HPLC法测定原位肝脏隔离灌注猪血浆中丝裂霉素的浓度

Quantitation of plasma mitomycine C content by high-performance liquid chromatography on isolated liver perfusion in vivo in pigs

目的 建立一种简单、快捷的反相高效液相色谱法 (RP HPLC)测定猪血浆中丝裂霉素C(MMC)浓度的方法。方法 用LC 6AHPLC仪 ,以 μBondapakC18column(5 μm ,3 9× 30 0mm)为分析柱 ;流动相为CH3 CN :H2 O(30∶70 ) ,柱温 2 5℃ ,检测波长36 5nm ,流速 0 5ml·min-1;血浆样品用乙酸乙酯分两次直接萃取。结果 线性范围为 0 1～ 8 0mg·L-1,r =0 9998,n =5 ;提取回收率为 (79 71± 2 97) % ;日内、日间变异均小于 10 % ;最低检测浓度为 0 0 5mg·L-1。结论 该法简便、快速、可靠、特异性强 ,适用于生物样品中MMC监测及药代动力学研究。

AIM To develop a high performance liquid chromatography(HPLC) method for determing the plasma mitomycince C(MMC) concentrations in pigs. METHODS The LC 6A HPLC system was used. The analytical column was a μ Bondapak C 18 column(5 μm, 3 9×300 mm). The mobile phase was the acetonitrile and the water with the rate of 30∶70(V/V). The oven temperature was 25℃. The detective UV wavelength was 365 nm. The flow rate was 0 5 ml·min -1 . The plasma was extracted two times with ethylacetate. RESULTS The liner range was 0 1～8 0 mg·L -1 , r =0 998, n=5 . The recovery was (79 71±2 97)%. The RSD was less than 10% in intraday and interday. The minimal detectable concentration of this method was 0 05 mg·L -1 . CONCLUSION This is a simple, rapid and specific method. It can used for determining the concentration of MMC in plasma and studying its pharmacokinetics.