液相法微量检测胶原肽清除羟基自由基能力

Microdetermination of hydroxyl radical scavenging activity of collagen peptide by HPLC

建立了一种微量检测样品清除羟基自由能力的液相分析方法。采用Inertsil ODS-SP C18色谱柱(4.6mm×150mm,5μm),流动相为v(甲醇):v(0.02mol/L乙酸铵水溶液)=5:95,流速1.0mL/min,柱温30℃,样品使用量为30μL,液相检测时间为6min。在上述条件下测得胶原肽清除羟基自由基的半抑制浓度(IC50值)为15mg/mL,与传统分光光度法测定值一致。该方法灵敏度高,准确度和重现性好,样品使用量少,并且不受样品本身颜色的干扰,可广泛应用于微量检测天然抗氧化剂对羟基自由基的清除能力。

A HPLC method was developed in this study for the microdetermination of hydroxyl radical scavenging activity by peptide using an Inertsil ODS-SP RP-18e column（4.6 mm×150 mm,5 μm）.The column temperature was 30 ℃,the mobile phase was a mixture of methanol and 0.02 mol/L ammonium acetate solution（5：95,v/v） pumped at a flow rate of 1.0 mL/min,and the sample usage amount was 30 μL.In this condition,the 50% radical scavenging activity（IC50） of collagen peptide in the scavenging of hydroxyl radicals was 15 mg/mL,which correlated well with that of spectrophotometry.This method is of high sensitivity,accuracy and reproducibility without the interference from the color of samples,as well as the sample usage amount is minute.It can be used widely for microdetermination of hydroxyl radical scavenging activities of natural antioxidants.