富含α-亚麻酸的偏甘油酯的酶法制备及其理化性质研究

发布时间：2018-06-18 00:40

本文选题：甘油二酯 + 单甘酯　；参考：《华南理工大学》2012年硕士论文

【摘要】：本论文研究了富含α-亚麻酸的偏甘油酯的检测、制备、分离纯化及其理化性质，为富含多不饱和脂肪酸的甘油二酯及单甘酯的开发与应用提供依据。通过HPLC-RID方法检测甘油酯混合样品中的甘油三酯、甘油二酯及单甘酯的质量浓度并计算百分含量；通过单因素和响应面优化实验设计获得了采用酶制剂Lipozyme435催化亚麻籽油甘油解制备甘油二酯和单甘酯的最优工艺条件；对最优反应条件下获得的甘油二酯和单甘酯反应产物采用分子蒸馏进行了分离纯化，获得了富含α-亚麻酸的高纯度甘油二酯与单甘酯产品，并对产品进行了理化指标检测与储存稳定性研究。 1.研究了甘油酯混合样品中各酯类组分的HPLC-RID检测及百分含量计算方法，并对方法的准确度和精密度进行了考察。获得了甘油三酯，α-亚麻酸甘油二酯和α-亚麻酸单甘酯的标准曲线线性回归方程，依次分别为y1=1.3423x1、y2=1.5722x2和y3=1.4055x3、相关系数R1、R2与R3依次分别为0.999、0.998和0.999。精密度与回收率研究结果表明：在适宜的浓度范围内，采用HPLC-RID检测混合甘油酯样品中甘油三酯、甘油二酯和单甘酯的质量浓度，依照质量浓度计算各酯类成分的百分含量的方法精密度与准确度良好。在计算混合甘油酯产品中的酯类成分百分含量时，采用质量浓度计算百分含量与直接采用峰面积计算百分含量相比，甘油三酯与单甘酯百分含量分别高0.81%和1.73%，甘油二酯百分含量低2.54%，两种计算方法获得的数据较为接近。 2.研究了无溶剂体系下采用酶制剂Lipozyme435催化亚麻籽油甘油解制备甘油二酯的工艺。在采用单因素实验设计考察了底物摩尔比、反应时间、加酶量和反应温度4个因素对反应产物中甘油二酯含量影响的基础上，运用响应面实验设计优化得到了甘油二酯制备的最佳工艺条件，即底物摩尔比亚麻籽油：甘油（mol/mol）5：3，加酶量880PLU/g亚麻籽油，反应温度58.3℃，反应时间9.1h。在此条件下反应所得产物中的甘油二酯含量为50.21%，理论纯度可达60.12%，亚麻酸含量为46.22%。 3.研究了叔丁醇溶剂体系下采用酶制剂Lipozyme435催化亚麻籽油甘油解制备单甘酯的工艺。在采用单因素实验设计考察了底物摩尔比、反应时间、加酶量、油醇比和反应温度5个因素对反应产物中单甘酯含量影响的基础上，运用响应面实验设计优化了单甘酯制备的最佳工艺条件，即底物摩尔比亚麻籽油：甘油（mol/mol）为1：3，加酶量为601PLU/g亚麻籽油，反应温度为56.1℃，油醇比为亚麻籽油占叔丁醇（m/v）的39.7%，反应时间10.5h。在此条件下反应所得产物中单甘酯含量为71.1%，α-亚麻酸的含量达51.36%。 4.研究确定了甘油二酯与单甘酯反应产物采用分子蒸馏分离纯化的温度条件，其中一级分子蒸馏温度应在120℃，二级分子蒸馏温度应在220℃，在此条件下对产物进行纯化后获得了纯度分别为56.5%的甘油二酯产品和82.3%的单甘酯产品。所得甘油二酯产品在25℃下澄清、透明；水分及挥发物含量为0.13%；游离甘油含量为0.17%；酸价和皂化值分别为2.10mg KOH/g和182.60mg KOH/g；过氧化值为2.76mmol KOH/g；色泽为Red值4.0Yellow值8.3（25.4mm）。所得单甘酯产品在25℃下呈乳白色膏状；水分及挥发物含量为0.10%；游离甘油含量为2.08%；酸价和皂化值分别为3.20mg KOH/g和168.21mg KOH/g；过氧化值为0.50mmol KOH/g；色泽为Red值0.1Yellow值10.0（25.4mm），甘油二酯与单甘酯产品分别符合亚麻籽油标准GBT8235-2008和食品添加剂单、双硬脂酸甘油酯标准GB1986-2007。产品的氧化稳定性研究结果表明：甘油二酯产品储藏稳定性略优于亚麻籽油，低于大豆油，单甘酯产品的储存稳定性较差，，甘油二酯和单甘酯产品在添加抗氧化剂后储藏稳定性明显增强。
[Abstract]:In this paper, the determination, preparation, purification and physicochemical properties of the partial glycerol rich in alpha linolenic acid were studied, and the basis for the development and application of diesters and monoglycerides rich in polyunsaturated fatty acids was provided. The mass concentration of glycerol three ester, diester and monoglyceride in the mixed glyceride samples was detected by HPLC-RID method. The optimum technological conditions for the preparation of glycerol diester and monoglyceride with Lipozyme435 catalyzed glycerol in linseed oil were obtained by single factor and response surface optimization. Molecular distillation was used to obtain the product of diester and monoglyceride obtained under the optimal reaction conditions. High purity diacylglycerol and monoglyceride products, which are rich in alpha linolenic acid, have been studied, and physicochemical indexes and storage stability of the products have been studied.
1. the HPLC-RID detection and percentage calculation method of each component of glyceride mixed samples were studied, and the accuracy and accuracy of the method were investigated. The standard curve regression equation of triglyceride, alpha linolenic acid diester and alpha linolenic monoglyceride was obtained, which were respectively y1=1.3423x1, y2=1.5722x2 and y3=1.40, respectively. 55X3, the correlation coefficient R1, R2 and R3 are respectively the results of 0.999,0.998 and 0.999. precision and recovery respectively. In the suitable concentration range, HPLC-RID is used to detect the quality concentration of triglyceride, diester and monoglyceride in the samples of mixed glyceride, and the method of calculating the percentage of each component according to the quality concentration is precise. When calculating the percentage content of the ester components in the mixed glyceride, the content of the content calculated by the mass concentration is 0.81% and 1.73% higher in the content of the triglyceride and the monoglyceride, and the content of the diester of the triglyceride is 2.54%, and the data obtained by the two methods are compared. To be close.
