CN113201035A - Method for extracting and separating stevioside from stevia rebaudiana Bertoni by using low co-soluble solvent (DES) - Google Patents
Method for extracting and separating stevioside from stevia rebaudiana Bertoni by using low co-soluble solvent (DES) Download PDFInfo
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- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 title claims abstract description 49
- 229940013618 stevioside Drugs 0.000 title claims abstract description 48
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 title claims abstract description 48
- 235000019202 steviosides Nutrition 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 43
- 244000228451 Stevia rebaudiana Species 0.000 title claims abstract description 22
- 235000006092 Stevia rebaudiana Nutrition 0.000 title claims abstract description 21
- 239000002904 solvent Substances 0.000 title abstract description 8
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 claims abstract description 43
- 238000000605 extraction Methods 0.000 claims abstract description 39
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
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- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims description 6
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- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical group [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims description 6
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims 1
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 claims 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims 1
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- 239000001512 FEMA 4601 Substances 0.000 abstract description 33
- HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 abstract description 33
- HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 0.000 abstract description 33
- 235000019203 rebaudioside A Nutrition 0.000 abstract description 33
- 239000003960 organic solvent Substances 0.000 abstract description 10
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 239000011347 resin Substances 0.000 description 6
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- 235000013305 food Nutrition 0.000 description 4
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- QSRAJVGDWKFOGU-WBXIDTKBSA-N rebaudioside c Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]1(CC[C@H]2[C@@]3(C)[C@@H]([C@](CCC3)(C)C(=O)O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)CC3)C(=C)C[C@]23C1 QSRAJVGDWKFOGU-WBXIDTKBSA-N 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
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- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
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- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
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- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- -1 glycoside compounds Chemical class 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
- A23L27/11—Natural spices, flavouring agents or condiments; Extracts thereof obtained by solvent extraction
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
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Abstract
The invention relates to the technical field of separation of natural active ingredients, and 5 low cosolvent solvents are synthesized and used for extracting stevioside. The low co-dissolving solvent has better biocompatibility, and the purity of Rebaudioside A (RA) is higher than that of the traditional extraction process when the stevioside in the stevia rebaudiana is extracted. The low cosolvent aqueous solution can be used for extracting stevia rebaudiana, and stevioside with high RA purity (extraction capacity of 286mg/g adsorbent) can be extracted. The method simplifies the traditional stevioside extraction process, greatly reduces the use amount of organic solvents, and has excellent ecological environmental protection and safety. The method provides a new high-efficiency way for extracting and preparing high-purity stevioside from stevia rebaudiana.
Description
Technical Field
The invention relates to the field of separation of natural active ingredients, in particular to preparation of a low cosolvent and application of the low cosolvent in extraction of stevioside.
Background
Stevia sugar, which is a natural high-potency sweetener extracted from the natural plant stevia rebaudiana, has been approved as a food additive in many countries, and has a sweetness 300 times that of sucrose but a calorie of 1/300. At present, the third generation sugar is used as a sweetness agent instead of cane sugar, and is first used by international famous companies. Natural Stevioside is a mixture of several glycoside compounds, all of which contain the same hydrophobic Stevioside units, except that the glycosyl substituents at position C13 and position C18 differ, and the main components are Rebaudioside A (RA), Stevioside (St), and Rebaudioside C (RC). The total content of the three components in the natural stevia rebaudiana is more than 90%. Wherein RA has the highest sweetness, the sweetness of which is similar to that of cane sugar, and the sweetener is a novel natural sweetener with high sweetness, low calorie, easy dissolution, heat resistance and stability, and the content or purity of the sweetener is also a main index for measuring the quality of stevioside. It has health promoting and therapeutic effects on obesity, hypertension, and hyperglycemia. Besides, the stevioside product also contains various amino acids and various trace elements, so the stevioside not only can be used as a food sweetener, but also can be used as a nutrient source food or an auxiliary treatment medicine.
