CN108033986B - Method for preparing steviosin - Google Patents
Method for preparing steviosin Download PDFInfo
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- CN108033986B CN108033986B CN201810121890.XA CN201810121890A CN108033986B CN 108033986 B CN108033986 B CN 108033986B CN 201810121890 A CN201810121890 A CN 201810121890A CN 108033986 B CN108033986 B CN 108033986B
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- steviosin
- stevia rebaudiana
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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 63
- 238000000034 method Methods 0.000 title claims abstract description 42
- 244000228451 Stevia rebaudiana Species 0.000 claims abstract description 31
- 235000006092 Stevia rebaudiana Nutrition 0.000 claims abstract description 31
- 238000000605 extraction Methods 0.000 claims abstract description 31
- 229940013618 stevioside Drugs 0.000 claims abstract description 27
- 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 claims abstract description 27
- 235000019202 steviosides Nutrition 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- 239000003480 eluent Substances 0.000 claims description 20
- 239000002994 raw material Substances 0.000 claims description 20
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 18
- 238000004587 chromatography analysis Methods 0.000 claims description 16
- 238000001914 filtration Methods 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 11
- 108010059820 Polygalacturonase Proteins 0.000 claims description 10
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 9
- 238000001179 sorption measurement Methods 0.000 claims description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000003463 adsorbent Substances 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 8
- 229940059442 hemicellulase Drugs 0.000 claims description 8
- 108010002430 hemicellulase Proteins 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 239000000741 silica gel Substances 0.000 claims description 8
- 229910002027 silica gel Inorganic materials 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 229940088598 enzyme Drugs 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 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 description 4
- 241000544066 Stevia Species 0.000 claims description 3
- 235000009508 confectionery Nutrition 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 230000007547 defect Effects 0.000 abstract description 5
- 239000012535 impurity Substances 0.000 abstract description 5
- 229910017053 inorganic salt Inorganic materials 0.000 abstract description 4
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 37
- 239000000047 product Substances 0.000 description 18
- 235000019441 ethanol Nutrition 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 239000012528 membrane Substances 0.000 description 5
- 229930182470 glycoside Natural products 0.000 description 4
- 150000002338 glycosides Chemical class 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000011347 resin Substances 0.000 description 4
- 229920005989 resin Polymers 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000007670 refining Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 description 2
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 2
- 229930002875 chlorophyll Natural products 0.000 description 2
- 235000019804 chlorophyll Nutrition 0.000 description 2
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 2
- 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 2
- 238000011160 research Methods 0.000 description 2
- 229930182490 saponin Natural products 0.000 description 2
- 150000007949 saponins Chemical class 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 238000000194 supercritical-fluid extraction Methods 0.000 description 2
- 239000001648 tannin Substances 0.000 description 2
- 235000018553 tannin Nutrition 0.000 description 2
- 229920001864 tannin Polymers 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- 241000208838 Asteraceae Species 0.000 description 1
- 239000001512 FEMA 4601 Substances 0.000 description 1
- 239000001776 FEMA 4720 Substances 0.000 description 1
- 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 description 1
- -1 and the like Chemical compound 0.000 description 1
- 229960000892 attapulgite Drugs 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000006189 buccal tablet Substances 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 230000001013 cariogenic effect Effects 0.000 description 1
- 235000015218 chewing gum Nutrition 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000012733 comparative method Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 238000004332 deodorization Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 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 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 230000016615 flocculation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000874 microwave-assisted extraction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229910052625 palygorskite Inorganic materials 0.000 description 1
- 235000019203 rebaudioside A Nutrition 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- 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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Seasonings (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention relates to a preparation method for extracting high-purity stevia rebaudiana from stevia rebaudiana. The method disclosed by the invention is simple in steps, the defect of water resource consumption caused by a large amount of water used in the traditional stevioside extraction method is avoided, the defects that a large amount of organic solvent or inorganic salt is used in the existing method, the subsequent impurity removal process is increased, the loss rate is high are also avoided, the production cost is further reduced, and the excellent extraction rate is obtained while the product purity is improved. The purity of the steviosin product prepared by the method is more than 98.5%, and the extraction rate is as high as more than 13.95%.
Description
Technical Field
The invention relates to a preparation method of stevioside, in particular to a preparation method for extracting high-purity stevia rebaudiana from stevia rebaudiana.
