CN102603832A - Production method of spinosin - Google Patents
Production method of spinosin Download PDFInfo
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- CN102603832A CN102603832A CN2011100262068A CN201110026206A CN102603832A CN 102603832 A CN102603832 A CN 102603832A CN 2011100262068 A CN2011100262068 A CN 2011100262068A CN 201110026206 A CN201110026206 A CN 201110026206A CN 102603832 A CN102603832 A CN 102603832A
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Abstract
The invention discloses a production method of spinosin, which comprises the following steps: breaking wild jujube seeds, extracting by hot water and extracting to obtain extracted liquid; making the extracted liquid pass through a macroporous resin column for chromatography; collecting the extracted liquid which enriches the target element; adding 4-6% diatomite for decolorization; concentrating the decolorized liquid to obtain crude extract; making the crude extract pass through a polyamide resin column for chromatography; eluting by low-carbon alcohol or low-carbon ketone solution with different concentrations to elute in turns; collecting the eluted solution, concentrating to obtain crude spinosin crystals; and performing reflux and dissolution to the crude spinosin crystals by acetone, cooling and crystallizing, filtering and drying to obtain the finished product. The invented production method uses the hot water to replace organic solvent to perform the extraction, uses the macroporous resin column to replace a silica gel column to perform the chromatography, and uses the low-carbon alcohol and the low-carbon ketone to replace poisonous solvent chloroform; the cost is reduced, the production safety is good, and the product purity is high.
Description
Technical field:
The invention belongs to the Chemistry for Chinese Traditional Medicine field, be specifically related to the plain working method of a kind of Si Pinuo.
Background technology:
Spina Date Seed is called white sour jujube, sour jujube pin, jujube kernel, jujube nucleus, is the dry mature seed of Rhamnaceae plant wild jujube Ziziphus jujubeMill.var.spinosa (Bunge) Hu ex H.F.Chou.Sweet, sour, flat, return liver, courage, the heart channel of Hang-Shaoyin.The effect of have the tonifying liver of nourishing heart, tranquillizing by calming the heart, arrest sweating, promoting the production of body fluid, it is thirsty to be used for restlessness of asrhenia type and insomnia, palpitation with fear dreaminess, the empty hidrosis of body, Tianjin wound.Spina Date Seed beginning is stated from Shennong's Herbal, main product in Hebei, Shaanxi, Liaoning, Henan.Its chemical ingredients mainly contains lipid acid, triterpenoid saponin, flavones, steroidal and alkaloid compound.Wherein flavonoid compound has Si Pinuo element, wild jujube flavine, 6 ' "-asafoetide acyl Si Pinuo element, 6 ' "-the different Si Pinuo element of asafoetide acyl, 6 ' "-to coumaric acyl Si Pinuo element, swertisin, puerarin etc.Pharmacological research shows that Spina Date Seed has tranquilizing soporific, anticonvulsion, analgesia and body-temp. reducing, anxiety, anti-heart disorder, Posterior Pituitary is caused that the rat heart muscle ischemic has provide protection, step-down and expands blood vessel, reducing blood-fat and prevent and treat the arteriosclerosis effect; Also have lipoid peroxidization resistant, decompression anoxia endurance is had provide protection.
Si Pinuo plain (spinosin) is one of main flavonoid compound in the Spina Date Seed, has another name called sour jujube glycosides, wild jujube element, is yellow
Si Pinuo plain (spinosin)
Powder, molecular formula C
28H
32O
15, molecular weight 608.54 is dissolved in methyl alcohol.Pharmacopeia in 2010 first with it as one of index property composition of Spina Date Seed, followingly will increase the plain demand of Si Pinuo.
Do not see at present the patent report that the plain preparation of high purity Si Pinuo is arranged as yet.Wang Hui etc. are through multiple chromatogram separation means such as silica gel column chromatography, MDS column chromatography, Sephadex LH-20, open ODS column chromatography, preparation HPLC chromatograms; From the Spina Date Seed methanol extract, separate obtaining, this method is only applicable to laboratory chemical ingredients isolation identification.Li Huijun, Li Ping adopt hplc simultaneous determination Si Pinuo element and 6 ' "-the plain content of asafoetide acyl Si Pinuo; extract through petroleum ether degreasing, methanol eddy; silicagel column separates, and the chloroform-methanol wash-out prepares need testing solution, and this method is used harmful reagent such as a large amount of chloroforms, methyl alcohol; preparation amount is little, is unwell to industrial production.
Therefore, set up simple to operate, environmental pollution is less, production is comparatively safe, can prepare the plain production technique of highly purified Si Pinuo in a large number has crucial social effect and economic worth.
Summary of the invention:
In order to solve the deficiency of prior art, the present invention aims to provide a kind of low cost, high safety, the plain method of production Si Pinuo that purity is high.
