HPLC法测定人血浆中环磷酰胺的浓度

Quantitation of Plasma Cyclophosphamide Content by High Performance Liquid Chromatography

目的 建立一种可靠、便捷的高效液相色谱法测定人血浆中环磷酰胺 (CTX)浓度的方法。方法 以异环磷酰胺 (Ifo)为内标 ,采用惠普 1 1 0 0型高效液相色谱仪 ,分析柱HPLichrosphereC8柱 (2 50mm× 4mm ,5μm) ,前加HPLicrosphere保护柱 ;流动相为 :乙腈∶水 =1 8∶82 (V/V) ,流速为 1 .5mL/min ;检测波长为 1 95nm ;室温2 5℃。血浆样品以甲醇沉淀蛋白。结果 色谱峰分离良好 ,无干扰。线性方程为 :Y =4.861X +0 .1 81 7,r =0 .9999;线性范围 :0 .5～ 50 μg/mL ;检测限 :0 .1 μg/mL。 结论 本法是一种可靠、便捷的检测血浆CTX浓度的方法 。

Purpose: To develop a high-performance liquid chromatography method to determine the concentration of cyclophospharmide(CIX) in human plasma. Methods: The HP1100 high performance liquid chromatography (HPLC) system was used, including a quaternary pump a manual injector, a variable wavelength detector and an LC 2D Chem Station. The analytical column was an HP Lichrosphere C 8 column(250 mmx 4 mm, 5 μm) with an HP Licrosphere guard column. The isocritic mobile phase was the acetonitrile and water with the rate of 18:82(V/V). The flow rate was 1.5 mL/min. The detective UV wavelength was 195 nm. The plasma samples, added ifosfamide(Ifo) as internal standard, were deposited down the protein with methanol. The supernatant was evaporated by N2 at 40 °C. The remains was dissolved with 100 μL, water. 20 μL was injected to the sampler. Results: The chromatography was good and not interfered by the components of the plasma. The liner equation was Y= 4. 861X + 0. 181 7, r = 0. 999 9(0.5 - 50 μg/mL). The RSD of intra-day<10%, and the RSD of inter-day <10%. Conclusions: This is a accurate and convenient method to determine the concentration of CTX in plasma.