Background
Steviosin, also known as stevioside and stevioside, is a natural, high-sweetness, low-calorie, non-cariogenic, safe and non-toxic sweetener, and is widely applied to products in the forms of food beverages, oral liquids, buccal tablets, chewing gums and the like. The steviosin is extracted from stevia rebaudiana Bertoni (stevia rebaudiana Bertoni) belonging to Compositae family. Among the glycosides contained in stevia rebaudiana Bertoni, the glycosides which are high in content and economically valuable include stevioside, rebaudioside A, rebaudioside C, and the like, and stevioside is a generic name for the above various sweet glycosides. The annual demand of steviosin worldwide exceeds 1 million tons, and the demand thereof is increasing at a rate of 30% per year; the annual output of China is 3000 tons, more than 80 percent of the annual output is exported to Japan, Korea, United states and other countries, and the market development requirements can not be met far. Therefore, the research and development of the steviosin extraction process are vigorously carried out, the development of the steviosin industry in China is driven, and the method has very important practical significance for pulling the national economy of China.
The stevia rebaudiana leaves contain a large amount of nonpolar substances such as oil, chlorophyll, essential oil and the like, and also contain a large amount of water-soluble substances such as protein, organic acid, tannin, saponin, pigment, inorganic salt and the like, and the existence of the substances has great influence on the extraction of the stevioside. The traditional method for extracting the stevioside usually uses a large amount of water for soaking and extracting, so that a large amount of water resource is consumed, and a stevioside product with higher purity cannot be obtained due to the interference of other components. In recent years, research on methods for preparing steviosin has been increasing, and various improvements in extraction, purification, impurity removal, modification, and the like have been proposed.
For example, CN107474086A discloses a method for preparing stevioside, which comprises the steps of pressurized countercurrent water extraction, activated carbon column or resin column decoloration and deodorization, industrial preparative chromatographic separation, plate-frame filtration and spray drying, wherein the purity of the product can reach more than 90%.
CN107118243A discloses an industrial preparation method of stevioside, which comprises the following steps: repeatedly soaking stevia rebaudiana leaf in acetic acid solution to obtain extractive solution, filtering, loading onto adsorption column, eluting with 0.5-1.0% sodium hydroxide, eluting with ethanol to obtain eluate, recovering ethanol, decolorizing with water to obtain sugar solution, decolorizing with decolorizing resin, and spray drying to obtain stevioside product with total glycoside content of above 88.9%, wherein RA component is above 52.7%.
CN106146574A discloses a method for compounding stevioside in stevia rebaudiana leaves by microwave-ultrasound, which comprises the following steps: washing stevia rebaudiana leaves, soaking in absolute ethyl alcohol, drying to obtain stevia rebaudiana leaf powder, adding water and a complex enzyme, uniformly mixing, performing microwave heating, performing ultrasonic treatment to obtain an extracting solution and filter residues, uniformly stirring the filter residues and diethyl ether, performing centrifugal separation, combining supernatant liquid and the extracting solution to obtain a total extracting solution, uniformly mixing an iron trichloride solution, a calcium hydroxide solution, a polyacrylamide solution and the total extracting solution to obtain a mixed solution, mixing the mixed solution with an aluminum sulfate solution, performing supercritical extraction, adsorbing the extracted solution by using macroporous resin, performing reduced pressure distillation, concentrating and drying to obtain a stevioside crude product, dissolving the stevioside crude product in water, adding attapulgite, refining, performing centrifugal concentration and drying to obtain the stevioside, wherein the purity can reach over 94.1%.
These methods have significant advantages over conventional extraction methods, but have certain obstacles in industrial production due to poor selectivity, loss rate, cost, poor continuous operability, or excessive loss rate. Therefore, aiming at the defects of the method for preparing the steviosin in the prior art, the invention aims to provide the method for preparing the steviosin, which has the advantages of simple process, high production efficiency and simple and convenient operation.
Disclosure of Invention
The invention provides a preparation method for extracting stevioside from stevia rebaudiana, which comprises the following steps:
(1) placing stevia rebaudiana leaf in an extraction tank, adding water 1-3 times of the weight of the raw materials, adding 1.0-3.0% of biological enzyme and stevioside 0.1-0.5% of the weight of the raw materials, extracting at normal temperature for 0.5-2h, and filtering to obtain an extracting solution;
(2) adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution;
(3) separating the treatment liquid in the step (2) by simulated moving bed chromatography to obtain a steviosin component;
(4) and (4) concentrating, centrifuging and drying the steviosin component in the step (3) to obtain the steviosin product.
