CN101983966A - Process for simultaneously preparing asiaticoside and madecassoside from low asiaticoside content total glycosides - Google Patents
Process for simultaneously preparing asiaticoside and madecassoside from low asiaticoside content total glycosides Download PDFInfo
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- CN101983966A CN101983966A CN 201010556169 CN201010556169A CN101983966A CN 101983966 A CN101983966 A CN 101983966A CN 201010556169 CN201010556169 CN 201010556169 CN 201010556169 A CN201010556169 A CN 201010556169A CN 101983966 A CN101983966 A CN 101983966A
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- 150000002338 glycosides Chemical class 0.000 title claims abstract description 318
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- WYQVAPGDARQUBT-FGWHUCSPSA-N Madecassol Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O)OC[C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)[C@]12CC[C@H]([C@@H]([C@H]1C=1[C@@]([C@@]3(CC[C@H]4[C@](C)(CO)[C@@H](O)[C@H](O)C[C@]4(C)[C@H]3CC=1)C)(C)CC2)C)C)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O WYQVAPGDARQUBT-FGWHUCSPSA-N 0.000 title claims abstract description 78
- WYQVAPGDARQUBT-XCWYDTOWSA-N asiaticoside Natural products O=C(O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@H](O)[C@H](O)[C@H](O[C@H]3[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)[C@@H](CO)O2)O1)[C@@]12[C@@H]([C@@H](C)[C@H](C)CC1)C=1[C@](C)([C@@]3(C)[C@@H]([C@@]4(C)[C@H]([C@@](CO)(C)[C@@H](O)[C@H](O)C4)CC3)CC=1)CC2 WYQVAPGDARQUBT-XCWYDTOWSA-N 0.000 title claims abstract description 77
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- BNMGUJRJUUDLHW-HCZMHFOYSA-N Madecassoside Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O)OC[C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)[C@]12CC[C@H]([C@@H]([C@H]1C=1[C@@]([C@@]3(C[C@@H](O)[C@H]4[C@](C)(CO)[C@@H](O)[C@H](O)C[C@]4(C)[C@H]3CC=1)C)(C)CC2)C)C)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O BNMGUJRJUUDLHW-HCZMHFOYSA-N 0.000 title abstract description 5
- BNMGUJRJUUDLHW-HLUHVYOBSA-N Madecassoside Natural products C[C@@H]1CC[C@@]2(CC[C@]3(C)C(=CC[C@@H]4[C@@]5(C)C[C@@H](O)[C@H](O)[C@@](C)(CO)[C@@H]5[C@H](O)C[C@@]34C)[C@@H]2[C@H]1C)C(=O)O[C@@H]6O[C@H](CO[C@@H]7O[C@H](CO)[C@@H](O[C@@H]8O[C@H](C)[C@H](O)[C@@H](O)[C@H]8O)[C@H](O)[C@H]7O)[C@@H](O)[C@H](O)[C@H]6O BNMGUJRJUUDLHW-HLUHVYOBSA-N 0.000 title abstract description 4
- 229940090813 madecassoside Drugs 0.000 title abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 237
- 239000011347 resin Substances 0.000 claims abstract description 212
- 229920005989 resin Polymers 0.000 claims abstract description 212
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- GRTOGORTSDXSFK-XJTZBENFSA-N ajmalicine Chemical compound C1=CC=C2C(CCN3C[C@@H]4[C@H](C)OC=C([C@H]4C[C@H]33)C(=O)OC)=C3NC2=C1 GRTOGORTSDXSFK-XJTZBENFSA-N 0.000 claims description 44
- 239000003463 adsorbent Substances 0.000 claims description 43
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 36
- 239000012141 concentrate Substances 0.000 claims description 24
- 238000005516 engineering process Methods 0.000 claims description 15
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 4
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Abstract
The invention discloses a process for simultaneously preparing asiaticoside and madecassoside from low asiaticoside content total glycosides. The process comprises the following steps: 1) heating and dissolving the low asiaticoside content total glycosides with methanol at the temperature of between 50 and 65 DEG C, and cooling and crystallizing to prepare solid object I and crystal mother solution I; 2) re-crystallizing the solid object I twice under the condition in the step 1), drying in vacuum to prepare high purity madecassoside; 3) purifying the crystal mother solution I through a macroporous adsorption resin, concentrating and drying in vacuum to prepare high asiaticoside content total glycosides, dissolving with solvent I, adding solvent II to crystallize, and drying in vacuum to prepare high purity asiaticoside; and 4) purifying the crystal mother solution through a macroporous adsorption resin, concentrating the collected solution enriching total glycosides, and drying in vacuum to be used as an initial raw material for recycle. The method has simple process, low production cost, small pollution, high product yield and purity, and is suitable for industrial production.
