本文選題：雷公藤多苷 + 體外代謝模型　； 參考：《中國藥房》2015年04期

【摘要】：目的:建立液相色譜-串聯(lián)質(zhì)譜(LC-MS/MS)法測定大鼠和人肝微粒體中1′-羥基咪達(dá)唑侖的含量,研究雷公藤多苷對體外大鼠與人肝微粒體細(xì)胞色素P45(0CYP)3A酶活性的抑制作用。方法:將不同質(zhì)量濃度的雷公藤多苷(0.5、1.0、2.0、5.0、10.0、50.0、100.0μg/ml)分別與大鼠和人肝微粒體(0.4 mg/ml)共同孵育20 min后終止反應(yīng),采用LC-MS/MS法測定孵育液中咪達(dá)唑侖的代謝產(chǎn)物1′-羥基咪達(dá)唑侖的濃度以評價(jià)CYP3A酶活性并計(jì)算酶活性抑制率,采用Graph Pad Prism 5.0軟件進(jìn)行擬合并計(jì)算半數(shù)抑制濃度(IC50)。色譜柱為Agilent Eclipse XBD-C18,流動(dòng)相為水-甲醇(梯度洗脫),流速為0.3 ml/min,柱溫為30℃,進(jìn)樣量為2μl。采用電噴霧電離(ESI),定量離子為m/z 342.01→324.1(1′-羥基咪達(dá)唑侖)、m/z 383.01→337.2(內(nèi)標(biāo)氯雷他定)。結(jié)果:1′-羥基咪達(dá)唑侖檢測濃度在0.01~3μmol/L范圍內(nèi)與其和內(nèi)標(biāo)物峰面積比值呈良好線性關(guān)系;精密度試驗(yàn)RSD均小于10%,提取回收率為97.92%~101.65%,方法回收率為96.70%~104.10%,穩(wěn)定性試驗(yàn)RSD均小于10%。雷公藤多苷對大鼠和人微粒體中CYP3A的IC50分別為(48.610±1.32)μg/ml和(4.754±1.12)μg/ml;在0.5~100.0μg/ml范圍內(nèi)雷公藤多苷對CYP3A酶活性有抑制作用,且與質(zhì)量濃度呈正相關(guān)。結(jié)論:所建立的LC-MS/MS法可用于檢測大鼠和人肝微粒體中1′-羥基咪達(dá)唑侖的含量。雷公藤多苷對體外大鼠和人肝微粒體藥物代謝酶CYP3A均存在抑制作用,且對人肝微粒體CYP3A的抑制作用明顯強(qiáng)于大鼠。
[Abstract]:Aim: to establish a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method for the determination of 1-hydroxy-midazolam in rat and human liver microsomes and to study the inhibitory effect of tripterygium wilfordii polyglycosides on cytochrome P45 (0CYP) 3A enzyme activity in rat and human liver microsomes in vitro. Methods: different concentrations of tripterygium wilfordii polyglucoside (0.51.02.00.0.0.0.0.100.0 渭 g/ml) were incubated with rat and human liver microsomes (0.4 mg/ml) for 20 min, respectively, and the reaction was terminated after incubating with different concentrations of tripterygium wilfordii for 20 min. In order to evaluate the activity of CYP3A and calculate the inhibition rate of CYP3A, LC-MS / MS method was used to determine the metabolite of midazolam in incubation solution. The half inhibitory concentration (IC50) of midazolam was calculated using Graph Pad Prism 5.0 software. The chromatographic column is Agilent Eclipse XBD-C18, the mobile phase is water-methanol (gradient elution), the flow rate is 0.3 ml / min, the column temperature is 30 鈩，

本文編號：2110724