Wherein, the biological enzyme in the step (1) is preferably 1.0-3.0% of pectinase and/or hemicellulase, and more preferably a mixture of 1:1 of pectinase and hemicellulase.
Wherein, in the step (4), the adsorbent filled by the simulated moving bed chromatography is silica gel, and the eluent is 15% ethanol solution; the flow rate of the adsorption area is 1-2 BV/h; the flow rate of an elution area is 1-3 BV/h; the switching time is 500-800 s; controlling the temperature to be 20-30 ℃; the pressure is controlled between 0.15MPa and 0.45 MPa;
the purity of the steviosin product prepared by the method is more than 98.5%, and the extraction rate is as high as more than 13.95%.
The method disclosed by the invention is simple in steps, the defect of water resource consumption caused by a large amount of water used in the traditional stevioside extraction method is avoided, the defects that a large amount of organic solvent or inorganic salt is used in the existing method, the subsequent impurity removal process is increased, the loss rate is high are also avoided, the production cost is further reduced, and the excellent extraction rate is obtained while the product purity is improved.
As indicated in the background art, stevia leaves contain a large amount of nonpolar substances such as oil, chlorophyll and essential oil, and also contain a large amount of water-soluble substances such as protein, organic acid, tannin, saponin, pigment and inorganic salt, and the presence of the substances has great influence on the extraction of the stevioside. The prior art carries out extensive research on the extraction, purification and refining of the steviosin, provides extraction schemes such as enzymolysis, ultrasonic extraction, microwave extraction, ultrahigh pressure extraction, supercritical extraction and the like, and also provides purification and refining methods such as flocculation, macroporous resin separation, membrane separation, recrystallization and the like. The inventor of the application conducts a large number of experiments on the basis of the prior art, and finds that a small amount of stevioside is added during extraction during the experiment, so that the extraction rate can be greatly improved, and an unexpected effect is obtained by adopting a calcium carbonate impurity removal method. The experimental results provided below are used to demonstrate the significant beneficial effects of the preparation method of the present invention. The method for measuring the purity and extraction rate of the stevioside in the product is a method known in the field.
Experimental materials: stevia rebaudiana, 15% ethanol, pectinase, hemicellulase, calcium carbonate, diethyl ether, aluminum sulfate, ferric trichloride, calcium hydroxide, polyacrylamide and 65% ethanol
The method comprises the following steps: according to the method of example 1, 100g of stevia is taken as a raw material.
Comparative method 1: CN106146574A the method of example 1, wherein 100g of stevia rebaudiana is taken as raw material.
Control method 2: CN107474086A, the method of example 1, 100g stevia rebaudiana Bertoni is taken as raw material.
Control method 3: placing 100g of stevia rebaudiana leaves in an extraction tank, adding water 2 times the weight of the raw materials, adding a mixture (1: 1) of pectinase and hemicellulase with the weight of 2.0% and stevioside with the weight of 0.2%, extracting for 1h at normal temperature, and filtering to obtain an extracting solution; removing impurities from the extracting solution through a primary membrane, and concentrating through a secondary membrane to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 1BV/h, the flow rate of the eluent in an elution area is 2BV/h, the switching time is 700s, the temperature is controlled at 30 ℃, and the pressure is controlled at 0.3MPa, so as to obtain a steviosin component, concentrating, centrifuging and drying to obtain a steviosin product.
Control method 4: placing 100g of stevia rebaudiana leaf in an extraction tank, adding water 2 times the weight of the raw materials, adding a mixture (1: 1) of pectinase and hemicellulase 2.0 wt%, extracting at normal temperature for 1h, and filtering to obtain an extract; adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 1BV/h, the flow rate of the eluent in an elution area is 2BV/h, the switching time is 700s, the temperature is controlled at 30 ℃, and the pressure is controlled at 0.3MPa, so as to obtain a steviosin component, concentrating, centrifuging and drying to obtain a steviosin product.
Control method 5: placing 100g of stevia rebaudiana leaves in an extraction tank, adding water 2 times of the weight of the raw materials, and obtaining an extracting solution by adopting a three-stage continuous countercurrent extraction method; adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 1BV/h, the flow rate of the eluent in an elution area is 2BV/h, the switching time is 700s, the temperature is controlled at 30 ℃, and the pressure is controlled at 0.3MPa, so as to obtain a steviosin component, and concentrating, centrifuging and drying to obtain the steviosin product.
