CN105949341A - Method for extracting momordica polysaccharide from ion liquid - Google Patents
Method for extracting momordica polysaccharide from ion liquid Download PDFInfo
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- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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Abstract
The invention discloses a method for extracting momordica polysaccharide from ion liquid, and belongs to the technical field of momordica polysaccharide preparation. According to the method, bitter gourds are dried, are subjected to water removal, and are crushed to obtain bitter gourd powder; then, ultrasonic extraction and filtration are performed to obtain filter liquid and filter slag; the filter slag is subjected to enzymolysis and centrifugation to obtain supernate; then, the supernate is mixed with filter liquid to obtain bitter gourd extraction liquid; 1-methyl-5-clemizole and hexafluorophosphoric acid solution are stirred to take a reaction; liquid separation is performed to obtain an organic phase; the organic phase is washed to the neutral state; distillation is performed to obtain imidazolium salt ion liquid; then, the imidazolium salt ion liquid and dipotassium phosphate are added into the bitter gourd extraction liquid; after the filtration, filter liquid is subjected to still standing for liquid separation; then, obtained materials take a reaction with maleic acid; after the temperature reduction treatment, filtration is performed to obtain a mixed solution; then, tetraethylammonium hydroxide is added; stirring is performed for reaction; filtration is performed after the temperature reduction treatment; imidazolium salt ion liquid containing polysaccharide is obtained; finally, diethyl ether is added for reaction; filtration and reduced pressure distillation are finally performed; momordica polysaccharide is obtained through freeze drying. Compared with a conventional method, the method has the advantages that the extraction rate is improved by 30 to 40 percent; the purity reaches 97 percent or higher.
Description
Technical field
A kind of method that the invention discloses ionic liquid extract bitter melon polysaccharide, belongs to bitter melon polysaccharide preparing technical field.
Background technology
Fructus Momordicae charantiae has another name called balsam pear, insane melon, insane Fructus Vitis viniferae, brocade Fructus Litchi etc., is the fruit of Cucurbitaceae Momordica plant.It has good edibility and obvious medicinal function, have the title of " medicinal vegetable ".
Fructus Momordicae charantiae is a kind of tame renewable green resource, and all there are plantation, aboundresources in all parts of the country.Good vegetables are not only by Fructus Momordicae charantiae, are also good medicine, and it is the dietetic therapy good merchantable brand of a kind of medicine-food two-purpose, the polysaccharide material extracted from Fructus Momordicae charantiae, there is the multiple effects such as reduction blood glucose, antibacterial anti-inflammatory, non-oxidizability, and immunity can be improved, be the environmental friendly product of a kind of health that is beneficial to man.Bitter melon polysaccharide product appearance is faint yellow to dissolve in hot water to brown solid powder, easy moisture absorption, is insoluble in the organic solvents such as alcohol, ketone, ether, chloroform.Polysaccharide has the multiple biological activitys such as the immunologic function of raising human body, and modern medicine study shows, bitter melon polysaccharide has the multiple effects such as reduction blood glucose, antibacterial anti-inflammatory, non-oxidizability.
The Vitamin C content of Fructus Momordicae charantiae is the highest; have inside the effect Fructus Momordicae charantiae such as prevention vitamin C deficiency, protection cell membrane, atherosclerosis, raising physical stress ability, cardioprotection containing a kind of composition; can effectively suppress the canceration of a normal bag and promote the recovery of the special born of the same parents that attenuate; there is certain antitumaous effect, but also possess the feature having blood fat reducing and blood pressure.Moreover, Fructus Momordicae charantiae also has the effects such as pre-preventing bone rarefaction, endocrine regulation, antioxidation, the most further strengthening body resistance, improves the response mechanism inside health.
Bitter melon polysaccharide is a kind of compound heteropolysaccharide, and research finds that it has many biological activitys.Existing bitter melon polysaccharide extracting method is the longest, and energy resource consumption is big and extraction ratio is the highest, is not suitable for industrialized production.
