高效液相色谱法测定微乳体系中胰岛素含量的前处理方法比较

Study on the Pre-treatment Methods of Determination of Insulin in Microemulsions by RP-HPLC

目的 :研究反相高效液相色谱测定微乳体系中胰岛素含量的前处理方法 ,并对 3种前处理方法进行了比较。方法 :采用TritonX 10 0破乳、盐酸酸解、水饱和正丁醇萃取 3种方法对胰岛素微乳体系进行前处理 ,用RP HPLC对处理后样品中胰岛素的含量进行测定。结果 :TritonX 10 0、酸解、萃取 3种方法回收率分别为98 7%～ 10 0 6 % ,94 1%～ 96 8% ,89 2 %～ 92 7% ;RSD分别为 1 5 8%～ 3 2 0 % ,2 0 8%～ 4 30 % ,4 0 7%～6 0 9%。结论 :3种方法中 ,以TritonX 10 0破乳法的结果较为精确 ,回收率较高 ;操作较为方便简单、快速可靠 ;对胰岛素稳定性没有影响 ,样品消耗量较少。

AIM:To investigate the three pre-treatment methods about the determination of insulin in microemulsions before RP-HPLC.METHOD:Three methods-acidolysis by HCl,demulsification by TritonX-100 and extraction by n-butyl alcohol were used to release the insulin which was enclosed,inserted,and absorbed in the oil phase of microemulsions.RESULT:The recoveries of three methods(acidolysis,Triton X and extraction) were 94.1%～96.8%,98.7%～100.6%,and 89.2%～92.7% respectively;and the RSDs were 2.08%～4.30%,1.58%～3.20%,4.07%～6.09% respectively according to above sequence.CONCLUSION:Among the three methods,the results from pretreated by Triton X-100 acid solution were the most accurate,and the recoveries were also the highest.The pre-treatment through Triton X-100 acid solution was the most convenient,simple,rapid,and the least sample-consumed method.What is more,it had little influence on the stability of insulin in microemulsions.