CN101633676B - Method for preparing high-purity stachyose by using plant chromatographic separation technology - Google Patents
Method for preparing high-purity stachyose by using plant chromatographic separation technology Download PDFInfo
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Abstract
The invention discloses a method for preparing high-purity stachyose by using a plant chromatographic separation technology, comprising the following steps: (1) squeezing juice: leaching dry Chinese ortichoke or silvery strips, squeezing juice, extracting residues, leaching and re-squeezing by adding water, and mixing juice twice and taking the juice as sugar liquid; (2) removing impurities; (3) decolorizing; (4) desalting; (5) filtering by a film; (6) concentrating; (7) separating a chromatogram: selecting porous gel resin as an industrial chromatogram fixing phrase, and separating sugar content in Brix sugar liquid of 40-60 DEG C by using a size exclusion principle; (8) concentrating and drying spray to obtain high-purity stachyose. The method applies a size exclusion chromatographic technology to purify functional oligosaccharide and can successfully obtain the high-purity stachyose from natural plants and provide a technical platform for purifying other kinds of functional oligosaccharide; the separated purity is higher than 90 percent (a radio of stachyose to total sugar), and the stachyose has higher yield and lower loss ratio.
Description
Technical field
The present invention relates to a kind of preparation method of stachyose, particularly relate to the method that the industrial chromatographic separation technology of a kind of usefulness prepares the high purity water threose.
Background technology
Stachyose is the naturally occurring functional oligose that can significantly promote human intestinal profitable strain propagation in the Labiatae betony, is described as " natural superpower bifidus factor ".The stachyose structure is clear and definite, molecular formula C
24H
42O
21, molecular weight 666.59, it is made up of fructose, glucose, semi-lactosi, semi-lactosi polymerization, and simple structural table is shown fructose-Glucose-Galactose-semi-lactosi.
The stachyose product is nearly product innovation that developed in 10 years, it be a kind of be the carbohydrate admixture of main component with the stachyose.The technology of extracting the stachyose product from raw materials such as Rhizome of Bear's-foot Fern is ripe, and the product stachyose content that can provide in the market (accounting for the total reducing sugar quality) is about 70%.
The final gained stachyose content of stachyose preparation technology mentioned in " research that stachyose prepares gordian technique " (the Liang Li's heart) that read up the literature be up to 73.10%, mono-and di-saccharides content reaches 15%.The stachyose content that document " extraction and separation process of oligosaccharides research in the glutinous rehmannia " (Zhang Ruxue etc.) adopts water extraction to obtain is up to 60.51%.
Chinese patent application numbers 200510071045.9 " based on the raffinose family oligose and the production method thereof of stachyose; disclosed be to glutinous rehmannia, Rhizome of Bear's-foot Fern, the water extracts of plants such as bamboo shoot cooperate clarify and decolorize with chitosan and gac; 1-4 group sun-anionite-exchange resin cooperates and carries out desalination, and finally obtaining stachyose content is 62-71%.Chinese patent application number 01109692.6 " a kind of method of utilizing gestumor piece root to produce two qi factor oligose ", disclosed is after utilizing gestumor to carry for raw material passes through water, again through remove slag, removal of impurities, decolouring, filtration, refining, concentrate and technological process such as spraying drying makes Icing Sugar, wherein stachyose content is about 75%.Patent " method of Nanofiltering membrane purifying soybean oligosaccharide (application number 01101966.2) " is mentioned and please be handled with nanofiltration membrane by water separating the big soya-bean milk of big soya-bean milk after please albumen, slough salt wherein, the content of stachyose and raffinose in the raising soybean oligosaccharide.
It is raw material that above technology all adopts plants such as glutinous rehmannia, Rhizome of Bear's-foot Fern, and the stachyose content that obtains accounts for the 60-75% of total reducing sugar, and mono-and di-saccharides content is 15-30%.Its functional oligose purity is not high, has limited audient crowd and Application Areas.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, provide the industrial chromatographic separation technology of a kind of usefulness to prepare the method for high purity water threose.
