CN102276515B - Method for extracting deoxynojirimycin - Google Patents
Method for extracting deoxynojirimycin Download PDFInfo
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Abstract
The invention discloses a method for extracting a deoxynojirimycin monomer. The method comprises the following steps of: soaking a mulberry raw material in acidic aqueous solution, and extracting for 1 to 3 times; adsorbing extracting solution on a cation exchange resin, washing, and eluting by using ammonia water; performing reverse osmosis pre-concentration on eluent, recovering ammonia, dissolving an intermediate in a hydrophilic solvent, performing catalytic conversion, allowing the solution to pass through a gel chromatographic column, and eluting the hydrophilic solvent; recovering the solvent, dissolving in a mixed solvent, allowing the mixed solvent to pass through the chromatographic column, and eluting the mixed solvent; and recovering the mixed solution to obtain pure deoxynojirimycin products. The method is low in energy consumption and environment-friendly, and the pure deoxynojirimycin products with various purity grades of 10 percent, 50 percent, 99 percent and the like.
Description
Technical field
The present invention relates to the biological extraction technology, particularly a kind of extracting method of S-GI monomer.
Background technology
1-S-GI (1-deoxynojirimycin, DNJ) contain the N compound for a kind of polarity, its chemical name is 3,4,5-trihydroxy--2-methylol tetrahydropyridine, molecular formula is C
6H
13NO
4, relative molecular mass is 163.DNJ has the effect of very strong inhibition α-glucuroide, is effective constituents hypoglycemic, the treatment diabetes
[1～4], have in addition antiviral
[5], anti metastasis
[6]Deng pharmacological action.In recent years, Chinese scholars has been carried out a large amount of research and probes to the source of DNJ, in vivo synthesis mechanism and chemical synthesis process, wishes to find a kind of stable, efficient, economic approach that obtains DNJ
[7]
Li Yuliang
[8]Method Deng having reported extraction S-GI from mulberry leaf mainly comprises extraction, and extract is crossed Zeo-karb, and the steps such as n-butanol extraction obtain the S-GI sterling.This technique need to be used n-butanol extraction; Propyl carbinol is more expensive, and production cost is higher.
The Liu Wei Lam
[9]Deng having reported a kind of method of extracting S-GI from mulberry leaf, comprise that mulberry leaf add 70% extraction using alcohol, pass through macroporous adsorbent resin after extracting solution is concentrated, effluent liquid by macroporous adsorbent resin, concentrated, by Zeo-karb, with the distillation washing, with 0.5mol/L ammoniacal liquor wash-out, collect elutriant, concentrated, concentrated solution will have the component of impurity to discard by silica gel purification, merge and contain the purer component of 1-DNJ, concentrated, vacuum-drying namely gets 1-DNJ (DNJ).This technique needs extraction using alcohol, needed by macroporous resin before upper cationic exchange quality, and 4 times concentrated; Process energy consumption is high, uses the ethanol cost also high.
Zhou Huiyan
[10]Deng having reported the method for utilizing reverse column chromatography technology to extract the purifying S-GI, the content of S-GI reaches 10.7%; This technology fails to obtain sterling.
Liu Shuxing
[11]Deng to adsorb under DXA-6 resin dress post, pH10, original liquid concentration 0.352 mg/mL, flow velocity 2.0 BV/h conditions, 80% ethanolic soln extracts S-GI with the flow velocity wash-out of 2.5 BV/h.Extraction process need to be used ethanol, and the purity of clear and definite final product not.
Chinese patent technology (03101988.9) has been reported a kind of preparation that contains mulberry leaf total alkali medicinal extract and preparation method thereof: mulberry leaf water or hydrophilic solvent extract, flocculation, and supernatant liquor ion-exchange, the hydrophilic solvent refluxing extraction is concentrated into the dried medicinal extract that obtains.The purity of product reaches more than 50%, and said preparation has significantly hypoglycemic and improve the sugar tolerance effect.The method needs flocculation; Need to be refluxed with hydrophilic solvent by the goods and materials of resin absorption elutes, some inconvenience in actually operating.
