CN106083985A - A kind of method that membrance separation is combined isolated and purified tea saponin with macroporous resin - Google Patents
A kind of method that membrance separation is combined isolated and purified tea saponin with macroporous resin Download PDFInfo
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- CN106083985A CN106083985A CN201610488522.XA CN201610488522A CN106083985A CN 106083985 A CN106083985 A CN 106083985A CN 201610488522 A CN201610488522 A CN 201610488522A CN 106083985 A CN106083985 A CN 106083985A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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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
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- 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
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Abstract
A kind of membrance separation and macroporous resin are combined the method for isolated and purified tea saponin, and tea seed cake is size-reduced, defat, dried, obtain tea saponin crude product by microwave-assisted ultrasonic extraction method.Crude product is after organic solvent dissolves, first through 50 nm ~ 0.2 μm ceramic membrane filters, then concentrate through 100 Da ~ 1000 Da NF membrane, finally use Amberlite XAD7HP macroporous resin adsorption and parsing, obtaining high-purity tea saponin, tea saponin purity after purification can reach more than 94%.The present invention have product purity height, favorable reproducibility, easy and simple to handle, pollute the advantages such as little, equipment one-time investment is little, operating cost is low, adapt to industrialized production and prepare.
Description
Technical field
The invention belongs to deep-processing technical field of agricultural products, relate to the isolation and purification method of tea saponin in tea seed cake.
Background technology
Tea saponin be a class with Triterpenoids sapogenins as parent nucleus, collateral branch is with different glucosides and the saponin of functional group
The general name of compound.Tea saponin not only has the biological activitys such as antibacterial, antiinflammatory, pain relieving, reduction lipid cholesterol, also has breast
The performances such as change, dispersion, moistening, foaming, steady bubble, are a kind of excellent natural surfactants, are widely used in building materials, change
The industries such as work, biological medicine and food, application prospect is the most wide.
Oil tea carries in the tea seed cake after oil rich in tea saponin, is a kind of good source extracting tea saponin.Pure tea soap
Element is in colorless prismatic crystal, and owing to tea saponin is of a great variety, its structure is complicated and similar, and isomers is many, isolates single
The tea saponin sterling of structure is abnormal difficult, causes the purity selling tea saponin product in current China market the highest so that it is
Application at Fine Chemicals such as medicine and cosmetics is subject to certain restrictions.Therefore, it is possible to efficiently prepare highly purified tea
The separating and purifying technology of Saponin is problem demanding prompt solution during camellia oleiferaindustry.
The isolation and purification method of common tea saponin mainly has n-butanol extraction, precipitation flocculence, salting out method, silica gel column layer
Analysis method, purification by macroporous resin method, membrane separation process etc..But these methods all come with some shortcomings.As used n-butanol extraction
The tea saponin impurity obtained is more；It is many, to environment that precipitation flocculence and salting out method consume reagent；Silica gel column chromatography
Tea saponin is easily caused dead absorption, yields poorly, be not suitable for commercial production.And membrane separation process has material and does not undergoes phase transition, divides
From effective, consume energy the advantages such as low, technique is simple, easy and simple to handle, cleanliness without any pollution；Resin purification method has stability height, choosing
Selecting property is good, mild condition, low power consuming, be prone to automatization, be conducive to the advantage such as environmental conservation.Membrance separation is tied with resin purification
Close, it is possible to obtain the tea saponin that purity is higher, and resin is reused number of times and is greatly improved, and can reduce purification cost.
Summary of the invention
The purpose of the present invention is to propose to a kind of method that membrance separation is combined isolated and purified tea saponin with macroporous resin.
The method that a kind of membrance separation of the present invention is combined isolated and purified tea saponin with macroporous resin, according to the following steps.
(1) preparation of tea saponin crude product: tea seed cake is pulverized, sieves, used 2 ~ 8 times of petroleum ether or normal hexane, reflux extraction 1 ~ 3
Hour extracting remove oil residues fat.Afterwards, using dry microwave to process defat tea seed cake, microwave power is 100 W ~ 800 W, the time
It is 1 min ~ 10 min.Being subsequently adding 5 ~ 30 times of mass percent 50 ~ 80% ethanol solution, supersound extraction, diverging power is 1200
W ~ 1500 W, temperature is 40 ~ 60 DEG C, supersound extraction 0.5 ~ 1 h.By the volatilization of extracting solution evaporation and concentration to ethanol completely, with 1 ~ 10
Times volume ethyl ketone: the concentrated solution of ether mixed liquor (1:1) washing tea saponin, discards organic layer.Water layer adds 1 ~ 10 times of volume
N-butanol extraction, makes tea saponin transfer in alcohol layer, adds activated carbon decolorizing, evaporation and concentration, is dried to obtain tea saponin crude product.