Description
Technical field
The present invention relates to the total glycosides of a kind of low centella asiatica glucoside content sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously.
Background technology
Herba Centellae [Centella asiatica (L.) UrBan] is a samphire, and how (Linnaeus Carolus) was included into Lawn Pennywort Herb in 1764 with Herba Centellae and belongs to (Hydrocotyle), and names the L. into Hydrocotyleasiatica the botanist woods.In 1879, with its independent from Lawn Pennywort Herb belongs to and set up Centella (Centella), be included into this genus and rename Centella asiatica (L.) Urb. as by Herba Centellae according to the rib number of the arrangement of petal and fruit for fritz UrBan.Herba Centellae mainly is distributed in the southern hemisphere and northern hemisphere torrid zone and subtropical zone, and main product ground is South Africa, India, Sri Lanka, Malaysia, Indonesia, Australia, Japan etc., and China also produces Herba Centellae.Herba Centellae for two thousand years, among the people is usually used in treating flu, tonsillitis, infectious hepatitis, dysentery, wound, furuncle swelling toxin, traumatic hemorrhage etc. at the medicinal history of China.Research of Herba Centellae modern pharmacology and application have had very great development, show that Herba Centellae can be used for treating dysthymia disorders, skin wound, stomach ulcer, infectious hepatitis, tetter and meningococcal meningitis etc.
The complex chemical composition of Herba Centellae mainly contains triterpenes, flavonoid, volatile oil, polyyne alkene class etc.Herba Centellae total glycosides is that Herba Centellae is carried effective component main in the thing, mainly by centella asiatica glucoside (Aisaticoside, CAS No.:16830-15-2), asiaticoside (Asaiticoside A; Madecassoside, CAS No.:34540-22-2) and centella asiatica glucoside B materials such as (Asaiticoside B, CAS No.:125265-68-1) composition.Wherein centella asiatica glucoside has been widely used in fields such as medicine, makeup, studies show that centella asiatica glucoside has the promotion wound healing, the pharmacological action of treatment scar.
The structural formula following (R=Glu-Glu-Rha) of centella asiatica glucoside (a), asiaticoside (b) and centella asiatica glucoside B (c):
Industrial, often the content of centella asiatica glucoside, asiaticoside and centella asiatica glucoside B has obvious variation to the asiatic centella total glycoside product that extracts from Herba Centellae because of the difference of the Herba Centellae place of production or technology causes wherein.In some Herba Centellae total glycosides centella asiatica glucoside content greater than 20% and asiaticoside/centella asiatica glucoside (w/w) less than 2, some then centella asiatica glucoside content be lower than 20% and asiaticoside/centella asiatica glucoside (w/w) greater than 2.