Chinese patent technology (02113004.3) has been reported Ramulus Mori extract and extracting method and new purposes: Ramulus Mori extract extracts more than 1 time or 1 time by single medicinal material ramulus mori with 0～95% aqueous ethanolic solution or chloroform, acetone, ethyl acetate, methyl alcohol equal solvent or being combined under 10-100 ℃ of temperature condition of they, united extraction liquid reclaims solvent and get final product; Or be further purified with following method: it is 7.5-9.5 that extracting solution is transferred pH, and it is 4-6 that centrifugal rear supernatant liquor is transferred pH, and precipitation is washed to neutrality, is drying to obtain; Extracting solution is through sherwood oil or gasoline degreasing and get; Extracting solution extracts respectively or successively and gets with chloroform, ethyl acetate, acetone, methyl alcohol or water after sherwood oil or gasoline degreasing; Extracting solution gets with 10-95% aqueous ethanolic solution desorption after absorption with macroporous adsorbent resin; Extracting solution after the absorption such as diatomite, aluminum oxide, after chloroform, ethyl acetate, acetone, methyl alcohol, ethanol equal solvent wash-out, concentrate drying and getting; Extracting solution is through column chromatography for separation such as polymeric amide, silica gel or ion exchange resin.Above purification process can be alone or be adopted the combination of several method.Contain a large amount of flavones ingredients in the Ramulus Mori extract of gained, comprise rutin, Quercetin, morin etc.The not clear and definite extract of this patented technology contains the purity of DNJ, and petroleum ether degreasing, macroporous resin adsorption etc. during purifying, and schedule of operation is complicated.
Chinese patent technology (200710060127.2) has been reported the method for continuously extracting alkaloids, flavones and polysaccharide active components from mulberry leaf: selecting solvent is 20%～90% ethanol or 10%～90% acetone refluxing extraction, obtains filtrate and filter residue; After filtrate concentrates, is dissolved in water, separate to such an extent that content is 30%～50% flavonoid compound by macroporous resin; Through the solution after macroporous resin, then separate to such an extent that content is 40%～60% alkaloid compound by ion exchange resin; Mulberry leaf filter residue, water decoct and decolour, and 10%～60% alcohol precipitation gets content and be 55%～85% acidic polysaccharose compounds.Ethanol or acetone that this Technology Need is a large amount of extract, and the Technology cost is high, and energy consumption is high, do not get the DNJ sterling.
Chinese patent technology (200510030333.X) has been reported the method for extracting effective components from mulberry leaf: have the resin of Folium Mori extract to be placed in extraction kettle absorption, pass into continuously CO
2Extract wash-out, collect the CO that flows out extraction kettle
2, unload and be depressed into normal pressure, CO
2Reuse after emptying or pressurization, the residue after the acquisition release is mulberry leaf effective constituent, CO in extraction kettle
2Be supercritical state.Method of the present invention; purge process is difficult for oxidation, is convenient to carry out that mass-producing is extracted from mulberry leaf, purification mulberry leaf deoxynojirimycin alkaloid and flavones in mulberry leaves, and selectivity is high; in product, the total content of S-GI alkaloid and flavones in mulberry leaves can reach 75～90%.This technology adopts supercritical extraction technique, and the finished product do not obtain the S-GI sterling yet.
Chinese patent technology (200710067498.3) has been reported a kind of method of extracting 1-DNJ from mulberry leaf: by add the mineral acid acidification in the extracting solution of mulberry leaf, through the cooling impurity of removing, then obtain through column chromatography.Can make Folium Mori extract content be not less than 10%, but not get sterling.
