Department of Food Chemistry, Technology and Biotechnology, Gdańsk University of Technology, 11/12 G. Narutowicza Str., 80-233 Gdańsk, Poland.
Phytochem Anal. 2011 Sep-Oct;22(5):392-402. doi: 10.1002/pca.1294. Epub 2011 Mar 4.
The wide application of natural and artificial antioxidants in the food, cosmetic and pharmaceutical industries as well as the recognition of the importance of food antioxidants for supporting human health have created a demand for reliable and industrially applicable methods of determining antioxidative activity. This requirement can be fulfilled with the recently proposed HPLC-post-column derivatisation approach, enabling rapid measurements of antioxidative potential along with profiling antioxidants in complex mixtures.
To employ the commercially available post-column derivatisation device for the on-line detection of antioxidants and to optimise analytical parameters enabling its application for routine profiling of antioxidants in complex mixtures such as those of plant or food origin.
The mixtures of standards and fruit extracts were resolved using an optimised HPLC method followed by the on-line derivatisation of analytes in a PCX post-column derivatisation instrument. Such parameters as the type of indicator reagent (ABTS, DPPH, Folin-Ciocalteu), its concentration and the temperature of the derivatisation reaction were investigated. The determinations of the Trolox equivalents (TE) values were compared with those obtained by corresponding bulk spectrophotometric tests.
The study confirms that the commercial HPLC-coupled post-column derivatisation instruments are suitable for routine on-line detection of antioxidants in complex mixtures and the determination of their TE values. The analytical parameters optimised here appear to represent a ready-to-use toolbox for the food and pharmaceutical industries, enabling the monitoring of bioactive substances along the production line and during storage, and the characterisation of plant material by creating chromatographic profiles supplemented with antioxidant fingerprints.
天然和人工抗氧化剂在食品、化妆品和制药行业的广泛应用，以及人们认识到食物抗氧化剂对支持人类健康的重要性，这两者都创造了对可靠且适用于工业的抗氧化活性测定方法的需求。这种需求可以通过最近提出的 HPLC 柱后衍生化方法来满足，该方法能够快速测量抗氧化潜力，并对复杂混合物中的抗氧化剂进行分析。
采用市售的柱后衍生化装置对抗氧化剂进行在线检测，并优化分析参数，使其能够用于复杂混合物（如植物或食物来源的混合物）中抗氧化剂的常规分析。
采用优化的 HPLC 方法分离标准品和水果提取物混合物，然后在 PCX 柱后衍生化仪器中对分析物进行在线衍生化。研究了指示剂试剂（ABTS、DPPH、Folin-Ciocalteu）的类型、浓度和衍生化反应温度等参数。并将 Trolox 当量（TE）值的测定结果与相应的比色分光光度法测试结果进行了比较。
研究证实，商用 HPLC 柱后衍生化仪器适用于复杂混合物中抗氧化剂的常规在线检测和 TE 值的测定。这里优化的分析参数似乎代表了食品和制药行业的即用型工具包，能够在生产线和储存过程中监测生物活性物质，并通过创建补充抗氧化指纹的色谱图谱来对植物材料进行特征描述。
