利用酸水解氨基酸法测定脑蛋白水解物片中肽的含量

摘要

Objective: To investigate the analytic method of peptide contents from Cerebroprotein Hydrolysate Tablets.Methods: Used Waters AccQ-Tag method by Kromasil 100-3 C18 column (250 mm×4.6 mm, 5μm) to analyze amino acids.Gradient eluents consisted of sodium acetate solution (pH=5.00) and 60％ acetonitrile.The detection wavelength was 248 nm, column temperature was 371, the sample size was 5 μl.The equation between the total amino acids contents of the hydrolytic and nun-hydrolytic sample was the contents of peptide.Results: The calibration curves for amino acids had good linearities (r=0.999 1-0.999 9, n=5).The mean recoveries of nun-hydrolytic was 100.6％ with RSD of 0.7％ (n=9); the mean recoveries of peptide was 100.4％ with RSD of 1.1％ (n=9).Conclusion: The method is suitable for the further determination of peptide contents in drug.%目的:探讨脑蛋白水解物片中肽含量的测定方法.方法:采用盐酸水解多肽方法,利用Waters AccQ-Tag氨基酸衍生技术,使用Kromasil 100-5 C18色谱柱(250.0 min×4.6 mm,5μm)进行液相色谱分析,以乙酸钠(pH=5.00)缓冲液为流动相A,60%乙腈为流动相B进行梯度洗脱,检测波长为248 nm,柱温为37℃,进样量为5μl.水解后游离氨基酸总量扣除水解前氨基酸总量即为肽含量.结果:各氨基酸测定结果具有良好线性(r=0.999 1～0.999 9,n=5),未水解氨基酸平均回收率为100.6%(RSD为0.7%,n=9),肽含量平均回收率为100.4%(RSD为1.1%,n=9).结论:本方法准确可行,可作为药品中肽含量测定的方法补充和完善.