CN102643322A - Method for simultaneously preparing pedunculoside and syringin - Google Patents
Method for simultaneously preparing pedunculoside and syringin Download PDFInfo
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Abstract
The invention discloses a method for simultaneously preparing high-purity syringin and pedunculoside. According to the method, the high-purity syringin and the high-purity pedunculoside are prepared by using holly barks as a raw material, adopting a solvent extraction method and a macromolecular resin adsorption method for extraction and separation and adopting re-crystallization, open ozone depleting substance (ODS) column chromatography or Se-phadex LH-20 column chromatography, liquid chromatography and the like. The purity of each of the syringin and the pedunculoside is over 90 percent, and the method is simple, feasible and suitable for industrialized production.
Description
Technical field
The present invention relates to a kind of method for preparing pedunculoside and Syringin simultaneously.
Background technology
Ovateleaf Holly Bark is the dry bark of holly plant Ilex rotunda Thunb. Ilex rotunda Thunb., and the beginning is stated from " south of the Five Ridges gather medicinal herbs record ", has another name called that Lignum Aquilariae Resinatum, sheep are not eaten, agalloch eaglewood etc. in vain.Sparse woods or ditch, small stream limit are distributed in Jiangsu, Anhui, Zhejiang, Jiangxi, Fujian, Taiwan, Hunan, Guangdong, Guangxi, Yunnan under the Ilex rotunda Thunb. growth Yushan Hill, are people side, China south herbal medicine commonly used.Have clearing heat and detoxicatingly, dampness removing lenitive effect is usually used in treating summer-heat and damp heating, swelling and pain in the throat, damp-heat dysentery, abdominal distention, rheumatic arthralgia, eczema, sore furuncle, wound.In addition, Ovateleaf Holly Bark extraction using alcohol position has stronger antibacterial and anti-inflammatory action [Zhang Rongwen, Huang Zhaosheng; Fan Qingya, etc. Ovateleaf Holly Bark antibacterial anti-inflammatory efficient part screening [J]. Chinese tcm-academic periodical, 2008; 26 (8): 1820-1822], and to the irritability Hypertensive Rats step-down and reducing heart rate effect [Liang Yanling, Dong Yanfen are arranged; Luo Jipeng. the Ovateleaf Holly Bark ethanol extraction is to the experimental study [J] of irritability Hypertensive Rats hypotensive effect. Chinese medicinal materials, 2005,28 (7): 582-584.]; The Ovateleaf Holly Bark n-butanol extract have arrhythmia and function of resisting myocardial ischemia [Chen Xiaoxia, what ice, Xu Yuanfen, etc. Ovateleaf Holly Bark n-butanol extract arrhythmia is studied [J] with function of resisting myocardial ischemia. Pharmacology and Clinics of Chinese Materia Medica, 1998,14 (4) .].The clinical application of China to these article focus mostly on to treatment of diseases such as pipe intestinal digesting system [Dai Weibo, Mei Quanxi, Ceng Congyan. Ovateleaf Holly Bark chemical constitution study and Clinical advances [J]. Asia-Pacific traditional medicine, 2008 (12): 137-139.].
Rescue will be mainly contains triterpenes, flavonoids and phenolic compounds [Guangzhou City Institute for Drug Control. Herbal save Bing experimental study [J]. New medicine communication, 1970 (2) :23-26.], And separated get pedunculoside [Zhu Renhong, Hongshan Hai, Wang Youhai. medicine will be saved in the glycoside [J]. Acta Chimica Sinica, 1956,22 (2) :128-133.], iron holly acid [Man Yuk, Chen Zhongliang. rescue will be the chemical constituents of (I) [J]. herbal medicine, 1991,22 (6) :246-248.], iron holly acid isopropylidene ketal, mustard aldehyde, lilac aldehyde 3 - acetyl oleanolic acid, stearic acid, syringin, mustard aldehyde glucoside, slope mold alkyd, bitter Dingdong Qing glycosides H ,3-O-α-L-arabinopyranosyl-3β, 19α-dihydroxy-oleanolic If the alkyl -12 - en-28 - acid-28-O-β-D-glucopyranosyl esters, clove Lignans 4'-O-β-D-glucopyranoside, caffeic acid, 4-O-β-D - glucopyranoside, vanillic acid 4-O-β-D-glucopyranoside, β-sitosterol, daucosterol, two cloves ether, friedelin 3 - hydroxy-oleanane, nonadecanoic acid .
