CN101081857A - Method for purifying forsythiaside A from forsythia extractive - Google Patents
Method for purifying forsythiaside A from forsythia extractive Download PDFInfo
- Publication number
- CN101081857A CN101081857A CN 200610027037 CN200610027037A CN101081857A CN 101081857 A CN101081857 A CN 101081857A CN 200610027037 CN200610027037 CN 200610027037 CN 200610027037 A CN200610027037 A CN 200610027037A CN 101081857 A CN101081857 A CN 101081857A
- Authority
- CN
- China
- Prior art keywords
- forsythiaside
- water
- ratio
- peak
- purifying
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Saccharide Compounds (AREA)
Abstract
The present invention relates to process of purifying forsythia extract to obtain forsythoside A. Specifically, forsythia phenols are further purified to obtain high purity forsythoside A in high extracting efficiency.
Description
Technical field
The present invention relates to a kind of purification process of Chinese medicine Fructus Forsythiae extract, belong to medical technical field.
Background technology
The capsule of weeping forsythia (Fructus Forsythiae) is the Oleaceae forsythia.In China's tradition ancient prescription, the fruit medicine of the capsule of weeping forsythia commonly used uses jointly with other antipyretic and antidotal type medicine, plays effect clearing heat and detoxicating, mass dissipating and swelling eliminating, is the version " conventional Chinese medicine that Chinese pharmacopoeia is recorded in 2000.Chinese scholars is more to capsule of weeping forsythia chemical ingredients and pharmacological research, has realized that the antimicrobial antiviral activity highly significant of the capsule of weeping forsythia.Its main antimicrobial antiviral activity composition is a Fructus Forsythiae ester glycoside, and in the Fructus Forsythiae ester glycoside almost all be forsythiaside A in action, it all has extremely strong restraining effect to 11 kinds of pathogenic bacterium such as streptococcus aureuses, also has very strong antimycotic, antiviral effect.Be in the past with phyllyrin as the index of investigating capsule of weeping forsythia extraction process, deeply find phyllyrin and do not have antibiotic activity along with the effective constituent research of the capsule of weeping forsythia in recent years.From the standard of gradually forsythiaside A being estimated as capsule of weeping forsythia quality of medicinal material, in Chinese patent medicine, be used as the index of control quality product.The structural formula of forsythiaside A is as follows:
Also put down in writing the preparation method of some forsythiaside As in the document.As adopt polymeric amide pressured column chromatography to carry out initial gross separation, and use the dextrane gel column purification again, with methyl alcohol and water system wash-out, perhaps the methyl alcohol of water, different concns, ethanolic soln are solvent, carry out methods such as supersound extraction.But the anti-microbial activity that to have mentioned a problem in the document repeatedly be exactly the capsule of weeping forsythia becomes the split pole instability, forsythiaside A wherein is caffeinic derivative, have ester bond and glycosidic link in the molecular structure, under acid, alkali and hot conditions, decompose easily, and the anti-microbial activity of the coffic acid, D-glucose, L-rhamnosyl and the secondary glycoside that produce after decomposing weakens greatly.Owing to failed to notice this problem in the past, so patent medicine such as antitoxic bolus of honeysuckle flower and forsythia, YINQIAO JIEDU PIAN only detect coffic acid, and hydrolysis may take place in forsythiaside A in the course of processing.Therefore prior art is to avoid the hydrolysis of forsythiaside A, all wants controlled temperature, avoids contacting strong acid and strong base, and this has limited to the extraction process of forsythiaside A to a great extent, has reduced extraction efficiency.
The contriver has summed up a kind of high temperature extraction method that can keep the Fructus Forsythiae extract anti-microbial activity, and has applied for patent through research repeatedly, and application number is 200510030815.5.The extract that this patent is carried be a kind of total aldehydes matter of the capsule of weeping forsythia, and the antimicrobial antiviral activity of this type of material is more inferior than forsythiaside A, but can be by the total phenols extract of the capsule of weeping forsythia being carried out purifies and separates, and then obtains a large amount of forsythiaside As.
Summary of the invention
Purpose of the present invention just provides a kind of from Fructus Forsythiae extract, mainly is the method that further separation and purification obtains forsythiaside A in the total aldehydes matter of the capsule of weeping forsythia.