Chinese patent technology (200810122998.7) has been reported a kind of method of separating the 1-DNJ monomer from total alkaloids of Folium Mori: mulberry leaf total alkali is water-soluble, ion exchange resin absorption, after absorption is completed, the distribution wash-out is also collected S-GI sterling part, concentrating under reduced pressure is dry, repeatedly recrystallization, obtain sterling.The method requires " total alkaloids " of raw material to contain S-GI more than 40%, and the stepwise elution agent is the alcoholic solution of different concns, and recrystallization is methyl alcohol and acetone mixing solutions; Need a large amount of testings during stepwise elution and collect the sterling position, operate cumbersomely, the mixed solvent of recrystallization is difficult to processing.
Chinese patent technology (200810236144.1) has been reported a kind of coproduction of mulberry tea flavone, polysaccharide and alkaloidal composite extracting technique, does not obtain the S-GI sterling but separate.
Chinese patent technology (200910192542.2) reported a kind of from mulberry leaf the method for successive extract 1-DNJ (DNJ), flavones: will mulberry leaf and ethanol extract after mixing, centrifuging adds the flocculation agent flocculation sediment after concentrated; After removing precipitation, filtrate is added a plurality of cation exchange resin columns of connection, and the macroporous adsorptive resins of flowing through and being chained together with this resin column; Filtrate adds and is washed till the macroporous resin column effluent liquid with pure water after finishing and it is disconnected when colourless, then with ammoniacal liquor or the ethanolic soln that contains ammonia wash-out 1-DNJ (DNJ) on the cationic resin column; Elutriant is gone up cationic resin column again, collects elutriant after repetitive operation, with spraying drying after 80D nanofiltration membrane thickening filtration, gets the Powdered 1-DNJ of brown color (DNJ).Washing with alcohol macroporous resin column with 3%～10% to be removing colloidality impurity, then uses 50%～70% ethanol elution flavones, collects the red-brown elutriant, and concentrating under reduced pressure reclaims spraying drying after ethanol, gets the Powdered total flavonoids of Folium Mori of yellowish brown.This technology raw material need to extract repeatedly with ethanol, and production cost is higher, and energy consumption is also higher.
Chinese patent technology (200710099911.4) has been reported a kind of method of spent ion exchange resin separating folium mori total.This technology does not obtain the S-GI monomer.
Chinese patent technology (201010130263.6) has been reported a kind of flavones, alkaloidal method of preparing from mulberry leaf: flavones is extracted in the buck heating, 90-95% ethanol and gac extract alkaloid again, macroporous resin column is crossed in the flavone extractive acid adjustment, acidizing crystal and 70-80% dissolve with ethanol recrystallization, alkaloids extract is concentrated, acetone dissolves again, peroxidation aluminium short wide column, ammonification crystallization and 85-90% dissolve with ethanol recrystallization.Present technique needs a large amount of organic solvents, and does not obtain the S-GI monomer.
Chinese patent technology (201110021131.4) has been reported the preparation method of 1-Deoxynojirimycin in Mulberry Leaves: adopt high-voltage pulse electric field technology to produce the method for 1-DNJ, owing to being extract at room temperature, improve simultaneously quality product and production efficiency, reduced production cost.This technology does not obtain the S-GI sterling.
Chinese patent technology (200910167602.5) has been reported and extracted the method for separating high-purity 1-DNJ in the natural product: raw material dry powder extracts with hydrophilic solvent, the extracting solution acidifying, soak with anionite-exchange resin after concentrated, use cation exchange column chromatography, organic solvent extraction, recrystallization, gel filtration chromatography purifying after effluent liquid and water lotion are concentrated, in products therefrom, 1-S-GI content is no less than 98%, and concrete enforcement has entered pilot scale.This Technology Need anionite-exchange resin and Zeo-karb need the multiple organic solvents such as alcohol, acetone, chloroform, butanols simultaneously, especially relate to chloroform, and toxicity is larger; Need in leaching process repeatedly to concentrate, energy consumption is higher.