In the more activeconstituents of Ovateleaf Holly Bark, higher and active outstanding with pedunculoside and Syringin content especially, the former belongs to pentacyclic triterpene, and the latter belongs to the phenylpropyl alcohol glycosides compound.Pedunculoside hyperlipidemia rats to diet induced when dosage is 40mg/kg has reducing total cholesterol, low density lipoprotein cholesterol and AI significantly, and the effect of high density lipoprotein increasing SUV; Effect [the Jahromi M A F that when dosage is 75mg/kg, Triton inductive hyperlipidemia rats is had remarkable reducing total cholesterol and total fat; Gupta M; Manickam M, et al.Hypolipidemic Activity of Pedunculoside, A Constitunt of Ilex Doniana [J] .Pharmaceutical Biology; 1999,37 (1): 37-41.].Discovery pedunculosides such as Li Chaosheng can obviously alleviate degree of myocardial ischemia and the scope (dosage is 40mg/kg) that the dog branch of coronary artery causes; Reduce myocardial oxygen consumption index (dosage is 40mg/kg); And can obviously resist bariumchloride (4mg/kg) inductive rat anti irregular pulse (dosage is 25-100mg/kg); Also can reduce the brain infarction area (dosage is 100mg/kg) of focal cerebral ischemia rat, show Folium Ilicis macrocarpae to cerebral ischemia and myocardial ischemia have the certain protection effect [Li Chaosheng. the preparation method of Peltatin glucoside and application [P] .CO7J63/00.200610017186.7.2008-08-01].Syringin adopts determination of tube method that the clotting time is shortened, and anastalsis is arranged; In the body test dog femoral artery is cut; Dog and rabbit lobe of the liver partly excise, dog spleen cross recess, and the rabbit ear and mesenteric vein cut; All can shorten bleeding stopping period; It is relevant that its hemostatic mechanism part and vascular smooth muscle shrink, this stripped rabbit ear blood vessel perfusion experiment can obtain proof [institute for drug control, Guangzhou. the experimental study of herbal medicine Ovateleaf Holly Bark [J]. new pharmaceutical communication, 1970 (2): 23-26.].Clinical observation confirms that the syringin injection liquid has anastalsis, and like more remarkable to the hemorrhage curative effect of urinary system; Intravenously administrable, curative effect is significantly improved, and have no side effect [Pang Xiaoxiong, Lv Huachong, Luo Jipeng. the separation of Pedunculoside II and evaluation [J] in the Ovateleaf Holly Bark. ACAD J GCP, 1997,13 (2) .].
At present, Shang Weiyou report of rapid extraction pedunculoside and two compositions of Syringin simultaneously from Ovateleaf Holly Bark.The method of existing extraction purifying pedunculoside and Syringin mainly is traditional ethanol-extracted, column chromatography purification.The tradition purification process is very long and the target compound loss is more, adopts repeatedly silica gel column chromatography, and operation steps is various and elutriant is big with toxicity such as chloroform, acetone, ETHYLE ACETATE, the higher reagent of price is main.Chinese patent " preparation method of Peltatin glucoside and application " (publication number CN101033245) adopts traditional alcohol extracting Ovateleaf Holly Bark method, and only obtaining pedunculoside and purity is 98%, has lost a large amount of Syringins.The separation of employing conventional post chromatography methods such as Pang Xiaoxiong obtains pedunculoside, and this method time is long, and the organic reagent consumption is big, and output is little.Syringin Chinese patent (CN101307079) adopts benzene to separate as solvent extraction with acetone and obtains Syringin, and this method solvent for use toxicity is bigger.
In sum, in the process of extraction separation pedunculoside and Syringin, loss, the purge process of ubiquity pedunculoside and Syringa oblata Lindl. is very long, dissolvent residual toxicity big, the cost problem of higher.Therefore, be necessary to seek a kind of not only easy but also extract the method for purifying pedunculoside and Syringin safely.
Summary of the invention
The object of the present invention is to provide a kind of method that from the Chinese medicine Ovateleaf Holly Bark, obtains high purity pedunculoside and Syringin in a large number, fast.
