甜菊糖的纯化研究

发布时间：2018-05-23 19:45

本文选题：甜菊糖苷 + 纯化　；参考：《南昌大学》2012年硕士论文

【摘要】：甜菊糖是一类糖苷物质,来源于甜叶菊叶片的提取物,它的纯度以及其主要成分甜菊苷和莱鲍迪甙A(RA)在甜菊糖中的比例直接影响着甜菊糖的品质。目前,国内对甜菊糖的深加工还有所欠缺,甜菊糖生产以初级加工为主,产品多作为原材料出口国外,并且生产过程中的能耗问题、环保问题和绿色安全问题比较突出。本研究以江西赣县的甜叶菊叶为原料,对甜菊糖的水提工艺进行了优化,采用较为环保的聚合氯化铝(PAC)对水提液进行絮凝除杂,运用乙醇重结晶甜菊糖来提高产品纯度和RA的含量,研究结果可为工业化生产提供理论支持和数据参考。主要研究内容如下： (1)研究了甜菊糖的测定方法。通过探讨甜菊糖的检测方法,确定了葸酮-硫酸法测定甜菊糖含量的方法为：1mL样液中加入4mL的3.3%葸酮-硫酸试剂,边加边混匀,立即置于冰水浴中,待所有样品加完后,一起置于100℃的水浴中7min,然后一起取出置于冰水浴中冷却10min,在620nm处测定吸光度；确定了分析甜菊糖苷混合物的液相色谱条件为：Inertsil NH2柱(4.6×250mm,5μm)流动相乙腈：水(70：30),流速1mL/min,进样量20gL,室温操作；并确定了甜菊糖苷的简单紫外检测法,可适合脱色好且胶质和蛋白质含量低的甜菊糖苷产品的总糖苷含量测定。 (2)优化了甜菊糖苷水提工艺。以甜叶菊糖苷的水提得率为评价标准,通过单因素和正交试验优化了甜菊糖苷的水提参数,得到水提的最优条件为：提取温度为70℃、提取时间为3h、料液比为1：15,在此条件下甜叶菊糖苷的水提得率可达到97%。 (3)确定了甜菊糖苷的絮凝条件。以甜菊糖苷损失率、水提液的脱色率和透光率为评价指标,对三种絮凝剂的絮凝效果进行比较,研究表明：PAC可用于甜叶菊水提液的澄清处理；对其澄清影响因素进行研究,确定最佳絮凝条件为：水浴温度65℃,絮凝剂添加量18mg/mL,辅助搅拌20min,作用时间1h,25℃离心。 (4)建立了甜菊糖苷的最佳重结晶响应面模型。在单因素的基础上,通过四因素三水平回归方程建立重结晶响应面模型。各响应值(Yi)对X1、X2、X3、X4的回归方程分别为：Y1(结晶率)=-282.71467+4.94347X1+0.92322X2+18.20933X3+11.45667X4-10-3X1X2-4×10-3X1X3-2×10-3X1X4-6.56882×10-16X2X3-1.66667×10-3X2X4+1.07692×10-14X3X4-0.02928X12-4.11111×10-3X22-2.172X32-0.543X42Y2(纯度)=-420.08083+9.709X1+1.61417X2-6.93167X3+2.4725X4+1.6667×10-3X1X2+0.155X1X3+0.081X1X4+0.10417X2X3+0.010417X2X4+0.3X3X4-0.068X12-0.010816X22-2.02X32-0.50938X42Y3(RA/Total)=-14.54122+0.27694X1+0.046239X2+0.12993X3+0.23999X4+2.30496×104X1X2+3.06965×10-3X1X3-1.37227×10-3X1X4+3.00937×10-4X2X3+5.43917×10-4X2X4+2.01963×10-3X3X4-1.82752×10-3X12-3.60353×10-4X22-0.048684X32-0.010284X42 研究表明,各因素对结晶率响应值影响的强弱依次为：X1(溶解温度)X4(乙醇含水量)X3(液固比)X2(重结晶时间)；对重结晶产品纯度响应值影响的强弱依次为：X3(液固比)X2(重结晶时间)X1(溶解温度)X4(乙醇含水量)；对RA/Total含量响应值影响的强弱依次为：X3(液固比)X4(乙醇含水量)X1(溶解温度)X2(重结晶时间)。
[Abstract]:Stevioside is a kind of glycoside, which is derived from the extract of leaves of Stevia. Its purity and the proportion of stevioside and A (RA) in stevioside directly affect the quality of stevioside. At present, the deep processing of stevioside is still lacking in our country. The production of stevioside is mainly in primary processing, and the products are used as raw materials. The problem of energy consumption, environmental protection and green safety are more prominent in the process of production. In this study, the water extraction process of steviodies was optimized by using chrysanthemum leaves in Ganxian, Jiangxi. The water extraction was flocculated by environmentally friendly polyaluminum chloride (PAC) and alcohol was recrystallized with steviodies. High product purity and RA content can provide theoretical support and data reference for industrial production.
(1) the method for determination of steviodies was studied. By exploring the method of detecting stevioside sugar, the method of determining the content of steviodie by acetone sulfuric acid method was as follows: the 4mL in 1mL sample was added with 3.3% ketone sulphuric acid reagents, and mixed evenly, and immediately placed in the ice water bath. After all the samples were added, it was placed together in a water bath of 100 degrees centigrade, and then taken together, and then taken together. In the ice water bath, the 10min was cooled