A new method based on HPLC-MS for the rapid screening and identification of antioxidants from methanol extract of Scutellariae Radix has been developed. An appropriate amount of diphenyl-1-picrylhydrazyl(DPPH. ) was added to the methanol extract of Scutellariae Radix and incubated in dark place at room temperature for half an hour while DPPH. was replaced by methanol in the control group. This two group samples were analyzed by HPLC-MS. Comparing the chromatograms of experiment and control group, the peak areas of antioxidants after the interaction with DPPH. had a decrease in the chromatogram while for those not responsible for DPPH. had no change in their peak areas. The relative binding affinities of antioxidants were calculated based on HPLC data while the structure identification was carried out based on MS data. Four antioxidants were screen out from methanol extract of Scutellariae Radix by this novel method and were identified as baicalin, Oroxylin A-7-O-glucuronide, wogonoside and oroxylin A based on HPLC and MS date. Because of the powerful ability of HPLC-MS for small molecule separation and identification, the method has the potential to be a high throughput tool for screening antioxidants from complicated systems such as herbal extracts while do not need laborious pre-separation and additional instrument improvement.%建立了高效液相色谱-质谱联用技术快速筛选和鉴定黄芩甲醇提取物中抗氧化活性成分的方法.在黄芩甲醇提物中加入适量的二苯基三硝基苯肼( DPPH)作为实验组,避光室温反应30 min后直接经液相色谱分析,并与不加入DPPH的空白组进行比较.有抗氧化活性的成分因会与DPPH反应而使峰面积减少,而不具有抗氧化活性的成分峰面积则不变.基于色谱数据可以获得有抗氧化活性组分的相对活性强度,基于质谱数据可获得活性组分的结构信息用于结构鉴定.本方法成功地从黄芩甲醇提取物中筛选并鉴定出了黄芩苷、千层纸素A-7-O-B-D-葡萄糖醛酸苷、汉黄芩苷和千层纸素A4种较强的抗氧化活性成分.本方法利用HPLC-MS技术对复杂体系(如中药提取物)中小分子的分离和鉴定优势,可以直接筛选出黄芩甲醇提取物中的抗氧化活性成分,而不需要繁琐的前期分离和纯化,并且不需要仪器改装工作,有利于实现复杂体系中抗氧化活性成分的高通量筛选.
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