Realize the technical scheme of invention: the present invention is the method that is purified into forsythiaside A from the total aldehydes matter of the capsule of weeping forsythia.The acquisition of the total aldehydes matter of the capsule of weeping forsythia can have a lot of methods, such as:
Hot formulation: with water extraction capsule of weeping forsythia starting material, filter, concentrating under reduced pressure adds diatomite and stirs; Diatomite after drying with the 70-80% ethanol ultrasonic extraction again merges alcohol extract, or with 70-80% ethanol and water elution; Last macroporous resin, ethanolic soln is collected in water elution, ethanolysis absorption, and concentrating under reduced pressure gets Fructus Forsythiae extract.
Cold formulation: with 8 times of 50% methyl alcohol cold soaking capsule of weeping forsythia starting material, spend the night, supersound extraction 20min collects alcoholic solution, and drying under reduced pressure promptly gets Fructus Forsythiae extract.
Said capsule of weeping forsythia starting material are part or all of capsule of weeping forsythia plants such as the fruit that comprises the capsule of weeping forsythia, leaf, seed.
Realize that technical scheme of the present invention is: thick silica gel dry method is loaded in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder, with thick silica gel with 1: the ratio of 2-5 mixing in mortar, sample on the solid, applied sample amount are 1: 30-60 is the moving phase wash-out with ethyl acetate-acetone-water or ethyl acetate-C1-C5 lower alcohol-water or chloroform-C1-C5 lower alcohol-water, ratio is 9-1: 0.5-5: 5-0.5, the control flow velocity is collected the forsythiaside A peak at 8-12ml/min, merges, concentrating under reduced pressure, vacuum-drying; Go up the RP-18 silicagel column again, applied sample amount is 1: 50-100, and purifying, the methanol-water wash-out, ratio is 1-4: 6-9, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
Preferably, technical scheme of the present invention is: thick silica gel dry method is loaded in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder, with thick silica gel with 1: the ratio of 2-5 mixing in mortar, sample on the solid, applied sample amount are 1: 30-60 is the moving phase wash-out with ethyl acetate-acetone-water, ratio is 8: 2: 0.5, the control flow velocity is collected the forsythiaside A peak at 8-12ml/min, merges, concentrating under reduced pressure, vacuum-drying; Go up the RP-18 silicagel column again, applied sample amount is 1: 50-100, and purifying, the methanol-water wash-out, ratio is 3: 7, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
Advantage of the present invention: 1. extraction efficiency improves greatly, can save better and utilize starting material, obtains highly purified forsythiaside A; 2. continue purifying from the basis of forsythol, needn't be subjected to the restriction of prior art said essential " controlled temperature ".
Description of drawings
Forsythiaside A purity detecting high-efficient liquid phase chromatogram
Embodiment
Embodiment 1
Thick silica gel dry method is loaded on (blade diameter length ratio is 1: 15) in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder 10 grams are with thick silica gel 20 grams of 100 orders mixing in mortar, sample on the solid, applied sample amount 1: 30, with ethyl acetate-acetone-water (8: 2: 0.5) is the moving phase wash-out, and control flow velocity 10ml/min collects the forsythiaside A peak, merge concentrating under reduced pressure, vacuum-drying; Go up RP-18 silicagel column (blade diameter length ratio 1: 12, applied sample amount 1: 50) purifying again, methanol-water (3: 7) wash-out is collected the forsythiaside A peak, merge, and concentrating under reduced pressure, vacuum-drying promptly gets the forsythiaside A product.