Summary of the invention
The technical problem that present invention mainly solves: the longest for existing bitter melon polysaccharide extracting method, energy resource consumption is big and extraction ratio is the highest, the problem being not suitable for industrialized production, a kind of method that ionic liquid extract bitter melon polysaccharide is provided, Fructus Momordicae charantiae is dried and obtains Fructus Momordicae charantiae powder except pulverizing after water by the method, carry out ultrasonic extraction the most again, filtrate and filtering residue is obtained after filtration, wherein filtering residue carries out enzymolysis, it is centrifuged and obtains supernatant, then filtrate is mixed with supernatant, obtain balsam pear extract, 1-methyl-5-chloro imidazoles reacts with hexafluorophosphoric acid solution stirring, separatory obtains organic facies, washing organic facies distills to obtain imidazolium ionic liquid to neutrality；Again imidazolium ionic liquid, dipotassium hydrogen phosphate are added in balsam pear extract, after filtration, filtrate is stood and obtain organic facies, react itself and maleic acid to filter after cooling processes and prepare mixed solution, add tetraethyl ammonium hydroxide stirring reaction in solution, cooling is filtered after processing, and must contain the imidazolium ionic liquid of polysaccharide；Finally adding ether reaction, finally filter, distillation of reducing pressure, lyophilizing obtains one bitter melon polysaccharide.The present invention relatively traditional method for extracting rate improves 30～40%, and purity has reached more than 97%, and the present invention is simple to operate, be prone to industrialized production.
In order to solve above-mentioned technical problem, the technical solution adopted in the present invention is:
(1) fresh bitter is washed with deionized 3～5 times, clean Fructus Momordicae charantiae is dried in 95～105 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 2～5%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 200～300mL deionized waters, beaker is placed in 180～220W sonic oscillation instrument ultrasonic 20～30min, filter, collect filtrate and filtering residue respectively；
(2) filtering residue is joined in enzymatic vessel, 200～300mL deionized waters, 0.3～0.5g cellulase, 0.3～0.5g pectase, 0.5～0.8g protease it is sequentially added in enzymatic vessel, control temperature at 45～55 DEG C, stirring reaction 20～30min, temperature is increased to 100～110 DEG C after terminating by reaction, stirring 5～10min, obtain enzymolysis solution, enzymolysis solution is placed in 6000～8000r/min centrifuges centrifugal 10～20min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；
(3) measure 100～200mL deionized waters and join in the there-necked flask with thermometer and reflux, 2～3g1-methyl-5-chloro imidazoles are added in flask, flask is placed in water-bath, controlling temperature is 0～5 DEG C, adding 100～200mL mass concentrations while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 1～2h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；
(4) above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps (2) is standby, stirring 5～10min, add 2～3g dipotassium hydrogen phosphates, stirring extraction 20～30min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 2～4g maleic acids are added in flask, stirring 1～2h, by on 0.3～0.5L liquid nitrogen spraying to flask, flask temperature is made to be down to-20～-30 DEG C, filter, obtain mixed solution, 3～5g tetraethyl ammonium hydroxides are added in solution, stirring reaction 20～30min, by on 0.1～0.3L liquid nitrogen spraying to flask, flask temperature is made to be down to-10～-20 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；
(5) the above-mentioned imidazolium ionic liquid containing polysaccharide is added 300～400mL ether, stir 10～20min, filter, obtain filtrate, by filtrate decompression distill, obtain solid, solid is placed in-10～-15 DEG C of vacuum freeze dryers in, lyophilizing 1～2h, obtains powder, i.e. bitter melon polysaccharide.
The application process of the present invention is: adds the bitter melon polysaccharide powder that the 5～10g present invention prepare in 300～500mL boiled water, directly takes after mixing it with water after stirring.After testing, hyperglycemic patients is taken every day after the meal, takes every day 2～3 times, takes continuously 10～15 days, from original 7.1～9.2mmol/L, blood glucose is reduced to 4.5～4.8mmol/L, and blood sugar decreasing effect is notable, can extensively apply.
The invention has the beneficial effects as follows:
(1) extraction process of the present invention is simple, and raw material is easy to get, and effectively reduces preparation cost, is suitable for large-scale production；
(2) present invention relatively traditional method for extracting rate improves 30～40%, and purity has reached more than 97%, and the present invention is simple to operate, be prone to industrialized production.
