CN107371672A - A kind of method for improving STEVIA REBAUDIANA REB D yield - Google Patents
A kind of method for improving STEVIA REBAUDIANA REB D yield Download PDFInfo
- Publication number
- CN107371672A CN107371672A CN201710444801.0A CN201710444801A CN107371672A CN 107371672 A CN107371672 A CN 107371672A CN 201710444801 A CN201710444801 A CN 201710444801A CN 107371672 A CN107371672 A CN 107371672A
- Authority
- CN
- China
- Prior art keywords
- stevia rebaudiana
- parts
- reb
- yield
- steviol glycoside
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Forests & Forestry (AREA)
- Ecology (AREA)
- Biochemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Saccharide Compounds (AREA)
- Seasonings (AREA)
Abstract
The invention discloses a kind of method for improving STEVIA REBAUDIANA REB D yield, mainly realized by the cultivation of STEVIA REBAUDIANA and steviol glycoside extracting method the two aspects, wherein in the cultivation of STEVIA REBAUDIANA, 2 months are needed before STEVIA REBAUDIANA is gathered in, spray including 0.01 0.05 parts of LREE, 0.1 0.2 parts of photosynthetic bacteria, 58 parts of salt, chelating 15 25 parts of conditioning agents in parts by weight of potassium to stimulate STEVIA REBAUDIANA blade face, the amount once, sprayed every time is sprayed within every 57 days to be defined by blade face moistening.Compared with prior art, the present invention makes Rebaudiodside A D content improve 25 times on the basis of in the past；2nd, boiled using two kinds of enzymolysis, heating, accelerate steviol glycoside from the rate of release in STEVIA REBAUDIANA blade, also eliminate the use of etoh solvent, accelerate the extraction rate of steviol glycoside.
Description
Technical field
The present invention relates to technical field of extraction of Chinese traditional medicine, and in particular to a kind of method for improving STEVIA REBAUDIANA REB-D yield.
Background technology
Sweet ingredient in STEVIA REBAUDIANA blade is referred to as steviol glycoside, is a kind of tetracylic diterpene derivatives, comprising but not
It is limited to stevioside, rebaudioside A, B, C, D, F, and exist with small concentration in the leaf of STEVIA REBAUDIANA.Its sugariness is about the 300 of sucrose
Times, research shows, steviol glycoside has the characteristics of high sugariness, low in calories, without obvious toxic-side effects, can suppress hyperglycaemia, high blood
Pressure, and by anti-inflammatory, antitumor, antidiarrheal diuresis and assist immunoregulatory effect.For convenience, Rebaudiodside A claims herein
For REB-A, REB-B, REB-C, REB-D etc., in Rebaudiodside A, REB-D is with than REB-A with more preferable sugared feature and more
Preferable taste.
On REB extraction, in the prior art generally by stevioside leaf crushed, extract, filtered, adsorbing separation,
The processes such as concentration, precipitation are come what is realized, and in the extraction process of reality, REB-D is to mix other Rebaudiodside As to be carried together
Take out.But because of the REB contents limitation of stevioside plant in itself, plus bad in terms of PROCESS FOR TREATMENT, carry in the prior art
The steviol glycoside total content got is not high, especially REB-D content, and extraction cost is high.
The content of the invention
For above-mentioned the shortcomings of the prior art, it is an object of the invention to provide one kind to improve STEVIA REBAUDIANA REB-D yield
Method, this method is more suitable for industrialized production.
