CN105063119B - The method of Geniposide is extracted from cape jasmine - Google Patents
The method of Geniposide is extracted from cape jasmine Download PDFInfo
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Abstract
The invention discloses the methods that Geniposide is extracted from cape jasmine, include the following steps:Prewet stacking, feedstock processing, digest, deoil for the first time, is second enzymatic hydrolysis, column chromatography, third time enzymatic hydrolysis, column chromatography, dry.The present invention first uses enzyme to extract vegetable fat, separates oil with non-oil component to affinity difference and the grease specific gravity difference of oil with water using non-oil component, increase product yield and improves conversion ratio.
Description
Technical field
The present invention relates to the extracting methods of field of biotechnology, and the method for Geniposide is extracted specifically from cape jasmine.
Background technique
Mast is the fruit of Rubiaceae (Rubiaceae) plant mast (Gardenia jasminoides Ellis), long
Ellipse or ellipse are in peony or yellowish red color, and the first batch of dual-purpose of drug and food resource that the Wei Zishu Ministry of Public Health promulgates, is in tradition
Medicine.Mast has purging intense heat relieving restlessness, and clearing heat and promoting diuresis, removing pattogenic heat from the blood and toxic material from the body, lidan liver, decompression, sterilize anti-inflammatory, the function of externally used detumescence analgesic
Effect.It is mainly used for treating pyreticosis, Jaundice Jaundice, bruise, hypertension, diabetes etc. on tcm clinical practice.Chemistry in mast
There are many ingredient, wherein mainly have crocin, the iridoids such as crocetin (Ichietal) and geniposide (Geniposide)
Compound, Geniposidic are its principle active components.Geniposide is a kind of Iridoidglucoside, soluble easily in water.
Geniposide can be hydrolyzed to Geniposide by R- glucosaccharase.Modern research shows that geniposide is to digestive system, cardiovascular system
System general the nervous system disease in has significant curative effect, in addition, there are also certain anti-inflammatory and treatment soft tissue injuries for geniposide
Effect.Geniposide is also widely used in other fields other than medicinal, such as can be used as plant promoter, life
Analyte detection agent etc. is a kind of important bioactive substance.It is widely used in medical treatment, field of health care products.
Separating-purifying after Geniposide (Genipin) is decomposed by geniposide and obtain, main application has:Geniposide can be with
Amino acid or albumen qualitative response, generate a kind of mast cyanine, and this natural pigment is used for food dye because of its nontoxicity
The production of material；Excellent natural biological crosslinking agent:Biological material can be made with crosslinkings such as protein, collagen, gelatin and chitosans
Material, such as artificial skelecton, wound dressing materials, toxicity is far below glutaraldehyde and other common chemical cross-linking agents；It can be used for controlling
Treat liver diseases, decompression, defaecation etc..The result of study of one medical centre of Israel shows that Geniposide can alleviate II type sugar
Urinate the symptom of disease；Prepare the crosslinking agent of immobilised enzymes；It is to inhibit mutagen activity most in the iridoid glycoside found so far
Strong substance；Also there is document report Geniposide and inhibit Apoptosis with antithrombotic；Generally speaking, Geniposide application is wide
It is general, there is potentiality to be exploited, vast market prospect and huge utility value.
Geniposide is primarily present in the natural plants such as mast, Cortex Eucommiae, is the desirable feedstock for extracting geniposide, domestic
That there is recovery rates in existing extraction process scheme is not high, product purity is low, energy consumption is big, largely uses organic solvent, useless
The problems such as water is big.
Summary of the invention
The object of the present invention is to provide the methods that Geniposide is extracted from cape jasmine, have simple process, are suitble to production, production
The advantages that at low cost.
