HPLC法同时测定大鼠血浆中3种CYP 450探针药物的浓度
发布时间：2018-11-20 04:24
【摘要】：建立了同时测定大鼠血浆中3种细胞色素P450(CYP450)亚型酶(CYP1A2、CYP2C9、CYP3A4)活性的Cocktail探针药物的高效液相色谱(HPLC)检测方法。分别取非那西丁,咪达唑仑和甲苯磺丁脲三种探针药物,制成混合探针溶液,用一种提取溶剂系统对3种探针药物同时提取,并用高效液相色谱法同时测定3种探针药物的血药浓度,并对其进行专属性、线性范围及检测限、精密度、回收率、稳定性的验证。色谱条件选用Diamonsil C18色谱柱(4.6 mm×250 mm,5μm);流动相为甲醇-50 m M磷酸盐缓冲对(45∶55,p H 3.2);检测波长230 nm;流速1.0 m L/min;柱温35℃。结果表明:非那西丁、咪达唑仑、甲苯磺丁脲的线性范围分别为0.10~10.0μg/m L(r=0.9993),0.05~5.0μg/m L(r=0.9999),0.10~50.0μg/m L(r=0.9991);方法回收率分别为79.28%~87.91%,76.43%~85.36%,82.79%~90.61%。该HPLC检测方法快捷,准确,灵敏度高,可以在同一次试验中,同时测定大鼠血浆中3种CYP450亚酶探针药物(非那西丁,甲苯磺丁脲,咪达唑仑)的浓度,各探针药物的吸收峰分离完全,无干扰,可用于开展某些受试药物对大鼠CYP450的3个主要亚型酶(CYP1A2,CYP2C9,CYP3A4)调控作用的相关研究,从而为相关肝药酶代谢研究提供方法学参考。
[Abstract]:A high performance liquid chromatographic (HPLC) method for the determination of the activity of three cytochrome P450 (CYP450) subtypes (CYP1A2,CYP2C9,CYP3A4) in rat plasma by high performance liquid chromatography (HPLC) was developed. Three probe drugs, phenacetin, midazolam and toluenesulfonylurea, were used to prepare the mixed probe solution, and the three probe drugs were extracted simultaneously by a solvent extraction system. The plasma concentrations of three probe drugs were determined by HPLC, and the specificity, linear range, detection limit, precision, recovery rate and stability were verified. The chromatographic conditions were as follows: Diamonsil C18 column (4.6 mm 脳 250 mm,5 渭 m);) as mobile phase, methanol-50 mm phosphate buffer pair (45: 55 渭 m);), detection wavelength of 230 nm;, flow rate of 1.0 mL / min, column temperature of 35 鈩，
本文编号：2343714
[Abstract]:A high performance liquid chromatographic (HPLC) method for the determination of the activity of three cytochrome P450 (CYP450) subtypes (CYP1A2,CYP2C9,CYP3A4) in rat plasma by high performance liquid chromatography (HPLC) was developed. Three probe drugs, phenacetin, midazolam and toluenesulfonylurea, were used to prepare the mixed probe solution, and the three probe drugs were extracted simultaneously by a solvent extraction system. The plasma concentrations of three probe drugs were determined by HPLC, and the specificity, linear range, detection limit, precision, recovery rate and stability were verified. The chromatographic conditions were as follows: Diamonsil C18 column (4.6 mm 脳 250 mm,5 渭 m);) as mobile phase, methanol-50 mm phosphate buffer pair (45: 55 渭 m);), detection wavelength of 230 nm;, flow rate of 1.0 mL / min, column temperature of 35 鈩，
本文编号：2343714
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