沙棘类胡萝卜素复合酶法提取及纯化的优化研究

Study on optimization of carotenoids complex-enzyme extraction and purification from Hippophae rhamnoides

以沙棘果实为原料,对复合酶法提取及纯化类胡萝卜素的最优条件进行研究。实验结果表明复合酶法提取沙棘类胡萝卜素的最佳条件为:纤维素酶:果胶酶2∶1(g∶g)、酶解温度30℃、酶解时间25 min、酶加入量0.20%、酶解pH7,此条件下提取率为89.88%。通过皂化法和硅胶柱层析法纯化类胡萝卜素粗提物,试验确定皂化最适宜条件为:碱加入量5%KOH乙醇液、皂化温度室温(20℃)、皂化时间1.5 h,皂化后类胡萝卜素含量从23.08%提高到69.53%,得率为82.34%。硅胶柱层析法进一步纯化皂化后的类胡萝卜素,条件为:洗脱剂为石油醚-丙酮梯度洗脱,洗脱速度10.0 mL/min,类胡萝卜素的含量从69.53%提高到84.36%,得率为61.93%。

The optimum conditions of complex enzyme extraction and purification of carotenoids from Hippophae rhamnoides were researched in this paper.The results showed the optimum conditions of complex exzyme extraction were： cellulose： pectinase 2∶1（g∶g）,30 ℃,25 min,enzyme addition 0.20%,pH7,under this conditions the extraction ratio was 89.88%.Then the thick extraction was purified by ways of saponification and silica gel column chromatography.The experiments showed that the optimized conditions of saponification could be obtained as following： 5% KOH alcohol liquor,room temperature（20 ℃）,1.5 h,under this conditions the content of carotenoids was enhanced from 23.08% to 69.53%,and the yield was 82.34%.Carotenoids could be further purified by Silica gel column chromatography,the optimum conditions were： gradient elution of sherwood oil and acetone,eluting velocity 10.0 mL/min.The content of cartenoids could be sublimated from 69.53% to 84.36%,and the yield was 61.93%.