Detailed description of the invention
First fresh bitter is washed with deionized 3～5 times, after Xi Jinging, Fructus Momordicae charantiae is dried in 95～105 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 2～5%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 200～300mL deionized waters, beaker is placed in 180～220W sonic oscillation instrument ultrasonic 20～30min, filter, collect filtrate and filtering residue respectively；Then filtering residue is joined in enzymatic vessel, 200～300mL deionized waters, 0.3～0.5g cellulase, 0.3～0.5g pectase, 0.5～0.8g protease it is sequentially added in enzymatic vessel, control temperature at 45～55 DEG C, stirring reaction 20～30min, temperature is increased to 100～110 DEG C after terminating by question response, stirring 5～10min, obtain enzymolysis solution, enzymolysis solution is placed in 6000～8000r/min centrifuges centrifugal 10～20min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；Measure 100～200mL deionized waters again and join in the there-necked flask with thermometer and reflux, 2～3g1-methyl-5-chloro imidazoles are added in flask, flask is placed in water-bath, controlling temperature is 0～5 DEG C, adding 100～200mL mass concentrations while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 1～2h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；Again above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps is standby, stirring 5～10min, add 2～3g dipotassium hydrogen phosphates, stirring extraction 20～30min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 2～4g maleic acids are added in flask, stirring 1～2h, again by 0.3～0.5L liquid nitrogen spraying to flask, flask temperature is made to be down to-20～-30 DEG C, filter, obtain mixed solution, 3～5g tetraethyl ammonium hydroxides are added in solution, stirring reaction 20～30min, by on 0.1～0.3L liquid nitrogen spraying to flask, flask temperature is made to be down to-10～-20 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；Finally the above-mentioned imidazolium ionic liquid containing polysaccharide is added 300～400mL ether, stir 10～20min, filter, obtain filtrate, by filtrate decompression distill, obtain solid, solid is placed in-10～-15 DEG C of vacuum freeze dryers in, lyophilizing 1～2h, obtains powder, i.e. bitter melon polysaccharide.
Example 1
First fresh bitter is washed with deionized 3 times, after Xi Jinging, Fructus Momordicae charantiae is dried in 95 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 2%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 200mL deionized water, beaker is placed in ultrasonic 20min in 180W sonic oscillation instrument, filter, collect filtrate and filtering residue respectively；Then filtering residue is joined in enzymatic vessel, in enzymatic vessel, be sequentially added into 200mL deionized water, 0.3g cellulase, 0.3g pectase, 0.5g protease, control temperature at 45 DEG C, stirring reaction 20min, temperature is increased to 100 DEG C after terminating by question response, stirs 5min, obtains enzymolysis solution, enzymolysis solution is placed in 6000r/min centrifuge centrifugal 10min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；Measure 100mL deionized water again and join in the there-necked flask with thermometer and reflux, in flask, add 2g1-methyl-5-chloro imidazoles, flask is placed in water-bath, controlling temperature is 0 DEG C, and adding 100mL mass concentration while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 1h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；Again above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps is standby, stirring 5min, add 2g dipotassium hydrogen phosphate, stirring extraction 20min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 2g maleic acid is added in flask, stirring 1h, again by 0.3L liquid nitrogen spraying to flask, flask temperature is made to be down to-20 DEG C, filter, obtain mixed solution, 3g tetraethyl ammonium hydroxide is added in solution, stirring reaction 20min, by on 0.1L liquid nitrogen spraying to flask, flask temperature is made to be down to-10 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；Finally the above-mentioned imidazolium ionic liquid containing polysaccharide is added 300mL ether, stir 10min, filter, obtain filtrate, filtrate decompression is distilled, obtains solid, solid is placed in-10 DEG C of vacuum freeze dryers, lyophilizing 1h, obtains powder, i.e. bitter melon polysaccharide.