(2) prepared by sample: weigh tea saponin dissolving crude product that a certain amount of step (1) obtains in mass percent 50 ~ 80%
In ethanol solution, it is configured to the tea saponin solution that mass percent is 1% ~ 10%.
(3) ceramic membrane filter: use the tea saponin solution that 50 nm ~ 0.2 μm ceramic membrane filter step (2) obtain.
(4) NF membrane concentrates: use the tea saponin solution that 100 Da ~ 1000 Da NF membrane concentration step (3) obtain.
(5) macroporous resin is further purified: first load in post by Amberlite XAD7HP macroporous resin, then with 0.3
~ 0.6 mL/min flow velocity, the tea saponin that step (4) is obtained with concentration 20 ~ 50 mg/mL loading, with 10%, 30 ~ 50%, 60 ~
Each 2 ~ 5 BV of ethanol of 80%, carry out gradient elution under flow velocity 0.5 ~ 1.5 mL/min, collect 30 ~ 50% and 60 ~ 80% ethanol
Eluent.
(6) eluent that step (5) is collected the ethanol obtained concentrates, is dried, and obtains high purity white powder tea soap
Element.
The present invention is mainly characterized by.
1, reasonable in design, easy and simple to handle, favorable reproducibility, tea saponin purity are up to more than 94%.
2, according to membrance separation principle, ceramic membrane can retain the macromole impurity such as most protein, purification effect
Good；And NF membrane can pass through the small-molecule substance such as water, inorganic salt, concentrated effect is good.
3, according to similar principle and the macroporous resin adsorption principle of mixing, the polar macroporous tree of Amberlite XAD7HP is selected
Fat, specific surface area is big, and average pore size is relatively big, and isolated and purified tea saponin is effective.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail, it should be understood that the scope of the present invention is non-is only limitted to this
The scope of a little embodiments.
Embodiment 1.
(1) preparation of tea saponin crude product: tea seed cake pulverized, crosses 40 mesh sieves, is heated to reflux defat 2 hours with 4 times of petroleum ether,
Precipitation is dried.Weighing defat tea seed cake 100 g, regulation microwave power is 1000 W, and the dry process time is 6 min.It is subsequently adding
The ethanol water of 700 mL 70%, the power that regulation dissipates is 1200 W, and temperature is 50 DEG C, ultrasonic and circulated extraction 1 h, filters
Obtain extracting solution.By the volatilization of extracting solution evaporation and concentration to ethanol completely, then with 1 times of volume ethyl ketone: ether mixed liquor (1:1) washs
The concentrated solution of tea saponin, discards organic layer.Water layer adds the n-butanol extraction of 3 times of volumes, makes tea saponin transfer to, in alcohol layer, add
Enter activated carbon decolorizing, evaporation and concentration, be dried to obtain tea saponin crude product.
(2) prepared by sample: weighs certain tea saponin crude product and is dissolved in the ethanol water of certain volume 70%, is configured to 1%
Tea saponin solution.
(3) ceramic membrane filter: by 1% tea saponin solution through 0.14 μm ceramic membrane, operation pressure is 3.5 Bar, carries out
Infinite reflux endless form is tested, and collects permeate.
(4) NF membrane concentrates: use 360 Da NF membrane to concentrate the permeate after ceramic membrane filter, operates pressure
It is 11 Bar, tea saponin permeate 2600 mL of ceramic membrane separation of learning from else's experience, it is concentrated to give concentrated solution 750 through 360 Da nanofiltrations
ML, cycles of concentration is 3.5 times, concentrates experiment and lasts 22 min.
(5) Amberlite XAD7HP purification by macroporous resin: weigh the Amberlite XAD7HP that 100 g pretreatment is good
Macroporous resin, wet method dress post, rushes post with 95% ethanol, then with distilled water punching to without alcohol taste.By above-mentioned the purest through membrance separation
Change the tea saponin obtained, be made into the aqueous solution that concentration is 30 mg/mL, with 0.5 mL/min speed loading 250 mL tea saponin sample
Product, stand 2 h after end of the sample so that it is absorption is completely.It is colourless for being washed till effluent with distilled water, with 10%, 40%, 70%
Each 3 BV of ethanol, carry out gradient elution under flow velocity 1 mL/min, collect the eluent of 40% and 70% ethanol, carry out tea saponin
It is further purified.Adsorption rate is conciliate adsorption rate and is up to 92% and 94% respectively, finally the eluent of ethanol is concentrated, is obtained after drying
White powder tea saponin.Final yield is 55.3%, and purity is 95%.
Embodiment 2.