The stevioside has a complex structure, a plurality of active sites, easy inactivation and poor stability, and is difficult to realize chemical total synthesis in industrial production, so the extraction from the stevia rebaudiana is a main way for obtaining the stevioside. At present, the industrial method for extracting the stevioside is to extract mixed liquid of the stevioside and other substances from stevia rebaudiana raw materials and then remove the other substances by adopting different separation methods and equipment with various characteristics, so that a high-purity stevioside product is obtained. The other substances in the extraction mixture are collectively called as impurities, at least comprise impurities such as protein, organic acid, islet glycoside, chlorophyll, inorganic salt and the like, the content of the impurities is 5-7 times of the content of glucoside, and the impurities are required to be purified, so that the components are complex. In order to remove these impurities, the industrial production usually adopts the technological process of multiple times using organic solvent precipitation, liquid-liquid extraction and column chromatography, as shown in figure 1. The process shown in the figure is only the extraction process flow part in the whole production process of the high-purity stevioside product (high-purity RA), and the extraction basically aims to obtain a stevioside crude product with certain purity (RA is more than 60 percent) as a raw material for further separation (RA is more than 80 percent) and purification (RA 95-99 percent). The extraction process has long flow, large use amount of organic solvents (diethyl ether and ethanol), and high difficulty in controlling the volatilization of the organic solvents and completely recovering organic components in the waste liquid, so that the requirements on ecological environmental protection and production safety are difficult to meet, and the cost is difficult to reduce.
In the development process of further improving the traditional extraction process, some new technologies are applied, for example, the technologies such as an ion precipitation method, a resin adsorption method, membrane separation, molecular imprinting and the like are combined, so that the use amount of an organic solvent can be reduced, and the traditional process flow can be shortened to a certain extent.
Nevertheless, there are still a number of problems. For example, the purification method mainly utilizes flocculation to remove impurities for centrifugal separation, and usually adopts inorganic low molecules such as iron salt or aluminum salt as a flocculating agent and lime as a coagulant aid for acid-base neutralization reaction to form flocculation precipitation. But the flocculation reaction is slow, and the stevioside is easy to decompose and damage under the condition; meanwhile, the consumption of the flocculating agent is large, and the flocculation reaction and filter-pressing clarification period is longer; the flocculated solution also needs further impurity removal. Resin adsorption processes use the adsorption characteristics of the resin and a selective eluent to effect separation from other leaching components. For example, studies on adsorption of rebaudioside a and stevioside by macroporous resin D392 [ J392 ] are disclosed]2012.23.021) has good adsorption selectivity to RA and St in water solution by using macroporous resin D392, and the purity of RA after one-time adsorption and resolution reaches 70.4%. However, the resin adsorption method has the defects of large using amount of column packing, high price and easy inactivation; the equipment investment and the operation cost of large-scale continuous production are high, and the process uses a plurality of volatile organic solvents of ethanol in the elution process, so that the ecological environment-friendly effect and the production safety are not good enough. In response to these problems, separation of products by utilizing the selectivity of membranes has been developed. For example, Yao et al (Yao Guohuaxin, Maobu, Wang Xu. Membrane technology for extracting stevioside [ J)]The results of the separation and purification of the extract of the stevia sugar water by using a microfiltration membrane and an ultrafiltration membrane and the final desalination and concentration by using a nanofiltration membrane show that the purity of RA reaches about 87 percent, but the attenuation of the membrane flux at the nanofiltration membrane stage gradually attenuates along with the extension of the filtration time, and the membrane flux is only 1L/(h.m) after 3h2). Due to the operationThe layer-by-layer operation in the process causes great increase of production cost and time, and is not beneficial to large-scale industrial use.
