漳平水仙茶饼多酚的纯化及其体外活性研究

Separation and Purification of Polyphenols from Zhangping Shuixian Tea Cake by Macroporous Resin and Its Vitro Activity

为了研究大孔树脂对水仙茶饼多酚的纯化效果,试验以水仙茶饼多酚粗提液为原料,比较了9种不同型号大孔树脂对水仙茶饼粗多酚的静态吸附和解吸性能,再以吸附率和解吸率为指标通过静态、动态吸附解吸试验。考察了各因素对LX-28树脂纯化水仙饼茶粗多酚的影响,最后探讨了纯化前后的水仙茶饼多酚的体外抗氧化和抑菌效果。结果表明:最佳纯化树脂LX-28,最佳吸附和解吸条件为1.5 mg/mL的质量浓度的水仙茶饼多酚粗提取液(pH4.0~5.0)以2.0 mL/min的流速上样95 mL,吸附平衡后去离子水冲洗至无色,再用70 mL,65%体积分数乙醇溶液(pH6.0)为洗脱剂,以3.0 mL/min流速洗脱,在此纯化条件下LX-28树脂对水仙茶饼多酚的吸附率和解吸率分别为(93.587±0.379)%和(95.330±1.282)%,树脂可重复使用4次,纯度提高了约3.04倍;对DPPH·和ABTS+·自由基清除率的IC50值分别从纯化前的1.279、3.682 mg/mL降低到0.295、1.525 mg/mL;对供试的大肠杆菌、金黄色葡萄球菌、米根霉和紫红曲霉均有不同程度的抑制作用,纯化后的水仙茶饼多酚的抑菌效果优于纯化前。

The present study focuses on the separation and purification technology of the crude polyphenol extracted from Zhangping Shuixian tea cake by using a macroporous adsorption resin method. Nine different types of macroporous resins were adopted. Through the comparison of the separation effect, LX-28 was selected as the most efficient adsorbent as it had strong adsorption ability and high desorption rate. Based on the static and dynamic experiments of resin adsorption-desorption, the impact of LX-28 macroporous resin when purifying the polyphenols of Zhangping Shuixian tea cake was investigated, with adsorption and desorption rate as the index. Results were as follows： the optimal experimental conditions for static adsorption and desorption were determined as sample concentration 1.5 mg/mL （pH4.0-5.0）, sample volume 95 mL, flow rate 2.0 mL/min, elution ethanol concentration 65% （pH6.0）, elution volume 70 mL, flow rate 3.0 mL/min was built and applied to separate the polyphenols of Zhangping Shuixian tea cake. Additionally, under the above conditions, the desorption and desorption rate was （93.587_＋0.379）%, （95.330＋1.282）%, LX-28 macroporous resin could be reused for 4 times, the purity rate was 3.04 times more than the original solution. Antioxidant test showed that the scavenging rates IC50 values to DPPH· and ABTS＋· radical reduced from 1.279, 3.682 mg/mL to 0.295 and 1.525 mg/mL after purification. Antibacterial test showed that the effect of purified polyphenols on Escherichia coli, Staphylococcus aureus, Rhizopus oryzae and Monascus purpureus was better than unpurified polyphenols.