CN104745287A - Extracting method for ginseng seed oil - Google Patents
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Abstract
本发明涉及一种从人参子中提取人参子油的制备方法，该方法包括以下步骤：（1）将人参子去壳后进行机械细粉碎；（2）将水加入人参子粉中，调节混合液的温度和pH，然后向混合液中加入酶进行酶解得到酶解液；（3）将酶解液灭酶后离心分离，得到游离油、乳状液、水解液和残渣。乳状液将破乳后得到游离油；（4）残渣反复水洗离心后，得到液面上的游离油和对乳状液破乳后得到的游离油；（5）将（3）和（4）所得到的游离油合并得到人参子油。本发明方法工艺路线简单，生产过程中能耗相对低,提取条件温和，操作简单，油的品质较好，油的提取率高。
The invention relates to a preparation method for extracting ginseng seed oil from ginseng seeds. The method comprises the following steps: (1) mechanically finely crushing ginseng seeds after shelling; (2) adding water to ginseng seed powder, adjusting the mixing The temperature and pH of the solution, and then add enzymes to the mixed solution for enzymolysis to obtain an enzymolysis solution; (3) Centrifuge the enzymolysis solution after deactivation to obtain free oil, emulsion, hydrolyzate and residue. The emulsion will be demulsified to obtain free oil; (4) After the residue is repeatedly washed and centrifuged, the free oil on the liquid surface and the free oil obtained after demulsification of the emulsion are obtained; (5) The oil obtained in (3) and (4) The obtained free oils were combined to obtain ginseng seed oil. The method of the invention has simple process route, relatively low energy consumption in the production process, mild extraction conditions, simple operation, good oil quality and high oil extraction rate.
Description
技术领域 technical field
本发明属于植物油脂的提取加工技术领域，具体涉及一种采用水酶法提取人参子油的方法。 The invention belongs to the technical field of extraction and processing of vegetable oils, and in particular relates to a method for extracting ginseng seed oil by using an aqueous enzymatic method.
背景技术 Background technique
人参(Panax ginseng Meyer)在亚洲作为中药和保健食品的历史已有1000多年。国内外对人参皂甙和人参多糖等成分的药效和保健作用研究较多。 Ginseng ( Panax ginseng Meyer ) has been used as a traditional Chinese medicine and health food in Asia for more than 1,000 years. There are many researches at home and abroad on the efficacy and health care of ingredients such as ginsenosides and ginseng polysaccharides.
人参子是人参果实内半圆形籽粒，7、8月间种籽成熟，种子与果实同时采集。据《本草纲目拾遗》记载：“人参子，如腰子式，生青熟红，如黄豆大小”。人参种子富含蛋白、油脂、碳水化合物等多种营养物质，但是人参子的功效成分尤其是参子油的相关研究较少。 Ginseng seeds are semicircular seeds in the ginseng fruit. The seeds mature in July and August, and the seeds and fruits are collected at the same time. According to "Compendium of Materia Medica Supplements" records: "Ginseng seeds, like kidneys, raw green and ripe red, the size of soybeans." Ginseng seeds are rich in protein, oil, carbohydrates and other nutrients, but there are few studies on the functional components of ginseng seeds, especially ginseng seed oil.
目前，植物油脂提取的方法主要采用超临界CO2萃取、传统的压榨法、有机溶剂浸提法、水代法等，这些传统的提取方法存在机械设备复杂、运行成本高、有机溶剂残留、提取率低等不足。水酶法从植物油料中提油是利用机械和酶解手段来破坏植物的细胞壁，使油脂游离出来,工艺简单，处理条件温和，操作安全，污染小，提取出的油品质较好，因此水酶法提油得到了广泛地应用。采用超临界CO2流体萃取人参子油工艺复杂和设备昂贵，人参子油提取率低较；采用冷榨制取人参子油的方法，需要两次压榨，压榨饼中残油较多。采用水酶法提取人参子油可以有效提高人参子油的提取率，分离油料中的油和蛋白质，固体残渣可用于生产人参子蛋白，原料综合利用率高，浪费少。 At present, the methods of vegetable oil extraction mainly adopt supercritical CO2 extraction, traditional pressing method, organic solvent extraction method, water substitution method, etc. These traditional extraction methods have complex mechanical equipment, high operating costs, organic solvent residues, and extraction rate. Low and insufficient. The aqueous enzymatic method of extracting oil from vegetable oils uses mechanical and enzymatic methods to destroy the cell walls of plants, so that the oil is released. The process is simple, the treatment conditions are mild, the operation is safe, the pollution is small, and the extracted oil is of good quality. Enzymatic oil extraction has been widely used. Extraction of ginseng seed oil by supercritical CO 2 fluid is complex and expensive equipment, and the extraction rate of ginseng seed oil is low; the method of producing ginseng seed oil by cold pressing requires two pressings, and there is more residual oil in the pressed cake. Extracting ginseng seed oil by water enzymatic method can effectively improve the extraction rate of ginseng seed oil, separate oil and protein in oil, and solid residue can be used to produce ginseng seed protein, with high comprehensive utilization rate of raw materials and less waste.