Thick silica gel dry method is loaded on (blade diameter length ratio is 1: 15) in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder 10 grams are with thick silica gel 20 grams of 100 orders mixing in mortar, sample on the solid, applied sample amount 1: 30, with ethyl acetate-acetone-water (9: 0.5: 0.5) is the moving phase wash-out, and control flow velocity 8ml/min collects the forsythiaside A peak, merge concentrating under reduced pressure, vacuum-drying; Go up RP-18 silicagel column (blade diameter length ratio 1: 12, applied sample amount are 500 grams) purifying again, the methanol-water wash-out, ratio is 4: 6, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
Embodiment 3
Thick silica gel dry method is loaded on (blade diameter length ratio is 1: 15) in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder 10 grams are with thick silica gel 50 grams of 200 orders mixing in mortar, sample on the solid, applied sample amount 1: 60, with ethyl acetate-methanol-water (1: 4.5: 4.5) is the moving phase wash-out, and control flow velocity 12ml/min collects the forsythiaside A peak, merge concentrating under reduced pressure, vacuum-drying; Go up RP-18 silicagel column (blade diameter length ratio 1: 12, applied sample amount are 1: 100) purifying again, the methanol-water wash-out, ratio is 1: 9, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
Thick silica gel dry method is loaded on (blade diameter length ratio is 1: 15) in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder 10 grams are with thick silica gel 35 grams of 200 orders mixing in mortar, sample on the solid, 1: 45 gram of applied sample amount, with chloroform-methanol-water (5: 2.5: 2.5) is the moving phase wash-out, and the control flow velocity is collected the forsythiaside A peak at 10ml/min, merge concentrating under reduced pressure, vacuum-drying; Go up RP-18 silicagel column (blade diameter length ratio 1: 12, applied sample amount are 1: 75) purifying again, the methanol-water wash-out, ratio is 2: 8, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
The discriminating of forsythiaside A
It is an amount of that precision takes by weighing the forsythiaside A that makes by the embodiment method, adds methyl alcohol and make the solution that every 1ml contains forsythiaside A 0.62mg; Accurately drawing above-mentioned solution 40 μ l, put on silica gel g thin-layer plate, is developping agent with chloroform-acetone-water (7.5: 1.5: 0.5), and exhibition distance: 8cm launches, and takes out, and dries; Spray is with 10% ethanol solution of sulfuric acid, and after drying, it is clear to be heated to the spot colour developing at 105 ℃, R
fValue is 0.50; Not having other spot on the used thin layer plate shows.
The forsythiaside A purity test
Measure according to high performance liquid chromatography (2005 editions appendix VID of the Pharmacopoeia of the People's Republic of China)
Chromatographic condition and system suitability test: octadecylsilane chemically bonded silica is a weighting agent, and methanol-water-Glacial acetic acid (33: 67: 0.2) is a moving phase; Detect wavelength 290nm; Flow velocity 1.0ml/min; 25 ℃ of column temperatures.Theoretical plate number is calculated by the forsythiaside A peak and is no less than 8000.
The preparation of reference substance solution: it is an amount of to get the forsythiaside A reference substance, adds 50% methyl alcohol and makes the solution that every 1ml contains forsythiaside A 0.03943mg, shakes up, promptly.
Assay method: the accurate reference substance solution 10 μ l that draw, inject liquid chromatograph, measurement result sees Table 13-1.
Table 13-1 forsythiaside A reference substance purity test data
| The peak numbering | Peak 1 | Peak 2 (forsythiaside A) | Peak 3 |
| Retention time (min) peak area integrated value peak area per-cent | 8.736 1.63 0.32 | 19.254 501.97 98.20 | 33.361 8.17 1.60 |
Attached liquid phase collection of illustrative plates
The spectral data of reference substance and parsing
1UV
max(MeOH/H
2O 1∶1)：220，292，336nm。
2IR (KBr) v
Max: 3400 (OH), 1697 (conjugation carbonyls), 1605 (carbon-carbon double bonds), 1522,1283,1159,1059,980,812cm
-1
3 ESIMS：623[M-H]
-
4
1H-NMR(CD
