性关系良好(r=0.9954),高、中、低三个浓度的提取回收率在103%–115%范围内,日内、日间精密度均小于15%,准确度在–6%~15%之间。该方法简单快速灵敏,适用于检测8,2′-二异戊烯基槲皮素-3-甲醚的血药浓度并进行大鼠体内的药物动力学研究。药物动力学试验结果表明,雌性大鼠口服灌胃给药100 mg/kg的8,2′-二异戊烯基槲皮素-3-甲醚,2小时达到血浆峰浓度,生物半衰期为6.79小时,20小时内能维持一定量的血药浓度,有利于药物在生物体内产生作用。