CN106967142B - Method that is a kind of while extracting momordica glycoside V, VI and 11-O base glycosides V - Google Patents
Method that is a kind of while extracting momordica glycoside V, VI and 11-O base glycosides V Download PDFInfo
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- 229930182470 glycoside Natural products 0.000 title claims abstract description 127
- 150000002338 glycosides Chemical class 0.000 title claims abstract description 127
- 235000009815 Momordica Nutrition 0.000 title claims abstract description 58
- 241000218984 Momordica Species 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 38
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 84
- 239000004952 Polyamide Substances 0.000 claims abstract description 42
- 229920002647 polyamide Polymers 0.000 claims abstract description 42
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 238000002425 crystallisation Methods 0.000 claims abstract description 33
- 230000008025 crystallization Effects 0.000 claims abstract description 33
- 241001409321 Siraitia grosvenorii Species 0.000 claims abstract description 29
- 239000012452 mother liquor Substances 0.000 claims abstract description 29
- 235000011171 Thladiantha grosvenorii Nutrition 0.000 claims abstract description 28
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 24
- LTDANPHZAHSOBN-IPIJHGFVSA-N (2R,3R,4S,5S,6R)-2-[[(2R,3S,4S,5R,6R)-6-[[(3S,8R,9R,10S,11R,13R,14S,17R)-17-[(2R,5R)-5-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-3-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-6-[[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl]oxy-6-hydroxy-6-methylheptan-2-yl]-11-hydroxy-4,4,9,13,14-pentamethyl-2,3,7,8,10,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-3,4-dihydroxy-5-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]methoxy]-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@H](CC[C@@H](C)[C@@H]1[C@]2(C[C@@H](O)[C@@]3(C)[C@@H]4C(C([C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]6[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O6)O)O5)O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)CC4)(C)C)=CC[C@@H]3[C@]2(C)CC1)C)C(C)(C)O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O LTDANPHZAHSOBN-IPIJHGFVSA-N 0.000 claims abstract description 20
- 239000013078 crystal Substances 0.000 claims abstract description 13
- 238000001816 cooling Methods 0.000 claims abstract description 12
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 238000004090 dissolution Methods 0.000 claims abstract 2
- 239000012141 concentrate Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 235000019441 ethanol Nutrition 0.000 claims description 23
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 230000001476 alcoholic effect Effects 0.000 claims description 15
- 238000010828 elution Methods 0.000 claims description 14
- 235000009508 confectionery Nutrition 0.000 claims description 12
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 8
- KTUQUZJOVNIKNZ-UHFFFAOYSA-N butan-1-ol;hydrate Chemical compound O.CCCCO KTUQUZJOVNIKNZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 235000013399 edible fruits Nutrition 0.000 claims description 2
- 244000269722 Thea sinensis Species 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 16
- 241001122767 Theaceae Species 0.000 description 11
- 229930189775 mogroside Natural products 0.000 description 9
- 239000012535 impurity Substances 0.000 description 7
- 238000000926 separation method Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 150000008442 polyphenolic compounds Chemical class 0.000 description 5
- 235000013824 polyphenols Nutrition 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 4
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 239000004615 ingredient Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 238000010812 external standard method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 150000001412 amines Chemical class 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229930003944 flavone Natural products 0.000 description 2
- 150000002213 flavones Chemical class 0.000 description 2
- 235000011949 flavones Nutrition 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920002292 Nylon 6 Polymers 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000018927 edible plant Nutrition 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 150000002191 fatty alcohols Chemical class 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000021022 fresh fruits Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical group 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 235000021096 natural sweeteners Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 description 1
- -1 triterpene glucoside Chemical class 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
- C07J17/005—Glycosides
Abstract
Method that is a kind of while extracting momordica glycoside V, VI and 11-O base glycosides V, comprising the following steps: (1) dissolve momordica glycoside V refinement mother liquor, extracting n-butyl alcohol obtains n-butanol layer；(2) it is concentrated, cooling, crystallization, filtering obtains crystal and crystallization mother liquor, and crystal is dry, obtains mogroside VI；(3) by crystallization mother liquor concentrations to no alcohol, polyamide chromatography column is crossed in dissolution, collects efflux, is concentrated, dry, obtains momordica glycoside V；(4) successively chromatographic column is eluted with low pure and mild height alcohol, collects height alcohol eluen, be concentrated, cooling, crystallization filters, and it is dry, obtain Siraitia grosvenorii 11-O base glycosides V.Purity >=97% of mogroside VI obtained by the method for the present invention, yield >=83%, purity >=89% of momordica glycoside V, purity >=96% of yield >=89%, 11-O base glycosides V, yield >=85%；The method of the present invention mild condition, safety and environmental protection are suitable for industrialized production.
