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HPLC法同時(shí)測定大鼠血漿中3種CYP 450探針?biāo)幬锏臐舛?/H1>
發(fā)布時(shí)間:2018-11-20 04:24
【摘要】:建立了同時(shí)測定大鼠血漿中3種細(xì)胞色素P450(CYP450)亞型酶(CYP1A2、CYP2C9、CYP3A4)活性的Cocktail探針?biāo)幬锏母咝б合嗌V(HPLC)檢測方法。分別取非那西丁,咪達(dá)唑侖和甲苯磺丁脲三種探針?biāo)幬?制成混合探針溶液,用一種提取溶劑系統(tǒng)對3種探針?biāo)幬锿瑫r(shí)提取,并用高效液相色譜法同時(shí)測定3種探針?biāo)幬锏难帩舛?并對其進(jìn)行專屬性、線性范圍及檢測限、精密度、回收率、穩(wěn)定性的驗(yàn)證。色譜條件選用Diamonsil C18色譜柱(4.6 mm×250 mm,5μm);流動相為甲醇-50 m M磷酸鹽緩沖對(45∶55,p H 3.2);檢測波長230 nm;流速1.0 m L/min;柱溫35℃。結(jié)果表明:非那西丁、咪達(dá)唑侖、甲苯磺丁脲的線性范圍分別為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);方法回收率分別為79.28%~87.91%,76.43%~85.36%,82.79%~90.61%。該HPLC檢測方法快捷,準(zhǔn)確,靈敏度高,可以在同一次試驗(yàn)中,同時(shí)測定大鼠血漿中3種CYP450亞酶探針?biāo)幬?非那西丁,甲苯磺丁脲,咪達(dá)唑侖)的濃度,各探針?biāo)幬锏奈辗宸蛛x完全,無干擾,可用于開展某些受試藥物對大鼠CYP450的3個(gè)主要亞型酶(CYP1A2,CYP2C9,CYP3A4)調(diào)控作用的相關(guān)研究,從而為相關(guān)肝藥酶代謝研究提供方法學(xué)參考。
[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,

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