The structural formula of said pedunculoside is suc as formula shown in the I:
(formula I)
The structural formula of said Syringin is suc as formula shown in the II:
(formula II)
Method provided by the present invention comprises the steps:
1) aqueous ethanolic solution with volume(tric)fraction 50%-70% is an extraction agent, and Ovateleaf Holly Bark is extracted, and collects extracting solution;
2) said extracting solution is carried out column chromatography for separation through macroporous adsorbent resin; With the alcohol-water is that eluent carries out gradient elution, and the collected volume mark is that the elution fraction and the volume(tric)fraction of 10%-30% aqueous ethanolic solution are the elution fraction of 40%-70% aqueous ethanolic solution respectively;
3) with said volume(tric)fraction be the elution fraction of 10%-30% aqueous ethanolic solution concentrate, dry, obtain the Syringin bullion, adopt recrystallization or column chromatography to carry out purifying said Syringin bullion, obtain the Syringin of purifying;
With said volume(tric)fraction be the elution fraction of 40%-70% aqueous ethanolic solution concentrate, dry, obtain the pedunculoside bullion, adopt recrystallization method or column chromatography to carry out purifying said pedunculoside bullion, obtain the pedunculoside of purifying.
Wherein, Ovateleaf Holly Bark described in the step 1) is dry bark or the stem skin of holly plant Ilex rotunda Thunb. Ilex rotunda Thunb..
The method of said extraction can be methods such as cold soaking extraction, supersound extraction or refluxing extraction, preferred refluxing extraction.
When adopting refluxing extraction, said refluxing extraction can be carried out 1-3 time, and each time of extracting is 30 minutes-90 minutes, and when extracting, the consumption of extraction agent and the proportioning of Ovateleaf Holly Bark are (8-16) L: 1kg at every turn.
With also needing before the macroporous adsorbent resin on the extracting solution of step 1) it is concentrated, the concentration of gained liquid concentrator is 0.04-0.16g crude drug/mL.
Step 2) macroporous adsorbent resin described in can be selected from the macroporous adsorbent resin of following any one model: HP20, NKA-II, HPD-100A, SP825, DM301, D101, AB-8 and S-8.
The actual conditions of gradient elution step 2) is following: use volume(tric)fraction to carry out wash-out as the 10%-30% aqueous ethanolic solution earlier, use volume(tric)fraction to carry out wash-out as the 40%-70% aqueous ethanolic solution again; But 2-5 times of column volume of each gradient wash-out.According to the Liquid Detection result, merge elutriant, reclaim solvent, concentrate, drying, the bullion of Syringin and pedunculoside.
Recrystallization described in the step 3) can carry out under heating, normal temperature or the state of cooling.Solvent used in the recrystallization of the present invention can be methyl alcohol, ethanol, acetone or their aqueous solution.
The method of said recrystallization is specific as follows: Syringin bullion or pedunculoside bullion are dissolved in the solvent; Place 50 ℃ of-70 ℃ of water-baths to make it reach supersaturated solution, leave standstill 5-60h in room temperature again, separate out crystallization; Filter; Drying can obtain highly purified Syringin and pedunculoside respectively, and purity is respectively more than 90% and 98%.
Column chromatography described in the step 3) can be selected from following at least a: open ODS column chromatography, Sephadex LH-20 column chromatography, preparative liquid chromatography.
Used eluent can be the methanol aqueous solution that volume(tric)fraction is 10%-70% in the said column chromatography.
In addition, the dry drying means of using always for this area described in the present invention as usually press dry dry, drying under reduced pressure, lyophilize.
The inventive method has following advantage:
1, the present invention can make full use of the Ovateleaf Holly Bark medicinal material, and its two big staple pedunculoside is carried out efficient and rational separating with Syringin.
2, this method is simple, and the cycle is short, can realize large-scale industrialization production.
3, the product yield is high, and purity is high.Wherein, the purity of Syringin is greater than 90%, and the purity of pedunculoside is greater than 98%.
Embodiment
Through specific embodiment method of the present invention is described below, but the present invention is not limited thereto.
Experimental technique described in the following embodiment like no specified otherwise, is ordinary method; Said reagent and material like no specified otherwise, all can obtain from commercial sources.
Used Chinese medicinal materials Ovateleaf Holly Bark meets the relevant regulations under a text Ovateleaf Holly Bark of Chinese Pharmacopoeia version in 2010 item among the following embodiment.Before feeding intake, be tested and appraised, medicinal material is in kind to conform to the quality conformance with standard with title.
