利用HPLC法检测活性肽HMG\|CoA还原酶抑制剂活性

浙江农业学报

利用HPLC法检测活性肽HMG\|CoA还原酶抑制剂活性

（1浙江省农业科学院农产品质量标准研究所，浙江杭州 3100212； 2 浙江省创意农业工程技术研究中心，浙江杭州 3100211； 3 丽水市农业局，浙江丽水 323000；4四川农业大学食品学院，四川雅安 625014）

出版日期:2015-04-25 发布日期:2015-04-30

Determination of HMG\|CoA reductase inhibitor activity of bioactive peptides by HPLC

（1 Institute of Quality and Standard for Agriculture Products， Zhejiang Academy of Agricultural Sciences， Hangzhou 310021， China； 2 Zhejiang Engineering Research Center for Creative Agriculture, Hangzhou 310021, China； 3 Lishui Municipal Bureal of Agriculture, Lishui 323000, China； 4 College of Food Science， Sichuan Agricultural University， Yaan 625014， China）

Online:2015-04-25 Published:2015-04-30

摘要/Abstract

摘要： 通过检测反应体系中NADPH的浓度变化，建立了一套测定HMG\|CoA还原酶抑制剂活性的HPLC法。以Symmetry C18色谱柱（5 μm，4.6 mm×250 mm）进行分离，流动相为：V（K2HPO4\|KH2PO4）∶V（甲醇）=85∶15，pH 7.2，流速1 mL·min-1；检测波长337 nm，进样量20 μL，柱温25℃。在此条件下，NADPH的线性范围为0.6～600 μmol·L-1，加标回收率为94.48%～101.81%，相对标准偏差（n=5）小于4.49%，最低检出限为0.01 μmol·L-1。用此方法对蚕蛹蛋白肽的HMG\|CoA还原酶抑制剂活性进行了测定，在酶解肽浓度为0.5 mg·mL-1时，其抑制率为28.70%。

关键词: 降血脂肽；HMG\, CoA还原酶；NADPH；液相色谱

Abstract: A HPLC method was established for the determination of HMG\|CoA reductase inhibitor activity in vitro by measuring the concentration of NADPH in the determination system. The chromatographic conditions were as follows: Symmetry C18 column （5 μm， 4.6 mm×250 mm）; mobile phase: V（K2HPO4\|KH2PO4）:V（methanol）=85∶15; pH 7.2; flow rate: 1 mL·min-1; detection wavelength: 337 nm; sample quantity: 20 μL; column temperature: 25℃. In this condition， the NADPH linear range was 0.6-600 μmol·L-1. The standard addition recovery rate was 94.48%-101.81%， and the relative standard deviation （n=5） was less than 4.49%. The lowest detection limit was 0.01 μmol·L-1. The inhibitor activity of silkworm chrysalis protein peptide HMG\|CoA reductase was measured by the above method， and the inhibition rate was 28.70% at the concentration of 0.5 mg·mL-1.

Key words: hypolipidemic peptide, HMG\, CoA reductase, NADPH, HPLC

王伟1，2，李官平3，张玉1，2，*，杨玉英4，张一帆1，邬应龙4. 利用HPLC法检测活性肽HMG\|CoA还原酶抑制剂活性[J]. 浙江农业学报.