For in above-mentioned two kinds of Herba Centellae total glycosidess the former, Zhejiang University's chemical industry is employing crystallization process that the Lv Xiuyang researcher seminar of pharmaceutical engineering institute green pharmaceutical engineering research department successfully develops prepares the pure product of centella asiatica glucoside from Herba Centellae total glycosides a technology (patent: CN2008-10162211), can prepare highly purified centella asiatica glucoside product efficiently, have energy-efficient, characteristics such as product purity is high, rate of recovery height.But but there is the problem that the centella asiatica glucoside rate of recovery is on the low side, product purity is not high in this process application when the latter.Simultaneously, in order from the surplus liquid of the crystallization of this technological process, further to reclaim residual centella asiatica glucoside and asiaticoside, be necessary very much to seek a kind of technology for preparing centella asiatica glucoside and asiaticoside from the low total glycosides of centella asiatica glucoside content simultaneously.
Summary of the invention
The purpose of this invention is to provide the total glycosides of a kind of low centella asiatica glucoside content sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously.
The step of technology is as follows:
1) the low total glycosides of centella asiatica glucoside content uses methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance I and crystalline mother solution I down at 50～65 ℃;
2) solid substance I continues to use methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance II and crystalline mother solution II down at 50～65 ℃;
3) solid substance II uses methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance III and crystalline mother solution III down at 50～65 ℃ once more;
4) solid substance III obtains the high purity asiaticoside after vacuum-drying;
5) crystalline mother solution I is through purification with macroreticular resin, and the collection liquid that is rich in total glycosides concentrates the final vacuum drying, obtains the total glycosides of high centella asiatica glucoside content;
6) the total glycosides of high centella asiatica glucoside content with solvent I dissolving, add the solvent II crystallization again and obtain solid substance IV and crystalline mother solution IV;
7) solid substance IV obtains high-purity asiaticoside after vacuum-drying;
8) merge crystalline mother solution II, III, IV through purification with macroreticular resin, the collection liquid that is rich in total glycosides concentrates the final vacuum drying and is back to step 1) as starting raw material.
Wherein solvent I is methyl alcohol or dimethyl formamide in the step 6), and add-on is 2.5～10mL/g Herba Centellae total glycosides.Solvent II is acetone, acetonitrile, ether, ethyl acetate or water in the step 6), and add-on is 0.5～10 times of solvent I add-on.Step 5), 8) the macroporous adsorbent resin model in is HPD100, HPD200, HPD300, HPD400, HPD500, HPD600, HPD700, X-5, AB-8 or D4020.
The total glycosides of low centella asiatica glucoside content among the present invention is meant centella asiatica glucoside, asiaticoside, centella asiatica glucoside B total content greater than 70%, centella asiatica glucoside content be lower than 20% and asiaticoside/centella asiatica glucoside (w/w) greater than 2 Herba Centellae total glycosides.The total glycosides of high centella asiatica glucoside content is meant centella asiatica glucoside, asiaticoside, centella asiatica glucoside B total content greater than 70%, centella asiatica glucoside content be higher than 20% and asiaticoside/centella asiatica glucoside (w/w) less than 2 Herba Centellae total glycosides.High-purity asiaticoside is meant purity greater than 90% centella asiatica glucoside product, and the high purity asiaticoside is meant that purity is greater than 90% asiaticoside product.The crystallisation by cooling process is at room temperature carried out, and crystallization time is usually more than 4 hours.
The present invention has that process is simple, production cost is low, pollutes less, product centella asiatica glucoside yield and purity advantages of higher, is suitable for suitability for industrialized production.
Description of drawings
Accompanying drawing is that the low total glycosides of centella asiatica glucoside content sets out and prepares the process flow diagram of centella asiatica glucoside and asiaticoside simultaneously.
Embodiment
Below with embodiment processing method of the present invention is further described.Protection scope of the present invention is not subjected to the restriction of embodiment, and protection scope of the present invention is determined by claims.
The Herba Centellae total glycosides raw material is provided by the natural pharmaceutcal corporation, Ltd of Guangxi Chang Zhou, and wherein the centella asiatica glucoside mass content is 15.0% after testing, and asiaticoside content is 33%, and centella asiatica glucoside B mass content accounts for 28%.