Technical scheme of the present invention is summarized as follows:
The industrial chromatographic separation technology of a kind of usefulness prepares the method for high purity water threose, comprises the steps:
(1) squeeze the juice: in mass ratio is 1: the ratio of 3-6, drying stachys sieboldii miq or argentate strip are added water in 50 ℃ of-90 ℃ of lixiviates 0.5 hour-1.5 hours, squeeze the juice, get residue, add 1 times-3 times 0.5 hour-1.5 hours multiple press for extracting juicees of 50 ℃ of-90 ℃ of lixiviates of water to the residue quality, mixing twice squeezeding juice is liquid glucose;
(2) removal of impurities: described liquid glucose is warming up to 50-100 ℃, heated 10-30 minute, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 5-12g: 1L, staticly settles 30-90 minute, gets supernatant liquor;
(3) decolouring: in the ratio of 1L: 4-10g, in supernatant liquor, add gac, be heated to 60-90 ℃, decoloured 30-60 minute, remove by filter gac;
(4) desalination: the liquid after the decolouring is cooled to 20-30 ℃, through Zeo-karb, anionite-exchange resin specific conductivity is reduced to below the 150 μ s/cm successively;
(5) membrane filtration: is that 5000～100000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.01-0.08MPa is concentrated into 40-60 ° of Brix liquid glucose with the liquid behind the membrane filtration at 40～80 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, utilizes the size exclusion principle that the sugar in the 40-60 ° of Brix liquid glucose is separated;
(8) concentrated and spraying drying: is spraying drying after 0.01-0.08MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 55-80 ℃ of vacuum tightness, obtains the high purity water threose.
Step (1) is preferably: in mass ratio is 1: 4 ratio, and drying stachys sieboldii miq or argentate strip are added water in 70 ℃ of lixiviates 1.0 hours, squeezes the juice, and gets residue, adds 2 times of 1.0 hours multiple press for extracting juicees of 70 ℃ of lixiviates of water to the residue quality, and mixing twice squeezeding juice is liquid glucose.
Step (2) is preferably: described liquid glucose is warming up to 80-90 ℃, heated 20 minutes, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 8g: 1L, gets supernatant liquor.
Step (3) is preferably: in the ratio of 1L: 7g, add gac in supernatant liquor, be heated to 80-90 ℃, decoloured 40-50 minute, remove by filter gac.
Step (6) is preferably: is that 0.06MPa is concentrated into 50 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 60 ℃, vacuum tightness.
Step (7) is preferably: selecting porous gel resin for use is the industrial chromatography stationary phase, and utilizing the size exclusion principle is that 25: the 1 pairs of sugars in 40-60 ° of Brix liquid glucose separate at the aspect ratio of 50 ℃ of column temperatures, pillar.
Advantage of the present invention:
1. the domestic purification that first the size exclusion chromatography technology is applied to functional oligose, and success from natural phant, obtained high purity stachyose, for the purification of other functional oligoses provides technology platform.
2. be the industrial chromatography stationary phase by selecting porous gel resin for use, separation obtains purity and is higher than more than 90% (stachyose accounts for the total reducing sugar ratio), and the stachyose yield is higher, rate of loss is low.
3. contain mono-and di-saccharides in the product hardly, further enlarged the crowd that benefits from and the Application Areas of stachyose.
4. improve the utility value of raw materials such as Rhizome of Bear's-foot Fern, argentate strip, increased farmers' income, created high social.
5. adopt the industrial chromatography partition method directly from natural phant, to extract high purity stachyose, Herba Lycopi's oligomeric galactose family functional oligose content such as stachyose is up to 98% (raffinose, stachyose and a hair stamen pentasaccharides account for the total reducing sugar ratio) in the product that obtains, but industrialized mass.
Description of drawings
Fig. 1 is a sugar color atlas before embodiment 1 chromatrographic separation step.
Fig. 2 is a sugar color atlas after embodiment 1 chromatrographic separation step.
