中文摘要：

目的建立灵敏、快速、稳定的测定大鼠血浆中CYP450探针药物/代谢产物,咖啡因/副黄嘌呤、奥美拉唑/5-羟基奥美拉唑、右美沙芬/右啡烷、咪达唑仑/1＇-羟基咪达唑仑的LC-MS/MS方法,并用于其在大鼠体内的药代动力学研究。方法 100μL血浆样品加入内标地西泮及乙酸铵溶液,乙酸乙酯进行液液萃取,离心取上清挥干复溶,LC-MS/MS检测。选用Agilent Eclipse Plus-C_（18）色谱柱（50 mm×2.1mm,3.5μm）,乙腈和0.01%甲酸（1 mmol·L^-1甲酸铵）作为流动相,梯度洗脱,流速为0.3 m L·min^-1,进样量10μL。采用电喷雾电离源（ESI）,扫描方式为多反应正离子监测模式（MRM＋）,咖啡因m/z：195.0/138.1;副黄嘌呤m/z：181.1/124.1;奥美拉唑m/z：346.1/198.1;5-羟基奥美拉唑m/z：362.1/214.1;右美沙芬m/z：272.2/147.1;右啡烷m/z：258.1/157.1;咪达唑仑m/z：326.1/291.1;1＇-羟基咪达唑仑m/z：342.1/324.1;内标地西泮m/z：285.1/154.0。结果 咖啡因、副黄嘌呤、奥美拉唑、5-羟基奥美拉唑、右美沙芬、右啡烷、咪达唑仑和1＇-羟基咪达唑仑的线性范围分别为1.95～2 000、0.98～250、0.48～2 000、0.98～250、0.98～2 000、0.48～125、1.95～2 000和1.95～250μg·L^-1RSD〈15%且无基质效应。结论 该方法快速、灵敏、重现性好,可用于血浆中上述探针药物/代谢产物浓度的测定,便于深入进行代谢酶CYP1A2、CYP2C19、CYP2D6和CYP3A4的代谢相关研究。

英文摘要：

Aim To develop a sensitive,rapid and ac-curate LC-MS /MS method for the simultaneous deter-mination of cytochrome P450 probe substrates,inclu-ding caffeine and its metabolite paraxanthine for CYP1 A2,omeprazole and its metabolite 5-hydroxyome-prazole for CYP2C1 9,dextromethorphan and its metab-olite dextrorphan for CYP2D6,midazolam and its me-tabolite 1 ′-hydroxymidazolam for CYP3A4.Methods Probe drugs with the IS diazepam were extracted using ethyl acetate.Gradient elution was performed on an Agilent Eclipse Plus-C1 8 column （50 mm ×2.1 mm, 3.5 μm）.The mobile phase consisted of 0.01 % for-mic acid（1 mmol·L^-1 ammonium formate）and aceto-nitrile.The flow rate was 0.3 mL·min^-1 ,and the in-jection volume was 1 0 μL.The analyte was detected u-sing electrospray ionization（ESI）in positive multiple reaction monitoring（MRM＋）mode.The reaction se-lected ions were 1 95.0 /1 38.1 m /z for caffeine, 1 81 .1 /1 24.1 m /z for paraxanthine,346.1 /1 98.1 m /z for omeprazole,362.1 /21 4.1 m /z for 5-hydroxyome-prazole, 272.2 /1 47.1 m /z for dextromethorphan, 258.1 /1 57.1 m /z for dextrorphan,326.1 /291 .1 m /z for midazolam,342.1 /324.1 m /z for 1 ′-hydroxymid-azolam and 285.1 /1 54.0 m /z for diazepam as internal standard.Results The linear ranges of caffeine,pa-raxanthine,omeprazole,5-hydroxyomeprazole,dextro-methorphan, dextrophan, midazolam and 1 ′-hydroxymidazolam were 1 .95 ～2 000,0.98 ～250, 0.48 ～2 000,0.98 ～250,0.98 ～2 000,0.48 ～1 25,1 .95 ～2 000 and 1 .95 ～250 μg·L^-1 respec-tively.The RSD of all probe drugs was less than 1 5%and matrix effects in plasma on the ionization of probe drugs were negligible.Conclusion This sensitive and rapid LC-MS /MS method is suitable for determination of the drug/metabolite concentrations in plasma,so as to study the metabolism of CYP1 A2, CYP2C1 9, CYP2D6 and CYP3A4 in depth.