Analysis condition used among the embodiment is as follows:
High performance liquid chromatography (HPLC) analysis condition: Agilent1100 liquid-phase chromatographic analysis system, chromatographic column is Synergi 4 μ Hydro-RP 80A reversed-phase column (4.6mm * 250mm, 4 μ m, Phenomenex), mobile phase methanol-water (60: 40), flow velocity 0.5mL/min, 25 ℃ of column temperatures, ultraviolet detection wavelength 205nm, centella asiatica glucoside standard reference material are available from Sigma company, and the asiaticoside standard reference material is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
The concrete steps of purification with macroreticular resin are as follows among the present invention:
1) crystalline mother solution is through concentrating the final vacuum drying, and water is dissolved to 1.5g/L concentration again;
2) the pretreated macroporous adsorbent resin of in chromatography column, packing into, aspect ratio is 3～5;
3) material solution flows through resin column with the flow velocity of 0.5～1.5BV/h, and total saponins penetrates to the effluent liquid;
4) the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow phase (the alcoholic acid volume percent is 30%) with 0.5～2BV/h, elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes.
Embodiment 1
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 93mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 28.1g, and adding 140.4mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 17.5g, yield 80.6%, purity 91.2%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 40.5g, makes total glycosides aqueous solution with the 27L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD100 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 16.7g, add 41.8mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 41.8mL water (solvent I add-on 1 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 6.8g, yield 69.6%, purity 91.4%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 12g, use the 8L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD100, resin column is high to be 9cm (aspect ratio is 3), and the bed volume of resin column is 63.6mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 2.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 8.3g, as the starting raw material reuse.
Embodiment 2
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 138mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 16.3g, and adding 40.7mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 12.6g, yield 58.7%, purity 92.5%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 44.4g, makes total glycosides aqueous solution with the 29.6L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD200 in internal diameter is the chromatography column of 3cm, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 7 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 20.6g, add 103.2mL dimethyl formamide (5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 51.6mL water (solvent I add-on 0.5 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.2g, yield 53.3%, purity 90.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 18.5g, use the 12.3L water dissolution, in being the chromatography column of 4cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD200, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 12.8g, as the starting raw material reuse.
Embodiment 3
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 450mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 60 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 18g, adds 135mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 7.7g, and adding 57.4mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 3.6g, yield 17.2%, purity 93.9%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 54.4g, makes total glycosides aqueous solution with the 36.2L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD300 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 30.6g, add 229.3mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 917.1mL water (solvent I add-on 4 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.1g, yield 52.3%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 27.5g, use the 18.3L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD300, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 19.1g, as the starting raw material reuse.
Embodiment 4
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 600mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 65 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 8.4g, adds 21mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 6.3g, and adding 31.7mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 4g, yield 18.7%, purity 93.9%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 54g, makes total glycosides aqueous solution with the 36L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD400 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 30.2g, add 302.5mL dimethyl formamide (10mL solvent I/g Herba Centellae total glycosides), dissolving back adds 605mL water (solvent I add-on 2 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.5g, yield 45.5%, purity 89.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 27.8g, use the 18.5L water dissolution, in being the chromatography column of 4cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD400, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 19.3g, as the starting raw material reuse.
Embodiment 5
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 93mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 28.1g, and adding 280.9mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 9.2g, yield 43.1%, purity 93.1%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 49.8g, makes total glycosides aqueous solution with the 33.2L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD500 in internal diameter is the chromatography column of 4cm, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 26g, add 65.1mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 651.1mL water (solvent I add-on 10 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 7.5g, yield 76.7%, purity 91.4%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 19.6g, use the 13.1L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD500, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 13.6g, as the starting raw material reuse.
Embodiment 6
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 138mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 16.3g, and adding 81.4mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 10.1g, yield 47.6%, purity 93%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 46.9g, makes total glycosides aqueous solution with the 31.2L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD600 in internal diameter is the chromatography column of 3cm, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 6 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 23.1g, add 115.3mL dimethyl formamide (5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 922.2mL water (solvent I add-on 8 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.9g, yield 60.9%, purity 90.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 20.1g, use the 13.4L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD600, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 14g, as the starting raw material reuse.
