CN102391347A - Joint preparation method for euscaphic acid and tormentic acid - Google Patents
Joint preparation method for euscaphic acid and tormentic acid Download PDFInfo
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- CN102391347A CN102391347A CN2011102259426A CN201110225942A CN102391347A CN 102391347 A CN102391347 A CN 102391347A CN 2011102259426 A CN2011102259426 A CN 2011102259426A CN 201110225942 A CN201110225942 A CN 201110225942A CN 102391347 A CN102391347 A CN 102391347A
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Abstract
The invention belongs to the field of separation of natural products, and provides a joint preparation method for euscaphic acid and tormentic acid. The preparation method comprises the following steps of: extracting with an ethanol liquid; treating with macroporous resin; eluting sugars and other impurities with water and a low-concentration aqueous solution serving as a water-soluble organic solvent; eluting with a high-concentration aqueous solution serving as a water-soluble organic solvent; concentrating the eluent under reduced pressure to dryness to obtain a total triterpene extract; purifying with an efficient liquid phase preparative chromatograph; segmentally collecting an euscaphic acid fraction and a tormentic acid fraction; and thermally refluxing and crystallizing with methanol to obtain high-purity euscaphic acid and tormentic acid. A product prepared with the method has high product purity and good quality.
Description
Technical field
The invention belongs to the Separation of Natural Products field, especially relate to a kind of method that adopts the high performance liquid preparative chromatography method from solid public fruit, to prepare Jacaric acid and Tormentic acid simultaneously.
Background technology
Gu public fruit
Rosa odorata Sweet var.giganteaRehd.et Wils. belongs to gul, and its root property is flat, flavor acid, has the effect of relieving diarrhea with astringents, is usually used in diseases such as treatment diarrhoea, bacillary dysentery, is the long Yi nationality, distributed over Yunnan, Sichuan and Guizhou's medicinal material among the people of applicating history simply.Modern study shows and contains multiple triterpenes composition in the Radix Rosae Giganteae, and this triterpenes components has analgesic and anti-inflammatory effects preferably, and Jacaric acid is wherein to measure three higher terpene components with committee water chestnut dish acid.In addition, research such as Liu Liang shows with the Tormentic acid to be that the Herba Potentillae Freynianae triterpene compound of staple has obvious antivirus action.In view of the special pharmacological effect of these two kinds of triterpene substances, its DEVELOPMENT PROSPECT is infinitely-great.
Jacaric acid (Rosolic acid), molecular formula: C
19H
14O
3Molecular weight: 290.31.
Tormentic acid (Tormentic acid), molecular weight: 488.71, physico-chemical property: colourless crystallization type powder, mp.260～262 ℃ (methanol-water); Mp273 ℃, [α] D-20 ° (pyridine).IR?γmaxcm\-1:3400(-OH)，1700(C=O)，835(C=C)。MSm/z：488(M+)，264。1HNMR。13CNMR。White rod brilliant (acetone), mp276～279 ℃.Medicine turns usefulness into: hypoglycemic activity and anti-microbial effect.
Correlative study report to Tormentic acid and Jacaric acid in the existing document is less relatively, does not see the method that has two kinds of materials to prepare simultaneously, and also not seeing has the related patent U.S. Patent No. report.
Summary of the invention
The technical problem that the present invention will solve provides the combined prepn process of a kind of Jacaric acid and Tormentic acid, and this method is simple to operate, makes the product content height.
The present invention realizes through following technical scheme:
The combined prepn process of a kind of Jacaric acid and Tormentic acid is characterized in that comprising following steps:
1) extract: get solid public fruit pulverizing medicinal materials, add 40 ~ 70% extraction using alcohols 2 ~ 3 times, medicinal material and solvent ratios are 1: (5 ~ 10), and united extraction liquid is condensed into medicinal extract;
2) macroporous resin removal of impurities: the said extracted thing is crossed macroporous resin treatment; Elder generation's water and water-miscible organic solvent low concentration aqueous solution wash-out sugar and other impurity; The aqueous solution with the water-miscible organic solvent higher concentration carries out wash-out again, and elutriant is evaporated to dried total-triterpene extract;
3) the further separation and purification of preparation performance liquid: above-mentioned total-triterpene extract is used dissolve with methanol, and membrane filtration carries out separation and purification with high performance liquid preparative chromatography to it, collects the target elutriant respectively, reclaims reagent to small volume, places crystallization;
4) recrystallization: above-mentioned crystallization is got Jacaric acid and Tormentic acid product 2 ~ 3 times with methyl alcohol thermal backflow crystallization.