This example operation is easy, during use, adds the bitter melon polysaccharide powder that the 5g present invention prepares, directly take after mixing it with water after stirring in 300mL boiled water.After testing, hyperglycemic patients is taken every day after the meal, takes every day 2 times, takes continuously 10 days, from original 7.1mmol/L, blood glucose is reduced to 4.5mmol/L, and blood sugar decreasing effect is notable, can extensively apply.
Example 2
First fresh bitter is washed with deionized 4 times, after Xi Jinging, Fructus Momordicae charantiae is dried in 100 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 3.5%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 250mL deionized water, beaker is placed in ultrasonic 25min in 200W sonic oscillation instrument, filter, collect filtrate and filtering residue respectively；Then filtering residue is joined in enzymatic vessel, in enzymatic vessel, be sequentially added into 250mL deionized water, 0.4g cellulase, 0.4g pectase, 0.65g protease, control temperature at 50 DEG C, stirring reaction 25min, temperature is increased to 105 DEG C after terminating by question response, stirs 8min, obtains enzymolysis solution, enzymolysis solution is placed in 7000r/min centrifuge centrifugal 15min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；Measure 150mL deionized water again and join in the there-necked flask with thermometer and reflux, in flask, add 2.5g1-methyl-5-chloro imidazoles, flask is placed in water-bath, controlling temperature is 3 DEG C, and adding 150mL mass concentration while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 1.5h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；Again above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps is standby, stirring 7min, add 2.5g dipotassium hydrogen phosphate, stirring extraction 25min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 3g maleic acid is added in flask, stirring 1.5h, again by 0.4L liquid nitrogen spraying to flask, flask temperature is made to be down to-25 DEG C, filter, obtain mixed solution, 4g tetraethyl ammonium hydroxide is added in solution, stirring reaction 25min, by on 0.15L liquid nitrogen spraying to flask, flask temperature is made to be down to-15 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；Finally the above-mentioned imidazolium ionic liquid containing polysaccharide is added 350mL ether, stir 15min, filter, obtain filtrate, filtrate decompression is distilled, obtains solid, solid is placed in-13 DEG C of vacuum freeze dryers, lyophilizing 1.5h, obtains powder, i.e. bitter melon polysaccharide.
This example operation is easy, during use, adds the bitter melon polysaccharide powder that the 8g present invention prepares, directly take after mixing it with water after stirring in 400mL boiled water.After testing, hyperglycemic patients is taken every day after the meal, takes every day 2 times, takes continuously 13 days, from original 8.1mmol/L, blood glucose is reduced to 4.7mmol/L, and blood sugar decreasing effect is notable, can extensively apply.
Example 3
First fresh bitter is washed with deionized 5 times, after Xi Jinging, Fructus Momordicae charantiae is dried in 105 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 5%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 300mL deionized water, beaker is placed in ultrasonic 30min in 220W sonic oscillation instrument, filter, collect filtrate and filtering residue respectively；Then filtering residue is joined in enzymatic vessel, in enzymatic vessel, be sequentially added into 300mL deionized water, 0.5g cellulase, 0.5g pectase, 0.8g protease, control temperature at 55 DEG C, stirring reaction 30min, temperature is increased to 110 DEG C after terminating by question response, stirs 10min, obtains enzymolysis solution, enzymolysis solution is placed in 8000r/min centrifuge centrifugal 20min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；Measure 200mL deionized water again and join in the there-necked flask with thermometer and reflux, in flask, add 3g1-methyl-5-chloro imidazoles, flask is placed in water-bath, controlling temperature is 5 DEG C, and adding 200mL mass concentration while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 2h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；Again above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps is standby, stirring 10min, add 3g dipotassium hydrogen phosphate, stirring extraction 30min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 4g maleic acid is added in flask, stirring 2h, again by 0.5L liquid nitrogen spraying to flask, flask temperature is made to be down to-30 DEG C, filter, obtain mixed solution, 5g tetraethyl ammonium hydroxide is added in solution, stirring reaction 30min, by on 0.3L liquid nitrogen spraying to flask, flask temperature is made to be down to-20 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；Finally the above-mentioned imidazolium ionic liquid containing polysaccharide is added 400mL ether, stir 20min, filter, obtain filtrate, filtrate decompression is distilled, obtains solid, solid is placed in-15 DEG C of vacuum freeze dryers, lyophilizing 2h, obtains powder, i.e. bitter melon polysaccharide.