The invention is realized by the following technical scheme, specifically comprises the following steps：
1) it prewets stacking：Cape jasmine is built into heap in the cool, is sprayed water, after water is from the outflow of heap bottom, is stacked naturally；
2) feedstock processing：The cape jasmine that step 1) is obtained crushes or is crushed, and addition is equivalent to the pure of 2-3 times of weight of raw material
Water is heated to 50-60 DEG C of maintenance 5-10 minutes, obtains paste serous material；
3) it digests for the first time：The paste serous material that step 2) is obtained, tune pH are 4-5, and paste serous material 0.05-0.1% is added
The pectin-cellulose complex enzyme of parts by weight and the mannitol of 0.001 parts by weight carry out enzymatic hydrolysis 40-60 minutes, obtain first time enzyme
Solve liquid；
4) it deoils：The first time enzymolysis liquid that step 3) is obtained is added the pure water of 2-3 times of weight, is heated to 60-70 DEG C
Extraction 60-120 minutes, obtains upper layer oil slick liquid and subnatant, slag mixture, separates；
5) it digests for second：Subnatant, the slag mixture that step 4) is obtained, adjusting pH is 4.5, and addition is equivalent to mixture
The cellulase of 0.1-0.2% parts by weight carries out enzymatic hydrolysis 40-60 minutes, obtains second of enzymolysis liquid, is repeated twice；
6) column chromatographs：Merge second of enzymolysis liquid, filter, concentration is chromatographed by column, inhaled with macroporous absorbent resin
Attached, it is colourless for cleaning resin to trickle with deionized water, then the ethanol solution for being 15-20% with concentration elutes, and collection is washed
De- liquid；
7) third time digests：The eluent that step 6) is obtained, adjusting pH is 4.5, and beta-glucosidase is added and is digested,
The dosage of enzyme is the 5-10% of eluent weight, and the concentration of enzyme is 1mg/ml, and hydrolysis temperature is 50-60 DEG C；
8) column chromatographs：Step 7) is obtained into enzymolysis liquid, is chromatographed by column, is adsorbed with macroreticular resin, deionized water is used
It is colourless for cleaning resin to trickle, then the ethanol solution for being 10-20% with concentration elutes, and collects eluent；
9) dry：The eluent concentration that step 8) is obtained, then by being dried to obtain powdered Geniposide.
The stacking prewetted described in step 1) of the present invention, preferably 1-2 days.
Pectin-cellulose complex enzyme described in step 3), preferably activity are ten thousand active unit of 1-3/gram (milliliter) solid-states
Or liquid pectin-fiber composite enzyme.
The preferred D101 type of macroreticular resin, DA201 type described in step 6) and step 8).
Compared with prior art, advantages of the present invention：
1, contain a large amount of grease in the plant cell of cape jasmine, be mainly made of the compound that ingredient is triglycerides mixed
Close object, these greases in the form of elaioleucite and oil body protoplast, in discontinuous graininess, together with granule protein body irregularly
Existence forms exist in the cell etc. for dispersion.Existing extracting method is led to after directly the raw materials such as cape jasmine are smashed or crushed
Cross water mention/modes such as alcohol extracting, enzymatic hydrolysis or Microwave Extraction directly extract, when extracting, the grease in cell can the same time-division
Separate out, the ingredients such as group and geniposide containing greasy, Geniposidic acid, pigment mix, thus influence it is subsequent effectively
The separation of ingredient increases the processing difficulty of process.The present invention first use enzyme extract vegetable fat, using non-oil component to oil and
The affinity difference and grease specific gravity of water are different and separate oil with non-oil component, increase product yield and improve extraction
Rate.
2, the present invention is prewetted stackings using cape jasmine, adds suitable quantity of water, is built heap and is fermented, with the rising of fermentation process temperature, Cape jasmine
Son gradually softens, and moisture enters inside naturally, conducive to the separation of vegetable fat, conducive to subsequent enzymatic hydrolysis, deoils.
3, the present invention first removes cell wall using pectin-cellulose complex enzyme, discharges the grease in plant cell, is added sweet
Reveal alcohol, osmotic pressure can be kept, conducive to the release of grease.
4, using method of the invention, geniposide high conversion rate, 80% or more.
Specific embodiment
With embodiment, the invention will be further described below, but the invention is not limited to these embodiments.