Description
Technical field
The present invention relates to a kind of methods for extracting mogroside, and in particular to a kind of to extract momordica glycoside V, VI simultaneously
With the method for 11-O base glycosides V.
Background technique
Siraitia grosvenorii is the traditional medicinal and edible plant in China, and mogroside is its main functional component, is a kind of high
The ideal natural sweetener of sugariness, low-calorie, by the concern of people due to it is with multiple biological activities.Mogroside
There are many, be triterpene alcohol with Aglycone research shows that mogroside is a kind of triterpene glucoside, have two by
The glucoside side chain of four or less glucose units composition, is connected with β-glycosidic bond with aglycon, the company between side chain glucose sugar
Connecing key is β -1,6 and β -1,2 glycosidic bond.Wherein, the content highest of momordica glycoside V, followed by 11-O base glycosides V and glycosides VI, three
The molecular formula of kind sweet tea glycosides is as follows:
Momordica glycoside V
Siraitia grosvenorii 11-O base glycosides V mogroside VI
The extraction of mogroside has the industrialization experience of many years at home, separation for momordica glycoside V and mentions
Pure research is than wide.
CN101029071A discloses a kind of method that high-purity Momordia grosvenori aglycone V is prepared from Siraitia grosvenorii, is with Siraitia grosvenorii
Fresh fruit is raw material, is extracted by alcohol reflux, ethyl acetate and extracting n-butyl alcohol, and polyamide and silica gel column chromatography obtain Siraitia grosvenorii
Glycosides V.But this method is excessively complicated, production cost is high, and can only obtain a kind of ingredient of momordica grosvenori glycoside V.
CN106074669A discloses a kind of extracting method of Siraitia grosvenorii polyphenol, is passed through using Siraitia grosvenorii pericarp as raw material
Dry, pulverize, EtOH Sonicate extract, polycaprolactam, elution, concentration and etc. obtain Siraitia grosvenorii polyphenol extract.But it should
Method is not directed to the separation method of mogroside.
CN1375499A discloses a kind of method that separation multiple components are extracted from Siraitia grosvenorii, is with Siraitia grosvenorii for original
Material, is extracted by buck, fatty alcohol precipitating, and the processing such as ion exchange resin, macroreticular resin, polyamide obtains Siraitia grosvenorii
Sweet tea glycosides, momordica grosvenori polysaccharide and grosvenor momordica flavonoid.But the mogroside of this method acquisition is the mixture of a variety of sweet tea glycosides, and
The high purity product of each sweet tea glycosides is not obtained.
CN101851265A discloses a kind of method that a variety of active ingredients is extracted from dried fructus momordicae, is first by dry arhat
Fruit stoning, then by adding water sudden strain of a muscle to mention, centrifugation, multiple polyamide chromatography, D213 decoloration, alcohol precipitation, enzymatic hydrolysis and etc., respectively obtain sweet tea
The ingredients such as glycosides, polyphenol, polysaccharide, oligosaccharide and amino acid.But the mogroside that this method obtains equally is a variety of sweet tea glycosides
Mixture does not obtain the high purity product of each sweet tea glycosides.
From the foregoing, it will be observed that the existing report about Siraitia grosvenorii 11-O base glycosides V and the separation of glycosides VI and purification is seldom.In actual production
Obtained Fructus Monordicae extract is usually the mixture of the total glycosides of Siraitia grosvenorii, therefore, there is an urgent need to find a kind of method, is realized single
The separation of ingredient, and realize industrialization.
Summary of the invention
The technical problem to be solved by the present invention is to overcome drawbacks described above of the existing technology, provide a kind of gained production
Product purity, high income, at low cost, process conditions are mild, safety and environmental protection, suitable for extracting mogroside while industrialized production
V, the method for VI and 11-O base glycosides V.