Chinese patent technology (201110038204.0) has been reported by extracting the method for separating high-purity 1-DNJ in mulberry leaf: raw material powder is extracted with water-containing solvent, filter the dregs of a decoction, material is centrifugal, gets extracting solution; Extracting solution is concentrated into without ethanol, ultrafiltration membrance filter; Transfer pH to 3.0-4.0 with mineral acid again, precipitation, centrifugal removal of impurities; Strong acidic ion resin post on the acidifying feed liquid, washing, then carry out the ammoniacal liquor wash-out; Macroporous resin column on feed liquid is washed, then uses the ethanol gradient elution of different concns, collects the target elutriant; Target elutriant concentrating under reduced pressure, drying obtain crude product; The crude product anhydrous alcohol solution, effluent liquid is collected in the removal of impurities of parlkaline adsorption column; The effluent liquid concentrating under reduced pressure carries out normal temperature crystallization and recrystallization, obtains the 1-DNJ highly finished product.In the whole technological process of the present invention, do not use other organic solvent except ethanol, operational safety, 1-DNJ content can reach 98.3%.This technique need to adopt ultra-filtration technique, ethanol distribution wash-out, and collecting the target elutriant needs constantly to detect outflow component etc., complicated operation; Energy consumption is higher.
Up to the present, on existing document and patented technology, extract the S-GI sterling or use too much organic solvent, especially using chloroform, causing security not high; Perhaps need repeatedly to concentrate, energy consumption is higher; Perhaps need to use too much step, perhaps need the substep detects and collects target component, because S-GI does not have uv-absorbing, detect cumbersome, complicated operation, industrial production be difficult to control; Perhaps can't obtain sterling etc.
[1]?Gross?V,?Andus?T,?Tran-ThiT?A,?et?al.?1-Deoxynojirimycin?in?pairs?oligo?saccharide?processing?of?α?
1-proteinase?inhibitor?and?inhibits?its?secretion?in?primary?cultures?of?rat?hepatocytes[J].?BiolChem,?1983,?258(20):12203-12209.
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[4]?Oku?T,?YamadaM,?Nakamura?M,?et?al.?Inhibitory?effects?of?extractives?from?leaves?of?Morus?alba?on?human?and?rat?small?intestinal?disaccharides?activity[J].?J?Nutr,?2006,?95(5):933-938.
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Summary of the invention
Deficiency for existing S-GI monomer extractive technique; the extracting method that the purpose of this invention is to provide a kind of S-GI (DNJ) monomer; the extraction that can realize DNJ separates, and technique is simple, save energy; pollution-free; recycle is implemented easily can adapt to large-scale production and with low cost; extraction efficiency is high simultaneously, good product purity.
The extracting method of DNJ monomer of the present invention comprises the steps:
(1) the mulberry raw material with the sour water of 2～10 times (w/v) (0～5%w/v) or aqueous solution soaking (room temperature～100 ℃) extract each 0.1～48 hour 1～3 time;
(2) extracting solution is with the cationic exchange resin adsorption of 0.1～1 times of (v/v) volume ratio, after using again the washing of 1～10 times of Zeo-karb volume (v/v), with ammoniacal liquor (0.05～2mol/L) wash-out of 1～10 times of Zeo-karb volume (v/v);
(3) with after 2～20 times of ammoniacal liquor elutriant reverse osmosis pre-concentration, Distillation recovery ammonia obtains intermediate A; The ammoniacal liquor that reclaims is used for the wash-out of (2);
(4) intermediate A is dissolved in hydrophilic solvent and is mixed with the solution of 1～20% (w/v), catalyzed conversion 1 hour, and according to gel: solution is gel chromatography column on the volume ratio of 1～10:1, the hydrophilic solvent wash-out;
(5) reclaim after solvent to get intermediate B, intermediate B is dissolved in mixed solvent, is mixed with the solution of 1～20% (w/v), and according to sorbent material: solution is chromatography column on the volume ratio of 1～10:1, solvent elution; The hydrophilic solvent that reclaims is used for the wash-out of (4);
(6) reclaim solvent, vacuum-drying namely gets the S-GI sterling; Recovered solvent is used for the wash-out of (5).
This Technology can obtain the S-GI sterling of the various purity grades such as 10%, 50%, 90%, 99%.