Embodiment 7
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 450mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 60 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 18g, adds 90mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 10.6g, and adding 26.6mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 8.2g, yield 38.6%, purity 93.3%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 49.8g, makes total glycosides aqueous solution with the 33.2L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD700 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 4 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 26g, add 195.1mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 390.1mL water (solvent I add-on 2 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.9g, yield 50.2%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 23.1g, use the 15.4L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD700, resin column is high to be 9cm (aspect ratio is 3), and the bed volume of resin column is 63.6mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 2.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 4 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 16g, as the starting raw material reuse.
Embodiment 8
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 600mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 65 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 8.4g, adds 21mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 6.3g, and adding 31.7mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 4g, yield 18.7%, purity 93.9%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 54g, makes total glycosides aqueous solution with the 36L water dissolution.Pack into through pretreated macroporous adsorbent resin X-5 in internal diameter is the chromatography column of 4cm, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 30.2g, add 75.6mL dimethyl formamide (2.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 37.8mL water (solvent I add-on 0.5 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 6.6g, yield 68.1%, purity 91.4%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 25.6g, use the 17.1L water dissolution, in being the chromatography column of 4cm, internal diameter packs into through pretreated macroporous adsorbent resin X-5, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 17.8g, as the starting raw material reuse.
Embodiment 9
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 372mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 9.7g, and adding 24.2mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 7.5g, yield 35.2%, purity 93.4%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 50.5g, makes total glycosides aqueous solution with the 33.7L water dissolution.Pack into through pretreated macroporous adsorbent resin AB-8 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 26.7g, add 133.7mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 133.7mL water (solvent I add-on 1 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.3g, yield 54.7%, purity 90.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 23.4g, use the 15.6L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin AB-8, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 16.3g, as the starting raw material reuse.
Embodiment 10
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 138mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 16.3g, and adding 81.4mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 10.1g, yield 47.6%, purity 93%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 47.9g, makes total glycosides aqueous solution with the 31.9L water dissolution.Pack into through pretreated macroporous adsorbent resin D4020 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 4 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 24.1g, add 180.4mL dimethyl formamide (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 1082.5mL water (solvent I add-on 6 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.2g, yield 53.6%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 20.8g, use the 13.9L water dissolution, in being the chromatography column of 4cm, internal diameter packs into through pretreated macroporous adsorbent resin D4020, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 14.4g, as the starting raw material reuse.
Embodiment 11
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 600mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 8.4g, adds 21mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 6.3g, and adding 31.7mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 4g, yield 18.7%, purity 93.9%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 55g, makes total glycosides aqueous solution with the 36.7L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD100 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 4 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 31.2g, add 156.2mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 156.2mL water (solvent I add-on 1 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.3g, yield 54.7%, purity 90.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 26.9g, use the 17.9L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD100, resin column is high to be 9cm (aspect ratio is 3), and the bed volume of resin column is 63.6mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 2.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 5 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 18.7g, as the starting raw material reuse.
Embodiment 12
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 450mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 18g, adds 90mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 10.6g, and adding 26.6mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 8.2g, yield 38.6%, purity 93.3%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 48.8g, makes total glycosides aqueous solution with the 32.5L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD200 in internal diameter is the chromatography column of 3cm, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 8 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 25g, add 250.1mL dimethyl formamide (10mL solvent I/g Herba Centellae total glycosides), dissolving back adds 125mL water (solvent I add-on 0.5 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.1g, yield 42.3%, purity 89.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 23.9g, use the 15.9L water dissolution, in being the chromatography column of 4cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD200, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 16.6g, as the starting raw material reuse.