The technical solution adopted by the present invention to solve the technical problems is as follows: a kind of to extract momordica glycoside V, VI and simultaneously
The method of 11-O base glycosides V, comprising the following steps:
(1) extracting n-butyl alcohol: momordica glycoside V refinement mother liquor is dissolved with water, and n-butanol is added and is extracted, water is separated
Layer, obtains n-butanol layer；
(2) crystallization is concentrated: n-butanol layer obtained by step (1) is concentrated under reduced pressure, cooling, stirring and crystallizing, filtering, obtain crystal and
N-butanol crystallization mother liquor, crystal is dry, obtain mogroside VI；
(3) polyamide chromatography: n-butanol crystallization mother liquor obtained by step (2) is concentrated under reduced pressure into no alcohol, by gained concentrate
It is dissolved with water, by polyamide chromatography column, collects upper prop efflux, be concentrated under reduced pressure, it is dry, obtain momordica glycoside V；
(4) gradient elution: successively use low alcoholic solution and height alcoholic solution to the polyamide chromatography column of step (3) Guo Zhuhou
It is eluted, collects height alcohol eluen, be concentrated, cooling, crystallization filters, and it is dry, obtain Siraitia grosvenorii 11-O base glycosides V.
Preferably, in step (1), in the momordica glycoside V refinement mother liquor, the mass content of mogroside VI is 10
~30%, the mass content of momordica glycoside V is that the mass content of 10~15%, 11-O base glycosides V is 5~20%.Sieve of the present invention
Chinese fruit sweet tea glycosides V refinement mother liquor is derived from using Fructus Monordicae extract as raw material, prepares high-purity momordica with organic solvent crystallization
Generated mother liquor when glycosides V.
Preferably, in step (1), the mass ratio of the momordica glycoside V refinement mother liquor, water and n-butanol is 1:5~10:
More preferable 1:5~10:5~15 4~30().The present inventor the study found that be added the water-soluble impurity of water dissolvable, pigment with
And inorganic salts etc., by being retained in remove in water layer, and the n-butanol being added is moderate to the solubility of total glycosides, and can be with moisture
Layer realizes the separation of impurity and total glycosides, this is that other solvents are irreplaceable.If the dosage of n-butanol is very few, the extraction of total glycosides
It takes and is not thorough；If the dosage of n-butanol is excessive, the task amount of concentration will be increased, and increase material loss.
Preferably, step (2), in (3), the temperature of the reduced pressure is 70~90 DEG C, the vacuum degree of reduced pressure is-
0.06~-0.09MPa, the mass concentration for being concentrated into concentrate is more preferable 25~30%) 20~35%(.If the concentration after concentration
Liquid concentration is too low, then the concentration of the crystallization of Momordia grosvenori aglycone VI is not achieved, and Momordia grosvenori aglycone VI is unable to fully be precipitated；If the concentration after concentration
Liquid excessive concentration, then impurity will also be precipitated, and reduce the purity of product.
Preferably, in step (2), the temperature of the stirring and crystallizing is 15~30 DEG C, and revolving speed is 60~120r/min, time
For 12~36h.Under the conditions of the crystallization, so that the crystal being precipitated is high-purity mogroside VI, it is other without being precipitated
Sweet tea glycosides or impurity.
Preferably, in step (3), the dosage of the water is 18~50 times (more preferable 20~30 times) of concentrate quality.
It is to chromatograph concentrate by polyamide chromatography column with the state of aqueous solution with the purpose that water dissolves, is more advantageous to absorption.Polyamides
The absorption of amine is " hydrogen bond principle ", and the substance ability that hydrogen bond is only formed with polyamide is adsorbable thereon, and resin adsorption
There is an optimal concentration, all will lead to adsorption rate decline higher or lower than this concentration, so, it is water-soluble if the dosage of water is very few
The excessive concentration of total material in liquid, then polyamide can not adsorb the impurity such as 11-O base glycosides V and flavones, polyphenol, lead to Siraitia grosvenorii
Sweet tea glycosides V can not be separated with 11-O base glycosides V and impurity；If the dosage of water is excessive, the time of upper prop will be extended, causes manpower, object
The waste of power.
Preferably, in step (3), the mesh number of the polyamide is 50~200 mesh, and dosage is the 4~20 of concentrate quality
Again (more preferable 5~10 times), the ratio of height to diameter of polyamide chromatography column is the more preferable 5~15:1 of 1~20:1().Use the mesh of polyamide
Be absorption 11-O base glycosides V, so that 11-O base glycosides V and momordica glycoside V be made to separate.If the dosage of polyamide is very few, or chromatography
The ratio of height to diameter of column is too small, and all will lead to that 11-O base glycosides V is unable to fully is adsorbed；If the dosage of polyamide is excessive or chromatographic column
Ratio of height to diameter it is excessive, all will increase elution difficulty and the time.
