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高效液相色谱荧光检测法测定人血浆中麦考酚酸的浓度
Determination of mycophenolic acid concentration in human plasma by high performance liquid chromatography with fluorescence detection
【摘要】 目的:建立高效液相色谱(HPLC)荧光检测法测定人血浆中麦考酚酸浓度的方法。方法:以萘普生为内标,血浆用甲醇蛋白沉淀后直接进样。采用KromasilC8(150mm×4.6mm,5μm),柱温:25°C;流动相∶乙腈:0.04mol·L-1甘氨酸缓冲液,pH9.2(20∶80);流速:1.0mL·min-1,荧光激发波长342nm,发射波长425nm。结果:血浆内源性杂质和常用合并用药对样品测定无干扰,麦考酚酸在0.05~40mg·L-1范围内线性关系良好(r=0.9999)。麦考酚酸的日内、日间RSD均小于7%,样品3次冻融及提取后在24h内稳定性良好。结论:HPLC法快速、灵敏、准确,可用于麦考酚酸的体内分析及常规的治疗药物监测。
【Abstract】 AIM: To determine the concentration of mycophenolic acid in human plasma by HPLC with fluorescence detection. METHODS: Using naproxen as internal standard, mycophenolic acid in plasma samples was determined by HPLC with protein precipitation. HPLC analytical column was created by Kromasil C 8 (150 mm ×4.6 mm, 5 μm). Separation was carried out at 25 °C. The mobile phase consisted of a mixture acetonitrile∶0.04 mol·L -1 glycine buffer, pH 9.2 ( 20∶80) pumped at a flow rate of 1.0 mL·min -1. A fluorescence detector was set at λ ex 342 nm, λ em 425 nm. RESULTS: The intrinsic impurity and the combination used drugs in plasma did not interfere with the determination of the sample tests. There was good linear relationships(1/C weighted) between peak area ratio of mycophenolic acid to internal standard and C(r=0.999 9) within the range of 0.05- 40 mg·L -1. The precision of within-run and between-run was good. CONCLUTION: The method established in this paper can be used to determine the concentrations of mycophenolic acid in human plasma. Method can also be applied to the clinical pharmacy research in vivo.
【Key words】 immunosuppressive agents; mycophenolic acid; chromatography, high pressure liquid; fluorescence detection;
- 【文献出处】 中国新药与临床杂志 ,Chinese Journal of New Drugs and Clinical Remedies , 编辑部邮箱 ,2005年01期
- 【分类号】R96
- 【被引频次】14
- 【下载频次】190