The objective of the invention is to realize through following proposal:
The working method that a kind of Si Pinuo is plain is characterized in that may further comprise the steps:
(1) extract: Spina Date Seed is broken, use hot water extraction, merging obtains extracting solution;
(2) resin column chromatography: with macroporous resin column chromatography on the extracting solution, use the cold water wash-out earlier, use the 50-95% ethanol elution again, collect the 50-95% ethanol eluate;
(3) decolour: above-mentioned elutriant is added the decolouring of 4-6% zeyssatite, filter, concentrate, get liquid concentrator;
(4) polyamide column chromatography: with polyamide column chromatography on the liquid concentrator,, use 75-90% low-carbon alcohol or low-carbon (LC) ketone wash-out again, collect the elutriant of 75-90% part, concentrated coarse-grain earlier with 20-40% low-carbon alcohol or low-carbon (LC) ketone wash-out;
(5) recrystallization: with acetone backflow dissolving, cooling crystallization filters with coarse-grain, and drying obtains highly purified finished product.
The said hot water temperature of step (1) is 60-100 ℃, and solid-liquid ratio is 1: 5-10 (L/kg), process for extracting are decocting method, extracting at constant temperature or ultrasonic extraction, and extraction time 0.5-3 hour, extraction time was 2-3 time.
The described macroporous resin of step (2) is a kind of in X-5 type macroporous resin, AB-8 type macroporous resin, the S-8 type macroporous resin.
The said cold water elution volume of step (2) consumption is a 1-2 times of column volume: said 50-95% ethanol elution volumetric usage is a 2-5 times of column volume.
The said polymeric amide of step (4) is the 50-100 order, and said eluent low-carbon alcohol is an ethanolic soln, and said low-carbon (LC) ketone is acetone soln, and the eluent consumption is a 2-5 times of column volume.
The invention has the beneficial effects as follows: adopt hot water extraction in the leaching process; Replaced original organic solvent extraction; Extraction yield is high, and Product Safety is high: adopt macroporous resin column chromatography and polyamide column chromatography lower, with short production cycle than existing silica gel column chromatography cost; Eluent has been avoided harmful reagent such as use chloroform, the recyclable utilization of solvent for use; Present method preparation amount is big, but suitability for industrialized production.
Embodiment:
To combine specific embodiment that the present invention is further described below, but embodiment of the present invention is not limited thereto.
HPLC method (document of delivering with reference to Xie Junbo etc. " HPLC method measure in the Spina Date Seed flavones sheet Si Pinuo plain ") is adopted in the plain detection of Si Pinuo among the following embodiment, and the method that the document adopts is:
Chromatographic condition: ODS C18 post (150mm * 3.9mm, 9 μ m);
Moving phase: acetonitrile-water (0.1% acetic acid), gradient elution, 0-5min, 10% acetonitrile, 5-20min, 17% acetonitrile, 20-30min, 23% acetonitrile, after the 30min, 35% acetonitrile;
Volumetric flow rate: 0.8ml/min:
Column temperature: 25 ℃;
Sample size: 20 μ l;
Detect wavelength: 335nm.
Embodiment 1:
Spina Date Seed is broken into meal, gets 1kg and place the 10L extractor, add 6L80-100 ℃ of hot water and decoct extraction, extract 2 times; Each 2 hours, filter, merge extracted twice liquid, with S-8 macroporous adsorptive resins on the extracting solution; With 3000ml cold water wash-out, use 60% ethanol elution of 6000ml then earlier, collect 60% ethanol eluate; Add 200g zeyssatite and stir decolouring, filter, destainer concentrate liquid concentrator; With polyamide column on the liquid concentrator, use the 1000ml30% ethanol elution earlier, use the 1250ml75% ethanol elution again; Collect 75% ethanol eluate, decompression recycling ethanol concentrates and obtains the plain coarse-grain of Si Pinuo; Coarse-grain with the dissolving of acetone reflux, is cooled to room temperature, leaves standstill crystallization; Leach crystal, vacuum-drying obtains the plain finished product 853mg of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.67%.
Embodiment 2:
Spina Date Seed is broken into meal, gets 1kg and place the 10L extractor, add 20L60-80 ℃ of hot water constant temperature soaking and extracting, extract 3 times; Each 2 hours, filter, merge No. three times extracting solution, with AB-8 macroporous adsorptive resins on the extracting solution; With 4L cold water wash-out, use 80% ethanol elution of 12L then earlier, collect 80% ethanol eluate; Add 500g zeyssatite and stir decolouring, filter, destainer concentrate liquid concentrator; With polyamide column on the liquid concentrator,, use 1500ml80% acetone wash-out more earlier with 1200ml20% acetone wash-out; Collect 80% acetone elutriant, reclaim under reduced pressure acetone concentrates and obtains the plain coarse-grain of Si Pinuo; Coarse-grain with the dissolving of acetone reflux, is cooled to room temperature, leaves standstill crystallization; Leach crystal, vacuum-drying obtains the plain finished product 827mg of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.28%.