This example operation is easy, during use, adds the bitter melon polysaccharide powder that the 10g present invention prepares, directly take after mixing it with water after stirring in 500mL boiled water.After testing, hyperglycemic patients is taken every day after the meal, takes every day 3 times, takes continuously 15 days, from original 9.2mmol/L, blood glucose is reduced to 4.8mmol/L, and blood sugar decreasing effect is notable, can extensively apply.
Claims (1)
1. the method for an ionic liquid extract bitter melon polysaccharide, it is characterised in that concrete extraction step is:
(1) fresh bitter is washed with deionized 3～5 times, clean Fructus Momordicae charantiae is dried in 95～105 DEG C of baking ovens to Fructus Momordicae charantiae moisture content be 2～5%, place in pulverizer and pulverize, obtain Fructus Momordicae charantiae powder, Fructus Momordicae charantiae powder is joined in beaker, in beaker, adds 200～300mL deionized waters, beaker is placed in 180～220W sonic oscillation instrument ultrasonic 20～30min, filter, collect filtrate and filtering residue respectively；
(2) filtering residue is joined in enzymatic vessel, 200～300mL deionized waters, 0.3～0.5g cellulase, 0.3～0.5g pectase, 0.5～0.8g protease it is sequentially added in enzymatic vessel, control temperature at 45～55 DEG C, stirring reaction 20～30min, temperature is increased to 100～110 DEG C after terminating by reaction, stirring 5～10min, obtain enzymolysis solution, enzymolysis solution is placed in 6000～8000r/min centrifuges centrifugal 10～20min, discard lower floor's impurity, obtain supernatant, supernatant is mixed with above-mentioned filtrate, obtain balsam pear extract, standby；
(3) measure 100～200mL deionized waters and join in the there-necked flask with thermometer and reflux, 2～3g1-methyl-5-chloro imidazoles are added in flask, flask is placed in water-bath, controlling temperature is 0～5 DEG C, adding 100～200mL mass concentrations while stirring in flask is 40% hexafluorophosphoric acid solution, stirring reaction 1～2h, after reaction terminates, stand, separatory, obtain organic facies, organic facies is washed with saturated sodium carbonate, until organic facies is neutrality, then distillation of reducing pressure, obtain colourless liquid, i.e. imidazolium ionic liquid；
(4) above-mentioned imidazolium ionic liquid is joined in the balsam pear extract that above-mentioned steps (2) is standby, stirring 5～10min, add 2～3g dipotassium hydrogen phosphates, stirring extraction 20～30min, filter, obtain filtrate, by filtrate stratification, separatory, obtain organic facies, organic facies is joined in there-necked flask, 2～4g maleic acids are added in flask, stirring 1～2h, by on 0.3～0.5L liquid nitrogen spraying to flask, flask temperature is made to be down to-20～-30 DEG C, filter, obtain mixed solution, 3～5g tetraethyl ammonium hydroxides are added in solution, stirring reaction 20～30min, by on 0.1～0.3L liquid nitrogen spraying to flask, flask temperature is made to be down to-10～-20 DEG C, filter, obtain the imidazolium ionic liquid containing polysaccharide；
(5) the above-mentioned imidazolium ionic liquid containing polysaccharide is added 300～400mL ether, stir 10～20min, filter, obtain filtrate, by filtrate decompression distill, obtain solid, solid is placed in-10～-15 DEG C of vacuum freeze dryers in, lyophilizing 1～2h, obtains powder, i.e. bitter melon polysaccharide.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109280091A (en) * | 2018-11-27 | 2019-01-29 | 苏州卫生职业技术学院 | A kind of biological enzyme extraction process of bitter melon polysaccharide |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109280091A (en) * | 2018-11-27 | 2019-01-29 | 苏州卫生职业技术学院 | A kind of biological enzyme extraction process of bitter melon polysaccharide |
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