Embodiment 13
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 60 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 207mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 11.7g, and adding 88mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 5.6g, yield 26.3%, purity 93.7%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 51.4g, makes total glycosides aqueous solution with the 34.3L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD300 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 27.6g, add 207.3mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 829mL water (solvent I add-on 4 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.1g, yield 52.3%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 25.5g, use the 17L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD300, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 17.7g, as the starting raw material reuse.
Embodiment 14
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 65 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 93mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 28.1g, and adding 140.4mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 17.5g, yield 80.6%, purity 91.2%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 40.5g, makes total glycosides aqueous solution with the 27L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD400 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 16.7g, add 167mL dimethyl formamide (10mL solvent I/g Herba Centellae total glycosides), dissolving back adds 334mL water (solvent I add-on 2 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.5g, yield 45.5%, purity 89.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 14.2g, use the 9.5L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD400, resin column is high to be 9cm (aspect ratio is 3), and the bed volume of resin column is 63.6mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 2.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 9.9g, as the starting raw material reuse.
Embodiment 15
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 600mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 8.4g, adds 21mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 6.3g, and adding 63.4mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 2.1g, yield 9.8%, purity 94.1%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 56.9g, makes total glycosides aqueous solution with the 38L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD500 in internal diameter is the chromatography column of 4cm, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 33.1g, add 82.8mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 828.3mL water (solvent I add-on 10 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 7.5g, yield 76.7%, purity 91.4%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 26.7g, use the 17.8L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD500, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 0.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 4 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 18.5g, as the starting raw material reuse.
Embodiment 16
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 450mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 18g, adds 90mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 10.6g, and adding 53.1mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 6.6g, yield 31.2%, purity 93.5%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 52.4g, makes total glycosides aqueous solution with the 34.9L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD600 in internal diameter is the chromatography column of 3cm, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 6 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 28.6g, add 71.5mL dimethyl formamide (2.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 571.7mL water (solvent I add-on 8 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 7.4g, yield 75.8%, purity 91.4%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 22.2g, use the 14.8L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD600, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 15.4g, as the starting raw material reuse.
Embodiment 17
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 60 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 138mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 16.3g, and adding 40.7mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 60 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 12.6g, yield 58.7%, purity 92.5%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 44.4g, makes total glycosides aqueous solution with the 29.6L water dissolution.Pack into through pretreated macroporous adsorbent resin HPD700 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 20.6g, add 154.8mL methyl alcohol (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 309.6mL water (solvent I add-on 2 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.9g, yield 50.2%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 18.7g, use the 12.5L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin HPD700, resin column is high to be 9cm (aspect ratio is 3), and the bed volume of resin column is 63.6mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 0.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 2.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 13g, as the starting raw material reuse.
Embodiment 18
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 65 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 93mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 28.1g, and adding 140.4mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 65 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 17.5g, yield 80.6%, purity 91.2%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 40.5g, makes total glycosides aqueous solution with the 27L water dissolution.Pack into through pretreated macroporous adsorbent resin X-5 in internal diameter is the chromatography column of 4cm, resin column is high to be 16cm (aspect ratio is 4), and the bed volume of resin column is 201.1mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 8.9L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 16.7g, add 125.3mL dimethyl formamide (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 62.6mL water (solvent I add-on 0.5 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 4.6g, yield 47.1%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 14.1g, use the 9.4L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin X-5, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 2BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 1 time, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 9.8g, as the starting raw material reuse.
Embodiment 19
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 300mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 50 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 27.6g, adds 276mL methyl alcohol (10mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 7.2g, and adding 17.9mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 50 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 5.5g, yield 26.2%, purity 93.7%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 51.5g, makes total glycosides aqueous solution with the 34.3L water dissolution.Pack into through pretreated macroporous adsorbent resin AB-8 in internal diameter is the chromatography column of 4cm, resin column is high to be 20cm (aspect ratio is 5), and the bed volume of resin column is 251.3mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 11.2L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 27.7g, add 138.3mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 138.3mL water (solvent I add-on 1 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.3g, yield 54.7%, purity 90.8%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 25.3g, use the 16.9L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin AB-8, resin column is high to be 15cm (aspect ratio is 5), and the bed volume of resin column is 106mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1.5BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 4.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 17.6g, as the starting raw material reuse.