To achieve the above object, the technical solution adopted by the present invention is：A kind of method for improving STEVIA REBAUDIANA REB-D yield,
Realized by the following aspects：
（1）The cultivation of STEVIA REBAUDIANA：2 months before STEVIA REBAUDIANA harvesting, spray including LREE 0.01-0.05 parts, photosynthetic bacteria
0.1-0.2 parts, salt 5-8 parts, the conditioning agent of potassium 15-25 parts in parts by weight is chelated to stimulate STEVIA REBAUDIANA blade face, it is every to spray within 5-7 days
Once, the amount sprayed every time is defined by blade face moistening；
（2）The extraction of steviol glycoside：A, blade is crushed to 200-300 mesh；B, plus after suitable quantity of water mixing, enzyme-added hydrolysis, enzyme used
For two kinds of enzymes of cellulase and neutral proteinase, the addition of its cellulase is the 2%-3% of substrate, preference temperature 40
DEG C, pH 4.5-6.5, vigor is 100,000 every gram of units, action time 1-1.5h；The addition of neutral proteinase is 0.05%-
0.25%, preference temperature is 35-55 DEG C, pH value 6-6.8, and vigor is 100,000 every gram of units, action time 0.5-0.8h；So
Heating is boiled afterwards, is maintained boil condition 1-2h, is filtered to obtain extract solution, filter residue, and wherein filter residue Repeat-heating boils 1-3 times, is collected
Whole filtrate, obtains total extract solution；C, by liquor ferri trichloridi and total extract solution that mass fraction is 20-30% according to 5-10:1-
3:50-80 volume ratio is well mixed, and adjusts pH value to 7-7.5 with calcium hydroxide, and is heated to 40-50 DEG C, is incubated 10-
25min, room temperature is subsequently cooled to, obtains mixed liquor；D, obtained with the HPD-392 purification with macroreticular resin steps c by pretreatment
The mixed liquor arrived, flow velocity during resin adsorption is 3-5BV/h, is eluted after adsorption saturation with ethanol solution, the speed of elution is 4-
6BV/h, obtain eluent；E, using the microporous barrier eluent that filtration step d is obtained again, medicinal decolouring aluminum oxide is then used
Decolourized, flow velocity 22.0-25.0mL/min, while be heated up to 40-45 DEG C, it is last it is filtered, concentrate, be dried to obtain stevia rebaudianum
Glucosides.
Preferably, above-mentioned conditioning agent also includes brassinosteroid 0.5-2 parts.
Preferably, above-mentioned conditioning agent is dissolved before spraying every time using 0-1 DEG C water 40-50 parts.
Preferably, ultrasonic wave is added while hydrolysis, the frequency of ultrasonic wave is 50KHZ.
Compared with prior art, the beneficial effects of the invention are as follows：Blade face is stimulated using conditioning agent when the 1st, cultivating, makes Lai Baodi
Glycosides D content improves 2-5 times on the basis of in the past；2nd, boiled using two kinds of enzymolysis, heating, accelerate steviol glycoside from sweetleaf
Rate of release in chrysanthemum blade, the use of etoh solvent is also eliminated, accelerate the extraction rate of steviol glycoside；3rd, using macropore
Resin adsorption, micro-pore-film filtration and aluminum oxide decolourize, and the yield of steviol glycoside can be made to reach more than 99.7%, cost reduces by 76%.
Embodiment
In conjunction with specific embodiment, next the present invention is described in further detail.Obviously, it is impossible to therefore limit the present invention
System is among described scope of embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making wound
The every other embodiment that the property made is obtained on the premise of working, all should belong to protection scope of the present invention.In following embodiments
The experimental method of unreceipted actual conditions, conventionally and condition, or selected according to catalogue.
Embodiment one
The method for improving STEVIA REBAUDIANA REB-D yield, is realized by the following aspects：
（1）The cultivation of STEVIA REBAUDIANA：STEVIA REBAUDIANA harvesting before 2 months, spray including 0.05 part of LREE, 0.2 part of photosynthetic bacteria,
8 parts of salt, 25 parts of conditioning agents in parts by weight of potassium are chelated to stimulate STEVIA REBAUDIANA blade face, spray the amount once, sprayed every time within every 5 days
By blade face, moistening is defined；Dissolved before spraying every time using 1 DEG C of 50 parts of water.