Embodiment 1：
The method that Geniposide is extracted from cape jasmine, specifically comprises the following steps：
1) it prewets stacking：Cape jasmine is built into heap in the cool, is sprayed water, after water is from the outflow of heap bottom, is stacked 1 day naturally；
2) feedstock processing：The cape jasmine that step 1) is obtained crushes, and the pure water for being equivalent to 2-3 times of weight of raw material is added, adds
Heat to 50 DEG C maintain 5 minutes, obtain paste serous material；
3) it digests for the first time：The paste serous material that step 2) is obtained, adjusting pH is 4, and 0.05% parts by weight of paste serous material are added
Activity be 10,000 active units/gram pectin-cellulose complex enzyme and 0.001 parts by weight mannitol carry out enzymatic hydrolysis 40 minutes, obtain
To first time enzymolysis liquid；
4) it deoils：The first time enzymolysis liquid that step 3) is obtained, is added the pure water of 2 times of weight, is heated to 60 DEG C of extractions
120 minutes, upper layer oil slick liquid and subnatant, slag mixture are obtained, is separated；
5) it digests for second：Subnatant, the slag mixture that step 4) is obtained, adjusting pH is 4.5, and addition is equivalent to mixture
The cellulase of 0.1% parts by weight carries out enzymatic hydrolysis 60 minutes, obtains second of enzymolysis liquid, is repeated twice；
6) column chromatographs：Merge second of enzymolysis liquid, filter, concentration, chromatographed by column, with D101 type macroporous absorbent resin into
Row absorption, it is colourless for cleaning resin to trickle with deionized water, then the ethanol solution for being 20% with concentration elutes, and collection is washed
De- liquid；
7) third time digests：The eluent that step 6) is obtained, adjusting pH is 4.5, and beta-glucosidase is added and is digested,
The dosage of enzyme is the 5% of eluent weight, and the concentration of enzyme is 1mg/ml, and hydrolysis temperature is 60 DEG C；
8) column chromatographs：Step 7) is obtained into enzymolysis liquid, is chromatographed by column, is adsorbed with D101 macroreticular resin, spend from
It is colourless that sub- water, which cleans resin to trickle, then the ethanol solution for being 20% with concentration elutes, and collects eluent；
9) dry：The eluent concentration that step 8) is obtained, then by being dried to obtain powdered Geniposide, geniposide turns
Rate is 81.1%.
Embodiment 2：
The method that Geniposide is extracted from cape jasmine, specifically comprises the following steps：
1) it prewets stacking：Cape jasmine is built into heap in the cool, is sprayed water, after water is from the outflow of heap bottom, is stacked 1 day naturally；
2) feedstock processing：The cape jasmine that step 1) is obtained is broken, and the pure water for being equivalent to 2-3 times of weight of raw material is added, adds
Heat to 60 DEG C maintain 8 minutes, obtain paste serous material；
3) it digests for the first time：The paste serous material that step 2) is obtained, adjusting pH is 4.5, and 0.1% parts by weight of paste serous material are added
Activity is that the mannitol of 20,000 active units/milliliter pectin-cellulose complex enzyme and 0.001 parts by weight carries out enzymatic hydrolysis 50 minutes,
Obtain first time enzymolysis liquid；
4) it deoils：The first time enzymolysis liquid that step 3) is obtained, is added the pure water of 3 times of weight, is heated to 70 DEG C of extractions
60 minutes, upper layer oil slick liquid and subnatant, slag mixture are obtained, is separated；
5) it digests for second：Subnatant, the slag mixture that step 4) is obtained, adjusting pH is 4.5, and addition is equivalent to mixture
The cellulase of 0.15% parts by weight carries out enzymatic hydrolysis 50 minutes, obtains second of enzymolysis liquid, is repeated twice；
6) column chromatographs：Merge second of enzymolysis liquid, filter, concentration is chromatographed by column, with DA201 type macroporous absorbent resin
It is adsorbed, it is colourless for cleaning resin to trickle with deionized water, then the ethanol solution for being 15% with concentration elutes, and collects
Eluent；
7) third time digests：The eluent that step 6) is obtained, adjusting pH is 4.5, and beta-glucosidase is added and is digested,
The dosage of enzyme is the 10% of eluent weight, and the concentration of enzyme is 1mg/ml, and hydrolysis temperature is 50 DEG C；
8) column chromatographs：Step 7) is obtained into enzymolysis liquid, is chromatographed by column, is adsorbed, is spent with DA201 type macroreticular resin
It is colourless that ionized water, which cleans resin to trickle, then the ethanol solution for being 15% with concentration elutes, and collects eluent；
9) dry：The eluent concentration that step 8) is obtained, then by being dried to obtain powdered Geniposide, geniposide turns
Rate is 82.3%.