Molecularly Imprinted Polymers (MIPs) have high "discriminatory" or specific selectivity for template molecules, and are functional polymer materials most likely to be used in "one-step" purification of RA. For example, Ye et al (Yanrejin, Yinyanye, Huaxiao, Tangle, Zhao Wei, Zhang wenbin) and rebaudioside A molecularly imprinted polymer preparation method [ P ]. Chinese patent application No. 201310120326.3) apply molecular imprinting technology to selectively extract RA in stevia rebaudiana aqueous solution, and finally the equilibrium adsorption capacity of the molecularly imprinted polymer to RA under equilibrium combination conditions can reach 138.1 μmoL/g (134mg rebaudioside A/g molecularly imprinted polymer), so that the performance of selectively adsorbing RA is very good, but for industrial preparation, the adsorption capacity of the adsorbent does not meet the requirement; in addition, RA and the molecular imprinting adsorbent are separated, and an organic solvent is required to be added for desorption to obtain the product.
If the stevioside can be selectively extracted without adsorbing other impurities and other organic solvents are not introduced in the separation process, the extraction and separation processes of the stevioside are greatly simplified, and the requirements of green, environment-friendly and clean processes are met. However, the main challenge is to have both good selectivity and sufficiently high adsorption or extraction capacity. Although a great deal of exploration is carried out at home and abroad, no actual breakthrough progress is seen.
The poorly co-soluble solvents (DES) are typically liquid materials formed from hydrogen bond acceptors (e.g., quaternary ammonium salts) and hydrogen bond donors (e.g., polyols, ureas, and carboxylic acids). The DES has low synthesis cost, and compared with the traditional solvent, the DES also has other special properties, such as wide liquid range, good thermal stability, strong designability, low vapor pressure, difficult volatilization, good recycling performance and the like, and is also called as novel ionic liquid.
In recent years, DES has attracted much attention in the fields of new materials, separation and purification, electrochemistry, catalysts and the like, and has been used in the processes of extraction, adsorption, catalysts and the like, thus showing good application prospects. In the field of food and drug applications, the safety of DES is of great importance. It is important to develop DESs with good biocompatibility, non-toxicity, and better safety. For example, Ahmad Ross Mansur et al (Ahmad Ross Mansura, Nho-Eul Song, Tae Gyu Nam et al, Optimizing the ultra-assisted specific solvent extraction of flavonoids in common buckwheat minerals food Chemistry,2019,438-445) developed a method for selective extraction and separation of flavonoids from common buckwheat sprouts using appropriately designed low co-solvents. The method proves that the used DES has good safety and application feasibility in the fields of biochemistry, pharmacy and food.
The technology develops a method for extracting stevioside by using a low cosolvent aqueous solution after exploring and researching the DES composition and performance in a large quantity, similar reports are not found at home and abroad, and the related DES stevioside extraction process route (extraction and separation) is shown as an attached figure 2.
Disclosure of Invention
Cleaning stevia rebaudiana Bertoni material, drying, pulverizing, and sieving to obtain stevia rebaudiana Bertoni powder. In the preparation process of the low co-solvent (DES), firstly, a proper amount of a mixture of a hydrogen bond acceptor and a hydrogen bond donor is added into a single-neck flask for standby, and the mixture is stirred for 4 hours at the temperature of 80 ℃. After the reaction is finished, colorless transparent liquid, namely the low cosolvent, is obtained. The DES can be stored in an environment lower than room temperature for standby, during the process of extracting stevioside by using a low cosolvent, a certain amount of crushed stevia rebaudiana is added into a DESS aqueous solution with a certain water content, extraction is carried out for a certain time at a certain temperature, solid-liquid separation is carried out after extraction is finished, and the liquid phase contains the stevioside with higher purity RA. The liquid phase can be further subjected to alcohol precipitation by using a certain amount of low-carbon alcohol, so that the RA purity in the stevioside product can be further improved.
The characteristics of the project are briefly described as follows
(1) A series of DES are designed and synthesized. The DES has good biocompatibility, thereby having good ecological environmental protection and safety.
