摘要
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 chromatographic method to determine angiotensin-converting enzyme inhibitor activity by Globin peptide was established. By using N-hippuryl-His-Leu tetrahydrate as the reaction substrate and hippuric acid as the reaction product. The optimal conditions of HPLC were as follows： the column of chromatogram Zorbax SB- C18（4.6mm× 150mm, 5μm）, column temperature 25℃ , mobile phase acetonitrile/water=25/75[V/V, both containing 0.05%（V/ V） trifluoroacetic acid], flow rate 1.0mL/min, UV-235nm detector, inject 101aL. The linear range was from 0.005 to hnmol/ L as the linear correlation coefficient was 0.9998, the minimum detection limit of melamine was determined to be 1μmol/L, the average recoveries of the method were 96.66% to 101.34%, and the RSD were 1.34%. It is a simple, short analyse time, accuracy and precise method for quality control of Globin production.
关键词
珠蛋白肽
高效液相色谱
血管紧张素转化酶
马尿酸
Globin peptide
high performance liquid chromatography
angiotensin-converting enzyme inhibitor
hippuric acid
