高选择性高效液相色谱-电化学检测法测定丹参中5种酚类物质

摘要

The determination of antioxidants continues to be interested, since the oxidative damage is thought to be one of the main mechanisms involved in nearly all chronic renal pathologies.A highly sensitive high performance liquid chromatography-electrochemical detection (HPLC-ECD) method was developed for evaluating the antioxidant properties of Salvia miltiorrhiza (Dan Shen).The method was optimized with respect to selectivity and sensitivity.Chromatographic conditions, including mobile phase pH value, buffer concentration, buffer type, organic solvent type, gradient profile and flow rate, were systematically investigated.Low pH value (2.8), low buffer concentration (20 mmol/L NaH2PO4), a shallow water-acetonitrile gradient, and a flow rate of 0.2 mL/min were the determined optimal conditions for the quantitative analysis of aimed five antioxidants from 14 batches of Dan Shen samples.The described method provided a good recovery (>95%), a very wide linear range (up to 104 for all analytes), a good precision (RSDs<4.01%), and a high sensitivity (LOQ of caffeic acid, 1.5 μg/L).Compared with UV detection, the described ECD method was also more effective for evaluating the antioxidant properties of Dan Shen, as it provided highly selective detection of electro-active antioxidants.%发展了一种高灵敏度的高效液相色谱-电化学检测(HPLC-ECD)方法,用于丹参中5种酚类物质的定量分析.为了获得较高的检测灵敏度,对流动相的pH、缓冲溶液类型和浓度、有机相种类和梯度以及流速做了系统研究.在较低的pH (2.8)、较低的缓冲盐浓度(20 mmol/L NaH2PO4)和较缓的乙腈梯度下,以0.2 mL/min流速可为5种酚类的分离检测提供较好的分离度和较高的检测灵敏度.在获得优化的分离参数后,将其用于14批丹参药材中5种酚类物质的定量分析.结果表明,该方法可获得较好的回收率(>95%)、较宽的线性范围(高达4个数量级)、较好的重复性(RSD<4.01%)和较高的灵敏度(咖啡酸的LOQ低至1.5 μg/L).与紫外检测方法相比,ECD检测方法具有更高的选择性,可减少非抗氧化活性的物质的干扰.