Mulberry raw material in this processing step (1) refers to one or more of ramulus mori, mulberry leaf, White Mulberry Root-bark etc. further; Sour water refers to sulfuric acid or aqueous hydrochloric acid; Immersion refers to soak under room temperature to 100 ℃.Due to soaking at room temperature, reduced energy consumption.
732,7320,001, D001,110,122, D113, D151, the various Zeo-karbs such as 724 Zeo-karb in this processing step (2) refers to common strongly-acid or weakly acidic cation-exchange resin further, comprising:.Extracting solution need not be processed directly upper Zeo-karb, significantly saves cost and has eliminated environmental issue.
Reverse osmosis technology pre-concentration ammoniacal liquor elutriant in this processing step (3) further; The reverse osmosis technology of this process using is common reverse osmosis technology.In the time of this technology concentrated liquor solution, with ammonia and DNJ enrichment, decrease the conventional concentrated energy consumption that consumes, be energy-conservation key.
Catalyzed conversion in this processing step (4), before referring to gel chromatography column, first use catalyzer (hydrochloric acid of 0.01～1%W/V or sulfuric acid) at 60～100 ℃ of lower catalyzed conversions further; Gel chromatography column refers to Superose, Sephdex LH-20, Hitrap SP, sephodex etc.; Described hydrophilic solvent refers to water, methyl alcohol, ethanol, acetone etc.The present invention has adopted catalyzed conversion, and it does not need to monitor the key of intermediate when being chromatography, is also the key that increases yield.
Chromatography column in this processing step (5) refers to calcium oxide, aluminium hydroxide, silica gel, gac, aluminium sesquioxide etc. further; Described solvent nail alcohol-ethyl acetate, ethanol-ethyl acetate, acetone-ethyl acetate etc.
The invention has the beneficial effects as follows:
At least a as raw material with in mulberry leaf, ramulus mori, White Mulberry Root-bark of the extracting method of DNJ monomer of the present invention, physics-chem characteristic in conjunction with the various compositions of DNJ, raw material is extracted under room temperature～100 ℃, extracting solution is directly gone up the Zeo-karb enrichment, DNJ ammonia soln after the ammoniacal liquor wash-out adopts reverse osmosis technology to carry out the pre-concentration enrichment, the decrease energy consumption; Intermediate carries out catalyzed conversion, directly collects product, the operation that need not to monitor during chromatography.Whole operating process energy consumption is very low, compares with traditional technology, has greatly saved the energy.
Ammonia in the ammonia soln that DNJ on wash-out Zeo-karb of the present invention uses, recycle after reclaiming; The hydrophilic solvent of wash-out gel column, and the mixed solvent of elution chromatography post etc., all recycle.Whole energy saving technology environmental protection, the recycle that has realized extracting solvent.
The eluting solvent that the present invention selects and intermediate catalytic conversion technique have been realized the classification wash-out automatically, and the separation each time in three separation need not middlely be monitored, and production technology is easily grasped.The product that obtains, its purity can be controlled at 10%, 50%, 90%, 99% 4 rank.Can satisfy the bulk drug of ofhypoglycemic medicine and cancer therapy drug, also can be used as reference substance and use, can also be as the use of other products materials.
Embodiment
Below will be described in detail the preferred embodiments of the present invention.Should be appreciated that preferred embodiment only for the present invention is described, rather than in order to limit protection scope of the present invention.
Embodiment 1
(1) mulberry branch 1Kg is with soaking 48 hours under the sulphuric acid soln room temperature of 10 liter 0.1% (w/v); Filter, obtain extracting solution (approximately 9 liters).
(2) 9 liters of extracting solutions with 0.9 liter of 732 Zeo-karb whip attachment that pre-treatment is good, then, with 0.9 premium on currency washing resin, then are used 0.9 liter of 2mol/L ammonia scrubbing resin.Approximately obtain 0.9 liter of ammoniacal liquor elutriant.
After 2 times of (3) 0.9 liters of ammoniacal liquor elutriant reverse osmosis concentration (approximately obtaining 0.45 liter of concentrated solution), Distillation recovery ammonia obtains intermediate A (approximately 10 grams); The ammoniacal liquor that reclaims is used for the wash-out of (2).