Embodiment 20
Get 60g Herba Centellae total glycosides raw material and add in the beaker, add 150mL methyl alcohol (2.5mL solvent I/g Herba Centellae total glycosides), be heated to the crystallization of 55 ℃ of dissolving postcooling and obtain solid substance I and crystalline mother solution I; Solid substance I weight after vacuum-drying is 37.2g, adds 186mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides), is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance II and crystalline mother solution II; Solid substance II weight after vacuum-drying is 21.9g, and adding 109.7mL methyl alcohol (5mL solvent I/g Herba Centellae total glycosides) is heated to the crystallization of 55 ℃ of dissolving postcooling and obtains solid substance III and crystalline mother solution III; Solid substance III is the asiaticoside product after vacuum-drying, and weight is 13.7g, yield 63.7%, purity 92.3%.
Crystalline mother solution I is through concentrating the final vacuum drying, and weight is 43.3g, makes total glycosides aqueous solution with the 28.9L water dissolution.Pack into through pretreated macroporous adsorbent resin D4020 in internal diameter is the chromatography column of 4cm, resin column is high to be 12cm (aspect ratio is 3), and the bed volume of resin column is 150.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 6.7L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 3 times, elutriant merges after HPLC analyzes, concentrate the final vacuum drying, weight is 19.5g, add 146.4mL dimethyl formamide (7.5mL solvent I/g Herba Centellae total glycosides), dissolving back adds 878.7mL water (solvent I add-on 6 times) and carries out crystallization, obtains solid substance IV and crystalline mother solution IV then after filtration; Solid substance IV is the high-purity asiaticoside product after vacuum-drying, weight is 5.2g, yield 53.6%, purity 90.3%.
Merge crystalline mother solution II, III, IV through concentrating the final vacuum drying, weight is to make total glycosides aqueous solution with water dissolution behind the 17.3g, use the 11.5L water dissolution, in being the chromatography column of 3cm, internal diameter packs into through pretreated macroporous adsorbent resin D4020, resin column is high to be 12cm (aspect ratio is 4), and the bed volume of resin column is 84.8mL; Total glycosides aqueous solution is crossed chromatography column with the flow velocity of 1BV/h, penetrates to the total glycosides of effluent liquid moderate snow grass, adds total glycosides aqueous solution 3.8L altogether; With the water washing resin post of 4 bed volume, then with the flow velocity wash-out asiatic centella total saponins of alcohol-water mixed flow with 1.5BV/h, the middle mutually alcoholic acid volume percent of alcohol-water mixed flow is 30% earlier.Above-mentioned purification with macroreticular resin process repeats 2 times, and elutriant merges the collection liquid that obtains being rich in total glycosides after HPLC analyzes, and collects liquid through concentrating the final vacuum drying, and weight is 12g, as the starting raw material reuse.
Claims (6)
1. the one kind low total glycosides of centella asiatica glucoside content sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, it is characterized in that the step of technology is as follows:
1) the low total glycosides of centella asiatica glucoside content uses methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance I and crystalline mother solution I down at 50～65 ℃;
2) solid substance I continues to use methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance II and crystalline mother solution II down at 50～65 ℃;
3) solid substance II uses methyl alcohol heating for dissolving, crystallisation by cooling to obtain solid substance III and crystalline mother solution III down at 50～65 ℃ once more;
4) solid substance III obtains the high purity asiaticoside after vacuum-drying;
5) crystalline mother solution I is through purification with macroreticular resin, and the collection liquid that is rich in total glycosides concentrates the final vacuum drying, obtains the total glycosides of high centella asiatica glucoside content;
6) the total glycosides of high centella asiatica glucoside content with solvent I dissolving, add the solvent II crystallization again and obtain solid substance IV and crystalline mother solution IV;
7) solid substance IV obtains high-purity asiaticoside after vacuum-drying;
8) merge crystalline mother solution II, III, IV through purification with macroreticular resin, the collection liquid that is rich in total glycosides concentrates the final vacuum drying and is back to step 1) as starting raw material.
