葡萄果皮花青素提取工艺优化研究

利用高效液相色谱(HPLC)对提取葡萄果皮中花青素的提取液、提取方法和色谱条件进行比较,采用正交实验设计选择最佳的提取工艺,利用5种花青素峰面积分析夏黑葡萄果皮所含花青素的含量。试验结果表明,提取液为甲醇：水=1:1,磷酸调pH=2.0,提取时超声辅助提取30min,为最佳的工艺提取出夏黑葡萄果皮中花青素的含量,色谱条件为流动相A：水/乙腈/甲酸= 40/50/10(v/v/v),流动相B：水/乙腈/甲酸= 87/3/10(v/v/v),柱温45℃,进样量20μL,检测波长520 nm,梯度洗脱条件为:(1)0-15 min,6% A-30% A；(2)15-30 min,30% A-50% A；(3)30-35 min,50% A-60% A；(4)35-40 min,60% A-6% A,呈现的色谱图分离度更好,总体效果更佳。

Optimizing the Extraction Process of Anthocyanin in Grape Peel

To study the effects of different conditions on the extraction of anthocyanins, an orthogonal experimental design L9(33) was used to optimize the condition of the high performance liquid chromatography (HPLC) assay of anthocyanin composition and content in peel of summer black grape. Three factors affecting extraction process of anthocyanin including, extraction solution, method and three chromatographic conditions were evaluated with an aim to establish a high- throughput extracting process of assay in anthocyanin composition and content in peel of grape. The results showed that the optimal process of assay in anthocyanin composition and content consisted of the extract solution of methanol: water=1:1, phosphoric acid pH value of 2.0, the extraction time of 30 min. The chromatographic condition included mobile phase A: water/ acetonitrile/ formic acid=40/50/10 (v/v/v) and mobile phase B: water/ acetonitrile/ formic acid=87/3/10 (v/v/v), column temperature of 45℃, injection volume of 20 μL, detection wavelength of 520 nm, and the gradient elution conditions were: (1) 0-15 min, 6%A→30%A; (2)15-30 min, 30%A→50%A; (3)30-35 min, 50%A→60%A; (4)35-40 min, 60%A→6%A. Under these conditions, elphinidin-3-O-glucoside, cyanidin-3-O-glucoside, pelargonidin-3-O-glucoside, peonidin-3-O-glucoside and malvidin-3-gluconside chloride were separated successfully.