Department of Physical and Analytical Chemistry, School of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Nagoya 463-8521, Japan.
Food Chem. 2012 Oct 15;134(4):2276-82. doi: 10.1016/j.foodchem.2012.02.219. Epub 2012 Mar 30.
We have proposed a novel and first strategy of high-speed countercurrent chromatography (HSCCC) purification for the efficient and effective discovery of antioxidant from natural product based on on-line HPLC method with radical scavenging assay. To achieve a strategy for HSCCC purification, the antioxidants in materials are identified by on-line HPLC with DPPH radical scavenging assay. Then, the optimal condition of target peaks would be investigated for the two-phase solvent system, and purified by HSCCC. In this study, the specific antioxidants in red cabbage, perilla and elderberry pigments were evaluated by on-line HPLC with DPPH radical scavenging assay, and purified by HSCCC technique. Specific antioxidants could be rapidly pinpointed in complex mixtures by on-line HPLC with DPPH radical scavenging assay. Then, the optimal two-phase solvent systems were investigated using these HPLC peaks. Finally, the purification of these nine antioxidants form three mixtures were performed by HSCCC. Using mass spectrometric analysis, these antioxidants were confirmed to cyanidin-based anthocyanin from red cabbage and elderberry pigments, and luteolin-based flavones from perrilla pigment. Due to the advantages derived from on-line HPLC with DPPH radical scavenging assay and HSCCC technique, a rapid, efficient and effective strategy has been developed for the discovery of antioxidants from natural products.
我们提出了一种基于在线 HPLC 方法与自由基清除试验的新型高速逆流色谱（HSCCC）纯化策略，用于从天然产物中高效、有效地发现抗氧化剂。为了实现 HSCCC 纯化策略，我们使用在线 HPLC 方法与 DPPH 自由基清除试验来鉴定材料中的抗氧化剂。然后，我们将针对目标峰的最佳条件进行双相溶剂系统的研究，并通过 HSCCC 进行纯化。在本研究中，我们使用在线 HPLC 方法与 DPPH 自由基清除试验评估了红色卷心菜、紫苏和接骨木浆果色素中的特定抗氧化剂，并通过 HSCCC 技术进行了纯化。通过在线 HPLC 方法与 DPPH 自由基清除试验，可以快速在复杂混合物中定位特定的抗氧化剂。然后，我们使用这些 HPLC 峰研究了最佳的两相溶剂系统。最后，通过 HSCCC 从三种混合物中纯化了这 9 种抗氧化剂。通过质谱分析，确认这些抗氧化剂为红色卷心菜和接骨木浆果色素中的花青素基类黄酮和紫苏色素中的木犀草素基类黄酮。由于在线 HPLC 方法与 DPPH 自由基清除试验和 HSCCC 技术的优势，我们从天然产物中发现抗氧化剂的策略已经得到了快速、高效和有效的发展。