Embodiment 1:
Get dry Ovateleaf Holly Bark 1Kg, adding 12 times of amounts (12L) volume(tric)fraction is 50% aqueous ethanolic solution refluxing extraction 1 time, and extraction time is 1h; Collect extracting solution, reclaim ethanol, be evaporated to 10L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated SP825 macroporous adsorbent resin (5L) for supernatant; Using 3 times of resin volumes, volume(tric)fractions earlier is 20% aqueous ethanolic solution wash-out, and using 4 times of resin volumes, volume(tric)fraction again is 50% aqueous ethanolic solution wash-out, detects (chromatographic condition: Agilent1200 high performance liquid chromatograph through the HPLC method; Shiseido UG120-C18 chromatographic column (4.6 * 150mm, 5 μ m), moving phase is acetonitrile and water; The gradient elution program is 0-10min, 10% acetonitrile; 10-20min, the 10%-40% acetonitrile; 20-30min, 40% acetonitrile; The detection wavelength is 210nm, and flow velocity is 1mL/min, and column temperature is 25 ℃), Syringin mainly concentrates on 20% ethanol elution part, and pedunculoside mainly concentrates on 50% ethanol elution part.
With 20% ethanol elution partially concd, drying gets the Syringin bullion.The Syringin bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 1.2g product.Be Syringin through detecting, purity is 90.2%.
With 50% ethanol elution partially concd, drying gets the pedunculoside bullion.The pedunculoside bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature again, place 48h, separate out crystallization, filter drying, 6.3g product.Be pedunculoside through detecting, purity is 98.5%.
Embodiment 2:
Get dry Ovateleaf Holly Bark 1Kg, adding 12 times of amounts (12L) volume(tric)fraction is 50% aqueous ethanolic solution refluxing extraction 2 times, extracts 1h at every turn; Merge No. 2 times extracting solution, reclaim ethanol, be evaporated to 12L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated SP825 macroporous adsorbent resin (5L) for supernatant; Using 2 times of resin volumes, volume(tric)fractions earlier is 10% aqueous ethanolic solution wash-out, and using 3 times of resin volumes, volume(tric)fraction again is 40% aqueous ethanolic solution wash-out, detects through the HPLC method; Syringin mainly concentrates on 10% ethanol elution part, and pedunculoside mainly concentrates on 40% ethanol elution part.
With 10% ethanol elution partially concd, drying gets the Syringin bullion.It is the dissolving of 10% methanol aqueous solution that bullion is added volume(tric)fraction, through the open ODS chromatographic column of having handled well, uses methanol-water system gradient elution then; Be that 10%, 20%, 30% methyl alcohol carries out wash-out with volume(tric)fraction respectively, 2 times of column volumes of each wash-out detect through the HPLC method; The stream part (volume(tric)fraction is the methanol-eluted fractions part of 10%-20%) that to contain Syringin merges; Reclaim solvent, drying under reduced pressure obtains the 1.1g product.Be Syringin through detecting, purity is 92.5%.
With 40% ethanol elution partially concd, drying gets the pedunculoside bullion.It is 40% methanol aqueous solution that bullion is added volume(tric)fraction, and through the open ODS chromatographic column of having handled well, using methanol-water system gradient elution then is that 40%, 50%, 60%, 70%, 95% methyl alcohol carries out wash-out with volume(tric)fraction respectively; 2 times of column volumes of each wash-out; Detect through the HPLC method, the stream part (volume(tric)fraction is the methanol-eluted fractions part of 50%-70%) that will contain pedunculoside merges, and reclaims solvent; Drying under reduced pressure obtains the 5.6g product.Be pedunculoside through detecting, purity is 98.6%.
Embodiment 3:
Get dry Ovateleaf Holly Bark 1Kg, adding 12 times of amounts (12L) volume(tric)fraction is 50% aqueous ethanolic solution refluxing extraction 3 times, extracts 1h at every turn; Merge No. 3 times extracting solution, reclaim ethanol, be evaporated to 8L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated HP20 macroporous adsorbent resin (5L) for supernatant; Using 4 times of resin volumes, volume(tric)fractions earlier is 30% aqueous ethanolic solution wash-out, and using 5 times of resin volumes, volume(tric)fraction again is 60% aqueous ethanolic solution wash-out, detects through the HPLC method; Syringin mainly concentrates on 30% ethanol elution part, and pedunculoside mainly concentrates on 60% ethanol elution part.
