Koleva I I, Niederländer H A, van Been T A
Laboratory of Organic Chemistry, Wageningen University, The Netherlands.
Anal Chem. 2000 May 15;72(10):2323-8. doi: 10.1021/ac9912451.
A rapid on-line method for screening of complex mixtures for radical scavenging components was developed using a methanolic solution of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) stable free radical. The HPLC-separated analytes react postcolumn with the DPPH solution, and the induced bleaching is detected as a negative peak by an absorbance detector at 517 nm. An optimized instrumental setup is presented. The method is suitable for both isocratic and gradient HPLC runs with mobile-phase compositions ranging from 10 to 90% organic solvent in water or buffer (pH 3-6). The method is simple, has a broad applicability, and uses common instruments, inexpensive and stable reagents, and a time-saving and nonlaborious experimental protocol. It can also be used for quantitative analysis. The method was applied to several pure natural antioxidants and plant extracts. The limits of detection were 0.33-94 microg/mL, depending on the compound tested.
开发了一种快速在线方法，用于筛选复杂混合物中的自由基清除成分，该方法使用2,2'-二苯基-1-苦基肼（DPPH）稳定自由基的甲醇溶液。经高效液相色谱（HPLC）分离的分析物在柱后与DPPH溶液反应，诱导的褪色通过517 nm处的吸光检测器检测为负峰。本文介绍了一种优化的仪器装置。该方法适用于等度和梯度HPLC运行，流动相组成范围为水或缓冲液（pH 3 - 6）中10%至90%的有机溶剂。该方法简单，适用性广，使用常见仪器、廉价且稳定的试剂以及省时省力的实验方案。它还可用于定量分析。该方法应用于几种纯天然抗氧化剂和植物提取物。检测限为0.33 - 94 μg/mL，具体取决于所测试的化合物。
