HPLC法测定血清肉碱的研究

目的:建立高效液相色谱(HPLC)测定血清中肉碱浓度的方法。方法:血清样品用乙腈、甲醇沉淀蛋白,加入肉碱衍生剂,60℃恒温水浴2h后进样10μl。色谱柱为AtlantisC18柱(4.6mm×150mm,5μm),流动相为甲醇∶异丙醇∶乙腈(35∶45∶20,v/v),流速1.0ml/min,紫外检测波长260nm。结果:在上述色谱条件下,血清肉碱分离良好,回归方程为Y=8490X-48200,r=0.9999,线性范围为12.5~400μmol/L。最低检测浓度为12.5μmol/L。日内精密度1.6%~13.2%,日间精密度(%)1.8%~12.1%,回收率98.5%。结论:通过加入肉碱衍生剂,建立了HPLC法测定血清中肉碱浓度的方法。此法操作简单,耗时少,有良好的准确度,适用于临床检测。

STUDY ON DETERMINATION OF CARNITINE IN SERUM BY HPLC

Objective: To develop a method for determination of carnitine in serum using HPLC. Methods: The serum proteins were precipitated by acetonitrile and methanol. The organic extract was added derivatizing reagent and the solution was warmed to 60℃ for 2 hours. Atlantis C18 column (4.6 mm×150 mm,5μm)was used as stationary phase and methanol-ispropand-acetonitrile (35∶45∶20,v/v) as mobile phase; flow rate 1.0ml/min, UV wavelengh at 260nm. Results: The typical chromatogram from serum samples showed clear separation of carnitine. Y= 8490X-48200, r= 0.9999. Intra-day RSD was 1.6%-13.2%, and inter-day 1.8%-12.1%. The recovery rate was 98.5%. Conclusion: A method for determination of carnitine in serum using HPLC by adding derivatizing reagent was developed and it is simple, rapid and precise, and can be used for clinical test.