(4) intermediate A is water-soluble and be mixed with 20% solution (approximately 50 milliliters), adds 100 ℃ of 0.01% hydrochloric acid (w/v) circulationization 1 hour next time, and upper 50 milliliters of Superose gel chromatography columns wash with water; Collect elutriant.
(5) get intermediate B after the recovery solvent, intermediate B (approximately 1 gram) is dissolved in 5 ml methanol-ethyl acetate (1:1) mixed solvent, and 5 gram silicagel columns on solution are with methyl alcohol-ethyl acetate (1:1) mixed solvent wash-out.
(6) reclaim solvent, vacuum-drying namely gets the S-GI sterling.S-GI purity 99%.
Embodiment 2
(1) mulberry leaf 1Kg soaked 0.1 hour with under 100 ℃ of the hydrochloric acid solns of 2 liter 5% (w/v), filtered, and obtained extracting solution (approximately 1 liter); Residue is used 2 liters of 0.5%(w/v again) 100 ℃ of hydrochloric acid solns under soaked 0.1 hour, filter, obtain extracting solution (approximately 1 liter); Residue with soaking 0.1 hour under 100 ℃, 2 premium on currency, filters again, obtains extracting solution (approximately 1 liter).Merge No. three times extracting solution, obtain approximately 3 liters of extracting solutions.
(2) 3 liters of extracting solutions with 3 liters of 001 Zeo-karb whip attachment that pre-treatment is good, then, with 30 premium on currency washing resins, then are used 30 liters of 0.05mol/L ammonia scrubbing resins.Approximately obtain 30 liters of ammoniacal liquor elutriants.
After 20 times of (3) 30 liters of ammoniacal liquor elutriant reverse osmosis concentration (approximately obtaining 1.5 liters of concentrated solutions), Distillation recovery ammonia obtains intermediate A (approximately 20 grams); The ammoniacal liquor that reclaims is used for the wash-out of (2).
(4) intermediate A is dissolved in methyl alcohol and is mixed with 1% solution (approximately 2000 milliliters), adds that under 60 ℃, 1% sulfuric acid (w/v), insulation transforms 1 hour, and upper 20000 milliliters of Sephdex LH-20 gel chromatography columns are used methanol-eluted fractions; Collect elutriant.
(5) get intermediate B after the recovery solvent, intermediate B (approximately 1 gram) is dissolved in 100 milliliters of ethanol-ethyl acetate (1:1), 1000 gram aluminium sesquioxide posts on solution, ethanol-ethyl acetate (1:1) wash-out.
(6) reclaim solvent, vacuum-drying namely gets the S-GI sterling.S-GI purity 50%.
Embodiment 3
(1) White Mulberry Root-bark 1Kg soaked 5 hours with under 60 ℃ of the sulphuric acid solns of 5 liter 1% (w/v), filtered, and obtained extracting solution (approximately 3 liters); Residue is used under 60 ℃ of the sulphuric acid solns of 5 liter 1% (w/v) again and was soaked 5 hours, filters, and obtains extracting solution (approximately 4 liters).Merge secondary raffinate, obtain approximately 7 liters of extracting solutions.
(2) 7 liters of extracting solutions with 3.5 liters of D151 Zeo-karb whip attachment that pre-treatment is good, then, with 7 premium on currency washing resins, then are used 7 liters of 0.5mol/L ammonia scrubbing resins.Approximately obtain 7 liters of ammoniacal liquor elutriants.
After 10 times of (3) 7 liters of ammoniacal liquor elutriant reverse osmosis concentration (approximately obtaining 0.7 liter of concentrated solution), Distillation recovery ammonia obtains intermediate A (approximately 30 grams); The ammoniacal liquor that reclaims is used for the wash-out of (2).