(1) preparation of tea saponin crude product: tea seed cake pulverized, crosses 40 mesh sieves, is heated to reflux defat 1.5 with 5 times of normal hexane little
Time, precipitation is dried.Weighing defat tea seed cake 100 g, regulation microwave power is 1000 W, and the dry process time is 5 min.Then add
Entering the ethanol water of 500 mL 60%, the power that regulation dissipates is 1300 W, and temperature is 60 DEG C, ultrasonic and circulated extraction 1 h, mistake
Filter to obtain extracting solution.By the volatilization of extracting solution evaporation and concentration to ethanol completely, then by 1 times of volume ethanol: ether mixed liquor (1:1) is washed
Wash the concentrated solution of tea saponin, discard organic layer.Water layer adds the n-butanol extraction of 4 times of volumes, makes tea saponin transfer in alcohol layer,
Add activated carbon decolorizing, evaporation and concentration, be dried to obtain tea saponin crude product.
(2) prepared by sample: weighs certain tea saponin crude product and is dissolved in the ethanol water of certain volume 80%, is configured to 5%
Tea saponin solution.
(3) ceramic membrane filter: by 5% tea saponin solution through 0.2 μm ceramic membrane, operation pressure is 4 Bar, entirely returns
Stream endless form experiment, collects permeate.
(4) NF membrane concentrates: using 360 Da NF membrane, operation pressure is 12 Bar, to passing through after ceramic membrane filter
Liquid concentrates.
(5) Amberlite XAD7HP purification by macroporous resin: weigh the Amberlite XAD7HP that 100 g pretreatment is good
Macroporous resin, wet method dress post, rushes post with 90% ethanol, then with distilled water punching to without alcohol taste.By above-mentioned the purest through membrance separation
Change the tea saponin obtained, be made into the aqueous solution that concentration is 40 mg/mL, with 0.7 ml/min speed loading 200 mL tea saponin sample
Product, stand 2 h after end of the sample so that it is absorption is completely.It is colourless for being washed till effluent with distilled water, with 10%, 45%, 75%
Each 3 BV of ethanol, carry out gradient elution under flow velocity 1 mL/min, collect the eluent of 45% and 75% ethanol, carry out tea saponin
It is further purified.Finally the eluent of ethanol concentrated, obtain white powder tea saponin after drying.Final yield is 53%, pure
Degree is 94.1%.
Embodiment 3.
(1) preparation of tea saponin crude product: tea seed cake pulverized, crosses 40 mesh sieves, is heated to reflux defat 2 hours with 3 times of petroleum ether,
Precipitation is dried.Weighing defat tea seed cake 100 g, regulation microwave power is 800 W, and the dry process time is 6 min.It is subsequently adding
The ethanol water of 800 mL 80%, the power that regulation dissipates is 1500 W, and temperature is 60 DEG C, ultrasonic and circulated extraction 0.5 h, mistake
Filter to obtain extracting solution.By the volatilization of extracting solution evaporation and concentration to ethanol completely, then with 1 times of volume ethyl ketone: ether mixed liquor (1:1) is washed
Wash the concentrated solution of tea saponin, discard organic layer.Water layer adds the n-butanol extraction of 4 times of volumes, makes tea saponin transfer in alcohol layer,
Add activated carbon decolorizing, evaporation and concentration, be dried to obtain tea saponin crude product.
(2) prepared by sample: weighs certain tea saponin crude product and is dissolved in the ethanol water of certain volume 75%, is configured to 2%
Tea saponin solution.
(3) ceramic membrane filter: by 2% tea saponin solution through 0.1 μm ceramic membrane, operation pressure is 3.5 Bar, carries out complete
Reflux cycle mode is tested, and collects permeate.
(4) NF membrane concentrates: using 150 Da NF membrane, operation pressure is 10 Bar, to passing through after ceramic membrane filter
Liquid concentrates.
(5) Amberlite XAD7HP purification by macroporous resin: weigh the Amberlite XAD7HP that 100 g pretreatment is good
Macroporous resin, wet method dress post, rushes post with 95% ethanol, then with distilled water punching to without alcohol taste.By above-mentioned the purest through membrance separation
Change the tea saponin obtained, be made into the aqueous solution that concentration is 30 mg/mL, with 0.5 ml/min speed loading 250 mL tea saponin sample
Product, stand 2 h after end of the sample so that it is absorption is completely.It is colourless for being washed till effluent with distilled water, with 10%, 40%, 75%
Each 4 BV of ethanol, carry out gradient elution under flow velocity 1 mL/min, collect the eluent of 40% and 75% ethanol, carry out tea saponin
It is further purified.Finally the eluent of ethanol concentrated, obtain white powder tea saponin after drying.Final yield is 56%, pure
Degree is 96.3%.