（2）The extraction of steviol glycoside：A, blade is crushed to 200 mesh；B, plus after suitable quantity of water mixing, enzyme-added hydrolysis, enzyme used
For two kinds of enzymes of cellulase and neutral proteinase, the addition of its cellulase is the 3% of substrate, and preference temperature is 40 DEG C, pH
For 4.5, vigor is 100,000 every gram of units, action time 1h；The addition of neutral proteinase is 0.05%, and preference temperature is 55 DEG C,
PH value is 6, and vigor is 100,000 every gram of units, action time 0.8h；Then heating is boiled, and is maintained boil condition 2h, is filtered
Extract solution, filter residue, wherein filter residue Repeat-heating boil 3 times, collect whole filtrate, obtain total extract solution；C, it is by mass fraction
30% liquor ferri trichloridi is with total extract solution according to 10:1:50 volume ratio is well mixed, and with calcium hydroxide adjust pH value to
7, and 50 DEG C are heated to, 10min is incubated, room temperature is subsequently cooled to, obtains mixed liquor；D, with the HPD-392 macropores by pretreatment
The mixed liquor that purification on adsorbent resins step c is obtained, flow velocity during resin adsorption is 3BV/h, is washed after adsorption saturation with ethanol solution
De-, the speed of elution is 4BV/h, obtains eluent；E, using the microporous barrier eluent that filtration step d is obtained again, then use
Medicinal decolouring is decolourized with aluminum oxide, flow velocity 23.0mL/min, while is heated up to 45 DEG C, last filtered, concentration, is dried
Obtain steviol glycoside.
Embodiment two
The method for improving STEVIA REBAUDIANA REB-D yield, is realized by the following aspects：
（1）The cultivation of STEVIA REBAUDIANA：STEVIA REBAUDIANA harvesting before 2 months, spray including 0.01 part of LREE, 0.1 part of photosynthetic bacteria,
0.5 part of 5 parts of salt, 15 parts of potassium of chelating, brassinosteroid conditioning agent in parts by weight stimulate STEVIA REBAUDIANA blade face, spray within every 7 days
Once, the amount sprayed every time is defined by blade face moistening；
（2）The extraction of steviol glycoside：A, blade is crushed to 300 mesh；B, plus after suitable quantity of water mixing, enzyme-added hydrolysis, while hydrolysis
Add ultrasonic wave, the frequency of ultrasonic wave is 50KHZ, and enzyme used is two kinds of enzymes of cellulase and neutral proteinase, its cellulase
Addition be the 2% of substrate, preference temperature is 40 DEG C, pH 6.5, and vigor is 100,000 every gram of units, action time 1.5h；In
Property protease addition be 0.25%, preference temperature be 35 DEG C, pH value 6.8, vigor is 100,000 every gram of units, action time
For 0.8h；Then heating is boiled, and is maintained boil condition 1h, is filtered to obtain extract solution, filter residue, wherein filter residue Repeat-heating boils 1-3
It is secondary, whole filtrate is collected, obtains total extract solution；C, by liquor ferri trichloridi and total extract solution that mass fraction is 20% according to 5:
3:80 volume ratio is well mixed, and adjusts pH value to 7.5 with calcium hydroxide, and is heated to 40 DEG C, is incubated 25min, Ran Houleng
But to room temperature, mixed liquor is obtained；D, the mixed liquor obtained with the HPD-392 purification with macroreticular resin steps c by pretreatment, tree
Flow velocity when fat adsorbs is 5BV/h, is eluted after adsorption saturation with ethanol solution, the speed of elution is 6BV/h, obtains eluent；e、
Using the microporous barrier eluent that filtration step d is obtained again, then decolourized using medicinal decolouring with aluminum oxide, flow velocity is
25.0mL/min, while be heated up to 40 DEG C, it is last it is filtered, concentrate, be dried to obtain steviol glycoside.
