HPLC法测定不同采收期的云南甜柿叶中三种黄酮的含量
Determination of three flavonoids in the leaves of Yunnan Diospyros kaki in different harvest periods by HPLC
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摘要: 利用回流提取法对不同采收期的云南甜柿叶进行预处理,采用高效液相色谱法测定样品中山奈酚-3-O-β-D-葡萄糖苷、山奈酚-3-O-β-D-半乳糖苷和槲皮素-3-O-β-D-葡萄糖苷的含量。具体方法为:提取条件为提取溶剂45%乙醇,液固比20∶1,提取4次,提取时间90 min;液相色谱条件为采用ZORBAX Eclipse XDB-C18（250 mm×4.6 mm,5μm）色谱柱分离,甲醇-0.4%磷酸溶液为洗脱系统进行梯度洗脱,检测波长265 nm;三种黄酮的相对标准偏差均小于1%,平均回收率均在99%以上。该方法具有较好的分离效果和较高的稳定性,适用于用来分析不同采收期的云南甜柿叶中上述三种黄酮的含量,测定结果表明三种黄酮在叶中积累具有规律性,随柿果实成长不断积累,并且果实成熟时达到最高。Abstract: A HPLC method was developed for determinating the contents of Kaempferol-3-O-β-D-glucopyranoside, Kaempferol-3-O-β-D-galactopyranoside, and Quercetin-3-O-β-D-glucopyranoside in the leaves of Yunnan Diospyros kaki in different harvest periods.The extraction conditions were 45% ethyl alcohol, the ratio of liquid-solid of 20∶ 1, extraction time of 90 min for 4 times.The HPLC elution was carried out using methanol-0.4% phosphate system at a flow rate of 1.0 m L/min, the injection volume was 10 μL, and the detector wavelength was set at 265 nm.The RSD values of the three flavonoids were all less than 1% and the average recoveries were all above 99%.This method showed good separation effect and high stability, and could be used for determining the contents of the three flavonoids in the leaves of Yunnan Diospyros kaki in different harvest periods.The result showed that the accumulation of them in leaves of Yunnan Diospyros kaki exhibited regularity, and the contents were increasing along with fruit growing.
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