反向模式-强阳离子交换-反向模式二维色谱法测定人血浆中甲氨蝶呤浓度

发布时间：2018-03-21 09:12

本文选题：二维色谱　切入点：甲氨蝶呤　出处：《分析化学》2015年01期 　论文类型：期刊论文

【摘要】：建立了反向模式-强阳离子交换-反向模式(Reversed phase-strong cation exchange-reversed phase)二维色谱平台测定人血浆中甲氨蝶呤浓度的方法。样品经三氯醋酸沉淀蛋白后,在ASTON C8一级柱(100 mm×4.6 mm,5μm)上完成初分离,通过六通阀切换,经ASTON SCX中间级(20 mm×4.6 mm,5μm)二次分离和储存,在SAC C8二级柱(100 mm×4.6 mm,5μm)上完成最后分离,并测定。一级柱流动相为10 mmol/L醋酸铵-乙腈(90∶10,V/V,以醋酸调至p H 3.8),流速为1.0 m L/min;中间级流动相为10 mmol/L H3PO4溶液(p H 3.0);二级柱流动相为50 mmol/L醋酸铵-乙腈(87∶13,V/V,以醋酸调至p H 5.2),流速为1.2 m L/min,检测波长306 nm。单次分析时间4 min,线性范围0.09~5.1μmol/L,检出限为0.005μmol/L,日内RSD小于1.8%,日间RSD小于3.5%,相对回收率99.1%~101.25%,绝对回收率85.7%~86.4%。本方法简便、准确,适合日常血药浓度监测和药代动力学研究。
[Abstract]:A two-dimensional chromatographic platform for the determination of methotrexate in human plasma by reverse mode, strong cation exchange and reverse mode was established. The sample was precipitated by trichloroacetic acid and separated on a ASTON C8 column of 100mm 脳 4.6mm ~ 5 渭 m. Through the six way valve switch, the final separation was completed on the SAC C8 secondary column (100mm 脳 4.6mm / 5 渭 m) by ASTON SCX intermediate stage 20mm 脳 4.6mm / 5 渭 m) secondary separation and storage, and the final separation was carried out on the SAC C8 second stage column (100mm 脳 4.6mm / 5 渭 m). The mobile phase was 10 mmol/L ammonium acetate-acetonitrile 90: 10 V / V, adjusted to pH 3.8U with acetic acid, the flow rate was 1.0 mL / min, the intermediate mobile phase was 10 mmol/L H _ 3PO _ 4 solution and the second stage column was 50 mmol/L ammonium acetate -acetonitrile 8713 V / V / V, adjusted to acetic acid. The detection wavelength was 306 nm 路min. The linear range was 0.09 渭 mol / L, the detection limit was 0.005 渭 mol / L, the detection limit was 0.005 渭 mol / L, the intra-day RSD was less than 1.8, the relative recovery rate was 99.1g / min, the absolute recovery was 85.7 渭 mol / L, and the absolute recovery was 86.40.The method was simple and convenient. It is accurate and suitable for daily blood concentration monitoring and pharmacokinetic study.
【作者单位】：中南大学湘雅医院药学部;中南大学药学院;
【分类号】：R96;O657.72

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