CN103030708B - A kind of Novel purification method of lycium barbarum polysaccharide - Google Patents
A kind of Novel purification method of lycium barbarum polysaccharide Download PDFInfo
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- CN103030708B CN103030708B CN201210595559.4A CN201210595559A CN103030708B CN 103030708 B CN103030708 B CN 103030708B CN 201210595559 A CN201210595559 A CN 201210595559A CN 103030708 B CN103030708 B CN 103030708B
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- polysaccharide
- methyl alcohol
- elutriant
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- lycium barbarum
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- 239000008518 lycium barbarum polysaccharide Substances 0.000 title claims abstract description 36
- 238000000746 purification Methods 0.000 title claims abstract description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 144
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 66
- 150000004676 glycans Polymers 0.000 claims abstract description 58
- 239000005017 polysaccharide Substances 0.000 claims abstract description 58
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 58
- 150000004804 polysaccharides Polymers 0.000 claims abstract description 58
- 238000000034 method Methods 0.000 claims abstract description 43
- 239000000706 filtrate Substances 0.000 claims abstract description 36
- 239000012535 impurity Substances 0.000 claims abstract description 25
- 238000001035 drying Methods 0.000 claims abstract description 20
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 17
- 239000011347 resin Substances 0.000 claims abstract description 15
- 229920005989 resin Polymers 0.000 claims abstract description 15
- 235000012054 meals Nutrition 0.000 claims abstract description 13
- 239000000047 product Substances 0.000 claims abstract description 13
- 239000000284 extract Substances 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 235000015468 Lycium chinense Nutrition 0.000 claims abstract description 6
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- 238000007670 refining Methods 0.000 claims abstract description 6
- 240000002012 Lycium chinense Species 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims abstract description 3
- 239000000203 mixture Substances 0.000 claims description 27
- 239000000243 solution Substances 0.000 claims description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 229910052799 carbon Inorganic materials 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 239000000741 silica gel Substances 0.000 claims description 8
- 229910002027 silica gel Inorganic materials 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 229960001866 silicon dioxide Drugs 0.000 claims description 8
- 238000000967 suction filtration Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 4
- 230000003213 activating Effects 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
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- 230000000694 effects Effects 0.000 abstract description 8
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- 238000000605 extraction Methods 0.000 description 3
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- XACHQDDXHDTRLX-GMPBGBGESA-N Physalien Natural products O=C(O[C@H]1CC(C)(C)C(/C=C/C(=C\C=C\C(=C/C=C/C=C(\C=C\C=C(/C=C/C=2C(C)(C)C[C@H](OC(=O)CCCCCCCCCCCCCCC)CC=2C)\C)/C)\C)/C)=C(C)C1)CCCCCCCCCCCCCCC XACHQDDXHDTRLX-GMPBGBGESA-N 0.000 description 2
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- PVNIIMVLHYAWGP-UHFFFAOYSA-N nicotinic acid Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
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- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
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- RKUNBYITZUJHSG-FXUDXRNXSA-N Hyoscyamine Chemical compound C1([C@@H](CO)C(=O)O[C@H]2C[C@H]3CC[C@@H](C2)N3C)=CC=CC=C1 RKUNBYITZUJHSG-FXUDXRNXSA-N 0.000 description 1
- 210000004185 Liver Anatomy 0.000 description 1
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- PNNNRSAQSRJVSB-BXKVDMCESA-N Rhamnose Chemical group C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
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- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
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- 240000002686 Solanum melongena Species 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-N Trimethylglycine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 1
- 206010047513 Vision blurred Diseases 0.000 description 1
- LRFVTYWOQMYALW-UHFFFAOYSA-N Xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 1
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- PYMYPHUHKUWMLA-WDCZJNDASA-N aldehydo-D-arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
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- 150000001746 carotenes Chemical class 0.000 description 1
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- 239000003610 charcoal Substances 0.000 description 1
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- 238000000151 deposition Methods 0.000 description 1
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
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- 235000019192 riboflavin Nutrition 0.000 description 1
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N β-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- DMASLKHVQRHNES-FKKUPVFPSA-N β-cryptoxanthin Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C DMASLKHVQRHNES-FKKUPVFPSA-N 0.000 description 1
Abstract
The present invention discloses a kind of Novel purification method of lycium barbarum polysaccharide, and the method is: Chinese wolfberry fruit powder is broken into meal, and the methanol eddy adding 15 times amount 85% extracts 120 minutes, filters; Filtrate, reclaims methyl alcohol, is concentrated into 3 Baumes, the hydrochloric acid by volume ratio 1: 1: concentrated solution pH value is adjusted to 2.5 by water, places 20 minutes, filters; Broad spectrum type macroporous resin adsorption post on filtrate, collects elutriant, and reclaim methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with the acetone of volume ratio 3: 1: methanol wash column impurity, continue with the acetone of volume ratio 7: 1: methanol-eluted fractions, collect elutriant; Carried out refining to obtain lycium barbarum polysaccharide finished product by collected polysaccharide elutriant, its content is more than 70%.The maximum feature of the inventive method utilizes that the coupling of macroporous adsorbent resin-silica gel column chromatography is convenient and swift, abundant raw material source, low production cost, separating effect are obvious, DNA purity is high.