Embodiment 3：
1) it prewets stacking：Cape jasmine is built into heap in the cool, is sprayed water, after water is from the outflow of heap bottom, is stacked 2 days naturally；
2) feedstock processing：The cape jasmine that step 1) is obtained crushes, and the pure water for being equivalent to 2-3 times of weight of raw material is added, adds
Heat to 55 DEG C maintain 10 minutes, obtain paste serous material；
3) it digests for the first time：The paste serous material that step 2) is obtained, adjusting pH is 5, and 0.08% parts by weight of paste serous material are added
Activity be 30,000 active units/gram pectin-cellulose complex enzyme and 0.001 parts by weight mannitol carry out enzymatic hydrolysis 60 minutes, obtain
To first time enzymolysis liquid；
4) it deoils：The first time enzymolysis liquid that step 3) is obtained, is added the pure water of 2 times of weight, is heated to 65 DEG C of extractions
100 minutes, upper layer oil slick liquid and subnatant, slag mixture are obtained, is separated；
5) it digests for second：Subnatant, the slag mixture that step 4) is obtained, adjusting pH is 4.5, and addition is equivalent to mixture
The cellulase of 0.2% parts by weight carries out enzymatic hydrolysis 40 minutes, obtains second of enzymolysis liquid, is repeated twice；
6) column chromatographs：Merge second of enzymolysis liquid, filter, concentration, chromatographed by column, with D101 type macroporous absorbent resin into
Row absorption, it is colourless for cleaning resin to trickle with deionized water, then the ethanol solution for being 20% with concentration elutes, and collection is washed
De- liquid；
7) third time digests：The eluent that step 6) is obtained, adjusting pH is 4.5, and beta-glucosidase is added and is digested,
The dosage of enzyme is the 8% of eluent weight, and the concentration of enzyme is 1mg/ml, and hydrolysis temperature is 55 DEG C；
8) column chromatographs：Step 7) is obtained into enzymolysis liquid, is chromatographed by column, is adsorbed, is spent with D101 type macroreticular resin
It is colourless that ionized water, which cleans resin to trickle, then the ethanol solution for being 10% with concentration elutes, and collects eluent；
9) dry：The eluent concentration that step 8) is obtained, then by being dried to obtain powdered Geniposide, geniposide turns
Rate is 82.5%.
Claims (3)
1. extracting the method for Geniposide from cape jasmine, which is characterized in that include the following steps：
1) it prewets stacking：Cape jasmine is built into heap in the cool, is sprayed water, after water is from the outflow of heap bottom, is stacked 1-2 days naturally；
2) feedstock processing：The cape jasmine that step 1) is obtained crushes or is crushed, and the pure water for being equivalent to 2-3 times of weight of raw material is added,
It is heated to 50-60 DEG C of maintenance 5-10 minutes, obtains paste serous material；
3) it digests for the first time：The paste serous material that step 2) is obtained, tune pH are 4-5, and paste serous material 0.05-0.1% weight is added
The pectin-cellulose complex enzyme of part and the mannitol of 0.001 parts by weight carry out enzymatic hydrolysis 40-60 minutes, obtain first time enzymolysis liquid；
4) it deoils：The pure water of 2-3 times of weight is added in the first time enzymolysis liquid that step 3) is obtained, and is heated to 60-70 DEG C of extraction
60-120 minutes, upper layer oil slick liquid and subnatant, slag mixture are obtained, is separated；
5) it digests for second：Subnatant, the slag mixture that step 4) is obtained, adjusting pH is 4.5, and addition is equivalent to mixture 0.1-
The cellulase of 0.2% parts by weight carries out enzymatic hydrolysis 40-60 minutes, obtains second of enzymolysis liquid, is repeated twice；
6) column chromatographs：Merge second of enzymolysis liquid, filter, concentration is chromatographed by column, adsorbed with macroporous absorbent resin, is used
It is colourless that deionized water, which cleans resin to trickle, then the ethanol solution for being 15-20% with concentration elutes, and collects eluent；
7) third time digests：The eluent that step 6) is obtained, adjusting pH is 4.5, and beta-glucosidase is added and is digested, enzyme
Dosage is the 5-10% of eluent weight, and the concentration of enzyme is 1mg/ml, and hydrolysis temperature is 50-60 DEG C；
8) column chromatographs：Step 7) is obtained into enzymolysis liquid, is chromatographed by column, is adsorbed with macroreticular resin, is cleaned with deionized water
Resin to trickle is colourless, then the ethanol solution for being 10-20% with concentration elutes, and collects eluent；
9) dry：The eluent concentration that step 8) is obtained, then by being dried to obtain powdered Geniposide.