3OD，400MHz)δ：7.59(1H，d，J＝16.0Hz，H-7’)，7.0(1H，d，J＝2.0 Hz，H-2’)，6.96(1H，dd，J＝8.2，2.0Hz，H-6’)，6.78(1H，d，J＝8.2Hz，H-5’)，6.68(1H，d，J＝8.2 Hz，H-3)，6.57(1H，dd，J＝8.2，2.0Hz，H-2)，6.29(1H，d，J＝16.0Hz，H-8’)，4.63(1H，d，J＝1.6 Hz，H-1)，4.36(1H，d，J＝7.8Hz，H-1”)，3.26-4.01(11H，m)，2.80(2H，m，H2-7)，1.19(3H，d，J＝6.3Hz，Me-6)。
5
13C-NMR(CD
3OD，100MHz)δ：168.8(C-9)，150.2(C-4’)，148.1(C-3’)，147.3(C-7’)，146.6(C-3)，145.2(C-4)，131.9(C-1)，128.2(C-1’)，123.6(C-6’)，121.8(C-6)，117.6(C-5)，117.0(C-5’)，116.8(C-2)，115.7(C-2’)，115.2(C-4)，105.0(C-1”)，102.8(C-1)，76.3(C-3”)，75.7(C-2”)，75.3(C-4”)，74.4(C-5)，72.9(C-8)，72.8(C-4)，72.8(C-3)，72.5(C-2)，70.4(C-5)，68.2(C-6”)，37.2(C-7)，1 8.5(C-6)。
Claims (2)
1. the method for a purifying forsythiaside A from Fructus Forsythiae extract is characterized in that: thick silica gel dry method is loaded in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder, with thick silica gel with 1: the ratio of 2-5 mixing in mortar, sample on the solid, applied sample amount are 1: 30-60 is the moving phase wash-out with ethyl acetate-acetone-water or ethyl acetate-C1-C5 lower alcohol-water or chloroform-C1-C5 lower alcohol-water, ratio is 9-1: 0.5-5: 5-0.5, the control flow velocity is collected the forsythiaside A peak at 8-12ml/min, merges, concentrating under reduced pressure, vacuum-drying; Go up the RP-18 silicagel column again, applied sample amount is 1: 50-100, and purifying, the methanol-water wash-out, ratio is 1-4: 6-9, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
2. as claimed in claim 1 a kind of from Fructus Forsythiae extract the method for purifying forsythiaside A, it is characterized in that: thick silica gel dry method is loaded in the glass column, and is connected on the ultraviolet chromatographic instrument.Fructus Forsythiae extract dry powder, with thick silica gel with 1: the ratio of 2-5 mixing in mortar, sample on the solid, applied sample amount are 1: 30-60 is the moving phase wash-out with ethyl acetate-acetone-water, ratio is 8: 2: 0.5, the control flow velocity is collected the forsythiaside A peak at 8-12ml/min, merges, concentrating under reduced pressure, vacuum-drying; Go up the RP-18 silicagel column again, applied sample amount is 1: 50-100, and purifying, the methanol-water wash-out, ratio is 3: 7, collects the forsythiaside A peak, merge, concentrating under reduced pressure, vacuum-drying promptly gets forsythiaside A.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006100270379A CN101081857B (en) | 2006-05-29 | 2006-05-29 | Method for purifying forsythiaside A from forsythia extractive |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006100270379A CN101081857B (en) | 2006-05-29 | 2006-05-29 | Method for purifying forsythiaside A from forsythia extractive |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101081857A true CN101081857A (en) | 2007-12-05 |
| CN101081857B CN101081857B (en) | 2010-06-16 |
Family
ID=38911672
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2006100270379A Active CN101081857B (en) | 2006-05-29 | 2006-05-29 | Method for purifying forsythiaside A from forsythia extractive |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101081857B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101215302B (en) * | 2008-01-08 | 2010-11-03 | 山西大学 | Method for preparing forsythiaside A |
| CN102050849A (en) * | 2010-12-09 | 2011-05-11 | 江苏天晟药业有限公司 | Forsythoside A and preparation method thereof |
| CN103193836A (en) * | 2013-04-19 | 2013-07-10 | 山东农业大学 | Method for separating and purifying monomers in forsythia suspensa flower |
| CN103450296A (en) * | 2013-08-29 | 2013-12-18 | 山西大学 | Method for rapidly preparing forsythin from forsythia suspense leaves |
| CN117229338A (en) * | 2023-11-13 | 2023-12-15 | 中泰(天津)生物科技有限公司 | Method for comprehensively extracting forsythin, forsythoside, rutin and polysaccharide from forsythia leaves and application of method |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1148375C (en) * | 2001-07-24 | 2004-05-05 | 山西大学 | Forsythigenol extracting process |
| CN100343204C (en) * | 2005-10-28 | 2007-10-17 | 上海玉森新药开发有限公司 | Forsythia seed extract and its prepn process |
-
2006
- 2006-05-29 CN CN2006100270379A patent/CN101081857B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101215302B (en) * | 2008-01-08 | 2010-11-03 | 山西大学 | Method for preparing forsythiaside A |
| CN102050849A (en) * | 2010-12-09 | 2011-05-11 | 江苏天晟药业有限公司 | Forsythoside A and preparation method thereof |
