Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH-HPLC-DAD-TOF/MS
Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH-HPLC-DAD-TOF/MS
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DOI：
10.1002/jssc.200800173
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发表时间：
2008-11-01
影响因子：
3.1
通讯作者：
Li, Ping
中科院分区：
文献类型：
--
作者：
Tang, Dan;Li, Hui-Jun;Li, Ping
建立了一种快速、简便的筛选和鉴定金银花中天然抗氧化剂的方法。假设与1，1-二苯基-2-苦肼基（DPPH）反应后，具有潜在抗氧化作用的化合物在HPLC色谱图中的峰面积（PAs）将显著减少或消失，并且可以通过HPLC-DAD-TOF/MS联用技术进行身份确认。使用所提出的方法，在FLJ提取物中发现约14种化合物具有潜在的抗氧化活性。分离得到的化合物分别为绿原酸（1）、1-O-咖啡酰奎宁酸（1（1-O-CQA，2）、咖啡酸（4）、4-O-CQA（5）、芦丁（7）、异槲皮苷（8）、木犀草素-7-O-葡萄糖苷（9）、忍冬苷（10）、4，5-O-二咖啡酰奎宁酸（4，5-O-diCQA，11）、3，5-O-diCQA（12）、1，3-O-diCQA（13）、3，4-O-diCQA（1,4）、1，4-O-diCQA（16）和毛地黄黄酮（17）。此外，还通过高效液相色谱法研究了现有鉴定化合物的自由基清除能力。结果表明，PAs含量显著降低的化合物为天然抗氧化剂，PAs含量不变的化合物为无活性化合物，表明该方法可广泛应用于中草药等复杂基质中天然抗氧化剂的快速筛选和鉴定。
A rapid and simple method has been developed for the screening and identification of natural antioxidants of Flos Lonicerae Japonicae (FLJ), derived from the flower buds of Lonicera japonica. The hypothesis is that upon reaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas (PAs) of compounds with potential antioxidant effects in the HPLC chromatograms will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC-DAD-TOF/MS hyphenated technique. Using the proposed approach, about 14 compounds in the FLJ extract were found to possess a potential antioxidant activity. They were identified as chlorogenic acid (1), 1-O-caffeoylquinic acid (1-O-CQA, 2), caffeic acid (4), 4-O-CQA (5), rutin (7), isoquercitrin (8), luteolin-7-O-glucoside (9), lonicerin (10), 4,5-O-dicaffeoylquinic acid (4,5-O-diCQA, 11), 3,5-O-diCQA (12), 1,3-O-diCQA (13), 3,4-O-diCQA (14), 1,4-O-diCQA (16), and luteolin (17). In addition, the free radical scavenging capacities of the available identified compounds were also investigated by HPLC assay. The results indicated that the compounds with PAs significantly decreasing were natural antioxidants, whereas those with PAs not changing presented no activities, which accordingly indicated that this newly proposed method could be widely applied for rapid screening and identification of natural antioxidants from complex matrices including Chinese herbal medicines.