With 30% ethanol elution partially concd, drying gets the Syringin bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 1.2g product.Be Syringin through detecting, purity is 90.5%.
With 60% ethanol elution partially concd, drying gets the pedunculoside bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 6.1g product.Be pedunculoside through detecting, purity is 98.4%.
Embodiment 4:
Get dry Ovateleaf Holly Bark 1Kg, adding 8 times of amounts (8L) volume(tric)fraction is 60% aqueous ethanolic solution refluxing extraction 1 time, and extraction time is 90min; Collect extracting solution, reclaim ethanol, be evaporated to 6L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated D101 macroporous adsorbent resin (5L) for supernatant; Using 3 times of resin volumes, volume(tric)fractions earlier is 30% aqueous ethanolic solution wash-out, and 3 times of resin volumes, volume(tric)fractions are 70% aqueous ethanolic solution wash-out again, detect through the HPLC method; Syringin mainly concentrates on 30% ethanol elution part, and pedunculoside mainly concentrates on 70% ethanol elution part.
With 30% ethanol elution partially concd, drying gets the Syringin bullion.Bullion is added dissolve with methanol, adopt the Agilent1200 preparative liquid chromatograph, the preparation separation obtains sample.Chromatographic condition is Mightysil RP-18GP 250 * 20mm (5 μ m) chromatographic column, and moving phase is that (10: 90, v/v), flow velocity was 1mL/min to acetonitrile-water.Press peak manual collection effluent, the effluent heating in water bath volatilizes, and obtains the 1.2g product.Be Syringin through detecting, purity is 91.3%.
With 70% ethanol elution partially concd, drying gets the pedunculoside bullion.Bullion is added dissolve with methanol, adopt the Agilent1200 preparative liquid chromatograph, the preparation separation obtains sample.Chromatographic condition is Mightysil RP-18GP 250 * 20mm (5 μ m) chromatographic column, and moving phase is that (40: 60, v/v), flow velocity was 1mL/min to acetonitrile-water.Press peak manual collection effluent, the effluent heating in water bath volatilizes, and obtains the 6.5g product.Be pedunculoside through detecting, purity is 98.8%.
Embodiment 5:
Get dry Ovateleaf Holly Bark 1Kg, adding 16 times of amounts (16L) volume(tric)fraction is 70% aqueous ethanolic solution refluxing extraction 3 times, and extraction time is 30min; Collect extracting solution, reclaim ethanol, be evaporated to 8L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated HPD-100A macroporous adsorbent resin (5L) for supernatant; Using 3 times of resin volumes, volume(tric)fractions earlier is 20% aqueous ethanolic solution wash-out, and using 4 times of resin volumes, volume(tric)fraction again is 50% aqueous ethanolic solution wash-out, detects through the HPLC method; Syringin mainly concentrates on 20% ethanol elution part, and pedunculoside mainly concentrates on 50% ethanol elution part.
With 20% ethanol elution partially concd, drying gets the Syringin bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 1.1g product.Be Syringin through detecting, purity is 90.8%.
With 50% ethanol elution partially concd, drying gets the pedunculoside bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 6.0g product.Be pedunculoside through detecting, purity is 98.3%.
Embodiment 6:
Get dry Ovateleaf Holly Bark 1Kg, adding 10 times of amounts (10L) volume(tric)fraction is that 70% aqueous ethanolic solution cold soaking extracts 2 times, and extraction time is 60min; Collect cooling bath, reclaim ethanol, be evaporated to 6L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated SP825 macroporous adsorbent resin (5L) for supernatant; Using 3 times of resin volumes, volume(tric)fractions earlier is 30% aqueous ethanolic solution wash-out, and using 4 times of resin volumes, volume(tric)fraction again is 50% aqueous ethanolic solution wash-out, detects through the HPLC method; Syringin mainly concentrates on 30% ethanol elution part, and pedunculoside mainly concentrates on 50% ethanol elution part.
With 30% ethanol elution partially concd, drying gets the Syringin bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 1.0g product.Be Syringin through detecting, purity is 90.5%.
