【摘要】 花色苷是植物中一类具有重要生物活性的次生代谢产物,因其含量较低,使得分离制备花色苷高纯品难度较大。本研究以茶树紫芽为实验材料,以花色苷得率与纯度为考察指标,优化筛选了应用高速逆流色谱结合半制备型液相色谱分离纯化花色苷高纯度的方法。结果表明,茶样经提取、脱脂、阳离子交换树脂分离等前处理后,总花色苷得率28.18 mg/g,纯度为20.38%;以高速逆流色谱分离的最优溶剂体系乙酸乙酯∶正丁醇∶乙腈∶0.1%三氟乙酸水溶液=8∶35∶13∶60(v/v/v/v),除去部分杂质后,总花色苷得率达到160.59 mg/g,纯度为43.64%,再结合半制备型液相色谱分离出5种花色苷组分,纯度最高达90.61%,其优化色谱条件为C18制备柱、波长280 nm、柱温30℃、流动相1%醋酸水溶液(A)-乙腈(B)、流速5 mL/min、进样量1 mL、梯度洗脱(0～55 min,10%～40%B)。研究表明高速逆流色谱结合半制备型液相色谱可有效地分离纯化茶树紫芽花色苷及其组分。更多还原

【Abstract】 Anthocyanins are a class of secondary metabolites in plants with important biological activities.it is difficult to separate and prepare high-purity anthocyanins from plants because of the low-content.the purple bud tea was used as experimental material in this study,taking the yield and the purity of anthocyanins as an indicators,the method of separation and purification of anthocyanins by high-speeed counter-current chromatography(HSCCC) coupled with semipreparative high performance liquid chromatography(PHPLC) were optimized and screened.The results showed that total anthocyanin yield was 28.18 mg/g and the purity of anthocyanin was 20.38% after sample pretreatment by extraction,degreasing and cation exchange resin.The optimal solvent system in high-speed counter-current chromatography was ethyl acetate: n-butanol: acetonitrile: 0.1% trifluoroacetic acid = 8:35:13:60(v/v/v/v),with most of the impurities were removed from samples,total anthocyanin yield reached 160.59 mg/g,the purity of anthocyanin was 43.64%,then,the five kinds of anthocyanin components was separated by semi-preparative high performance liquid chromatography with a purity of up to 90.61%,the optimized conditions were C18 preparative column,280 nm wavelength,column temperature was fixed at30 ℃,the mobile phase was 1% acetic acid(A)-acetonitrile(B),flow rate was 5 mL/min,injection volume was 1 mL with headspace sampling,condition of gradient elution: 0～55 minutes,B10% → 40%.These data indicated that high-speed counter-current chromatography coupled with semi-preparative high performance liquid chromatography can effectively separate and purify the anthocyanins and their components.更多还原