Preferably, in step (3), the flow velocity of upper prop is 0.5~2.0BV/h.Column bed body in 1BV=1 described in the method for the present invention
Product.
Preferably, in step (4), the alcohol in the minuent alcoholic solution and height alcoholic solution is methanol, ethyl alcohol, isopropanol
Or one or more of n-butanol etc.；The volumetric concentration of the minuent alcoholic solution is more preferable 15~30%) 10~40%(, institute
The volumetric concentration for stating height alcoholic solution is more preferable 70~80%) 50~85%(.Polarity using low pure and mild height alcohol is different,
Carry out gradient elution, gradient elution first removes the impurity such as flavones, polyphenol with low alcohol elution, then with height alcohol by target product
11-O base glycosides V is eluted and is collected.
Preferably, in step (4), the dosage of the minuent alcoholic solution is 2~5BV, the dosage of height alcoholic solution is 2~
5BV；The flow velocity of the elution is 0.5~2.0BV/h.
Preferably, in step (4), the temperature of the concentration is 60~80 DEG C, and the mass concentration for being concentrated into concentrate is 25
~45%.
Preferably, in step (4), the temperature of the crystallization is 15~30 DEG C, and the time is 12~36 h.
The principle of the method for the present invention are as follows: the ingredient of momordica glycoside V refinement mother liquor in addition to containing a small amount of momordica glycoside V it
Outside, main ingredient is 11-O base glycosides V and mogroside VI.Mogroside VI, momordica grosvenori glycoside V and 11-O are extracted at the same time
When base glycosides V, since proportion is high in mother liquor for mogroside VI, and its polarity is all smaller than other two kinds of sweet tea glycosides, can be just
It crystallizes and is precipitated in butanol, so extracting mogroside VI first；11-O base glycosides V can be with polyamide shape because of its distinctive structure
At hydrogen bond, therefore it can be adsorbed in polyamide chromatography column, and momordica glycoside V cannot form hydrogen bond with polyamide, so polyamides
Amine does not adsorb momordica glycoside V, thus is separated.
The method of the present invention has the beneficial effect that:
(1) momordica glycoside V obtained by the method for the present invention, VI and 11-O base glycosides V are white powder, through high performance liquid chromatography
Method detection, purity >=97% of mogroside VI, yield >=83%, purity >=89% of momordica glycoside V, yield >=89%, 11-
Purity >=96% of O base glycosides V, yield >=85%；Realize at the same from momordica glycoside V refinement mother liquor extract momordica glycoside V,
VI and 11-O base glycosides V, improves the utilization rate of Siraitia grosvenorii resource, increases value-added content of product；
(2) separation and purification mode mild condition that the method for the present invention uses, safety and environmental protection, to equipment without particular/special requirement,
Suitable for industrial production.
Specific embodiment
Below with reference to embodiment, the invention will be further described.
Momordica glycoside V refinement mother liquor used in the embodiment of the present invention is limited from Hunan China really living resources share
Company is using Fructus Monordicae extract as raw material, generated mother liquor when preparing high-purity momordica glycoside V with organic solvent crystallization, warp
High-performance liquid chromatography detection, wherein the mass content of mogroside VI is 20.1%, and the quality of momordica glycoside V contains
Amount is 10.9%, and the mass content of Siraitia grosvenorii 11-O base glycosides V is 15.5%；Polyamide (mesh used in the embodiment of the present invention
Number is 50~100 mesh) it is purchased from Solution on Chemical Reagents in Shanghai company, Chinese Medicine group；Raw material and chemistry used in the embodiment of the present invention
Reagent is obtained by routine business approach unless otherwise specified.