2. the total glycosides of a kind of low centella asiatica glucoside content according to claim 1 sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, it is characterized in that solvent I is methyl alcohol or dimethyl formamide in the described step 6).
3. the total glycosides of a kind of low centella asiatica glucoside content according to claim 1 sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, and the add-on that it is characterized in that solvent I in the described step 6) is 2.5～10mL/g Herba Centellae total glycosides.
4. the total glycosides of a kind of low centella asiatica glucoside content according to claim 1 sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, it is characterized in that solvent II is acetone, acetonitrile, ether, ethyl acetate or water in the described step 6).
5. the total glycosides of a kind of low centella asiatica glucoside content according to claim 1 sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, and the add-on that it is characterized in that solvent II in the described step 6) is 0.5～10 times of solvent I add-on.
6. the total glycosides of a kind of low centella asiatica glucoside content according to claim 1 sets out and prepares the technology of centella asiatica glucoside and asiaticoside simultaneously, it is characterized in that the macroporous adsorbent resin model in described step 5), the step 8) is HPD100, HPD200, HPD300, HPD400, HPD500, HPD600, HPD700, X-5, AB-8 or D4020.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109001307A (en) * | 2018-06-08 | 2018-12-14 | 桂林三金药业股份有限公司 | The HPLC characteristic spectrum and its construction method of three gold preparations |
| CN112773745A (en) * | 2021-02-06 | 2021-05-11 | 广州贝塔健康生物科技有限公司 | Preparation method of asiaticoside extract and application of asiaticoside extract in cosmetics |
| CN113801184A (en) * | 2020-06-16 | 2021-12-17 | 盈科瑞（天津）创新医药研究有限公司 | Method for purifying madecassoside |
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| CN101323637A (en) * | 2008-07-29 | 2008-12-17 | 卢照凯 | Asiaticoside and preparation thereof |
| CN101407536A (en) * | 2008-11-27 | 2009-04-15 | 浙江大学 | Process for preparing high-purity asiaticoside by solvent crystallization |
| CN101652143A (en) * | 2007-02-12 | 2010-02-17 | Indus生物技术私人有限公司 | Be used for compositions and method thereof that the selectivity serotonin reuptake transporter suppresses |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101652143A (en) * | 2007-02-12 | 2010-02-17 | Indus生物技术私人有限公司 | Be used for compositions and method thereof that the selectivity serotonin reuptake transporter suppresses |
| CN101323637A (en) * | 2008-07-29 | 2008-12-17 | 卢照凯 | Asiaticoside and preparation thereof |
| CN101407536A (en) * | 2008-11-27 | 2009-04-15 | 浙江大学 | Process for preparing high-purity asiaticoside by solvent crystallization |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001307A (en) * | 2018-06-08 | 2018-12-14 | 桂林三金药业股份有限公司 | The HPLC characteristic spectrum and its construction method of three gold preparations |
| CN109001307B (en) * | 2018-06-08 | 2021-05-04 | 桂林三金药业股份有限公司 | HPLC (high Performance liquid chromatography) characteristic spectrum of Sanjin preparation and construction method thereof |
| CN113801184A (en) * | 2020-06-16 | 2021-12-17 | 盈科瑞（天津）创新医药研究有限公司 | Method for purifying madecassoside |
| CN112773745A (en) * | 2021-02-06 | 2021-05-11 | 广州贝塔健康生物科技有限公司 | Preparation method of asiaticoside extract and application of asiaticoside extract in cosmetics |
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