CN102432664A - Method for extracting theasaponin from tea seed pulp - Google Patents
Method for extracting theasaponin from tea seed pulp Download PDFInfo
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- CN102432664A CN102432664A CN2011104520904A CN201110452090A CN102432664A CN 102432664 A CN102432664 A CN 102432664A CN 2011104520904 A CN2011104520904 A CN 2011104520904A CN 201110452090 A CN201110452090 A CN 201110452090A CN 102432664 A CN102432664 A CN 102432664A
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Abstract
The invention relates to a method for extracting theasaponin from tea seed pulp. In the method, theasaponin is extracted by enzymolysis and ultrasonic treatment. The method is technically characterized by comprising the following steps: crushing the tea seed pulp, degreasing with petroleum ether, dissolving in water, adding single or composite bioenzyme to carry out enzymolysis reaction, filtering, ultrasonically leaching the filter residue, combining water extract, flocculating, precipitating with calcium oxide, dissolving in ethanol, decolorizing and drying to obtain higher- purity theasaponin. By adopting the method provided by the invention, the consumption amount of the organic solvent is less, the cost is low, the extraction yield is high, and the requirements of national clean production are conformed.
Description
Technical field
The invention belongs to agriculture and forestry by-product deep processing field, be specifically related to a kind of through enzymolysis and the ultrasonic auxiliary method of from the tea seed, extracting theasaponin.
Background technology
The theasaponin that from the depleted leached tea oil slag, extracts has good surface properties, has obtained in fields such as agricultural, aquaculture, daily-use chemical industry, medicine, building materials using widely.And traditional organic solvent extraction method (Yang Kunguo; Huang Mingquan; With the absolute ethyl alcohol is the tea saponin Study on Extraction Method of extraction agent. Hubei Institute For Nationalities's journal natural science edition, 2000.18 (3): 19-21) have problems such as organic solvent consumption is big, cost is high, seriously polluted.In recent years, constantly have the scholar that the process for extracting of theasaponin is improved: people such as the Liu Changsheng of Inst. of Oil Crops, Chinese Academy of Agriculture adopt supersonic method to assist ethanol lixiviate theasaponin (Liu Changsheng, Huang Fenghong; Xia Fujian, etc., the technical study of ultrasonic extraction method tea saponin; China's oil crops journal, 2006,28 (2): 203-206); Shortened extraction time, yield also improves a lot.But its used extraction solvent is an ethanol still, and cost is higher, and has potential safety hazard.Wang Gu very utilize the weak ammonia extraction theasaponin (Wang Guqi. ammonia process extracts tea saponin. natural generation research and development, 1991,3 (4): 94.97), extraction yield improves a lot, but ammoniacal liquor has pungency, foreign matter content is higher in the products obtained therefrom simultaneously.
Summary of the invention
The objective of the invention is to solve industry and go up problems such as the organic solvent consumption that exists in the extraction theasaponin process is big, cost is high, seriously polluted, it is high to have proposed a kind of not only extraction yield, and the method for from leached tea oil slag, extracting theasaponin of extraction process cleaning.
For achieving the above object, the method that the present invention adopts is following:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.0～5.5 backs are 10 to wherein adding AMS, the 100～200ml vigor that 50～80g vigor is 4000u/g
5The saccharifying enzyme of u/ml and 10～30g vigor are 10
4In the cellulase of u/g one or more; Stir enzymolysis 2～5h at 40～60 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out, filter and obtain filter residue and enzymolysis filtrating; In filter residue, add 50L water; At 50～70 ℃, 15～35kHz supersound extraction, 1～2h, get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 20～50g Tai-Ace S 150, at 30～50 ℃ of flocculation 0.5～2.0h, remove water-soluble protein and tannin impurity in the extract that anhydrates, flocculation then uses rotating speed to obtain clarifying upper water extract as the whizzer of 4000r/min is centrifugal;
4) in clarifying upper water extract, add 500～600g quicklime and stir, in 45 ℃ of reaction 1～3h, generate white precipitate, after the filtration, it is 60～85% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got white theasaponin powder.
