固相萃取-高效液相色谱法测定葡萄酒中白藜芦醇及其糖苷异构体

建立了固相萃取-高效液相色谱(SPE-HPLC)测定葡萄酒中白藜芦醇及其糖苷异构体的方法,并用此方法测定了市售的15种葡萄酒中顺反式白藜芦醇及其糖苷的含量。实验确定了葡萄酒样品中的白藜芦醇及其糖苷的固相萃取方法,通过优化萃取条件,有效地去除了葡萄酒中大量干扰成分。采用Hypersil C18柱(4.6 mm i.d.×250 mm,5μm)分离，以乙腈-水为流动相,线性梯度洗脱，用二极管阵列检测器分别于288 nm和306 nm波长下检测，4种组分分离良好,提高了分析结果的准确性。采用外标法定量,4种组分的质量浓度与其峰面积在一定的范围内有良好的线性关系,相关系数为0.9973～0.9999。4种组分的平均加标回收率为97.4%～98.8%(相对标准偏差为2.1%~3.2%)，检测限为0.002~0.005 mg/L。对市售15种葡萄酒的检测结果表明,白藜芦醇作为葡萄酒中重要的生物活性成分,其含量与葡萄酒的酿造方式、葡萄品种、葡萄产地有密切的关系。该方法还可用于其他天然产物中白藜芦醇及其糖苷异构体的测定。

Determination of the Contents of Resveratrols and Piceids in Wines by High Performance Liquid Chromatography with Solid Phase Extraction

A method has been developed for the determination of the contents of cis- and trans-resveratrols and piceids in wines by high performance liquid chromatography (HPLC) with solid phase extraction (SPE). The techniques of SPE including the activation of the SPE column, elimination of interference and elution conditions of samples were confirmed. The analytes were separated by using a Hypersil C18 column (4.6 mm i. d. x 250 mm, 5 microm) and acetonitrile-water as mobile phase with linear gradient elution. A photodiode array detector monitored at the detection wavelengths of 288 nm and 306 nm for cis- and trans-isomers, respectively. The external standard calibration curves were used in quantitation. There were good linear correlations between the concentrations and peak areas of the four components with the correlation coefficients of 0.9973-0.9999. The average recoveries were 97.4%-98.6% with relative standard deviations of 2.1%-3.2%. The detection limits were 0.002-0.005 mg/L. The method of SPE-HPLC eliminated a great deal of interferences in wines effectively by optimizing extraction conditions. The four isomers were well separated and the analytical results were improved well and truly. The results for determining 15 market wines showed that as an important bioactive substance, the resveratrol content has close relationships with brewing methods, varieties of grape and producing areas. The results also showed that it is a convenient, rapid, sensitive and accurate method. It can be used to analyze resveratrols and piceids in other natural samples.