Embodiment 1
(1) extracting n-butyl alcohol: by 100g momordica glycoside V refinement mother liquor with 500g water dissolve, be added 500g n-butanol into
Row extraction, separates water layer, obtains n-butanol layer；
(2) crystallization is concentrated: by n-butanol layer obtained by step (1) in the case where 80 DEG C, vacuum degree are -0.07MPa, being concentrated under reduced pressure into
The mass concentration of concentrate is 25%, cooling, and under 15 DEG C, revolving speed 120r/min, stirring and crystallizing 12h, filtering obtains crystal and just
Butanol crystallization mother liquor, crystal is dry, obtain 18.2g mogroside VI；
(3) polyamide chromatography: by n-butanol crystallization mother liquor obtained by step (2) in the case where 80 DEG C, vacuum degree are -0.07MPa, subtract
Pressure is concentrated into no alcohol, and gained 70g concentrate 1750g water is dissolved, and passes through polyamide chromatography column (wherein, with the flow velocity of 1BV/h
The dosage of polyamide is 420g, and the ratio of height to diameter of chromatographic column is 10:1), upper prop efflux is collected, be in 80 DEG C, vacuum degree-
Under 0.07MPa, it is concentrated under reduced pressure, it is dry, obtain 10.8g momordica glycoside V；
(4) gradient elution: the ethanol solution for being first 20% with 2BV volumetric concentration, with the flow velocity of 0.5BV/h to step (3) mistake
Polyamide chromatography column after column is eluted, then the ethanol solution for being 80% with 4BV volumetric concentration, is carried out with the flow velocity of 0.5BV/h
Elution, the eluent for the ethanol solution that collected volume concentration is 80%, at 65 DEG C, the mass concentration for being concentrated into concentrate is
30%, cooling, at 25 DEG C, 24 h of crystallization is filtered, dry, obtains 13.8g Siraitia grosvenorii 11-O base glycosides V.
Momordica glycoside V obtained by the present embodiment, VI and 11-O base glycosides V are white powder, through high performance liquid chromatography external standard
Method detection, the purity of gained mogroside VI are 97.5%, yield 88.28%；The purity of momordica glycoside V is 91.9%, is received
Rate is 91.06%；The purity of Siraitia grosvenorii 11-O base glycosides V is 98.2%, yield 87.43%.
Embodiment 2
(1) extracting n-butyl alcohol: 200g momordica glycoside V refinement mother liquor 1600g water is dissolved, 2400g n-butanol is added
It is extracted, separates water layer, obtain n-butanol layer；
(2) crystallization is concentrated: by n-butanol layer obtained by step (1) in the case where 85 DEG C, vacuum degree are -0.08MPa, being concentrated under reduced pressure into
The mass concentration of concentrate is 30%, cooling, and under 30 DEG C, revolving speed 80r/min, stirring and crystallizing 36h, filtering obtains crystal and positive fourth
Alcohol crystallization mother liquor, crystal is dry, obtain 34.3g mogroside VI；
(3) polyamide chromatography: by n-butanol crystallization mother liquor obtained by step (2) in the case where 85 DEG C, vacuum degree are -0.08MPa, subtract
Pressure is concentrated into no alcohol, and gained 135g concentrate 2700g water is dissolved, and passes through polyamide chromatography column (its with the flow velocity of 0.5BV/h
In, the dosage of polyamide is 1350g, and the ratio of height to diameter of chromatographic column is 8:1), upper prop efflux is collected, be in 85 DEG C, vacuum degree-
Under 0.08MPa, it is concentrated under reduced pressure, it is dry, obtain 20.8g momordica glycoside V；
(4) gradient elution: the methanol solution for being first 15% with 3BV volumetric concentration crosses column to step (3) with the flow velocity of 1BV/h
Polyamide chromatography column afterwards is eluted, then the methanol solution for being 75% with 5BV volumetric concentration, is washed with the flow velocity of 1BV/h
De-, the eluent for the methanol solution that collected volume concentration is 75%, at 60 DEG C, the mass concentration for being concentrated into concentrate is 35%,
Cooling, at 15 DEG C, 18 h of crystallization is filtered, dry, obtains 27.3g Siraitia grosvenorii 11-O base glycosides V.
Momordica glycoside V obtained by the present embodiment, VI and 11-O base glycosides V are white powder, through high performance liquid chromatography external standard
Method detection, the purity of gained mogroside VI are 98%, yield 83.62%；The purity of momordica glycoside V is 93.5%, yield
It is 89.21%；The purity of Siraitia grosvenorii 11-O base glycosides V is 96.9%, yield 85.33%.