Because the present invention adopts water to extract as solvent, and is less relatively to the influence that environment causes.The glycase that adopts simultaneously can make solution viscosity reduce, and cellulase, ultrasonication can destroy the cell walls of tea seed, and these effects all help the stripping of theasaponin, makes extraction yield be tending towards maximization.
Embodiment
Embodiment 1:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.5 backs are 10 to wherein adding AMS and the 100ml vigor that the 50g vigor is 4000u/g
5The saccharifying enzyme of u/ml stirs enzymolysis 2h at 50 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out; Filtration obtains filter residue and enzymolysis filtrating, in filter residue, adds 50L water, at 70 ℃, 15kHz supersound extraction 1h; Get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 20g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 30 ℃ of flocculation 0.5h;
4) in clarifying upper water extract, add the 580g quicklime and stir, in 45 ℃ of reaction 1h, generate white precipitate, after the filtration, it is 70% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 842g white theasaponin powder.
Embodiment 2:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.0 backs are 10 to wherein adding the 25g vigor
4The cellulase of u/g stirs enzymolysis 5h at 45 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out; Filtration obtains filter residue and enzymolysis filtrating, in filter residue, adds 50L water, at 70 ℃, 20kHz supersound extraction 2h; Get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 30g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 40 ℃ of flocculation 1h;
4) in clarifying upper water extract, add the 520g quicklime and stir, in 45 ℃ of reaction 1h, generate white precipitate, after the filtration, it is 75% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 698g white theasaponin powder.
Embodiment 3:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.3 backs are 10 to wherein adding AMS, the 150ml vigor that the 60g vigor is 4000u/g
5The saccharifying enzyme of u/ml and 10g vigor are 10
4In the cellulase of u/g one or more; Stir enzymolysis 4h at 50 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out, filter and obtain filter residue and enzymolysis filtrating; In filter residue, add 50L water; At 65 ℃, 30kHz supersound extraction 2h, get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 50g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 45 ℃ of flocculation 1.5h;
4) in clarifying upper water extract, add the 600g quicklime and stir, in 45 ℃ of reaction 3h, generate white precipitate, after the filtration, it is 85% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 822g white theasaponin powder.
Embodiment 4:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.4 backs are to wherein adding the AMS that the 80g vigor is 4000u/g, at 60 ℃ of stirring enzymolysis 5h; Behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out; Filtration obtains filter residue and enzymolysis filtrating, in filter residue, adds 50L water, at 50 ℃, 35kHz supersound extraction 1h; Get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 40g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 50 ℃ of flocculation 1h;
4) in clarifying upper water extract, add the 540g quicklime and stir, in 45 ℃ of reaction 2.5h, generate white precipitate, after the filtration, it is 80% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 795g white theasaponin powder.
Embodiment 5:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.2 backs are 10 to wherein adding the 200ml vigor
5The saccharifying enzyme of u/ml and 30g vigor are 10
4The cellulase of u/g stirs enzymolysis 5h at 40 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out; Filtration obtains filter residue and enzymolysis filtrating, in filter residue, adds 50L water, at 70 ℃, 30kHz supersound extraction 2h; Get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 35g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 30 ℃ of flocculation 1.5h;
4) in clarifying upper water extract, add the 545g quicklime and stir, in 45 ℃ of reaction 1h, generate white precipitate, after the filtration, it is 65% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 772g white theasaponin powder.
Embodiment 6:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.1 backs are 10 to wherein adding the 180ml vigor
5The saccharifying enzyme of u/ml stirs enzymolysis 4.5h at 60 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out; Filtration obtains filter residue and enzymolysis filtrating, in filter residue, adds 50L water, at 60 ℃, 20kHz supersound extraction 2h; Get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 45g Tai-Ace S 150,, remove water-soluble protein and tannin impurity in the extract that anhydrates, use rotating speed to obtain clarifying upper water extract after the flocculation as the whizzer of 4000r/min is centrifugal at 40 ℃ of flocculation 2h;
4) in clarifying upper water extract, add the 500g quicklime and stir, in 45 ℃ of reaction 2h, generate white precipitate, after the filtration, it is 60% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got 748g white theasaponin powder.
