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目的采用反相高效液相色谱法(RF-HPLC)技术,建立人免疫球蛋白中甘氨酸含量的测定方法。方法采用RF-HPLC,以邻苯二甲醛(OPA)为柱前衍生剂,采用自动进样器的自动衍生程序进行衍生化反应,在338nm波长处检测,用外标法计算甘氨酸的含量。结果当甘氨酸衍生化的取样范围为0.05～0.15 g/L时,直线相关系数为0.999 6～1.000 0,甘氨酸的回收率为96%～101%,日内及日间重复测定的相对标准偏差(RSD)分别为2.3%和1.5%,2位操作人员测定结果经T检验P值为0.77。结论 RF-HPLC测定结果稳定、准确,可用于人免疫球蛋白中甘氨酸含量的测定。
Objective To establish a method for the determination of glycine in human immunoglobulin by reverse-phase high-performance liquid chromatography (RF-HPLC). Methods The derivatization reaction was carried out by auto-derivatization with an autosampler using orthophthalaldehyde (OPA) as a pre-column derivatization reagent. The concentration of glycine was calculated at 338 nm. Results When the sampling range of glycine derivatization was 0.05-0.15 g / L, the linear correlation coefficient was 0.999 6-1.000 0 and the recovery of glycine was 96% -101%. The relative standard deviation (RSD) ) Were 2.3% and 1.5%, respectively, two operators of the test results by the T test P value of 0.77. Conclusion The results of RF-HPLC are stable and accurate and can be used for the determination of glycine in human immunoglobulin.