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
Embodiment 1
(1) squeeze the juice: select for use 10kg not have and add 80 ℃ of lixiviates of 40L water 1 hour after mildew and rot hay cadbait is cleaned, get residue after squeezing the juice and add 2 times of 1 hour multiple press for extracting juicees of 80 ℃ of lixiviates of water to the residue quality, mixing twice squeezeding juice is that liquid glucose is standby;
(2) removal of impurities: described liquid glucose is warming up to 90 ℃, heated 20 minutes, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 6g: 1L, staticly settles 60 minutes, gets supernatant liquor;
(3) decolouring: in the ratio of 1L: 6g, in supernatant liquor, add gac, be heated to 90 ℃, decoloured 40 minutes, remove by filter gac;
(4) desalination: the liquid after the decolouring is cooled to 25 ℃, through Zeo-karb, anionite-exchange resin, Zeo-karb, anionite-exchange resin specific conductivity is reduced to below the 50 μ s/cm successively;
(5) membrane filtration: is that 5000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.06MPa is concentrated into 50 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 60 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, utilize the size exclusion principle that the sugar in 50 ° of Brix liquid glucoses is separated, wherein pillar is selected the stainless steel column of diameter 4cm, high 100cm for use, and input speed is 5ml/min, and feeding temperature is 50 ℃;
(8) concentrated and spraying drying: is spraying drying after 0.05MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 70 ℃ of vacuum tightnesss, obtains the high purity water threose.The purity of stachyose product is 95% (accounting for the total reducing sugar ratio) after testing, and yield is 61.9%.
Embodiment 2
The industrial chromatographic separation technology of a kind of usefulness prepares the method for high purity water threose, comprises the steps:
(1) squeeze the juice: in mass ratio is 1: 4 ratio, and drying stachys sieboldii miq is added water in 70 ℃ of lixiviates 1.0 hours, squeezes the juice, and gets residue, adds 2 times of 1.0 hours multiple press for extracting juicees of 70 ℃ of lixiviates of water to the residue quality, and mixing twice squeezeding juice is liquid glucose;
(2) removal of impurities: described liquid glucose is warming up to 80 ℃, heated 20 minutes, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 8g: 1L, staticly settles 60 minutes, gets supernatant liquor;
(3) in 1L: the ratio of 7g, in supernatant liquor, add gac, be heated to 80 ℃, decoloured 50 minutes, remove by filter gac;
(4) desalination: the liquid after the decolouring is reduced to 20 ℃, through Zeo-karb, anionite-exchange resin specific conductivity is reduced to below the 100 μ s/cm successively;
(5) membrane filtration: is that 10000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.05MPa is concentrated into 50 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 60 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, and utilizing the size exclusion principle is that 30: the 1 pairs of sugars in 50 ° of Brix liquid glucoses separate at the aspect ratio of flow velocity 6ml/min, 40 ℃ of column temperatures, pillar;
(8) concentrated and spraying drying: is spraying drying after 0.08MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 60 ℃ of vacuum tightnesss, obtains the high purity water threose.The purity of stachyose product is 94.1% (accounting for the total reducing sugar ratio) after testing, and yield is 60.1%.
Embodiment 3
The industrial chromatographic separation technology of a kind of usefulness prepares the method for high purity water threose, comprises the steps:
(1) squeeze the juice: in mass ratio is 1: 3 ratio, and dry argentate strip is added water in 50 ℃ of lixiviates 1.5 hours, squeezes the juice, and gets residue, adds 1 times of 1.5 hours multiple press for extracting juice of 50 ℃ of lixiviates of water to the residue quality, and mixing twice squeezeding juice is liquid glucose;
(2) removal of impurities: described liquid glucose is warming up to 50 ℃, heated 30 minutes, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 5g: 1L, staticly settles 90 minutes, gets supernatant liquor;
(3) decolouring: in the ratio of 1L: 4g, in supernatant liquor, add gac, be heated to 60 ℃, decoloured 60 minutes, remove by filter gac;
(4) desalination: the liquid after the decolouring is reduced to 30 ℃, through Zeo-karb, anionite-exchange resin specific conductivity is reduced to below the 150 μ s/cm successively;
(5) membrane filtration: is that 50000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.08MPa is concentrated into 60 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 40 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, utilizes the size exclusion principle for 125cm (series connection of two posts) sugar in 60 ° of Brix liquid glucoses to be separated for 8cm, height at flow velocity 40ml/min, 20 ℃ of column temperatures, pillar diameter;
(8) concentrated and spraying drying: is spraying drying after 0.01MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 55 ℃ of vacuum tightnesss, obtains the high purity water threose.The purity of stachyose product is 94.9% (accounting for the total reducing sugar ratio) after testing, yield 59.9%.