Claims (3)
- A kind of 1. method for improving STEVIA REBAUDIANA REB-D yield, it is characterised in that realized by the following aspects：（1）The cultivation of STEVIA REBAUDIANA：2 months before STEVIA REBAUDIANA harvesting, spray including LREE 0.01-0.05 parts, photosynthetic bacteria 0.1-0.2 parts, salt 5-8 parts, the conditioning agent of potassium 15-25 parts in parts by weight is chelated to stimulate STEVIA REBAUDIANA blade face, it is every to spray within 5-7 days Once, the amount sprayed every time is defined by blade face moistening；（2）The extraction of steviol glycoside：A, blade is crushed to 200-300 mesh；B, plus after suitable quantity of water mixing, enzyme-added hydrolysis, enzyme used For two kinds of enzymes of cellulase and neutral proteinase, the addition of its cellulase is the 2%-3% of substrate, preference temperature 40 DEG C, pH 4.5-6.5, vigor is 100,000 every gram of units, action time 1-1.5h；The addition of neutral proteinase is 0.05%- 0.25%, preference temperature is 35-55 DEG C, pH value 6-6.8, and vigor is 100,000 every gram of units, action time 0.5-0.8h；So Heating is boiled afterwards, is maintained boil condition 1-2h, is filtered to obtain extract solution, filter residue, and wherein filter residue Repeat-heating boils 1-3 times, is collected Whole filtrate, obtains total extract solution；C, by liquor ferri trichloridi and total extract solution that mass fraction is 20-30% according to 5-10:1- 3:50-80 volume ratio is well mixed, and adjusts pH value to 7-7.5 with calcium hydroxide, and is heated to 40-50 DEG C, is incubated 10- 25min, room temperature is subsequently cooled to, obtains mixed liquor；D, obtained with the HPD-392 purification with macroreticular resin steps c by pretreatment The mixed liquor arrived, flow velocity during resin adsorption is 3-5BV/h, is eluted after adsorption saturation with ethanol solution, the speed of elution is 4- 6BV/h, obtain eluent；E, using the microporous barrier eluent that filtration step d is obtained again, medicinal decolouring aluminum oxide is then used Decolourized, flow velocity 22.0-25.0mL/min, while be heated up to 40-45 DEG C, it is last it is filtered, concentrate, be dried to obtain stevia rebaudianum Glucosides.
- 2. the method according to claim 1 for improving STEVIA REBAUDIANA REB-D yield, it is characterised in that above-mentioned conditioning agent also wraps Include brassinosteroid 0.5-2 parts.
- 3. the method according to claim 1 or 2 for improving STEVIA REBAUDIANA REB-D yield, above-mentioned conditioning agent before spraying every time Dissolved using 0-1 DEG C water 40-50 parts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710444801.0A CN107371672A (en) | 2017-06-14 | 2017-06-14 | A kind of method for improving STEVIA REBAUDIANA REB D yield |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710444801.0A CN107371672A (en) | 2017-06-14 | 2017-06-14 | A kind of method for improving STEVIA REBAUDIANA REB D yield |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107371672A true CN107371672A (en) | 2017-11-24 |
Family
ID=60331818
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710444801.0A Pending CN107371672A (en) | 2017-06-14 | 2017-06-14 | A kind of method for improving STEVIA REBAUDIANA REB D yield |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107371672A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105294790A (en) * | 2015-12-07 | 2016-02-03 | 湖南华诚生物资源股份有限公司 | Method for extracting high-purity steviol glycosides from stevia rebaudiana |
| CN105801639A (en) * | 2016-03-23 | 2016-07-27 | 卞毓平 | Method for extracting stevioside from stevia rebaudiana and stevioside product |
| CN106046075A (en) * | 2016-06-17 | 2016-10-26 | 蚌埠市华东生物科技有限公司 | Method for extracting stevioside from stevia rebaudiana leaves |
