高效液相色谱法筛选甘草甜味素中抗氧化成分

Screening and Identification of Antioxidants in Glycyrrhiza Extract Rich in Glycyrrhizine by High Performance Liquid Chromatography Coupled with Post Column DPPH Assay

为明确甘草甜味素中的抗氧化成分,将高效液相色谱(HPLC)与DPPH快速筛选自由基清除剂的方法与电喷雾串联质谱方法结合,建立甘草甜素中抗氧化活性成分在线筛选与鉴别的方法。HPLC色谱柱分离后,利用二极管阵列检测器得到各活性组分的紫外光谱图,流出液与DPPH在柱后反应管中混合反应,实现在线筛选自由基清除剂的作用。从甘草甜素中筛选出8个具有明显抗氧化活性的化合物,通过电喷雾质谱分析和紫外光谱图,实现了对各化合物的初步识别。8种抗氧化化合物分别为刺芒柄花素、南酸枣苷、甘草苷元-4'-芹糖葡萄糖苷、甘草苷元、甘草苷、刺毛甘草查耳酮、光果甘草酮和甘草查耳酮C。

A new method based on HPLC online scavenging DPPH radical activity and HPLC-MS/MS method for the identification of antioxidants from glycyrrhiza extract rich in glycyrrhizine was developed. The glycyrrhiza extract was separated by HPLC and the outlet of HPLC photo diode array detector flow cell was connected to a DPPH flow. Antioxidants were reacted with DPPH in post-column reaction coil and monitored by UV detector at 517 nm. The results showed that eight compounds with scavenging radical activity were found in the glycyrrhiza extract, which were identified as formononetin, choerospondin, apioliquiritin, liquiritigenin, liquiritin, echinatin, glyyunnansapogenin F, glabrone and licochalcone C by MS/MS and UV spectrometry.