2. the method according to claim 1 for extracting Geniposide from cape jasmine, it is characterised in that：Fruit described in step 3)
Glue-cellulose complex enzyme, for activity be ten thousand active unit of 1-3/gram solid-state or 1-3 ten thousand active units/milliliter liquid pectin-
Fiber composite enzyme.
3. the method according to claim 1 for extracting Geniposide from cape jasmine, it is characterised in that：Macropore described in step 6)
Absorption resin model is D101 type, DA201 type.
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| CN101029066A (en) * | 2007-04-13 | 2007-09-05 | 桂林莱茵生物科技股份有限公司 | Method for extracting genipin and geniposide from gardenia jasminoides |
| CN101899484A (en) * | 2009-12-28 | 2010-12-01 | 抚州市临川之信生物科技有限公司 | Preparation method of genipin |
| CN103305555A (en) * | 2013-06-28 | 2013-09-18 | 西安纽赛生物科技有限公司 | Method for converting geniposide into genipin |
| CN103695494A (en) * | 2013-12-11 | 2014-04-02 | 中华全国供销合作总社南京野生植物综合利用研究所 | Method for integral preparation of products such as crocetin, genipin, gardenia blue and the like |
| CN103788152A (en) * | 2012-10-26 | 2014-05-14 | 湖北老龙洞杜仲开发有限公司 | Method for preparing geniposide in eucommia leaf |
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2015
- 2015-08-31 CN CN201510541035.0A patent/CN105063119B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101029066A (en) * | 2007-04-13 | 2007-09-05 | 桂林莱茵生物科技股份有限公司 | Method for extracting genipin and geniposide from gardenia jasminoides |
| CN101899484A (en) * | 2009-12-28 | 2010-12-01 | 抚州市临川之信生物科技有限公司 | Preparation method of genipin |
| CN103788152A (en) * | 2012-10-26 | 2014-05-14 | 湖北老龙洞杜仲开发有限公司 | Method for preparing geniposide in eucommia leaf |
| CN103305555A (en) * | 2013-06-28 | 2013-09-18 | 西安纽赛生物科技有限公司 | Method for converting geniposide into genipin |
| CN103695494A (en) * | 2013-12-11 | 2014-04-02 | 中华全国供销合作总社南京野生植物综合利用研究所 | Method for integral preparation of products such as crocetin, genipin, gardenia blue and the like |
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Effective date of registration: 20200527 Address after: 541100 shop No.25 # 13, Guoao community, northwest Jiaojiao, Xinzhong Road, Lingui Town, Lingui District, Guilin City, Guangxi Zhuang Autonomous Region Patentee after: Guangxi youshike Food Co., Ltd Address before: 541004, fourth floor, 32 complex, Luan Luan Road, Seven Star District, Guilin, the Guangxi Zhuang Autonomous Region. Patentee before: Guangxi red Yao Pharmaceutical Co.,Ltd. |
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