不同血样处理对高效液相色谱法测定血清丝裂霉素含量的影响

Determination of mytomycin C in human serum by RAM-HPLC with different processing methods

目的:考查不同的血样处理方法对血清中丝裂霉素(mitomycin C,MMC)含量的影响。方法:使用Waters HPLC仪,配有限进填料(RAM)直接进样固定相,流动相:甲醇水(10∶90);流速:0.8 mL·min-1;紫外二极管阵列检测器,比较了血清样品不同处理方法的重现性、稳定性,并与血清直接进样进行比较。结果:不同的血清样品处理方法对MMC含量的影响不同,在萃取法中用氯仿异丙醇较醋酸乙酯萃取率高,沉淀法用乙腈沉淀对MMC含量影响较小。直接进样中MMC与血清中的蛋白质有良好的分离。结论:在MMC的血清样品处理方法中,用RAM HPLC法直接进样,样品不经前处理,简单快速,精密度好,但对一些取样量较大且含量较低的样品,萃取法为一种有效的方法。

OBJECTIVE To study the mitomycin C(MMC)concentration by different processing methods in human serum. METHODS A direct injection method for determination of MMC in serum by restricted-access media (RAM)-HPLC was developed. The mobile phase consisted of menthol-water(10∶90) at a flow-rate of 0.8 mL·min -1 . The HPLC was using equipped with aphotodiode array detector UV detection at 365 nm,the repecatabilit and stability of different methods were compared. RESULTS The results of different methods were different, the extraction using chloroform and isopropanol is better and the precipitation by acetonitrile is better than methanol,MMC and protein in bio-fluid were well separated by direct injection. CONCLUSION In different processing methods, direct injection assay is simple,reliable,high sensitive,makes it suitable for large scale routine sample analysis while this extraction are method is speciaug useful for large quantity of sample with low concentration.