Embodiment 3
(1) extracting n-butyl alcohol: by 150g momordica glycoside V refinement mother liquor with 900g water dissolve, be added 900g n-butanol into
Row extraction, separates water layer, obtains n-butanol layer；
(2) crystallization is concentrated: by n-butanol layer obtained by step (1) in the case where 75 DEG C, vacuum degree are -0.06MPa, being concentrated under reduced pressure into
The mass concentration of concentrate is 30%, cooling, and under 25 DEG C, revolving speed 60r/min, 24 h of stirring and crystallizing, filtering obtains crystal and just
Butanol crystallization mother liquor, crystal is dry, obtain 25.8g mogroside VI；
(3) polyamide chromatography: by n-butanol crystallization mother liquor obtained by step (2) in the case where 75 DEG C, vacuum degree are -0.06MPa, subtract
Pressure is concentrated into no alcohol, and gained 100g concentrate 2000g water is dissolved, and passes through polyamide chromatography column (its with the flow velocity of 1.5BV/h
In, the dosage of polyamide is 500g, and the ratio of height to diameter of chromatographic column is 15:1), upper prop efflux is collected, be in 75 DEG C, vacuum degree-
Under 0.06MPa, it is concentrated under reduced pressure, it is dry, obtain 16.9g momordica glycoside V；
(4) gradient elution: the aqueous isopropanol for being first 30% with 3BV volumetric concentration, with the flow velocity of 1.5BV/h to step (3)
Polyamide chromatography column after crossing column is eluted；The aqueous isopropanol that 5BV volumetric concentration is 70% is added, with the stream of 1.5BV/h
Speed is eluted, and the eluent for the aqueous isopropanol that collected volume concentration is 70% is concentrated into the quality of concentrate at 75 DEG C
Concentration is 40%, cooling, and at 30 DEG C, 36 h of crystallization is filtered, dry, obtains 20.2g Siraitia grosvenorii 11-O base glycosides V.
Momordica glycoside V obtained by the present embodiment, VI and 11-O base glycosides V are white powder, through high performance liquid chromatography external standard
Method detection, the purity of gained mogroside VI are 97.1wt%, yield 83.09%；The purity of momordica glycoside V is 89.6%,
Yield is 92.61%；The purity of Siraitia grosvenorii 11-O base glycosides V is 97.9wt%, yield 85.06%.
Claims (12)
1. a kind of method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously, which comprises the following steps:
(1) extracting n-butyl alcohol: momordica glycoside V refinement mother liquor is dissolved with water, and n-butanol is added and is extracted, water layer is separated,
Obtain n-butanol layer；
(2) crystallization is concentrated: n-butanol layer obtained by step (1) is concentrated under reduced pressure, cooling, stirring and crystallizing, filtering obtains crystal and positive fourth
Alcohol crystallization mother liquor, crystal is dry, obtain mogroside VI；
(3) polyamide chromatography: n-butanol crystallization mother liquor obtained by step (2) is concentrated under reduced pressure into no alcohol, by gained concentrate phase
When collecting upper prop efflux by polyamide chromatography column in 18~50 times of its quality of water dissolution, reduced pressure is dry, obtains sieve
Chinese fruit sweet tea glycosides V；
In step (2), (3), the temperature of the reduced pressure is 70~90 DEG C, the vacuum degree of reduced pressure is -0.06~-
0.09MPa, the mass concentration for being concentrated into concentrate is 20~35%；
(4) gradient elution: successively the polyamide chromatography column of step (3) Guo Zhuhou is carried out with low alcoholic solution and height alcoholic solution
Height alcohol eluen is collected in elution, is concentrated, cooling, and crystallization filters, dry, obtains Siraitia grosvenorii 11-O base glycosides V；The minuent alcohol is molten
Alcohol in liquid and height alcoholic solution is one or more of methanol, ethyl alcohol, isopropanol or n-butanol；The minuent alcoholic solution
Volumetric concentration be 10~40%, the volumetric concentration of the height alcoholic solution is 50~85%；The dosage of the minuent alcoholic solution is 2
~5BV, the dosage of height alcoholic solution are 2~5BV；The flow velocity of the elution is 0.5~2.0BV/h.
2. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 1, it is characterised in that: step
Suddenly in (1), in the momordica glycoside V refinement mother liquor, the mass content of mogroside VI is 10~30%, momordica glycoside V
Mass content be 10~15%, 11-O base glycosides V mass content be 5~20%.
3. method that is according to claim 1 or claim 2 while extracting momordica glycoside V, VI and 11-O base glycosides V, it is characterised in that:
In step (1), the mass ratio of the momordica glycoside V refinement mother liquor, water and n-butanol is 1:5~10:4~30.