Description
Technical field:
The invention belongs to field of natural organic chemistry, relate to a kind of purification process of lycium barbarum polysaccharide, particularly relate to a kind of novel method utilizing macroporous adsorbent resin-silica gel column chromatography coupled method purified wolfberry fruit polysaccharide.
Background technology:
Matrimony vine, formal name used at school: Lyciumchinense, another name Gou Qizi, matrimony vine red real, beet is sub, FRUCTUS LYCII BARBARI, dog milk, dark purple green pepper, Chinese holly hoof, FRUCTUS LYCII, frame, Chinese holly eggplant eggplant, red ear pendant, blood Chinese holly, Chinese holly ground bud, matrimony vine beans, blood matrimony vine, fructus lycii sinensis, be the fruit of Solanaceae Lycium matrimony vine.Containing trimethyl-glycine (betane), coromegine (aTCMLIBopine), l-tropine tropate (gyoscyamine), lycium barbarum polysaccharide, again containing Zea mays xanthin, physalin, kryptoxanthin (cryptosxanthin), Scopoletin (scopo-letin), carotene, VitB1, riboflavin, nicotinic acid, vitamins C etc., there is enriching yin improving eyesight, except irascibility, be beneficial to sleep, establishing-Yang, beauty face-whitening-nourishing, the effect such as delay senility.
Lycium barbarum polysaccharide is a kind of water-soluble polysaccharide extracting from matrimony vine and obtain.Clearly this polysaccharide system protein-polysaccharide, is made up of pectinose, glucose, semi-lactosi, seminose, wood sugar, rhamnosyl 6 kinds of monosaccharide components.The main active ingredient show after deliberation, lycium barbarum polysaccharide is wolfberry fruit immunity moderation, delaying senility, can improve the symptoms such as the elderly's fatiguability, poor appetite and blurred vision, and have reducing blood-fat, anti-fatty liveranti-fatty liver, the anti-ageing effect of waiting for a long time.In recent years, it is found that in natural phant matrimony vine containing a large amount of lycium barbarum polysaccharides, because its raw material matrimony vine source is wider, therefrom extract lycium barbarum polysaccharide and achieve encouraging progress, name is called in the patent application of " a kind of extraction and separation method of lycium barbarum polysaccharide " and just discloses by matrimony vine with drying, pulverizing, sieve process, and ultrasound assisted extraction also refines the method for lycium barbarum polysaccharide.Lycium barbarum polysaccharide extraction process in above-mentioned patent documentation is mainly refined through steps such as raw materials pretreatment, ultrasound-assisted extraction, alcohol deposition method separation and is formed, although last lycium barbarum polysaccharide purity is very high, but its technique itself expends greatly, practical application effect is nothing like the effect that macroporous adsorbent resin coupling silica gel chromatography is separated.
Summary of the invention
The object of the invention is the Novel purification method providing a kind of lycium barbarum polysaccharide; The object of the invention is also to provide a kind of convenient and swift, abundant raw material source, low production cost, separating effect are obvious, DNA purity is high method utilizing macroporous adsorbent resin-silica gel column chromatography coupled method enriching and purifying lycium barbarum polysaccharide.
The present invention seeks to be achieved through the following technical solutions.
