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目的建立人血浆中左卡尼汀浓度的高效液相色谱-质谱(HPLC-MS)测定法。方法以米屈肼作为内标,血浆样品经蛋白沉淀法处理,用氰基色谱柱(Ultimate XB-CN,4.6mm×250mm,5μm)进行分离,以乙腈-30mmol·L-1醋酸铵水溶液(含0.8%甲酸)(80∶20)为流动相,柱温40℃,流速1.0mL·min-1,柱后分流比为4∶1,HPLC-MS/ESI+法选择性监测准分子离子峰[M+H]+(左卡尼汀:m/z=162.1,IS:m/z=147.1)。结果左卡尼汀及内标米屈肼在氰基柱上保留较好,分离完全,左卡尼汀在0.4~12.8mg·L-1内线性关系良好,r=0.9994,平均萃取回收率大于80%,方法回收率大于92%,小于114%,日内日间RSD均小于12%。结论本方法简便快速、灵敏准确,适用于人血浆中左卡尼汀临床治疗监测和药物研究的需要。
Objective To establish a high performance liquid chromatography-mass spectrometry (HPLC-MS) method for the determination of levocarnitine in human plasma. Methods Meldrum hydrazide was used as an internal standard. The plasma samples were treated with protein precipitation method and separated on a Cyanometric column (Ultimate XB-CN, 4.6 mm × 250 mm, 5 μm). The samples were eluted with acetonitrile-30 mmol·L-1 ammonium acetate aqueous solution 0.8% formic acid) (80:20) was used as the mobile phase. The column temperature was 40 ℃, the flow rate was 1.0 mL · min-1, the post-column split ratio was 4:1, and the excimer ion peak was selectively monitored by HPLC-MS / ESI + M + H] + (L-carnitine: m / z = 162.1, IS: m / z = 147.1). RESULTS: L-carnitine and internal standard Metsium hydrazide retained better on the cyano-column with good separation. L-carnitine showed a good linearity in the range of 0.4-12.8 mg · L-1, r = 0.9994. The average extraction recovery was greater than 80%. The recovery rate of the method was more than 92% and less than 114%. The intraday RSDs were less than 12%. Conclusion The method is rapid, sensitive and accurate and suitable for the clinical monitoring and drug research of L-carnitine in human plasma.