发明内容 Contents of the invention
本发明的目的是为了解决人参子油提取方面存在的机械设备复杂、运行成本高、有机溶剂残留、提取率低等问题，提供一种采用水酶法制取人参子油工艺方法。 The purpose of the present invention is to provide a process for preparing ginseng seed oil by using an aqueous enzymatic method to solve the problems of complex mechanical equipment, high operating cost, residual organic solvent, and low extraction rate in the extraction of ginseng seed oil.
本发明提供的工艺路线见工艺流程图1。 The process route provided by the present invention is shown in process flow diagram 1.
本发明提供的技术方案包括以下步骤。 The technical solution provided by the invention includes the following steps.
1. 将人参子去壳后进行机械细粉碎，使人参子粉达到40-200目。 1. After the ginseng seeds are dehulled, they are mechanically crushed to make the ginseng seed powder reach 40-200 mesh.
2. 将水加入人参子粉中，人参子粉与水的比例为1:5-15。然后向混合液中加入酶包括纤维素酶、果胶酶、复合纤维素酶、蛋白酶、α-淀粉酶等或者它们的混合物进行酶解，酶解过程中不断搅拌；控制酶解温度为30-80℃，酶解pH为3.0-11.0，酶解时间为2-8h。 2. Add water to ginseng seed powder, the ratio of ginseng seed powder to water is 1:5-15. Then add enzymes to the mixed solution including cellulase, pectinase, complex cellulase, protease, α-amylase, etc. or their mixtures for enzymolysis, stirring constantly during the enzymolysis process; controlling the enzymolysis temperature to 30- 80°C, enzymolysis pH 3.0-11.0, enzymolysis time 2-8h.
3. 将酶解液灭酶后离心分离，离心转速为4000-12000r/min，离心30-90min得到游离油、乳状液、水解液和残渣。乳状液将破乳后得到游离油。 3. Centrifuge the enzymatic hydrolyzate after inactivating the enzyme. The centrifugal speed is 4000-12000r/min, and centrifuge for 30-90min to obtain free oil, emulsion, hydrolyzate and residue. The emulsion will break to give free oil.
4. 残渣反复水洗离心后，得到液面上的游离油和对乳状液破乳后得到的游离油。 4. After the residue is repeatedly washed and centrifuged, the free oil on the liquid surface and the free oil obtained after breaking the emulsion are obtained.
5. 将3和4所得到的游离油合并得到人参子油。 5. The free oils obtained in 3 and 4 are combined to obtain ginseng seed oil.
6. 人参子油的GC-MS分析。 6. GC-MS analysis of ginseng seed oil.
色谱条件：色谱柱：WAX（60m-250mm-0.25μm）；进样口温度：250℃；升温程序：从100℃开始，保持1min，再以5℃/min升至250℃，保持20min；载气为高纯度氦气；进样方式为分流进样，分流比为50：1。质谱条件：离子源温度230℃；四级杆温度150℃；倍增电压1047V；质量扫描范围m/z20-500。 Chromatographic conditions: chromatographic column: WAX (60m-250mm-0.25μm); inlet temperature: 250°C; heating program: start at 100°C, keep for 1min, then rise to 250°C at 5°C/min, and keep for 20min; The gas is high-purity helium; the sampling method is split injection, and the split ratio is 50:1. Mass spectrometry conditions: ion source temperature 230°C; quadrupole temperature 150°C; multiplier voltage 1047V; mass scan range m/z20-500.
本发明的有益效果： Beneficial effects of the present invention:
首次采用水酶法提取人参子油，该技术处理条件温和，人参子油的提取率高且质量较好；工艺路线简单，生产过程能耗相对低，能够灵活控制生产量；可以同时提取人参子油和人参子中的蛋白质；本发明所得的人参子油提取率为72.10-80.41%。 For the first time, the water enzymatic method was used to extract ginseng seed oil. This technology has mild treatment conditions, high extraction rate and good quality of ginseng seed oil; the process route is simple, the energy consumption in the production process is relatively low, and the production volume can be flexibly controlled; ginseng seed oil can be extracted at the same time protein in oil and ginseng seed; the extraction rate of ginseng seed oil obtained in the present invention is 72.10-80.41%.