The Novel purification method of lycium barbarum polysaccharide of the present invention comprises the steps:
Chinese wolfberry fruit powder is broken into meal, and 85% ~ 100% methanol eddy adding meal quality 15 ~ 20 times amount extracts 120 ~ 180 minutes, filters; Filtrate is concentrated into 3 ~ 5 Baumes after reclaiming methyl alcohol and obtains concentrated solution, then uses the hydrochloric acid of volume ratio 1: 1: concentrated solution pH value is adjusted to 2.5 ~ 3 by the aqueous solution, places after 20 ~ 40 minutes and filters; HPD400, HPD100, DM-130 or AB-8 broad spectrum type macroporous resin adsorption post on filtrate, adsorb and completely wash impurity with water, use the methanol wash column impurity of 5% ~ 20% again, then the methanol-eluted fractions of 50% ~ 80% is used, obtain polysaccharide work in-process elutriant, after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol is 1 ~ 3: 1 third with washing impurity with carbinol mixture, then uses acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 5 ~ 7: 1, obtains polysaccharide elutriant; Carried out refining to obtain lycium barbarum polysaccharide by polysaccharide elutriant, its content is more than 70%.
The Novel purification method of lycium barbarum polysaccharide of the present invention comprises the steps:
Take the FRUCTUS LYCII being ground into meal, 85% methanol eddy adding meal quality 20 times amount extracts 180 minutes, filters; Filtrate is concentrated into 5 Baumes after reclaiming methyl alcohol and obtains concentrated solution, then uses the hydrochloric acid of volume ratio 1: 1: concentrated solution pH value is adjusted to 3 by the aqueous solution, places after 40 minutes and filters; HPD400 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 5%, then use the methanol-eluted fractions of 50%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol be 1: 1 acetone and carbinol mixture wash impurity, then use acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 5: 1, obtains polysaccharide elutriant; Polysaccharide elutriant is carried out refining to obtain lycium barbarum polysaccharide.
Described macroporous resin adsorption step adopts wet method dress post, retains liquid level, by the filtrate after adjust pH by adsorption column, and flow velocity per minute 0.05ml/g resin.
Described silica gel column chromatography step adopts wet method dress post, retains liquid level, and work in-process are mixed activation after silica gel by weight at 1: 1, and by the mixture mixed as above silicagel column, elution flow rate per minute 0.1ml/g silica gel.Silica gel activating condition is, 110 degrees Celsius 30 minutes.
Adopt activated carbon decolorizing in the present invention, recrystallization method refines.Add injection active carbon by 4g/100ml elutriant, 25 ~ 40 degrees Celsius are stirred 20 ~ 40 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate; Intermediate 7 ~ 9 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product.
In the present invention, treating process comprises the steps:
Add injection active carbon by 4g/100ml elutriant, 25 degrees Celsius are stirred 40 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate; Intermediate 9 times of methyl alcohol, 40 degrees Celsius of thermosols, filter; Filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product.
Finished product 7 times of recrystallizing methanol in treating process in the present invention, filtrated stock, reclaim methyl alcohol, concentrate drying obtains polysaccharide work in-process.
The present invention's application macroporous adsorbent resin-silica gel column chromatography coupled method enriching and purifying lycium barbarum polysaccharide, good to the adsorption selectivity of lycium barbarum polysaccharide, absorption is fast resolves also soon, and loading capacity is larger; Abundant raw material source, low production cost, separating effect is obvious, and DNA purity is high, can obtain the polysaccharide work in-process of content more than 45% and the polysaccharide finished product of content more than 70%, overcome the shortcoming that traditional extraction rate is relatively low, DNA purity is low.
The present invention selects that stable in physicochemical property, surface-area are comparatively large, exchange velocity is very fast, physical strength is high, contamination resistance is strong, the macroporous adsorbent resin of Heat stability is good, be not dissolved in acid, alkali and organic matchmaker, better to organism selectivity, not by the impact that inorganic salts and strong ion low molecular compound exist, from solution, polysaccharide can be adsorbed selectively by physical adsorption, absorption is fast, parsing is also fast, and loading capacity is larger.
The present invention selects silica gel chromatography purifying and binding activities charcoal and recrystallizing and refining, and separating effect is better.
