RP-HPLC法测定酪蛋白水解物中ACE抑制肽的含量

赵勇; 胡志和; 孙振刚; 武文起; 冯永强; 薛璐

doi:10.13386/j.issn1002-0306.2015.10.002

RP-HPLC法测定酪蛋白水解物中ACE抑制肽的含量

Quantitative determination of ACE inhibitory peptides in casein hydrolyzate by RP-HPLC

摘要

摘要: 目的:为了建立定量检测酪蛋白水解物中的血管紧张素转换酶（ACE）抑制肽的反相高效液相色谱（RP-HPLC）方法。方法:酪蛋白先经过胃蛋白酶水解、再经过胰蛋白酶水解,用风味蛋白酶和离子交换树脂对水解物先后进行脱苦和脱盐处理。建立标准品十肽酪氨酸-谷氨酰胺-赖氨酸-苯丙氨酸-脯氨酸-谷氨酰胺-酪氨酸-亮氨酸-谷氨酰胺-酪氨酸（YQKFPQYLQY）的RP-HPLC检测方法,运用外标法测定酪蛋白水解物中的ACE抑制肽YQKFPQYLQY的含量。结果:标准品十肽YQKFPQYLQY质量浓度在2.5⁴.5mg/m L（R2=0.95496）范围内与峰面积呈良好的线性关系。酪蛋白水解物中所含十肽质量分数为4.96694%±0.00131%。结论:本液相检测方法操作简便、快速,适用于ACE抑制肽YQKFPQYLQY的定量检测分析。

Abstract: The purpose was to establish a method for quantitative determination angiotensin converting enzyme (ACE) inhibitory peptides in hydrolyzate of casein using reversed-phase high performance liquid chromatography ( RP-HPLC) . Casein was hydrolyzed by pepsin, and then was hydrolyzed by trypsin. With flavourzyme for debittering of casein hydrolyzate. Salts of casein hydrolyzate were removed by ion exchange resin. The RP-HPLC method to detect decapeptide YQKFPQYLQY was established. Using external standard method to determine YQKFPQYLQY of casein hydrolyzate. Results showed that, the linear range of decapeptide was 2.5⁴.5mg/m L (R2=0.95496) . The decapeptide concentration in the casein hydrolyzate was 4.96694%±0.00131%.Therefore, the RP-HPLC was simple and rapid, suitable for the quantitative detection of ACE inhibitory peptides YQKFPQYLQY.