Embodiment 4
The industrial chromatographic separation technology of a kind of usefulness prepares the method for high purity water threose, comprises the steps:
(1) squeeze the juice: in mass ratio is 1: 6 ratio, and dry argentate strip is added water in 90 ℃ of lixiviates 0.5 hour, squeezes the juice, and gets residue, adds 3 times of 0.5 hour multiple press for extracting juicees of 90 ℃ of lixiviates of water to the residue quality, and mixing twice squeezeding juice is liquid glucose;
(2) removal of impurities: described liquid glucose is warming up to 100 ℃, heated 10 minutes, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 12g: 1L, staticly settles 30 minutes, gets supernatant liquor;
(3) decolouring: in the ratio of 1L: 10g, in supernatant liquor, add gac, be heated to 90 ℃, decoloured 30 minutes, remove by filter gac;
(4) desalination: the liquid after the decolouring is reduced to 25 ℃, makes specific conductivity below 50 μ s/cm through Zeo-karb, anionite-exchange resin, Zeo-karb, anionite-exchange resin successively;
(5) membrane filtration: is that 100000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.01MPa is concentrated into 40 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 80 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, and utilizing the size exclusion principle is that 32: the 1 pairs of sugars in 40 ° of Brix liquid glucoses separate at the aspect ratio of flow velocity 120ml/min, 60 ℃ of column temperatures, pillar;
(8) concentrated and spraying drying: is spraying drying after 0.06MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 80 ℃ of vacuum tightnesss, obtains the high purity water threose.The purity of stachyose product is 94.4% (accounting for the total reducing sugar ratio) after testing, yield 60.2%.
Claims (6)
1. one kind prepares the method for high purity water threose with industrial chromatographic separation technology, it is characterized in that comprising the steps:
(1) squeeze the juice: in mass ratio is 1: the ratio of 3-6, drying stachys sieboldii miq is added water in 50 ℃ of-90 ℃ of lixiviates 0.5 hour-1.5 hours, squeeze the juice, get residue, add 1 times-3 times 0.5 hour-1.5 hours multiple press for extracting juicees of 50 ℃ of-90 ℃ of lixiviates of water to the residue quality, mixing twice squeezeding juice is liquid glucose;
(2) removal of impurities: described liquid glucose is warming up to 50-100 ℃, heated 10-30 minute, filter, disgorging joins finings sodium hydroxide in the filtrate in the ratio of 5-12g: 1L, staticly settles 30-90 minute, gets supernatant liquor;
(3) decolouring: in the ratio of 1L: 4-10g, in supernatant liquor, add gac, be heated to 60-90 ℃, decoloured 30-60 minute, remove by filter gac;
(4) desalination: the liquid after the decolouring is cooled to 20-30 ℃, through Zeo-karb, anionite-exchange resin specific conductivity is reduced to below the 150 μ s/cm successively;
(5) membrane filtration: is that 5000～100000 ultra-filtration membrane filters with the liquid after the desalination by molecular weight cut-off, removes the impurity in the liquid glucose;
(6) concentrate: is that 0.01-0.08MPa is concentrated into 40-60 ° of Brix liquid glucose with the liquid behind the membrane filtration at 40～80 ℃, vacuum tightness;
(7) chromatographic separation: selecting porous gel resin for use is the industrial chromatography stationary phase, utilizes the size exclusion principle that the sugar in the 40-60 ° of Brix liquid glucose is separated;
(8) concentrated and spraying drying: is spraying drying after 0.01-0.08MPa carries out vacuum concentration through the liquid glucose after the chromatographic separation 55-80 ℃ of vacuum tightness, obtains the high purity water threose.
2. the industrial chromatographic separation technology of a kind of usefulness according to claim 1 prepares the method for high purity water threose, it is characterized in that described step (1) is: the ratio that in mass ratio is 1: 4, drying stachys sieboldii miq is added water in 70 ℃ of lixiviates 1.0 hours, squeeze the juice, get residue, add 2 times of 1.0 hours multiple press for extracting juicees of 70 ℃ of lixiviates of water to the residue quality, mixing twice squeezeding juice is liquid glucose.
