柱前衍生化RP-HPLC法测定肌肽的含量

Content determination of carnosion by pre-column derivatization with PR-HPLC

目的建立简单、快速测定肌肽含量的方法。方法采用2，4-二硝基氯苯柱前衍生化RP-HPLC法。衍生化试剂：2，4-二硝基氯苯；色谱柱：Diamonsil^TMC18柱（200mm×4．6mm，5μm）；流动相：乙腈-0．15mol·L^-1乙酸钠缓冲溶液（体积比1：5）；流速：1．0mL·min^-1；检测波长：360nm。结果肌肽质量浓度在2．0～20．0mg·L^-1（r=0．9994）内与其衍生化物峰面积呈良好的线性关系，平均回收率为96．8％，RSD为0．8％（n=9）。结论2，4-二硝基氯苯柱前衍生化RP-HPLC法可用于测定肌肽含量。

Objective To estabish a pre-column derivatization with RP-HPLC method for the determination of content of carnosion. Methods 2,4-Dinitrochlorobenzene was used as derivatization regent. The reversed phase HPLC system consisted of a Diamonsil^TM C18 column（250 mm × 4.6 mm,5 μm） and a mixture of acetonitrile：Sodium acetate solution（ V ： V= l：5）as mobile phase. The flow rate was 1.0mg·L^-1. The UV detection wavelength was 360nm. Results A linearity between major derivative peak area and tiopronin concentration was achieved in the range of 2.0 - 20.0mg·L^-1, with r=0.999 4. The average recovery was 96. 8% and RSD was 0.8% （n =9）. Conclusions The method can be used for determination of content of camosion.