School of Biotechnology and Health Sciences & International Healthcare Innovation Institute (Jiangmen), Wuyi University, Jiangmen 529020, China; School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China; Guangdong Food Green Processing and Nutrition Regulation Technologies Research Center, Guangzhou 510640, China.
School of Biotechnology and Health Sciences & International Healthcare Innovation Institute (Jiangmen), Wuyi University, Jiangmen 529020, China.
Food Chem. 2024 Mar 15;436:137670. doi: 10.1016/j.foodchem.2023.137670. Epub 2023 Oct 5.
Food-derived antioxidant peptides have become the focus of research due to their high safety and low cost. However, the discovery is suffering from a low efficient and empirical approach, involving multi-step off-line separation and identification. In this work, an on-line stop-flow RPLC × SEC-MS/DPPH radical scavenging activity analysis system was developed. For optimization, the conditions: 10 m reaction loop, 200 μM DPPH radical concentration, 40℃ temperature and 0.06 % formic acid were recommended. The system was fully validated by its application in glutathione analysis. The system was further applied in analysis of complex mixed standards, and the dipeptides GC (Gly-Cys) and CW (Cys-Trp) with relatively strong DPPH radical scavenging activity were validated. Maize protein hydrolysates were used for tests and the peptide AC (Ala-Cys) of high probability with strong DPPH radical scavenging activity was identified, demonstrating a high potential of the system. This would help to facilitate the discovery of antioxidative peptides in the future.
由于其安全性高、成本低，食源性抗氧化肽已成为研究的焦点。然而，目前的发现方法效率低下且具有经验性，涉及多步离线分离和鉴定。本研究开发了一种在线停流反相高效液相色谱-体积排阻色谱-二苯代苦味肼基自由基清除活性分析系统。为了优化系统，推荐使用 10 m 反应环、200 μM DPPH 自由基浓度、40℃温度和 0.06%甲酸的条件。该系统通过应用于谷胱甘肽分析进行了充分验证。该系统进一步应用于复杂混合标准品的分析，验证了具有相对较强 DPPH 自由基清除活性的二肽 GC（Gly-Cys）和 CW（Cys-Trp）。还使用玉米蛋白水解物进行了测试，并鉴定出具有强 DPPH 自由基清除活性的高概率肽 AC（Ala-Cys），表明该系统具有很大的潜力。这将有助于促进未来抗氧化肽的发现。