Process for extracting in the said step 1) can be selected heating reflux method or ultrasonic extraction for use, extracts temperature and is controlled at 60 ~ 100 ℃.
Said step 2) water-miscible organic solvent in can be a kind of in methyl alcohol, ethanol, the propyl alcohol etc.
Filter membrane aperture in the said step 3) is 0.2 ~ 0.45 μ m.
High performance liquid preparative chromatography in the said step 3) is to be stationary phase with the reversed-phase bonded silica, is moving phase with acetonitrile-0.3% trifluoroacetic acid (30 ~ 38:62 ~ 70), and the target flow point is collected in the UV-detector monitoring.
Described reversed-phase bonded silica is C-8 or C-18 bonded silica gel.
The wavelength set of described UV-detector is 210nm.
Positively effect of the present invention is:
1) adopt macroporous resin to remove polyose impurity, simple to operate, the macroporous resin treatment amount is big, and is reusable;
2) preparation performance liquid further separation and purification: used reversed phase high efficiency liquid phase preparative hplc, advantage such as have separating size height, favorable reproducibility, disengaging time is short, preparation amount is big isolatingly can reach more than 98% to product purity;
3) solvent systems of the present invention's use is comparatively simple, also has low cost, the advantage that the recovery is high.
To combine embodiment to further specify the present invention below, but the scope that the present invention requires to protect is not limited to following embodiment.
Embodiment
Embodiment 1:
Get solid public fruit medicinal material 1kg, pulverize, add the thermal backflow of 10L40% ethanol liquid and extract united extraction liquid 2 times; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and the desugar of 3BV30% ethanol aqueous wash and other impurity, is carried out wash-out with the 5BV65% aqueous ethanolic solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.45 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (30:70) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 3 times, leach crystal, get highly purified Jacaric acid 1.2g (content 98.2%) and Tormentic acid 0.7g (content 98.7%).
Embodiment 2:
Get solid public fruit medicinal material 1Kg, pulverize, add the 5L70% ethanol ultrasonic extraction 3 times, united extraction liquid; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and 3BV30% methanol aqueous solution wash-out sugar and other impurity, is carried out wash-out with the 5BV70% methanol aqueous solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.22 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (32:68) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 2 times, leach crystal, get highly purified Jacaric acid 1.1g (content 98.3%) and Tormentic acid 0.8g (content 98.5%).
Embodiment 3:
Get solid public fruit medicinal material 5Kg, pulverize, add the 40L50% ethanol ultrasonic extraction 2 times, united extraction liquid; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and 3BV25% aqueous propanol solution wash-out sugar and other impurity, is carried out wash-out with the 5BV70% aqueous propanol solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.22 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (35:65) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 3 times, leach crystal, get highly purified Jacaric acid 5.4g (content 98.5%) and Tormentic acid 3.1g (content 99.1%).
Embodiment 4:
Get solid public fruit medicinal material 5Kg, pulverize, add the 35L55% alcohol heating reflux and extract united extraction liquid 3 times; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and the desugar of 3BV30% ethanol aqueous wash and other impurity, is carried out wash-out with the 5BV75% aqueous ethanolic solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.22 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (33:67) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 3 times, leach crystal, get highly purified Jacaric acid 4.1g (content 99.2%) and Tormentic acid 2.8g (content 99.3%).
Embodiment 5:
Get solid public fruit medicinal material 10Kg, pulverize, add the 60L65% ethanol ultrasonic extraction 2 times, united extraction liquid; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and the desugar of 3BV35% ethanol aqueous wash and other impurity, is carried out wash-out with the 5BV75% aqueous ethanolic solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.45 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (38:62) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 2 times, leach crystal, get highly purified Jacaric acid 9.8g (content 98.5%) and Tormentic acid 5.9g (content 98.6%).