| CN106146574A (en) * | 2016-06-17 | 2016-11-23 | 蚌埠市华东生物科技有限公司 | A kind of method of stevioside in microwave ultrasound compound collecting leaf of Folium Stevlae Rebaudianae |
| CN106631356A (en) * | 2016-11-03 | 2017-05-10 | 明光市大全甜叶菊专业合作社 | Method for increasing content of steviol glycoside in stevia rebaudiana |
-
2017
- 2017-06-14 CN CN201710444801.0A patent/CN107371672A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105294790A (en) * | 2015-12-07 | 2016-02-03 | 湖南华诚生物资源股份有限公司 | Method for extracting high-purity steviol glycosides from stevia rebaudiana |
| CN105801639A (en) * | 2016-03-23 | 2016-07-27 | 卞毓平 | Method for extracting stevioside from stevia rebaudiana and stevioside product |
| CN106046075A (en) * | 2016-06-17 | 2016-10-26 | 蚌埠市华东生物科技有限公司 | Method for extracting stevioside from stevia rebaudiana leaves |
| CN106146574A (en) * | 2016-06-17 | 2016-11-23 | 蚌埠市华东生物科技有限公司 | A kind of method of stevioside in microwave ultrasound compound collecting leaf of Folium Stevlae Rebaudianae |
| CN106631356A (en) * | 2016-11-03 | 2017-05-10 | 明光市大全甜叶菊专业合作社 | Method for increasing content of steviol glycoside in stevia rebaudiana |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2020048049A1 (en) | Method for preparing luo han guo sweetener composition from siraitia grosvenorii and use thereof | |
| CN103461459B (en) | A kind of method taking day lily as raw material and produce dehydrated vegetables | |
| CN101177444A (en) | Method for extracting momordica grosvenori flavone glycoside from momordica grosvenori | |
| CN108752231A (en) | The method extracted theanine from Sweet tea and extract Rubusoside and tea polyphenols simultaneously | |
| CN109651480A (en) | A method of separation momordica glycoside V | |
| CN111269171B (en) | Preparation method of high-purity 1-deoxynojirimycin | |
| CN107079936A (en) | Prevent and treat the method for treating seeds of cucurbita pepo seedling rot | |
| CN104262446B (en) | The method of mogroside Ⅴ is extracted from Grosvenor Momordica | |
| CN101862010A (en) | Preparation method of fruit juice for reducing organic acid content | |
| CN109966337B (en) | Method for extracting sennoside from senna leaf | |
| WO2020155494A1 (en) | Sweetener composition and preparation method and use thereof | |
| KR100954217B1 (en) | Method for production of the extract containing L-arabinose by using organic acid | |
| CN108617819A (en) | A kind of honey grapefruit tea preparation method of dispelling fatigue | |
| CN112851722B (en) | Preparation method of geniposide and gardenia yellow pigment | |
| CN107371672A (en) | A kind of method for improving STEVIA REBAUDIANA REB D yield | |
| CN101597635B (en) | Method for preparing high purity stachyose | |
| CN106924304A (en) | The extracting method of Sweet tea active ingredient | |
| CN111153943A (en) | Method for extracting stevioside by recrystallization method | |
| CN105154485A (en) | Method for extracting compound amino acid from polygonatum odoratum through enzymolysis and ultrasonic treatment | |
| CN115403647A (en) | Method for industrially producing mogroside, momordica fructose/wine and mannitol by using fresh momordica grosvenori | |
| CN102603450A (en) | Continuous extraction method of amino acid and pectin in watermelon peel | |
| CN111393490B (en) | Method for extracting linarin from buddleja officinalis | |
| CN106966794A (en) | A kind of preparation method for improving fruit sweetness crop nutrient solution | |
| CN107897475A (en) | A kind of oligomeric candy of probiotics and preparation method thereof | |
| CN112717009B (en) | Preparation method of gardenia extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171124 |
|
| RJ01 | Rejection of invention patent application after publication |