附图说明 Description of drawings
图1：工艺流程图。 Figure 1: Process flow diagram.
图2：各因素及交汇作用对提油率的影响；（a）加酶量与料液比，（b）pH与料液比，（c）pH与加酶量。 Figure 2: Effects of various factors and confluence on oil extraction rate; (a) enzyme amount and solid-liquid ratio, (b) pH and solid-liquid ratio, (c) pH and enzyme amount.
图3：水酶法提取人参子油的总离子流图。 Figure 3: Total ion chromatogram of ginseng seed oil extracted by aqueous enzymatic method.
具体实施方式 Detailed ways
以下实施例进一步说明发明的内容，但不应理解为对本发明的限制。 The following examples further illustrate the content of the invention, but should not be construed as limiting the invention.
实施例1。 Example 1.
材料：人参子，由通化市蚂蚁河东北特产销售有限公司提供；纤维素酶、果胶酶、复合纤维素酶、Alcalase蛋白酶、α-淀粉酶由诺维信公司提供；氢氧化钠、石油醚等均为分析纯。 Materials: Ginseng seeds, provided by Tonghua Ant Hebei Specialty Sales Co., Ltd.; cellulase, pectinase, compound cellulase, Alcalase protease, α-amylase provided by Novozymes; sodium hydroxide, petroleum ether etc. are analytically pure.
主要仪器设备：FZ102 型微型植物粉碎机 天津市泰斯特仪器有限公司；GB1302 电子精密天平 梅特勒-托利多仪器(上海)有限公司；PHS-3D pH计 上海精科仪器有限公司；HH-2 恒温水浴锅 常州澳华仪器有限公司；KDC-1042 离心机 安徽中科中佳科学仪器有限公司；101A-2E 电热鼓风干燥箱 上海实验仪器厂有限公司； GCMS-5975-6890N气相色谱质谱仪 美国安捷伦公司。 Main instruments and equipment: FZ102 miniature plant pulverizer Tianjin Test Instrument Co., Ltd.; GB1302 electronic precision balance Mettler-Toledo Instruments (Shanghai) Co., Ltd.; PHS-3D pH meter Shanghai Jingke Instrument Co., Ltd.; HH- 2 Constant temperature water bath Changzhou Aohua Instrument Co., Ltd.; KDC-1042 centrifuge Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.; 101A-2E electric blast drying oven Shanghai Experimental Instrument Factory Co., Ltd.; GCMS-5975-6890N Gas chromatography mass spectrometer Agilent Corporation of America.
实验方法：取等量的人参子粉，每组料液比为1:10，调节每组的温度和pH至适当条件，将相应量的纤维素酶、果胶酶、复合纤维素酶、Alcalase、α-淀粉酶分别加入每组中，放入恒温水浴锅中酶解，酶解过程中需经常搅拌。酶解4h后，于100℃的沸水中灭酶5min，4000r/min离心20min，保留上层的游离油， 乳状液破乳得游离油，将残渣用蒸馏水清洗，再4000r/min离心20min，保留上层的游离油， 乳状液破乳得游离油，反复多次，将游离油合并得到人参子油。 Experimental method: Take the same amount of ginseng seed powder, the ratio of solid to liquid in each group is 1:10, adjust the temperature and pH of each group to appropriate conditions, and add corresponding amounts of cellulase, pectinase, compound cellulase, Alcalase , α-amylase were added to each group, and put into a constant temperature water bath for enzymolysis. During the enzymolysis process, frequent stirring was required. After 4 hours of enzymatic hydrolysis, inactivate the enzyme in boiling water at 100°C for 5 minutes, centrifuge at 4000r/min for 20min, retain the free oil in the upper layer, break the emulsion to obtain free oil, wash the residue with distilled water, centrifuge at 4000r/min for 20min, and retain the upper layer The free oil of the emulsion is demulsified to obtain the free oil, which is repeated several times, and the free oil is combined to obtain the ginseng seed oil.
结果：使用Alcalase的提油率最高，为75.34%。其次是α-淀粉酶，提油率为74.52%。提油率最低的是果胶酶，仅为67.73%。其中α-淀粉酶与Alcalase的提油效果相近，但α-淀粉酶提油时乳化严重。 Results: The oil extraction rate of Alcalase was the highest, which was 75.34%. Followed by α-amylase, the oil extraction rate is 74.52%. The lowest oil extraction rate is pectinase, only 67.73%. Among them, the oil extraction effect of α-amylase and Alcalase is similar, but the emulsification is serious when α-amylase extracts oil.