3. the industrial chromatographic separation technology of a kind of usefulness according to claim 1 prepares the method for high purity water threose, it is characterized in that described step (2) is: described liquid glucose is warming up to 80-90 ℃, heated 20 minutes, filter, disgorging, ratio in 8g: 1L joins finings sodium hydroxide in the filtrate, staticly settles 60 minutes, gets supernatant liquor.
4. the industrial chromatographic separation technology of a kind of usefulness according to claim 1 prepares the method for high purity water threose, it is characterized in that described step (3) is: in the ratio of 1L: 7g, in supernatant liquor, add gac, be heated to 80-90 ℃, decoloured 40-50 minute, and removed by filter gac.
5. the industrial chromatographic separation technology of a kind of usefulness according to claim 1 prepares the method for high purity water threose, it is characterized in that described step (6) is: is that 0.06MPa is concentrated into 50 ° of Brix liquid glucoses with the liquid behind the membrane filtration at 60 ℃, vacuum tightness.
6. the industrial chromatographic separation technology of a kind of usefulness according to claim 1 prepares the method for high purity water threose, it is characterized in that described step (7) is: selecting porous gel resin for use is the industrial chromatography stationary phase, utilize the size exclusion principle to be 25-40: under 1 the condition, the sugar in the 40-60 ° of Brix liquid glucose to be separated at column temperature 20-60 ℃, the aspect ratio of pillar.
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| CN102174117B (en) * | 2011-01-14 | 2012-11-14 | 南京农业大学 | Method for preparing Stachys floridana schuttl.Exbent. polysaccharide with antitumor activity |
| CN102226223B (en) * | 2011-03-29 | 2013-07-10 | 河南飞天农业开发股份有限公司 | Novel starch sugar mother liquor pretreatment technology |
| CN103265583B (en) * | 2013-03-05 | 2016-01-13 | 中国食品发酵工业研究院 | A kind of preparation method of stachyose crystal |
| JP2016531584A (en) * | 2013-09-05 | 2016-10-13 | ダウ グローバル テクノロジーズ エルエルシー | Chromatographic separation of sugars using a cation exchange resin formulation. |
| CN105315314A (en) * | 2014-07-31 | 2016-02-10 | 中国食品发酵工业研究院 | Industrial production method for extracting stachyose from radix rehmanniae |
| CN104262418B (en) * | 2014-09-26 | 2017-04-19 | 何龙 | Production method of crystallized water threose |
| CN106543238A (en) * | 2016-09-23 | 2017-03-29 | 合肥信达膜科技有限公司 | Stachyose film extraction process in a kind of Rhizoma Humatae Tyermanni |
| CN106632527A (en) * | 2016-12-19 | 2017-05-10 | 西安源森生物科技有限公司 | Stachyose preparation method |
| CN106636250B (en) * | 2017-03-09 | 2021-03-30 | 无限极（中国）有限公司 | Method for preparing stachyose |
| CN107286208A (en) * | 2017-08-20 | 2017-10-24 | 合肥信达膜科技有限公司 | A kind of extracting method for stachyose |
| CN114213475B (en) * | 2021-12-29 | 2023-01-31 | 山东百龙创园生物科技股份有限公司 | Preparation method of stachyose |
| CN115844012A (en) * | 2022-11-24 | 2023-03-28 | 浙江工业大学 | Composition with constipation relieving function, preparation method and equipment |
| CN116837064B (en) * | 2023-09-04 | 2023-11-28 | 成都速攻痛风病研究集团有限公司 | Preparation method of silkworm chrysalis protein peptide |
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Application publication date: 20100127 Assignee: Chengde Jingtian Food Technology Co.,Ltd. Assignor: China National Academy of Food & Fermentation Industries Contract record no.: 2013990000229 Denomination of invention: Method for preparing high-purity stachyose by using plant chromatographic separation technology Granted publication date: 20110608 License type: Common License Record date: 20130513 |
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Application publication date: 20100127 Assignee: Shangshi (Guangdong) big data service Co.,Ltd. Assignor: CHINA NATIONAL RESEARCH INSTITUTE OF FOOD & FERMENTATION INDUSTRIES Co.,Ltd. Contract record no.: X2021440000217 Denomination of invention: Preparation of high purity stachyose by industrial chromatographic separation technology Granted publication date: 20110608 License type: Common License Record date: 20211111 |
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