In order to verify feasibility of the present invention, carried out following experiment:
1. get equivalent de-oiling leached tea oil slag; Experimentize with traditional ethanolic soln extraction method and method of the present invention respectively, not only extraction yield is high by 15～20% than alcohol extracting method to find method of the present invention, and purity also improves a lot; Avoid the mass consumption organic solvent simultaneously, fundamentally alleviated pollution.
2. traditional ethanolic soln extraction method and the resulting theasaponin product of method of the present invention are carried out structure detection; Find both composition basically identicals; Explain that the theasaponin structure is not destroyed in the leaching process, numerous surfactivitys that theasaponin had have simultaneously also obtained maintenance.
This shows that the present invention has also further improved the extraction yield of theasaponin when reducing production costs, alleviated pollution, can be used for suitability for industrialized production.
Claims (1)
1. method of from leached tea oil slag, extracting theasaponin is characterized in that:
1) get the 5kg leached tea oil slag, be crushed to 40～60 orders with kibbler, remaining tea oil is removed with sherwood oil in dry back, obtains the de-oiling leached tea oil slag;
2) in the de-oiling leached tea oil slag, add 40L water, and adjusting pH value is that 5.0～5.5 backs are 10 to wherein adding AMS, the 100～200ml vigor that 50～80g vigor is 4000u/g
5The saccharifying enzyme of u/ml and 10～30g vigor are 10
4In the cellulase of u/g one or more; Stir enzymolysis 2～5h at 40～60 ℃, behind the enzymolysis temperature is risen to 90 ℃ of enzyme 5min that go out, filter and obtain filter residue and enzymolysis filtrating; In filter residue, add 50L water; At 50～70 ℃, 15～35kHz supersound extraction, 1～2h, get ultrasonic filtrating after the filtration, merge enzymolysis filtrating and ultrasonic filtrating and obtain the water extract;
3) in the water extract, add 20～50g Tai-Ace S 150, at 30～50 ℃ of flocculation 0.5～2.0h, remove water-soluble protein and tannin impurity in the extract that anhydrates, flocculation then uses rotating speed to obtain clarifying upper water extract as the whizzer of 4000r/min is centrifugal;
4) in clarifying upper water extract, add 500～600g quicklime and stir, in 45 ℃ of reaction 1～3h, generate white precipitate, after the filtration, it is 60～85% dissolve with ethanol that deposition is used volumetric concentration, crosses and filters the theasaponin ethanolic soln;
5) in the theasaponin ethanolic soln, add gac, boil 30min, filter, the solution for vacuum concentration that obtains, spraying drying are got white theasaponin powder.