The results of the experiments are shown in the following table.
Therefore, the method provided by the invention has the advantages of outstanding beneficial effects, high product purity and high extraction rate. The comparison method 3 adopts a membrane treatment purification method, and achieves the effect similar to that of the method, but the membrane treatment method has complex cleaning process, prolongs the whole production period, greatly increases the production cost, is not as simple and easy as the method of the application, and is beneficial to industrialized mass production.
Detailed Description
The present invention will be described in more detail with reference to the following embodiments.
Example 1
Placing stevia rebaudiana leaves in an extraction tank, adding water 2 times the weight of the raw materials, adding a mixture (1: 1) of pectinase and hemicellulase 2.0 wt% and stevioside 0.2 wt%, extracting at normal temperature for 1h, and filtering to obtain an extract; adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 1BV/h, the flow rate of the eluent in an elution area is 2BV/h, the switching time is 700s, the temperature is controlled at 30 ℃, and the pressure is controlled at 0.3MPa, so as to obtain a steviosin component, and concentrating, centrifuging and drying to obtain the steviosin product. The purity of the steviosin product reaches 99.3%, and the extraction rate reaches 14.01%.
Example 2
Placing stevia rebaudiana leaves in an extraction tank, adding water which is 3 times of the weight of the raw materials, adding 3.0 weight percent of pectinase and 0.1 weight percent of stevioside, extracting for 1 hour at normal temperature, and filtering to obtain an extracting solution; adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 2BV/h, the flow rate of the eluent in an elution area is 3BV/h, the switching time is 700s, the temperature is controlled at 25 ℃, and the pressure is controlled at 0.35MPa, so as to obtain a steviosin component, and concentrating, centrifuging and drying to obtain the steviosin product. The purity of the steviosin product reaches 98.8%, and the extraction rate reaches 13.99%.
Example 3
Placing stevia rebaudiana leaves in an extraction tank, adding water which is 1 time of the weight of the raw materials, adding 2.5 weight percent of pectinase and 0.35 weight percent of stevioside, extracting for 2 hours at normal temperature, and filtering to obtain an extracting solution; adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution; and (3) separating the treated liquid by virtue of a simulated moving bed chromatography, wherein an adsorbent filled in the simulated moving bed chromatography is silica gel, an eluent is 15% ethanol solution, the flow rate of the eluent in an adsorption area is 1BV/h, the flow rate of the eluent in an elution area is 1BV/h, the switching time is 700s, the temperature is controlled at 30 ℃, and the pressure is controlled at 0.3MPa, so as to obtain a steviosin component, and concentrating, centrifuging and drying to obtain the steviosin product. The purity of the steviosin product reaches 99.1%, and the extraction rate reaches 14.02%.
Claims (5)
1. A method for extracting steviosin from sweet stevia comprises the following steps:
(1) placing stevia rebaudiana leaf in an extraction tank, adding water 1-3 times of the weight of the raw materials, adding 1.0-3.0% of biological enzyme and stevioside 0.1-0.5% of the weight of the raw materials, extracting at normal temperature for 0.5-2h, and filtering to obtain an extracting solution;
(2) adding calcium carbonate powder in the same amount as the stevia rebaudiana raw material into the extracting solution, stirring, centrifuging, standing, and filtering to obtain a treatment solution;
(3) separating the treatment liquid in the step (2) by simulated moving bed chromatography to obtain a steviosin component;
(4) and (4) concentrating, centrifuging and drying the steviosin component in the step (3) to obtain a steviosin product.
2. The method according to claim 1, wherein the biological enzyme in the step (1) is 1.0-3.0% of pectinase and/or hemicellulase.
3. The method according to claim 2, wherein the biological enzyme is a 1:1 mixture of pectinase and hemicellulase.
4. The process according to claim 1, wherein in the step (4), the adsorbent packed by the simulated moving bed chromatography is silica gel, and the eluent is 15% ethanol solution; the flow rate of the adsorption area is 1-2 BV/h; the flow rate of an elution area is 1-3 BV/h; the switching time is 500-800 s; controlling the temperature to be 20-30 ℃; the pressure is controlled between 0.15MPa and 0.45 MPa.
5. The preparation method of claim 1, wherein the obtained steviosin product has a purity of above 98.5% and an extraction rate of above 13.95%.
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