目的 建立液相色谱-串联质谱法(LC-MS/MS)检测大鼠血浆中白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ浓度,研究苍术-香附提取物灌胃后大鼠体内3种活性成分的药动学特征。方法 以苯海拉明为内标,血浆样品采用乙酸乙酯液-液萃取法处理,采用ACQUITY UPLC BEH-C18色谱柱(2.1 mm×50 mm,1.7 μm),流动相为乙腈-体积分数0.05%甲酸水,通过电喷雾离子源进行正离子扫描,多反应监测技术(multiple reaction monitoring,MRM)检测模式。检测离子:m/z 231.2→185.1(白术内酯Ⅰ)、m/z 233.2→187.1(白术内酯Ⅱ)、m/z 249.2→231.1(白术内酯Ⅲ)、m/z 256.2→167.1(苯海拉明)。利用DAS 3.2.2软件计算药动学参数。结果 白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ的线性范围分别为0.315 0~20.00、0.315 0~20.00、3.150~200.0 ng·mL-1,方法学的各项指标均符合要求。苍术-香附提取物灌胃给药后的3种活性成分的药-时曲线呈双峰现象,t1/2分别为(12.593±3.27) (14.868±5.51) (9.736±4.08) h;tmax分别为(0.458±0.29) (0.389±0.13) (0.347±0.08) h;ρmax分别为(7.719±4.93) (13.43±3.50) (153.702±23.51)ng·mL-1;AUC0-t分别为(42.592±19.45) (64.132±15.12) (1 042.156±168.43)ng·h·mL-1。结论 该LC-MS/MS操作简便,灵敏度高,满足了严格的方法学验证,可用于苍术-香附提取物在大鼠体内药动学研究。
Abstract
OBJECTIVE To establish an LC-MS/MS method for determination of atractylenolide Ⅰ, atractylenolide Ⅱ and atractylenolide Ⅲ in rat plasma, and study the pharmacokinetics of the three active components of Atractylodes Rhizoma-Cyperus Rhizoma extract in rats. METHODS Diphenhydramine was used as the internal standard, and the plasma samples were treated by liquid-liquid extraction with ethyl acetate. ACQUITY UPLC BEH-C18 column (2.1 mm×50 mm, 1.7 μm) was used, and acetonitrile-0.05% formic acid water was used as the mobile phase. Positive ion scanning was carried out by electrospray ion source (ESI). Multiple reaction monitoring (MRM) was used to monitor precursor to product ion transition of m/z 231.2→185.1 for atractylenolide I, m/z 233.2→187.1 for atractylenolide Ⅱ, m/z 249.2→231.1 for Ⅲ, and m/z 256.2→167.1 for IS (diphenhydramine). Pharmacokinetic parameters were calculated by DAS 3.3.2 software. RESULTS The linear ranges of atractylenolide Ⅰ, atractylenolide Ⅱ and atractylenolide Ⅲ were 0.315 0-20.00, 0.315 0-20.00 and 3.150-200.0 ng mL-1, respectively. Indicators of the method validation were in line with requirements. The drug concentration-time curves of the three active components of the extract showed double peaks, and the pharmacokinetic parameters were as follows: t1/2: (12.593±3.27) (14.868±5.51) (9.736±4.08) h, tmax: (0.458±0.29) (0.389±0.13) (0.347±0.08) h, ρmax: (7.719±4.93) (13.43±3.50) (153.702±23.51) ng·mL-1, AUC0-t: (42.592±19.45) (64.132±15.12) (1 042.156±168.43) ng·h·mL-1. CONCLUSION The LC-MS/MS method is simple and sensitive and can be used to study the pharmacokinetics of Atractylodes Rhizoma-Cyperus Rhizoma extract in rats.
关键词
苍术-香附药对 /
白术内酯 /
药动学 /
液相色谱-串联质谱法
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Key words
Atractylodes Rhizoma-Cyperi Rhizoma drug pair /
atractylenolide /
pharmacokinetics /
LC-MS/MS
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中图分类号：
R969.1
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脚注
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基金
江苏省自然科学基金面上项目资助(BK20211394);江苏省中医药领军人才项目资助(SLJ0208);江苏省中医药科技发展计划面上项目资助(MS2021011);江苏省研究生科研与实践创新计划项目资助(KYCX22_1917)
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