(4) intermediate A is dissolved in ethanol and is mixed with 10% solution (approximately 300 milliliters), adds that under 75 ℃, 0.1% sulfuric acid (w/v), insulation transforms 1 hour, and upper 1500 milliliters of Hitrap SP gel chromatography columns are used ethanol elution; Collect elutriant.
(5) after the recovery solvent, intermediate B (approximately 5 grams) is dissolved in 50 milliliters of acetone-ethyl acetate (1:1), 250 gram activated carbon columns on solution, acetone-ethyl acetate (1:1) wash-out.
(6) reclaim solvent, vacuum-drying namely gets the S-GI sterling.S-GI purity 10%.
Embodiment 4
(1) mulberry branch 1Kg with soaking 12 hours under 90 ℃, 6 premium on currency, filters, and obtains extracting solution (approximately 4 liters); Residue with soaking 12 hours under 90 ℃, 4 premium on currency, filters again, obtains extracting solution (approximately 3 liters).Merge secondary raffinate, obtain approximately 7 liters of extracting solutions.
(2) 7 liters of extracting solutions with 1.4 liters of 122 Zeo-karb whip attachment that pre-treatment is good, then, with 2.8 premium on currency washing resins, then are used 2.8 liters of 0.1mol/L ammonia scrubbing resins.Approximately obtain 2.8 liters of ammoniacal liquor elutriants.
After 5 times of (3) 2.8 liters of ammoniacal liquor elutriant reverse osmosis concentration (approximately obtaining 0.6 liter of concentrated solution), Distillation recovery ammonia obtains intermediate A (approximately 10 grams); The ammoniacal liquor that reclaims is used for the wash-out of (2).
(4) intermediate A is dissolved in acetone and is mixed with 5% solution (approximately 200 milliliters), adds that under 60 ℃ of 0.2% hydrochloric acid (w/v), insulation transforms 1 hour, and upper 200 milliliters of sephodex gel chromatography columns are used the acetone wash-out; Collect elutriant.
(5) get intermediate B after the recovery solvent, intermediate B (approximately 2 grams) is dissolved in 40 ml methanol-ethyl acetate (1:1), 80 gram calcium oxide posts on solution, methyl alcohol-ethyl acetate (1:1) wash-out.
(6) reclaim solvent, vacuum-drying namely gets the S-GI sterling.S-GI purity 90%.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although with reference to preferred embodiment, the present invention is had been described in detail, those of ordinary skill in the art is to be understood that, can modify or be equal to replacement technical scheme of the present invention, and not breaking away from aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of claim scope of the present invention.
Claims (8)
1. the extracting method of a S-GI monomer, is characterized in that: comprise the steps:
(1) the mulberry raw material extracts 1～3 time with aqueous acid or the water soaking of 2～10 times of w/v, each 0.1～48 hour;
(2) with the cationic exchange resin adsorption of extracting solution with 0.1～1 times of volume; After using again the washing of 1～10 times of Zeo-karb volume, use at last the ammoniacal liquor wash-out of 1～10 times of Zeo-karb volume;
(3) with after 2～20 times of ammoniacal liquor elutriant reverse osmosis pre-concentration, Distillation recovery ammonia obtains intermediate A;
(4) the water-soluble or hydrophilic solvent of intermediate A and be mixed with the solution of 1～20%w/v uses the hydrochloric acid of 0.01～1%w/v or sulfuric acid catalysis to transform 1 hour, and according to gel: solution is gel chromatography column on the volume ratio of 1～10:1, water or hydrophilic solvent wash-out;
(5) get intermediate B after the described water of recovery or hydrophilic solvent, intermediate B is dissolved in the mixed solvent of methyl alcohol-ethyl acetate or Ethanol-Acetic Acid ethyl ester or acetone and ethyl acetate, be mixed with the solution of 1～20%w/v, according to sorbent material: solution is chromatography column on the volume ratio of 1～10:1, described mixed solvent wash-out;
(6) reclaim described mixed solvent, vacuum-drying namely gets the S-GI product.