With 50% ethanol elution partially concd, drying gets the pedunculoside bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 5.8g product.Be pedunculoside through detecting, purity is 98.6%.
Embodiment 7:
Get dry Ovateleaf Holly Bark 1Kg, adding 10 times of amounts (10L) volume(tric)fraction is 70% aqueous ethanolic solution supersound extraction 2 times, and each time is 30min; Collect extracting solution, reclaim ethanol, be evaporated to 10L; Centrifugal, (ID10cm * H70cm) is behind the end of the sample through pretreated SP825 macroporous adsorbent resin (5L) for supernatant; Using 3 times of resin volumes, volume(tric)fractions earlier is 30% aqueous ethanolic solution wash-out, and using 4 times of resin volumes, volume(tric)fraction again is 50% aqueous ethanolic solution wash-out, detects through the HPLC method; Syringin mainly concentrates on 30% ethanol elution part, and pedunculoside mainly concentrates on 50% ethanol elution part.
With 30% ethanol elution partially concd, drying gets the Syringin bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 1.3g product.Be Syringin through detecting, purity is 91.4%.
With 50% ethanol elution partially concd, drying gets the pedunculoside bullion.Bullion is added dissolve with methanol, place 60 ℃ of water-baths to make it reach supersaturated solution, leave standstill in room temperature, place 48h, separate out crystallization, filter, drying obtains the 6.3g product.Be pedunculoside through detecting, purity is 98.5%.
Claims (10)
1. a method of extracting pedunculoside and Syringin simultaneously comprises the steps:
1) aqueous ethanolic solution with volume(tric)fraction 50%-70% is an extraction agent, and Ovateleaf Holly Bark is extracted, and collects extracting solution;
2) said extracting solution is carried out column chromatography for separation through macroporous adsorbent resin; With the alcohol-water is that eluent carries out gradient elution, and the collected volume mark is that the elution fraction and the volume(tric)fraction of 10%-30% aqueous ethanolic solution are the elution fraction of 40%-70% aqueous ethanolic solution respectively;
3) with said volume(tric)fraction be the elution fraction of 10%-30% aqueous ethanolic solution concentrate, dry, obtain the Syringin bullion, adopt recrystallization or column chromatography to carry out purifying said Syringin bullion, obtain the Syringin of purifying;
With said volume(tric)fraction be the elution fraction of 40%-70% aqueous ethanolic solution concentrate, dry, obtain the pedunculoside bullion, adopt recrystallization method or column chromatography to carry out purifying said pedunculoside bullion, obtain the pedunculoside of purifying.
2. method according to claim 1 is characterized in that: the method for extracting described in the step 1) is cold soaking extraction, supersound extraction or refluxing extraction.
3. method according to claim 2 is characterized in that: the method for said extraction is a refluxing extraction; Said refluxing extraction is carried out 1-3 time, and each time of extracting is 30 minutes-90 minutes, and when extracting, the consumption of extraction agent and the proportioning of Ovateleaf Holly Bark are (8-16) L: 1kg at every turn.
4. according to each described method among the claim 1-3, it is characterized in that: in step 2) the preceding extracting solution that step 1) is obtained that also comprises carries out spissated step; The concentration of gained liquid concentrator is 0.04-0.16g crude drug/mL.
5. according to each described method among the claim 1-4, it is characterized in that: step 2) described in macroporous adsorbent resin be selected from the macroporous adsorbent resin of following any one model: HP20, NKA-II, HPD-100A, SP825, DM301, D101, AB-8 and S-8.
6. according to each described method among the claim 1-5; It is characterized in that: step 2) described in the condition of gradient elution following: use volume(tric)fraction to carry out wash-out earlier, use volume(tric)fraction to carry out wash-out again as the 40%-70% aqueous ethanolic solution as the 10%-30% aqueous ethanolic solution; Each gradient elution 2-5 times column volume.
7. according to each described method among the claim 1-6, it is characterized in that: described in the step 3) in the recrystallization method used solvent be methyl alcohol, ethanol, acetone or their aqueous solution.
8. method according to claim 7; It is characterized in that: the method for recrystallization described in the step 3) is specific as follows: Syringin bullion or pedunculoside bullion are dissolved in the solvent, place 50 ℃ of-70 ℃ of water-baths to make it reach supersaturated solution, leave standstill 5-60h in room temperature again; Separate out crystallization; Filter, drying obtains the Syringin bullion or the pedunculoside of purifying.