2. the preparation of glycerol diester from linseed oil catalyzed by Lipozyme435 in solvent free system was studied. On the basis of 4 factors, such as substrate molar ratio, reaction time, enzyme amount and reaction temperature, the response surface experiment was designed on the basis of the response surface experiment. The optimum process conditions for the preparation of diacylglycerol were obtained, namely the substrate mole ratio flax seed oil: glycerol (mol/mol) 5:3, and the amount of 880PLU/g linseed oil added to the enzyme, the reaction temperature was 58.3 C, the reaction time was 9.1h., the content of the diester in the product was 50.21%, the physical purity was 60.12%, the linolenic acid content was 46.22%.
3. the process of preparing monoglyceride in linseed oil catalyzed by enzyme preparation Lipozyme435 under the solvent system of tert butyl alcohol was studied. On the basis of the effect of 5 factors on the content of monoglyceride in the product, the effect of the ratio of substrate molar ratio, reaction time, enzyme amount, oil alcohol ratio and reaction temperature on the content of monoglyceride in the reaction products, the response surface experiment was used. The optimum process conditions for the preparation of monoglyceride, namely, the substrate mole ratio flax seed oil: glycerol (mol/mol) was 1:3, and the amount of enzyme was 601PLU/g flax seed oil, the reaction temperature was 56.1, and the oil and alcohol ratio was 39.7% of the linseed oil (m/v), and the reaction time 10.5h. was 71.1%, alpha subtermate. The content of hemp acid reached 51.36%.
4. the temperature conditions for the separation and purification of diacylglycerol and monoglyceride reaction products by molecular distillation are determined, in which the first order molecular distillation temperature should be 120, and the two stage molecular distillation temperature should be at 220. Under this condition, the product is purified and the purity of the product is 56.5%, and the product of 82.3% monoglyceride, respectively, is obtained. The diacylglycerol products were clarified and transparent at 25 C; the content of water and volatiles was 0.13%, the content of free glycerol was 0.17%; the acid value and saponification value were 2.10mg KOH/g and 182.60mg KOH/g, the peroxide value was 2.76mmol KOH/g; the color was Red value 4.0Yellow value 8.3 (25.4mm). The monoglyceride products were milk white paste at 25 degrees C; water The content and volatiles content are 0.10%, free glycerol content is 2.08%, acid value and saponification value are 3.20mg KOH/g and 168.21mg KOH/g, peroxidation value is 0.50mmol KOH/g, Red value 0.1Yellow value 10 (25.4mm), and glycerol diester and monoglyceride products are in line with linseed oil standard GBT8235-2008 and food additive list, distearic acid, respectively. The results of the oxidation stability of the glyceryl ester standard GB1986-2007. product show that the storage stability of the diester product is slightly better than the linseed oil. The storage stability of the product is lower than that of the soybean oil. The stability of the diester and monoglyceride products is obviously enhanced after the addition of antioxidants.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R151

【参考文献】