Many natural products are rich in antioxidants which play an important role in preventing or postponing a variety of diseases, such as cardiovascular and inflammatory disease, diabetes as well as breast cancer. In this paper, an automatic on-line 2,2-diphenyl-1-picrylhydrazyl-high performance liquid chromatography (DPPH-HPLC) method was established for antioxidants screening with nine standards including organic acids (4-hydroxyphenylacetic acid, p-coumaric acid, ferulic acid, and benzoic acid), alkaloids (coptisine and berberine), and flavonoids (quercitrin, astragalin, and quercetin). The optimal concentration of DPPH was determined, and six potential antioxidants including 4-hydroxyphenylacetic acid, p-coumaric acid, ferulic acid, quercitrin, astragalin, and quercetin, and three non-antioxidants including benzoic acid, coptisine, and berberine, were successfully screened out and validated by conventional DPPH radical scavenging activity assay. The established method has been applied to the crude samples of Saccharum officinarum rinds, Coptis chinensis powders, and Malus pumila leaves, consecutively. Two potential antioxidant compounds from Saccharum officinarum rinds and five potential antioxidant compounds from Malus pumila eaves were rapidly screened out. Then these seven potential antioxidants were purified and identified as p-coumaric acid, ferulic acid, phloridzin, isoquercitrin, quercetin-3-xyloside, quercetin-3-arabinoside, and quercetin-3-rhamnoside using countercurrent chromatography combined with mass spectrometry and their antioxidant activities were further evaluated by conventional DPPH radical scavenging assay. The activity result was in accordance with that of the established method. This established method is cheap and automatic, and could be used as an efficient tool for high-throughput antioxidant screening from various complex natural products.

许多天然产品富含抗氧化剂，这些抗氧化剂在预防或推迟各种疾病（例如心血管疾病和炎性疾病，糖尿病以及乳腺癌）中起着重要作用。本文建立了一种自动在线2,2,2-二苯基-1-吡啶并肼基高效液相色谱（DPPH-HPLC）方法，以9种标准品（包括有机酸（4-羟苯基乙酸，对香豆酸）进行抗氧化剂筛选，阿魏酸和苯甲酸），生物碱（可替宁和小ber碱）和类黄酮（槲皮素，黄芪素和槲皮素）。确定了DPPH的最佳浓度，并确定了六种潜在的抗氧化剂，包括4-羟苯基乙酸，p通过常规DPPH自由基清除活性试验成功筛选出了香豆酸，阿魏酸，槲皮素，黄芪素和槲皮素，以及三种非抗氧化剂，包括苯甲酸，黄连和小ber碱，并对其进行了验证。所建立的方法已被应用到的粗样品甘蔗皮，黄连粉剂和海棠荔叶，连续。快速筛选出了来自糖皮果皮的两种潜在的抗氧化剂化合物和来自苹果属屋檐的五个潜在的抗氧化剂化合物。然后将这七种潜在的抗氧化剂纯化并鉴定为p-香豆酸，阿魏酸，Phloridzin，异槲皮苷，槲皮素-3-木糖苷，槲皮素-3-阿拉伯糖苷和槲皮素-3-鼠李糖苷的组合使用逆流色谱法，并通过常规DPPH自由基清除试验进一步评估了它们的抗氧化活性。活性结果与所建立的方法一致。这种建立的方法便宜且自动，并且可以用作从各种复杂的天然产物中高通量抗氧化剂筛选的有效工具。