9. according to each described method among the claim 1-6, it is characterized in that: column chromatography described in the step 3) is selected from following at least a: open ODS column chromatography, Sephadex LH-20 column chromatography, preparative liquid chromatography;
Used eluent is that volume(tric)fraction is the methanol aqueous solution of 10-70% in the said column chromatography.
10. according to each described method among the claim 1-9, it is characterized in that: the purity of the Syringin of said purifying is greater than 90%, and the purity of the pedunculoside of said purifying is greater than 98%.
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| CN103351419A (en) * | 2013-07-09 | 2013-10-16 | 陕西中药研究所 | Two-step simultaneous preparation method for pedunculoside and syringin |
| CN104558067A (en) * | 2014-12-30 | 2015-04-29 | 广州白云山汉方现代药业有限公司 | Method for extracting and separating syringin from medicinal material of ovateleaf holly bark |
| CN104764828A (en) * | 2015-04-20 | 2015-07-08 | 广州白云山星群(药业)股份有限公司 | Construction method of fingerprint of Ilex rotunda thunb medicinal material and detection method of Ilex rotunda thunb medicinal material |
| CN105646638A (en) * | 2016-01-20 | 2016-06-08 | 中山大学 | Preparation method of pedunculoside |
| CN106589016A (en) * | 2016-12-05 | 2017-04-26 | 贵州省烟草科学研究院 | Method for extracting syringin compound from tobaccos |
| CN108912186A (en) * | 2018-09-16 | 2018-11-30 | 湖州展舒生物科技有限公司 | The preparation method of Syringin |
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| CN103351419B (en) * | 2013-07-09 | 2015-10-07 | 陕西中药研究所 | Two-step approach prepares the method for pedunculoside and Syringin simultaneously |
| CN103351419A (en) * | 2013-07-09 | 2013-10-16 | 陕西中药研究所 | Two-step simultaneous preparation method for pedunculoside and syringin |
| CN104558067B (en) * | 2014-12-30 | 2017-09-29 | 广州白云山汉方现代药业有限公司 | A kind of method that separating syringin is extracted in the medicinal material from iron holly bark |
| CN104558067A (en) * | 2014-12-30 | 2015-04-29 | 广州白云山汉方现代药业有限公司 | Method for extracting and separating syringin from medicinal material of ovateleaf holly bark |
| CN104764828A (en) * | 2015-04-20 | 2015-07-08 | 广州白云山星群(药业)股份有限公司 | Construction method of fingerprint of Ilex rotunda thunb medicinal material and detection method of Ilex rotunda thunb medicinal material |
| CN104764828B (en) * | 2015-04-20 | 2017-01-11 | 广州白云山星群(药业)股份有限公司 | Construction method of fingerprint of Ilex rotunda thunb medicinal material and detection method of Ilex rotunda thunb medicinal material |
| CN105646638A (en) * | 2016-01-20 | 2016-06-08 | 中山大学 | Preparation method of pedunculoside |
| CN106589016A (en) * | 2016-12-05 | 2017-04-26 | 贵州省烟草科学研究院 | Method for extracting syringin compound from tobaccos |
| CN108912186A (en) * | 2018-09-16 | 2018-11-30 | 湖州展舒生物科技有限公司 | The preparation method of Syringin |
| CN109970838A (en) * | 2019-03-19 | 2019-07-05 | 中山大学 | A kind of preparation method of pedunculoside |
| CN109970838B (en) * | 2019-03-19 | 2021-06-15 | 中山大学 | Preparation method of pedunculoside |
| CN114644675A (en) * | 2020-12-21 | 2022-06-21 | 广西大学 | Method for separating compass herb glycoside B from ovate leaf holly bark |
| CN114644677A (en) * | 2020-12-21 | 2022-06-21 | 广西大学 | Novel compound separated from ovate leaf holly bark and separation method thereof |
| CN114644677B (en) * | 2020-12-21 | 2023-05-12 | 广西大学 | New compound separated from cortex Ilicis Rotundae and separation method thereof |
| CN114644675B (en) * | 2020-12-21 | 2023-05-16 | 广西大学 | Method for separating Luo Pancao glycoside B from holly bark |
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