实施例2。 Example 2.
响应面法优化人参子油的提取工艺。 Optimization of extraction process of ginseng seed oil by response surface methodology.
1模型建立及显著性检验。 1 Model establishment and significance test.
在单因素试验的基础上，确定各因素的最佳范围，以料液比（X1）、加酶量（X2）、pH（X3）为个3因素为自变量，以人参子油的提取率（Y）为响应值， 应用 Design-Expert 6.0 软件设计响应面实验进行优化，采用响应面中心组合实验设计，进行三因素三水平的响应面分析试验。因素水平编码表见表1。 On the basis of the single factor test, the optimal range of each factor was determined, with three factors as the independent variable, the ratio of material to liquid (X 1 ), the amount of enzyme added (X 2 ), and the pH (X 3 ), and the ginseng seed oil The extraction rate (Y) is the response value. Design-Expert 6.0 software is used to design the response surface experiment for optimization, and the response surface center combination experiment design is adopted to conduct a three-factor and three-level response surface analysis experiment. See Table 1 for the coding table of factor levels.
表1 因素水平编码表 Table 1 Factor level coding table
表2 响应面实验方案及结果 Table 2 Response surface experiment scheme and results
根据表2，利用Design Expert 8.0.6软件对实验数据进行回归分析，由此得到水酶法提取人参子油得率对料液比、加酶量、PH的二次多项回归方程为：Y =79.56-0.68X1+1.29X2+0.69X32-0.082X1X3-0.32X2X3-0.60X1 2-0.598X2 2-4.56X3 2 According to Table 2, using Design Expert 8.0.6 software to carry out regression analysis on the experimental data, the quadratic polynomial regression equation of the yield of ginseng seed oil extracted by aqueous enzymatic method to the ratio of solid to liquid, the amount of enzyme added, and PH is obtained: Y =79.56-0.68X 1 +1.29X 2 +0.69X 32 -0.082X 1 X 3 -0.32X 2 X 3 -0.60X 1 2 -0.598X 2 2 -4.56X 3 2
表3 响应面方程方差分析。 Table 3 Response surface equation analysis of variance.
由表3可知，该模型回归（p＜0.0001）极显著，且失拟项（p＞0.05）不显著，且R2=99.98%，R2 Adj=99.96%，说明该模型与实验拟合的良好。该方程可以较为准确地分析和预测酶解反应条件与提油率之间的关系。其中A、B、C、BC、A2、B2、C2的影响是显著的，而AB、AC不显著。在此基础上优化最佳酶解条件编码值为：料液比-0.69，酶的添加量0.86，酶解pH0.04，预测提油率为80.43%，考虑到实际生产采用编码值为料液比-1，酶的添加量+ 1酶解pH 0，与第三组相同，结果为80.41，两者相差不大。各因素及交汇作用对提油率的影响见图2。 It can be seen from Table 3 that the model regression (p<0.0001) is extremely significant, and the lack of fit item (p>0.05) is not significant, and R 2 =99.98%, R 2 Adj =99.96%, indicating that the model fits well with the experiment good. This equation can accurately analyze and predict the relationship between enzymatic hydrolysis reaction conditions and oil extraction rate. Among them, the influence of A, B, C, BC, A 2 , B 2 , and C 2 is significant, while that of AB, AC is not significant. On this basis, optimize the coding value of the best enzymatic hydrolysis conditions: solid-liquid ratio -0.69, enzyme addition amount 0.86, enzymatic hydrolysis pH 0.04, predicted oil extraction rate is 80.43%, considering the actual production code value is material liquid Ratio -1, the amount of enzyme added + 1 enzymolysis pH 0, the same as the third group, the result is 80.41, there is little difference between the two. The influence of various factors and their confluence on the oil recovery rate is shown in Fig. 2.
实施例3。 Example 3.
采用GC-MS法对水酶法提取的人参子油进行分析，得其总离子流图见图3。经质谱数据库检索和人工谱图解析，鉴定出的人参子油的化学成分见表4 The ginseng seed oil extracted by aqueous enzymatic method was analyzed by GC-MS method, and the total ion chromatogram is shown in Figure 3. The chemical composition of the identified ginseng seed oil is shown in Table 4 through mass spectrometry database retrieval and manual spectrum analysis.
表4水酶法提取人参子油的GC-MS分析 Table 4 GC-MS analysis of ginseng seed oil extracted by aqueous enzymatic method
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