4. method that is according to claim 1 or claim 2 while extracting momordica glycoside V, VI and 11-O base glycosides V, it is characterised in that:
In step (2), the temperature of the stirring and crystallizing is 15~30 DEG C, and revolving speed is 60~120r/min, and the time is 12~36h.
5. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 3, it is characterised in that: step
Suddenly in (2), the temperature of the stirring and crystallizing is 15~30 DEG C, and revolving speed is 60~120r/min, and the time is 12~36h.
6. method that is according to claim 1 or claim 2 while extracting momordica glycoside V, VI and 11-O base glycosides V, it is characterised in that:
In step (3), the mesh number of the polyamide is 50~200 mesh, and dosage is 4~20 times of concentrate quality, polyamide chromatography column
Ratio of height to diameter be 1~20:1；The flow velocity of upper prop is 0.5~2.0BV/h.
7. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 3, it is characterised in that: step
Suddenly in (3), the mesh number of the polyamide is 50~200 mesh, and dosage is 4~20 times of concentrate quality, polyamide chromatography column
Ratio of height to diameter is 1~20:1；The flow velocity of upper prop is 0.5~2.0BV/h.
8. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 4, it is characterised in that: step
Suddenly in (3), the mesh number of the polyamide is 50~200 mesh, and dosage is 4~20 times of concentrate quality, polyamide chromatography column
Ratio of height to diameter is 1~20:1；The flow velocity of upper prop is 0.5~2.0BV/h.
9. method that is according to claim 1 or claim 2 while extracting momordica glycoside V, VI and 11-O base glycosides V, it is characterised in that:
In step (4), the temperature of the concentration is 60~80 DEG C, and the mass concentration for being concentrated into concentrate is 25~45%；The crystallization
Temperature is 15~30 DEG C, and the time is 12~36 h.
10. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 3, it is characterised in that: step
Suddenly in (4), the temperature of the concentration is 60~80 DEG C, and the mass concentration for being concentrated into concentrate is 25~45%；The temperature of the crystallization
Degree is 15~30 DEG C, and the time is 12~36 h.
11. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 4, it is characterised in that: step
Suddenly in (4), the temperature of the concentration is 60~80 DEG C, and the mass concentration for being concentrated into concentrate is 25~45%；The temperature of the crystallization
Degree is 15~30 DEG C, and the time is 12~36 h.
12. the method for extracting momordica glycoside V, VI and 11-O base glycosides V simultaneously according to claim 6, it is characterised in that: step
Suddenly in (4), the temperature of the concentration is 60~80 DEG C, and the mass concentration for being concentrated into concentrate is 25~45%；The temperature of the crystallization
Degree is 15~30 DEG C, and the time is 12~36 h.
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| CN107629105B (en) * | 2017-11-02 | 2020-02-07 | 湖南华诚生物资源股份有限公司 | Method for purifying flavor mogroside V |
| CN110357936B (en) * | 2019-07-16 | 2020-10-16 | 湖南华诚生物资源股份有限公司 | Semi-synthesis method of mogroside V |
| CN110343141B (en) * | 2019-07-22 | 2021-09-07 | 湖南艾达伦科技有限公司 | Preparation method and application of high-content mogroside monomer product |
| CN111333691B (en) * | 2020-04-24 | 2021-05-14 | 永州华茂生物科技有限责任公司 | Preparation method of mogroside V |
| CN113461765B (en) * | 2021-08-06 | 2022-07-19 | 湖南华诚生物资源股份有限公司 | Separation method of mogroside V and rare mogroside substances |
| CN113773360B (en) * | 2021-09-13 | 2022-05-31 | 湖南华诚生物资源股份有限公司 | Method for separating mogrol from fructus momordicae |
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Denomination of invention: A method for simultaneous extraction of Mogroside V, VI and 11-o-glycoside V from Siraitia grosvenorii Effective date of registration: 20210422 Granted publication date: 20190702 Pledgee: CITIC Bank Changsha branch Pledgor: HUNAN HUACHENG BIOTECH, Inc. Registration number: Y2021980002898 |
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Denomination of invention: A method for simultaneous extraction of mogroside V, VI and 11-O-glycoside V from Siraitia grosvenorii Granted publication date: 20190702 Pledgee: Agricultural Bank of China Limited Changsha Wangcheng District sub branch Pledgor: HUNAN HUACHENG BIOTECH, Inc. Registration number: Y2024980000707 |