利用HPLC法检测活性肽HMG-CoA还原酶抑制剂活性

Determination of HMG-CoA reductase inhibitor activity of bioactive peptides by HPLC

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（5μm,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20μL,柱温25℃。在此条件下,NADPH的线性范围为0.6-600μmol·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0.01μmol·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0.5mg·mL^-1时,其抑制率为28.70%。

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 （ 5μm,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 p, mol· 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.