Embodiment 6:
Get solid public fruit medicinal material 10Kg, pulverize, add 70L40% methyl alcohol heating and refluxing extraction 2 times, united extraction liquid; Be concentrated into low alcohol, last macroporous resin adsorption with 2BV water and the desugar of 3BV30% ethanol aqueous wash and other impurity, is carried out wash-out with the 5BV70% aqueous ethanolic solution earlier again; Elutriant is evaporated to dried total-triterpene extract, and the total triterpene of gained is dissolved with small amount of methanol, and the moving phase dilution filters through 0.45 μ m filter membrane; Adopting anti-phase to prepare the further separation and purification of performance liquid again, is stationary phase with the C-18 bonded silica gel, and acetonitrile-0.3% trifluoroacetic acid (30:70) solution is moving phase; Flow velocity is set at 30ml/min, sets UV-detector wavelength 210nm, Fractional Collections Jacaric acid flow point and Tormentic acid flow point; Reclaim reagent to small volume, place crystallization, leach crystallisate and dissolve with the methyl alcohol reflux; Put cold analysis brilliant 3 times, leach crystal, get highly purified Jacaric acid 10.2g (content 98.1%) and Tormentic acid 7.1g (content 98.1%).
Claims (5)
1. the combined prepn process of Jacaric acid and Tormentic acid is characterized in that comprising following steps:
1) extract: get solid public fruit pulverizing medicinal materials, add 40 ~ 70% extraction using alcohols 2 ~ 3 times, medicinal material and solvent ratios are 1: (5 ~ 10), united extraction liquid;
2) macroporous resin removal of impurities: the said extracted thing is crossed macroporous resin treatment; Elder generation's water and water-miscible organic solvent low concentration aqueous solution wash-out sugar and other impurity; The aqueous solution with the water-miscible organic solvent higher concentration carries out wash-out again, and elutriant is evaporated to dried total-triterpene extract;
3) the further separation and purification of preparation performance liquid: with above-mentioned total-triterpene extract, use dissolve with methanol, membrane filtration carries out separation and purification with high performance liquid preparative chromatography to it, collects the target elutriant respectively, reclaims reagent to small volume, places crystallization;
4) recrystallization: above-mentioned crystallization is got Jacaric acid and Tormentic acid product 2 ~ 3 times with methyl alcohol thermal backflow crystallization.
2. the combined prepn process of a kind of Jacaric acid as claimed in claim 1 and Tormentic acid is characterized in that the process for extracting in the said step 1) can be selected heating reflux method or ultrasonic extraction for use, extracts temperature and is controlled at 60 ~ 100 ℃.
3. the combined prepn process of a kind of Jacaric acid as claimed in claim 1 and Tormentic acid is characterized in that said step 2) in water-miscible organic solvent can be a kind of in methyl alcohol, ethanol, the propyl alcohol etc.
4. the combined prepn process of a kind of Jacaric acid as claimed in claim 1 and Tormentic acid is characterized in that the filter membrane aperture in the said step 3) is 0.2 ~ 0.45 μ m.
5. the combined prepn process of a kind of Jacaric acid as claimed in claim 1 and Tormentic acid; It is characterized in that the high performance liquid preparative chromatography in the said step 3) is to be stationary phase with the reversed-phase bonded silica; With acetonitrile-0.3% trifluoroacetic acid (30 ~ 38:62 ~ 70) is moving phase, and the target flow point is collected in the UV-detector monitoring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102259426A CN102391347A (en) | 2011-08-09 | 2011-08-09 | Joint preparation method for euscaphic acid and tormentic acid |
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|---|---|---|---|
| CN2011102259426A CN102391347A (en) | 2011-08-09 | 2011-08-09 | Joint preparation method for euscaphic acid and tormentic acid |
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| CN102391347A true CN102391347A (en) | 2012-03-28 |
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| CN2011102259426A Pending CN102391347A (en) | 2011-08-09 | 2011-08-09 | Joint preparation method for euscaphic acid and tormentic acid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106222A (en) * | 2015-09-14 | 2015-12-02 | 新乡医学院 | Application of tormentic acid in preparation of medicine for treating or preventing Alzheimer disease caused by estrogen lack |
| CN111548386A (en) * | 2020-05-28 | 2020-08-18 | 广西中医药大学第一附属医院 | Method for extracting tormentic acid and application thereof |
-
2011
- 2011-08-09 CN CN2011102259426A patent/CN102391347A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106222A (en) * | 2015-09-14 | 2015-12-02 | 新乡医学院 | Application of tormentic acid in preparation of medicine for treating or preventing Alzheimer disease caused by estrogen lack |
| CN105106222B (en) * | 2015-09-14 | 2020-05-08 | 新乡医学院 | Application of emetic acid in preparing medicament for treating or preventing Alzheimer disease caused by estrogen deficiency |
| CN111548386A (en) * | 2020-05-28 | 2020-08-18 | 广西中医药大学第一附属医院 | Method for extracting tormentic acid and application thereof |
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Application publication date: 20120328 |