清见蜜柑果皮中多甲氧基黄酮的提取分离

Extraction and Separation of Polymethoxyflavones in Kiyomi Tango Peels

清见蜜柑具有甜橙和宽皮柑的营养、风味和易剥皮的特点,而其果皮中多甲氧基黄酮的成分还未见报道,研究具有多种生物活性的多甲氧基黄酮对改良柑橘品种及食品、保健品开发具有重要意义。本文比较了清见蜜柑、芦柑、赣南脐橙果皮中多甲氧基黄酮的成分,结果表明蜜柑果皮显著不同于芦柑皮,接近于橙皮。采用超声提取,回流提取的方法提取清见蜜柑果皮中的多甲氧基黄酮,结果表明超声提取的最优方法为乙酸乙酯超声30 min,料液比1∶30,提取两次;回流提取的最优方法为乙酸乙酯90℃,回流4 h,提取两次。对乙酸乙酯提取物采用二氯甲烷-甲醇、正己烷-乙酸乙酯两种洗脱体系进行正相硅胶柱层析分离,通过TLC及HPLC分析表明正己烷-乙酸乙酯的洗脱体系的分离效果最好,且多甲氧基黄酮主要集中于1∶1及甲醇洗脱组分中,继而采用半制备型HPLC获得多甲氧基黄酮纯品。本文对清见蜜柑果皮中多甲氧基黄酮的提取分离,对开发利用蜜柑副产物资源具有重要意义。

Kiyomi tango possesses the characteristics of nutrition, flavor and easy peeling of Sweet orange and Citrus reticulate Blanco, while the components of Kiyomi tango peels have not been reported. It has great significance to investigate polymethoxyflavones from Kiyomi tango peels for improving citrus varieties and promoting the development of food and health care products. In this study, the polymethoxyflavones of Kiyomi tango, Lo tangerine and Gannan naval orange peels were compared, indicating that the polymethoxyflavones of Kiyomi tango peels were significantly different from those in Lo tangerine peels. The optimal method of ultrasonic extraction for polymethoxyflavones in Kiyomi tango peels was ethyl acetate as solvent, liquid to material ratio 1 ∶30, ultrasonic time 30 min and twice extraction, while the optimal method of reflux extraction was ethyl acetate as solvent, liquid to material ratio 1∶30, reflux time 4 h and twice extraction. Furthermore, ethyl acetate extracts were separated on column chromatography eluted with two differnet elution systems, methylene chloride-methanol and N-hexane-ethyl acetate. The separation effect of N-hexane-ethyl acetate system was better than that of methylene chloride-methanol, with the enrichment of polymethoxyflavones in the 1 ∶1 and pure methanol fractions, which were confirmed by TLC and HPLC detection. Eventually, 6 pure polymethoxyflavones were obtained by semi-preparative HPLC. In this study, the extraction and separation of polymethoxyflavones in Kiyomi tango peel were designed and optimized, which was important for developing and utilizing the by-products from Kiyomi tango peels.