(2) Extracting with DES water solution to obtain crude stevioside with RA concentration of 68.6%The traditional process (RA purity is mainly about 46 percent) improves by nearly 50 percent, is obviously superior to the product quality in the current extraction process, and the product yield (the ratio of stevioside to stevia rebaudiana dry leaves) is about 13.8 percent, which is about 12.5 percent of the extraction yield of the traditional process and about 12.9 percent of the extraction yield of the traditional process by adopting a microfiltration membrane-ultrafiltration membrane-nanofiltration membrane combined process or is slightly superior. Furthermore, it is comparable to the column chromatography results (70% RA). Proved to be capable of completely replacing the conventional process (shown in figure 1) of diethyl ether defatting, 70 percent ethanol extraction, liquid-liquid separation, Ca (OH)2+KAl(SO4)2Flocculation precipitation, centrifugal filtration and the like. Therefore, the technology can completely shorten the traditional extraction process of the stevioside greatly and simplify the operation process obviously.
(3) On the basis of the DES extraction process, crude stevioside with RA concentration as high as 84% can be obtained by adopting a single alcohol precipitation process of the technology, the column chromatography effect (70% RA) in the traditional process is improved by about 20%, and the technical effect (87% RA) of a microfiltration membrane, an ultrafiltration membrane and a nanofiltration membrane in a literature report is similar. The method is characterized in that on the basis of DES extraction, simple alcohol precipitation separation can be adopted to completely replace technologies such as column chromatography, membrane separation combination and the like which have high operation technical requirements and high equipment and operation costs in the existing technological process.
(4) The extraction process uses water as a solvent, the DES is difficult to volatilize and biocompatible feasible, no volatile organic solvent is used, and all the DES and water can be recovered and recycled. So the method completely meets the requirements of green, environmental protection and clean process.
(5) After the stevioside is extracted by the DES aqueous solution, in order to further separate and improve the content of RA in the stevioside, ethanol used in the current industrial production is adopted, the using amount is small, and relevant regulations or requirements at home and abroad are met. If the cost is not remarkably reduced, a nanofiltration membrane separation technology with higher cost can be adopted without using alcohol precipitation, so that the environmental protection and the safety can be further improved.
(6) After the content of the product RA in the extraction and separation processes is increased, better raw materials are provided for further producing the commercialized high-purity stevioside (95% -99%), the yield can be obviously increased, the cost can be reduced, and the economic effect is obvious.
Drawings
To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, and it should be understood that the following drawings only illustrate some examples of the present invention and therefore should not be considered as limiting the scope.
High performance liquid chromatography conditions: the detection wavelength is 210nm, the mobile phase is 68% acetonitrile/32% water (phosphate buffer pH is 2.60), the flow rate is 1mL/min, and the injection volume is 20 mu L; the column temperature was 40 ℃. HPLC analysis used a C18 column (Waters, Massachusetts, USA), SPD-M20A PDA detector (Shimadzu, Kyoto, Japan), column oven HCT-360LC (Hengao Tech & Dev, Tianjin, China), and workstation Class-VP (Shimadzu, Kyoto, Japan).
FIG. 1 is a conventional organic solvent extraction route; FIG. 2 is a DES extraction route provided by the present invention; FIG. 3 is a DES synthesis route provided by the present invention; FIG. 4 shows DES provided by the present invention[ Choline chloride][ ethylene glycol]The characteristic peaks of choline chloride and ethylene glycol show a certain red shift after DES is synthesized; FIG. 5 is a high performance liquid chromatogram of stevia sugar extracted by conventional method; FIG. 6 is a high performance liquid chromatogram of the aqueous solutions of DESS provided by the present invention for extracting stevioside.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially. The method for preparing DES and extracting stevioside provided by the embodiment of the invention is specifically explained below.
Example 1DES Synthesis (note: this example uses only hydrogen bond acceptor [ choline chloride ], hydrogen bond donor [ ethylene glycol ] as reactants, and the remaining DES synthesis conditions are exactly the same as in this example, only the difference in hydrogen bond donor in the product structure.)
Adding a proper amount of choline chloride and ethylene glycol into a single-neck flask for later use, weighing and preparing into choline chloride: mixing ethylene glycol at a molar ratio of 1:2 for later use; the mixture was stirred at 80 ℃ for 4 hours to give a colorless transparent liquid, i.e., a low co-solvent (DES). And (3) storing the DES in a room temperature environment for later use, wherein the moisture content is within 4 percent.