2. the extracting method of S-GI monomer according to claim 1, it is characterized in that: the mulberry raw material in step (1) refers to one or more in ramulus mori, mulberry leaf, White Mulberry Root-bark; Sour water refers to sulfuric acid or aqueous hydrochloric acid, and its quality concentration of volume percent is〉0%, and≤5%; Immersion refers to soak under room temperature to 100 ℃.
3. the extracting method of S-GI monomer according to claim 1, it is characterized in that: the Zeo-karb in step (2) refers to common strongly-acid or weakly acidic cation-exchange resin, selection 732,7320,001, D001,110,122, D113, D151,724.
4. the extracting method of S-GI monomer according to claim 1 is characterized in that: in step (2), the concentration of ammoniacal liquor is 0.05～2mol/L.
5. the extracting method of S-GI monomer according to claim 1, it is characterized in that: the hydrochloric acid of the use 0.01～1%w/v in step (4) or sulfuric acid catalysis transform, before referring to gel chromatography column, first make catalyzer at 60～100 ℃ of lower catalyzed conversions with hydrochloric acid or the sulfuric acid of 0.01～1%w/v; Gel chromatography column is selected from Superose, Sephdex LH-20, Hitrap SP, sephodex; Described hydrophilic solvent nail alcohol, ethanol or acetone.
6. the extracting method of S-GI monomer according to claim 1, it is characterized in that: the chromatography column in step (5) refers to calcium oxide, aluminium hydroxide, silica gel, gac or aluminium sesquioxide.
7. the extracting method of the described S-GI monomer of according to claim 1-6 any one is characterized in that: the ammoniacal liquor that step (3) reclaims is used for the wash-out of second step (2); The water that step (5) reclaims or hydrophilic solvent are used for the wash-out in (4) step; The described mixed solvent that step (6) reclaims is used for the wash-out in (5) step.
8. the extracting method of the described S-GI monomer of according to claim 1-6 any one, it is characterized in that: this extracting method can obtain the S-GI product of 10%, 50%, 90%, 99% purity.
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| CN103755623B (en) * | 2014-01-03 | 2015-12-30 | 广州军区广州总医院 | The acid ethanol solution that a kind of response phase method is optimized extracts the method for 1-Deoxynojirimycin in Mulberry Leaves |
| CN109170796B (en) * | 2018-09-28 | 2021-07-20 | 湖南华诚生物资源股份有限公司 | Method for extracting thaumatin from African arrowroot |
| CN112300058A (en) * | 2020-12-03 | 2021-02-02 | 中国热带农业科学院热带作物品种资源研究所 | Method for extracting 1-deoxynojirimycin from fresh mulberry leaves |
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| CN1762428A (en) * | 2005-10-09 | 2006-04-26 | 上海现代药物制剂工程研究中心有限公司 | Method for extracting efficient component from mulberry leaf |
| CN101020655A (en) * | 2007-03-15 | 2007-08-22 | 浙江大学 | Process of extracting 1-deoxy nojirimycin |
| CN101314586A (en) * | 2008-06-20 | 2008-12-03 | 江苏大学 | Method for separating 1-deoxidized nojirimycin monomer from mulberry leaf total alkaloid |
| CN101654428A (en) * | 2009-09-11 | 2010-02-24 | 成都市金医生科技健康产业有限公司 | Method for extracting and separating 1-deoxynojirimycin with high purity from natural products |
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| CN1762428A (en) * | 2005-10-09 | 2006-04-26 | 上海现代药物制剂工程研究中心有限公司 | Method for extracting efficient component from mulberry leaf |
| CN101020655A (en) * | 2007-03-15 | 2007-08-22 | 浙江大学 | Process of extracting 1-deoxy nojirimycin |
| CN101314586A (en) * | 2008-06-20 | 2008-12-03 | 江苏大学 | Method for separating 1-deoxidized nojirimycin monomer from mulberry leaf total alkaloid |
| CN101654428A (en) * | 2009-09-11 | 2010-02-24 | 成都市金医生科技健康产业有限公司 | Method for extracting and separating 1-deoxynojirimycin with high purity from natural products |
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