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目的探讨氨基酸分析的方法对蛋白含量国家标准品定量的可行性。方法采用常规酸水解的方法,通过Hitachi L8800型氨基酸分析仪对白蛋白标准品进行分析。先确定最佳水解时间,再按照最佳水解时间对样品进行水解,通过氨基酸分析,采用氨基酸加合法测定其含量。并验证方法的精密性和准确性。结果经7次测定,求得现行蛋白含量国家标准品[(2005)国生标字0009]的蛋白含量值为(95.02±3.99)mg/100 mg,相对标准偏差(relative standard deviation,RSD)为4.20%。多次试验中,标准品中各氨基酸的回收率在93.56%~105.12%之间。推算标准品中白蛋白含量值为20.52 mg,是凯氏定氮测定值的103.48%。结论氨基酸分析法测定蛋白含量自动化程度高,重复性好,可用于重组蛋白含量的分析。
Objective To investigate the feasibility of amino acid analysis for the quantification of national standards of protein content. Methods The conventional method of acid hydrolysis was used to analyze albumin standards by Hitachi L8800 amino acid analyzer. First determine the optimal hydrolysis time, and then hydrolysis of the sample according to the optimal hydrolysis time, amino acid analysis by amino acid determination of its content. And verify the accuracy and accuracy of the method. Results The protein content of the national standard [(2005) Guoji Dianzi 0009] was (95.02 ± 3.99) mg / 100 mg and the relative standard deviation (RSD) was 4.20 after seven determinations. %. In many experiments, the recovery rate of each amino acid in the standard ranged from 93.56% to 105.12%. Calculated standard protein content of 20.52 mg of albumin, Kjeldahl determination of 103.48%. Conclusion Amino acid analysis of protein content with high degree of automation, repeatability, and can be used for the analysis of recombinant protein content.