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建立了测定珠蛋白肽对血管紧张素转化酶抑制活性的高效液相色谱分析方法。以马尿酰-组胺酰-亮氨酸为底物,血管紧张素转化酶为催化剂,反应产物马尿酸为检测指标,未加珠蛋白肽的反应为空白对照。最佳色谱条件为:ZorbaxSB-C18(4.6mm×260mm,粒径5μm),柱温25℃,流动相为乙腈/水=25/75[体积比,各含0.05%三氟乙酸(体积分数)],流速1.0mL/min,紫外检测器,检测波长为228nm,进样量10μL。马尿酸浓度为0.005～1mmol/L,线性相关系数为0.9998,最低检测浓度为1μmol/L,回收率在96.66%～101.34%之间,相对标准偏差为1.34%之间。该方法操作简单、分析时间短、准确性和精密度高,利于对珠蛋白肽生产进行质量控制。
A high performance liquid chromatography method for the determination of inhibitory activity of globin peptides on angiotensin converting enzyme was established. Using hippuryl-histidyl-leucine as substrate and angiotensin converting enzyme as catalyst, the reaction product hippuric acid was the detection index, and the reaction without adding globin peptide was blank control. The optimal chromatographic conditions were as follows: Zorbax SB-C18 (4.6 mm × 260 mm, particle size 5 μm) with a column temperature of 25 ° C and a mobile phase of acetonitrile / water = 25/75 [ ], Flow rate 1.0mL / min, UV detector, detection wavelength of 228nm, injection volume 10μL. The concentration of hippuric acid was 0.005 ~ 1mmol / L, the linear correlation coefficient was 0.9998, the lowest detection concentration was 1μmol / L, the recovery was between 96.66% and 101.34%, and the relative standard deviation was 1.34%. The method is simple in operation, short in analysis time, high in accuracy and precision, which is good for quality control of globin peptide production.