and the absorbance was measured at 620nm. The liquid chromatography conditions for the analysis of the mixture of stevioside were determined as: Inertsil NH2 column (4.6 x 250mm, 5 uh m) mobile phase acetonitrile: water (70:30), velocity 1mL/min, sample volume 20gL, room temperature operation, and a simple UV detection method for stevioside, which was suitable for decolorization. Determination of total glucoside content of stevioside products with low colloid and protein content.
(2) optimize the water extraction process of stevioside. With the water extraction rate of stevioside as the evaluation standard, the water extraction parameters of stevioside were optimized by single factor and orthogonal test. The optimum conditions of the water extraction were as follows: the extraction temperature was 70, the extraction time was 3h, the ratio of the material to liquid was 1:15, and the water extraction rate of stevioside could reach 9 under this condition. 7%.
(3) the flocculation conditions of stevioside were determined. With the loss rate of stevioside, the decolorization rate and the transmittance of the water as the evaluation index, the flocculation effect of the three flocculants was compared. The study showed that PAC could be used in the clarification treatment of the extract of the sweet leaf chrysanthemum, and the influence of the factors on the clarification was studied and the optimum flocculation condition was to determine the temperature of the water bath. 65 18mg/mL, 20min, 1H and 25 C centrifugation.
(4) the optimum recrystallization response surface model of stevioside was established. On the basis of single factor, the recrystallized response surface model was established by four factors and three level regression equations. The regression equations of each response value (Yi) to X1, X2, X3 and X4 were Y1 (crystallization rate) =-282.71467+ 4.94347X1+0.92322X2+18.20933X3+11.45667X4-10-3X1X2-4 x 10-3X1X3-2 x 1, respectively. 0-3X1X4-6.56882 * 10-16X2X3-1.66667 * 10-3X2X4+1.07692 x 10-14X3X4-0.02928X12-4.11111 x 10-3X22-2.172X32-0.543X42Y2 (purity) =-420.08083+9.709X1+1.61417X2-6.93167X3+2.4725X4+1.6667 * 10-3X1X2+0.155X1X3+0.081X1X4+0.10417X2X3+0.010417X2X4+ 0.3X3X4-0.068X12-0.010816X22-2.02X32-0.50938X42Y3 (RA/Total) =-14.54122 +0.27694X1+0.046239X2+0.12993X3+0.23999X4+2.30496 * 104X1X2+3.06965 * 10-3X1X3-1.37227 x 10-3X1X4+3.00937 * 10-4X2X3+5.43917 * 10-4X2X4+2.01963 x 10-3X3X4-1.82752 x 10-3X12-3.60353 * 10-4X22-0.048684X32-0.010284X42
The results show that the strong and weak factors affecting the response value of crystallization rate are X1 (dissolved temperature) X4 (ethanol water content) X3 (liquid solid ratio) X2 (recrystallization time), and the intensity of the response to the purity of the recrystallized products is X3 (liquid-solid ratio) X2 (recrystallization time) X1 (dissolution temperature) X4 (ethanol water content), and the response to RA/Total content in response value. The intensity is X3 (liquid to solid ratio) X4 (water content of ethanol) X1 (dissolution temperature) X2 (recrystallization time).
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R151

【参考文献】