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102942604A (en) * | 2012-11-19 | 2013-02-27 | 浙江理工大学 | Method for separation and purification of tea saponin through foam |
| CN102952171A (en) * | 2012-11-06 | 2013-03-06 | 安徽新世界绿洲茶油有限公司 | Method for extracting camellia oleifera seed protein from camellia oleifera seed meal |
| CN106083984A (en) * | 2016-06-26 | 2016-11-09 | 周荣 | A kind of method from extracting tea saponin from tea seed meal |
| CN106967144A (en) * | 2017-03-28 | 2017-07-21 | 钦州市桂耘贸易有限公司 | A kind of method that Tea Saponin is extracted from tea bran |
| CN107056876A (en) * | 2017-05-22 | 2017-08-18 | 江西乔盛茶皂素科技有限公司 | A kind of method that high-purity tea saponin is extracted from camellia seed meal |
| CN107334678A (en) * | 2017-07-18 | 2017-11-10 | 安徽省华银茶油有限公司 | A kind of method that antibacterial wet tissue is prepared using camellia seed dregs of rice extract |
| CN107625688A (en) * | 2017-10-18 | 2018-01-26 | 湖南大三湘茶油电子商务有限公司 | Camellia seed extract solution and preparation method thereof |
| CN108774283A (en) * | 2018-04-13 | 2018-11-09 | 华侨大学 | A kind of method that high-pressure pulse electric coupling biological enzymolysis prepares Tea Saponin |
| CN110141591A (en) * | 2019-05-23 | 2019-08-20 | 佛山市欧若拉生物科技有限公司 | A kind of saikoside preparation method |
| CN111449090A (en) * | 2020-04-13 | 2020-07-28 | 湖南润农生态茶油有限公司 | Preparation method of tea seed cake extract |
| CN113024629A (en) * | 2021-01-05 | 2021-06-25 | 贵州乔盛生物科技有限公司 | Tea saponin purification method |
| CN113801172A (en) * | 2021-09-14 | 2021-12-17 | 广州市润研基因科技有限公司 | Method for separating protein in soapberry saponin water extract |
| CN115286680A (en) * | 2022-07-08 | 2022-11-04 | 南昌大学 | Method for extracting tea saponin from tea seed meal |
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Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102952171A (en) * | 2012-11-06 | 2013-03-06 | 安徽新世界绿洲茶油有限公司 | Method for extracting camellia oleifera seed protein from camellia oleifera seed meal |
| CN102942604A (en) * | 2012-11-19 | 2013-02-27 | 浙江理工大学 | Method for separation and purification of tea saponin through foam |
| CN106083984A (en) * | 2016-06-26 | 2016-11-09 | 周荣 | A kind of method from extracting tea saponin from tea seed meal |
| CN106083984B (en) * | 2016-06-26 | 2018-02-13 | 福建三九军大生物科技有限公司 | A kind of method from extracting tea saponin from tea seed meal |
| CN106967144A (en) * | 2017-03-28 | 2017-07-21 | 钦州市桂耘贸易有限公司 | A kind of method that Tea Saponin is extracted from tea bran |
| CN107056876B (en) * | 2017-05-22 | 2019-05-24 | 江西乔盛茶皂素科技有限公司 | A method of extracting high-purity tea saponin from camellia seed meal |
| CN107056876A (en) * | 2017-05-22 | 2017-08-18 | 江西乔盛茶皂素科技有限公司 | A kind of method that high-purity tea saponin is extracted from camellia seed meal |
| CN107334678A (en) * | 2017-07-18 | 2017-11-10 | 安徽省华银茶油有限公司 | A kind of method that antibacterial wet tissue is prepared using camellia seed dregs of rice extract |
| CN107625688A (en) * | 2017-10-18 | 2018-01-26 | 湖南大三湘茶油电子商务有限公司 | Camellia seed extract solution and preparation method thereof |
| CN108774283A (en) * | 2018-04-13 | 2018-11-09 | 华侨大学 | A kind of method that high-pressure pulse electric coupling biological enzymolysis prepares Tea Saponin |
| CN110141591A (en) * | 2019-05-23 | 2019-08-20 | 佛山市欧若拉生物科技有限公司 | A kind of saikoside preparation method |
| CN111449090A (en) * | 2020-04-13 | 2020-07-28 | 湖南润农生态茶油有限公司 | Preparation method of tea seed cake extract |
| CN113024629A (en) * | 2021-01-05 | 2021-06-25 | 贵州乔盛生物科技有限公司 | Tea saponin purification method |
| CN113801172A (en) * | 2021-09-14 | 2021-12-17 | 广州市润研基因科技有限公司 | Method for separating protein in soapberry saponin water extract |
| CN113801172B (en) * | 2021-09-14 | 2022-09-06 | 广州市润研基因科技有限公司 | Method for separating protein in soapberry saponin water extract |
| CN115286680A (en) * | 2022-07-08 | 2022-11-04 | 南昌大学 | Method for extracting tea saponin from tea seed meal |
| CN115286680B (en) * | 2022-07-08 | 2024-03-29 | 南昌大学 | Method for extracting tea saponin from tea seed meal |
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Application publication date: 20120502 |