| CN103193836A (en) * | 2013-04-19 | 2013-07-10 | 山东农业大学 | Method for separating and purifying monomers in forsythia suspensa flower |
| CN103193836B (en) * | 2013-04-19 | 2015-09-09 | 山东农业大学 | The separation purification method of monomeric compound in a kind of Flos Forsythiae |
| CN103450296A (en) * | 2013-08-29 | 2013-12-18 | 山西大学 | Method for rapidly preparing forsythin from forsythia suspense leaves |
| CN117229338A (en) * | 2023-11-13 | 2023-12-15 | 中泰(天津)生物科技有限公司 | Method for comprehensively extracting forsythin, forsythoside, rutin and polysaccharide from forsythia leaves and application of method |
| CN117229338B (en) * | 2023-11-13 | 2024-03-08 | 中泰(天津)生物科技有限公司 | Method for comprehensively extracting forsythin, forsythoside, rutin and polysaccharide from forsythia leaves and application of method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101081857B (en) | 2010-06-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101081855B (en) | Method for purifying forsythiaside A from forsythia extractive | |
| CN101081857B (en) | Method for purifying forsythiaside A from forsythia extractive | |
| CN105175239B (en) | Sesquiterpenoid compound capable of inhibiting activity of tobacco mosaic virus in tobacco, preparation method and applications thereof | |
| CN101612173B (en) | Glabrous sarcandra herb extract, quality control method for extract preparation and application of thereof | |
| CN102302685B (en) | Common lophatherum herb extract and preparation method and application thereof | |
| CN105440092B (en) | The fast preparation method of flavonoid glycoside in a kind of Extracted From Oil-tea-cake | |
| CN103554077B (en) | Chromone compound as well as preparation method and application thereof | |
| CN101096378A (en) | Method for abstracting and separating iridoid compound monomer from oldenlandia diffusa | |
| CN108191931A (en) | Three kinds of noval chemical compounds and its health care and medical usage in root of Paeonia sinjiangensis | |
| CN105061451A (en) | Highly oxidized diterpenoid compound and preparation method and medical application thereof | |
| CN101081856B (en) | Method for purifying forsythiaside A from forsythia extractive | |
| CN101347482B (en) | Ainsliaea fragrans extract and active component thereof as well as their quality control method | |
| CN102329355B (en) | The preparation method of a kind of Myricitroside and pharmaceutical composition thereof | |
| CN109265494B (en) | Method for extracting kaempferol glucoside compounds from camellia reticulata | |
| CN102068649A (en) | Quality control method for gastric condition-regulating pill as traditional Chinese preparation | |
| CN103896755B (en) | A kind of chalcone compounds preparation method | |
| CN102772501A (en) | Rheum emodi Wall. extract and its preparing method | |
| CN110272459A (en) | Two kinds of noval chemical compounds and its antioxidant activity position in root of Paeonia sinjiangensis | |
| CN111848709B (en) | Novel isopentene-based flavonoid compound and preparation method and application thereof | |
| CN103554071B (en) | Amidpulver compound and preparation method and application thereof | |
| CN104650053B (en) | Flavonoids compound, as well as preparation method and applications thereof | |
| CN108440612A (en) | The isolation and purification method of three kinds of iridoid constituents in a kind of radix scrophulariae | |
| CN109879920B (en) | Preparation method and application of coumarin glucoside compound | |
| CN105061212A (en) | Preparation method of neochlorogenic acid | |
| CN103922913B (en) | A kind of Chalcone Compounds and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: SHANDONG XINSHIDAI PHARMACEUTICAL INDUSTRY CO., LT Free format text: FORMER OWNER: SHANGHAI YOUSEEN NEW PHARMACEUTICAL DEVELOPMENT CO., LTD Effective date: 20100806 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 201203 NO.351, GUOSHOUJING ROAD, ZHANGJIANG HIGH-TECHNOLOGY PARK, PUDONG NEW DISTRICT, SHANGHAI CITY TO: 273400 NO.1, NORTH OUTER RING ROAD, FEIXIAN COUNTY, SHANDONG PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20100806 Address after: 273400 Shandong province Feixian County North Ring Road No. 1 Patentee after: Shandong Xinshidai Pharmaceutical Industry Co., Ltd. Address before: 201203 Shanghai Guo Shou Jing Road, Pudong New Area Zhangjiang hi tech Park No. 351 Patentee before: Shanghai Youseen New Medicine Exploitation Co., Ltd. |