Claims (1)
1. the method that membrance separation is combined isolated and purified tea saponin with macroporous resin, is characterized in that according to the following steps:
(1) tea seed cake pulverized, sieve, use 2 ~ 8 times of petroleum ether or normal hexane, backflow extraction extracting in 1 ~ 3 hour to remove oil residues
Fat；Afterwards, using dry microwave to process defat tea seed cake, microwave power is 100 W ~ 800 W, and the time is 1 min ~ 10 min；So
5 ~ 30 times of mass percent 50 ~ 80% ethanol solution of rear addition, supersound extraction, diverging power is 1200 W ~ 1500 W, and temperature is
40 ~ 60 DEG C, supersound extraction 0.5 ~ 1 h；By the volatilization of extracting solution evaporation and concentration to ethanol completely, 1:1 volume is pressed with 1 ~ 10 times of volume
Than ethyl ketone and the concentrated solution of the mixed liquor washing tea saponin of ether of mixing, discard organic layer；Water layer adds 1 ~ 10 times of volume
N-butanol extraction, makes tea saponin transfer in alcohol layer, adds activated carbon decolorizing, evaporation and concentration, is dried to obtain tea saponin crude product；
(2) weigh tea saponin dissolving crude product that step (1) obtains in the ethanol solution of mass percent 50 ~ 80%, be configured to matter
Amount percentage ratio is the tea saponin solution of 1% ~ 10%；
(3) the tea saponin solution that 50 nm ~ 0.2 μm ceramic membrane filter step (2) obtain is used；
(4) the tea saponin solution that 100 Da ~ 1000 Da NF membrane concentration step (3) obtain is used；
(5) first Amberlite XAD7HP macroporous resin is loaded in post, then with 0.3 ~ 0.6 mL/min flow velocity, by step
(4) tea saponin obtained is with concentration 20 ~ 50 mg/mL loading, and each 2 ~ 5 BV of ethanol with 10%, 30 ~ 50%, 60 ~ 80%, at flow velocity
Carry out gradient elution under 0.5 ~ 1.5 mL/min, collect the eluent of 30 ~ 50% and 60 ~ 80% ethanol；
(6) eluent that step (5) is collected the ethanol obtained concentrates, is dried, and obtains high purity white powder tea saponin.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383153A (en) * | 2017-08-20 | 2017-11-24 | 合肥信达膜科技有限公司 | A kind of extracting method of high-purity tea saponin |
| CN108003215A (en) * | 2017-12-25 | 2018-05-08 | 华远医药研究院有限公司 | The cleaning extracting method of Tea Saponin |
| CN108299542A (en) * | 2018-03-03 | 2018-07-20 | 广西师范大学 | A method of from the withered middle comprehensive extraction saponin of oil tea and polyphenol |
| CN109824638A (en) * | 2019-04-01 | 2019-05-31 | 山东笑康生物科技有限公司 | A method of the Ti Qu Ash skin ketone from cortex dictamni |
| CN110794075A (en) * | 2019-11-21 | 2020-02-14 | 华侨大学 | Analysis method of oil-tea camellia seed oil saponin compounds |
| CN110934794A (en) * | 2019-12-18 | 2020-03-31 | 宁夏艾伊生物工程研发有限公司 | Shampoo special for industrial enterprises |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383153A (en) * | 2017-08-20 | 2017-11-24 | 合肥信达膜科技有限公司 | A kind of extracting method of high-purity tea saponin |
| CN108003215A (en) * | 2017-12-25 | 2018-05-08 | 华远医药研究院有限公司 | The cleaning extracting method of Tea Saponin |
| CN108299542A (en) * | 2018-03-03 | 2018-07-20 | 广西师范大学 | A method of from the withered middle comprehensive extraction saponin of oil tea and polyphenol |
| CN108299542B (en) * | 2018-03-03 | 2020-12-08 | 广西师范大学 | Method for comprehensively extracting saponin and polyphenol from camellia oleifera abel cake |
| CN109824638A (en) * | 2019-04-01 | 2019-05-31 | 山东笑康生物科技有限公司 | A method of the Ti Qu Ash skin ketone from cortex dictamni |
| CN110794075A (en) * | 2019-11-21 | 2020-02-14 | 华侨大学 | Analysis method of oil-tea camellia seed oil saponin compounds |
| CN110794075B (en) * | 2019-11-21 | 2022-03-15 | 华侨大学 | Analysis method of oil-tea camellia seed oil saponin compounds |
| CN110934794A (en) * | 2019-12-18 | 2020-03-31 | 宁夏艾伊生物工程研发有限公司 | Shampoo special for industrial enterprises |
| CN110934794B (en) * | 2019-12-18 | 2021-07-16 | 宁夏艾伊生物工程研发有限公司 | Shampoo special for industrial enterprises |
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Application publication date: 20161109 |