Example 2 use of glycol-like DESS for extraction of stevia sugar
Cleaning stevia rebaudiana, naturally airing, crushing, sieving with a 200-mesh sieve, adding 5g of stevia rebaudiana into a DES (DES: water ═ 1: 20) g/mL water solution with the water content of 20%, placing into a 250mL single-neck flask, performing an extraction experiment at 60 ℃, wherein the solid-liquid ratio is 1:20(g/mL), the extraction time is 180min, and filtering and separating a solid-liquid phase to obtain an extracting solution containing stevioside (64.5% of RA); further separating, precipitating with 80% ethanol to obtain higher content of stevioside (81.2% of RA), with unit DES stevioside extraction rate of 182 mg/g.
Examples 3-11 DES other examples for extraction of stevia sugar
Examples 7-15, in which DES was applied to the extraction of stevioside under other different conditions, are shown in the following table. The extraction conditions not mentioned in the table are the same as in example 2.
In conclusion, the invention provides a preparation method of a low cosolvent and selective adsorption of the low cosolvent on stevioside, and the method is used for extracting the stevioside by using DES for the first time, so that a novel method for DES selective adsorption separation is established.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A method for selectively extracting stevia sugar components by using a low cosolvent aqueous solution is characterized by comprising the following steps:
(1) cleaning stevia rebaudiana Bertoni material, drying, pulverizing, and sieving to obtain stevia rebaudiana Bertoni powder;
(2) during the preparation process of the low co-solvent (DES), firstly, adding a proper amount of a mixture of a hydrogen bond acceptor and a hydrogen bond donor into a single-neck flask for later use, stirring the mixture for 4 hours at 80 ℃, obtaining colorless transparent liquid after the reaction is finished, namely the low co-solvent, and storing the DES in an environment lower than room temperature for later use;
(3) in the process of extracting stevioside by using a low cosolvent, adding a certain amount of crushed stevia rebaudiana into a DESS aqueous solution with a certain water content, and extracting for a certain time at a certain temperature; and (3) performing solid-liquid separation after extraction, wherein the liquid phase contains the stevioside with higher purity RA, and the liquid phase can be further subjected to alcohol precipitation by using a certain amount of low-carbon alcohol, so that the RA purity in the stevioside product can be further improved.
2. The extraction process of stevia sugar from stevia rebaudiana Bertoni according to claim 1, wherein in step (1), the stevia rebaudiana Bertoni is pulverized and sieved with 100-300 meshes.
3. The method of claim 1, wherein in step (2), the hydrogen bond acceptor of the low cosolvent is choline chloride or betaine, and the hydrogen bond donor is ethylene glycol, 1, 3-propanediol, glycerol, 1, 3-butanediol, or glucose.
4. The method of claim 1, wherein in step (3), the ratio of the low cosolvent to water in the low cosolvent aqueous solution is from 1:5g/mL to 1:30 g/mL.
5. The extraction process method of stevia sugar component in stevia according to claim 1, wherein in step (3), the extraction is performed at an extraction temperature of 40 ℃ to 80 ℃ for 30min to 180min at a solid-to-liquid ratio of 1:10g/mL to 1:70 g/mL.
6. The method for extracting stevia sugar from stevia rebaudiana Bertoni according to claim 1, wherein the lower alcohol used in the alcohol precipitation in step (3) is 50 to 90% methanol or ethanol.
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| CN102199178A (en) * | 2011-04-10 | 2011-09-28 | 胡玮 | Extraction process of rebaudioside A (RA) |
| US20130108718A1 (en) * | 2010-07-16 | 2013-05-02 | Justbio Inc. | Extraction method for providing an organic certifiable stevia rebaudiana extract |
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| US20130108718A1 (en) * | 2010-07-16 | 2013-05-02 | Justbio Inc. | Extraction method for providing an organic certifiable stevia rebaudiana extract |
| CN102199178A (en) * | 2011-04-10 | 2011-09-28 | 胡玮 | Extraction process of rebaudioside A (RA) |
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