Embodiment:
Embodiment 1:
Take FRUCTUS LYCII 10kg, be ground into meal, the methanol eddy adding 18 times amount 90% extracts 140 minutes, filters; Filtrate, reclaims methyl alcohol, is concentrated into 4 Baumes, the hydrochloric acid by volume ratio 1: 1: concentrated solution pH value is adjusted to 2.8 by the aqueous solution, places 40 minutes, filters; HPD100 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 15%, then use the methanol-eluted fractions of 70%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol be 2: 1 acetone and carbinol mixture wash impurity, then use acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 6: 1, obtains polysaccharide elutriant 2L; Add 80g injection active carbon, 30 degrees Celsius are stirred 30 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate 35g; Intermediate 8 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product 20g, content 78%.
Embodiment 2:
Take FRUCTUS LYCII 15kg, be ground into meal, the methanol eddy adding 20 times amount 95% extracts 160 minutes, filters; Filtrate, reclaims methyl alcohol, is concentrated into 5 Baumes, the hydrochloric acid by volume ratio 1: 1: concentrated solution pH value is adjusted to 3 by the aqueous solution, places 40 minutes, filters; HPD400 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 10%, then use the methanol-eluted fractions of 60%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol be 1: 1 acetone and carbinol mixture wash impurity, then use acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 7: 1, obtains polysaccharide elutriant 3.5L; Add 140g injection active carbon, 40 degrees Celsius are stirred 40 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate 60g; Intermediate 9 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product 32g, content 76.3%.
Embodiment 3:
Take FRUCTUS LYCII 20kg, be ground into meal, the methanol eddy adding 15 times amount 87% extracts 120 minutes, filters; Filtrate, reclaims methyl alcohol, is concentrated into 3 Baumes, the hydrochloric acid by volume ratio 1: 1: concentrated solution pH value is adjusted to 2.5 by the aqueous solution, places 40 minutes, filters; AB-8 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 5%, then use the methanol-eluted fractions of 80%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol be 3: 1 acetone and carbinol mixture wash impurity, then use acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 5: 1, obtains polysaccharide elutriant 5L; Add 200g injection active carbon, 25 degrees Celsius are stirred 20 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate 70g; Intermediate 8 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product 36g, content 77.5%.
Embodiment 4:
Take FRUCTUS LYCII 5kg, be ground into meal, the methanol eddy adding 20 times amount 98% extracts 160 minutes, filters; Filtrate, reclaims methyl alcohol, is concentrated into 3 Baumes, the hydrochloric acid by volume ratio 1: 1: concentrated solution pH value is adjusted to 2.6 by the aqueous solution, places 40 minutes, filters; DM-130 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 5%, then use the methanol-eluted fractions of 50%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol be 1: 1 acetone and carbinol mixture wash impurity, then use acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 7: 1, obtains polysaccharide elutriant 1200ml; Add 48g injection active carbon, 40 degrees Celsius are stirred 25 minutes, filter; Filtrate, reclaim mix reagent, concentrate drying obtains intermediate 20g; Intermediate 7 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product 12g, content 78.2%.
Claims (5)
1. a Novel purification method for lycium barbarum polysaccharide, is characterized in that the method comprises the steps:
Chinese wolfberry fruit powder is broken into meal, and 85% ~ 100% methanol eddy adding meal quality 15 ~ 20 times amount extracts 120 ~ 180 minutes, filters; Filtrate is concentrated into 3 ~ 5 Baumes after reclaiming methyl alcohol and obtains concentrated solution, then uses the hydrochloric acid of volume ratio 1:1: concentrated solution pH value is adjusted to 2.5 ~ 3 by the aqueous solution, places after 20 ~ 40 minutes and filters; HPD400, HPD100, DM-130 or AB-8 broad spectrum type macroporous resin adsorption post on filtrate, adsorb and completely wash impurity with water, use the methanol wash column impurity of 5% ~ 20% again, then the methanol-eluted fractions of 50% ~ 80% is used, obtain polysaccharide work in-process elutriant, after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol is that the acetone of 1 ~ 3:1 and carbinol mixture wash impurity, then uses acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 5 ~ 7:1, obtains polysaccharide elutriant; Polysaccharide elutriant is carried out refining to obtain lycium barbarum polysaccharide;
Wherein, described silica gel column chromatography step adopts wet method dress post, retains liquid level, by work in-process by weight 1:1 mix silica gel after activation, and the mixture mixed is placed in above silicagel column, elution flow rate per minute 0.1ml/g silica gel, silica gel activating condition is, 110 degrees Celsius 30 minutes; Employing activated carbon decolorizing, recrystallization method are refined, and add injection active carbon by 4g/100ml polysaccharide elutriant, and 25 ~ 40 degrees Celsius are stirred 20 ~ 40 minutes, filter; After filtrate reclaims mix reagent, concentrate drying obtains intermediate; Intermediate 7 ~ 9 times of methyl alcohol, 40 degrees Celsius of thermosols, filter: filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product.
2. the Novel purification method of lycium barbarum polysaccharide as claimed in claim 1, is characterized in that the method comprises the steps:
Take the FRUCTUS LYCII being ground into meal, 85% methanol eddy adding meal quality 20 times amount extracts 180 minutes, filters; Filtrate is concentrated into 5 Baumes after reclaiming methyl alcohol and obtains concentrated solution, then uses the hydrochloric acid of volume ratio 1:1: concentrated solution pH value is adjusted to 3 by the aqueous solution, places after 40 minutes and filters; HPD400 type macroporous resin adsorption post on filtrate, adsorbs and completely washes impurity with water, then use the methanol wash column impurity of 5%, then use the methanol-eluted fractions of 50%, obtain polysaccharide work in-process elutriant, and after polysaccharide work in-process elutriant reclaims methyl alcohol, concentrate drying obtains polysaccharide work in-process; By silica gel column chromatography on gained polysaccharide work in-process, and with acetone: the volume ratio of methyl alcohol is that the acetone of 1:1 and carbinol mixture wash impurity, then uses acetone: the volume ratio of methyl alcohol is acetone and the carbinol mixture wash-out of 5:1, obtains polysaccharide elutriant; Polysaccharide elutriant is carried out refining to obtain lycium barbarum polysaccharide.
3. the Novel purification method of lycium barbarum polysaccharide as claimed in claim 1 or 2, is characterized in that the step of macroporous resin adsorption described in the method adopts wet method dress post, retains liquid level, by the filtrate after adjust pH by adsorption column, and flow velocity per minute 0.05ml/g resin.
4. the Novel purification method of lycium barbarum polysaccharide as claimed in claim 1, is characterized in that in the method, treating process comprises the steps:
Add injection active carbon by 4g/100ml elutriant, 25 degrees Celsius are stirred 40 minutes, filter; Filtrate reclaims mix reagent, and concentrate drying obtains intermediate; Intermediate 9 times of methyl alcohol, 40 degrees Celsius of thermosols, filter; Filtrate cools crystallize out, and suction filtration obtains lycium barbarum polysaccharide finished product.
5. the Novel purification method of lycium barbarum polysaccharide as claimed in claim 4, is characterized in that finished product 7 times of recrystallizing methanol in the method, filtrated stock, reclaims methyl alcohol, and concentrate drying obtains the whole finished product of polysaccharide.
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| CN1670036A (en) * | 2005-03-18 | 2005-09-21 | 天津科技大学 | Chinese wolfberry polysaccharide extracting method and weight reduction product containing Chinese wolfberry polysaccharide |
| CN101812137A (en) * | 2009-12-28 | 2010-08-25 | 大兴安岭科丽尔生物工程有限责任公司 | Method for enriching agaric polysaccharide in purified black agaric by using macroporous absorption resin |
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| CN1670036A (en) * | 2005-03-18 | 2005-09-21 | 天津科技大学 | Chinese wolfberry polysaccharide extracting method and weight reduction product containing Chinese wolfberry polysaccharide |
| CN101812137A (en) * | 2009-12-28 | 2010-08-25 | 大兴安岭科丽尔生物工程有限责任公司 | Method for enriching agaric polysaccharide in purified black agaric by using macroporous absorption resin |
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| "宁夏枸杞新品种"宁杞1号"干果中总糖及多糖类物质的系统分析";高向东等;《药物生物技术》;19940623;第1卷(第2期);第43页 * |
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