Embodiment 3:
Spina Date Seed is broken into meal, gets 2kg and place the 20L extractor, add 16L70-90 ℃ of hot water supersound extraction, extract 3 times; Each 0.5 hour, filter, merge No. three times extracting solution, with X-3 macroporous adsorptive resins on the extracting solution; With 10L cold water wash-out, use 75% ethanol elution of 15L then earlier, collect 75% ethanol eluate, add 750g zeyssatite and stir decolouring; Filter, destainer concentrate liquid concentrator: with polyamide column on the liquid concentrator, use the 2L20% ethanol elution earlier, use the 3L90% ethanol elution again; Collect 90% ethanol eluate, decompression recycling ethanol concentrates and obtains the plain coarse-grain of Si Pinuo, and coarse-grain is dissolved with the acetone reflux; Be cooled to room temperature, leave standstill crystallization, leach crystal, vacuum-drying obtains the plain finished product 1.67g of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.44%.
Embodiment 4:
Spina Date Seed is broken into meal, gets 5kg and place the 50L extractor, add 35L80-90 ℃ of hot water constant temperature soaking and extracting, extract 2 times; Each 3 hours, filter, merge extracted twice liquid, with S-8 macroporous adsorptive resins on the extracting solution; With 45L cold water wash-out, use 95% ethanol elution of 60L then earlier, collect 95% ethanol eluate; Add and to stir decolouring on the 3.3kg diatom, filter, destainer concentrate liquid concentrator; With polyamide column on the liquid concentrator, use the 5L20% ethanol elution earlier, use the 6.5L55% ethanol elution again; Collect 55% ethanol eluate, decompression recycling ethanol concentrates and obtains the plain coarse-grain of Si Pinuo; Coarse-grain with the dissolving of acetone reflux, is cooled to room temperature, leaves standstill crystallization; Leach crystal, vacuum-drying obtains the plain finished product 4.1g of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.06%.
Embodiment 5:
Spina Date Seed is broken into meal, gets 10kg and place the 100L extractor, add 50L65-85 ℃ of hot water supersound extraction, extract 3 times; Each 1 hour, filter, merge No. three times extracting solution, with AB-8 macroporous adsorptive resins on the extracting solution; With 30L cold water wash-out, use 50% ethanol elution of 45L then earlier, collect 50% ethanol eluate, add 2.2kg zeyssatite and stir decolouring; Filter, destainer concentrate liquid concentrator: with polyamide column on the liquid concentrator, use the 8L25% ethanol elution earlier, use the 12L85% ethanol elution again; Collect 85% ethanol eluate, decompression recycling ethanol concentrates and obtains the plain coarse-grain of Si Pinuo, and coarse-grain is dissolved with the acetone reflux; Be cooled to room temperature, leave standstill crystallization, leach crystal, vacuum-drying obtains the plain finished product 8.3g of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.67%.
Embodiment 6:
Spina Date Seed is broken into meal, gets 25kg and place the 200L extractor, add 150L70-85 ℃ of hot water and decoct extraction, extract 2 times; Each 3 hours, filter, merge extracted twice liquid, with X-5 macroporous adsorptive resins on the extracting solution; With 60L cold water wash-out, use 60% ethanol elution of 80L then earlier, collect 60% ethanol eluate; Add and to stir decolouring on the 3.2kg diatom, filter, destainer concentrate liquid concentrator; With polyamide column on the liquid concentrator, use the 20L30% ethanol elution earlier, use the 30L75% ethanol elution again; Collect 75% ethanol eluate, decompression recycling ethanol concentrates and obtains the plain coarse-grain of Si Pinuo; Coarse-grain with the dissolving of acetone reflux, is cooled to room temperature, leaves standstill crystallization; Leach crystal, vacuum-drying obtains the plain finished product 20.6g of high purity Si Pinuo.Measuring the plain finished product purity of Si Pinuo through the HPLC method is 98.71%.
Claims (5)
1. the plain working method of Yi Zhong Si Pinuo is characterized in that may further comprise the steps:
(1) extract: Spina Date Seed is broken, use hot water extraction, merging obtains extracting solution;
(2) resin column chromatography: with macroporous resin column chromatography on the extracting solution, use the cold water wash-out earlier, use the 50-95% ethanol elution again, collect the 50-95% ethanol eluate:
(3) decolour: above-mentioned elutriant is added the decolouring of 4-6% zeyssatite, filter, concentrate, get liquid concentrator;
(4) polyamide column chromatography: with polyamide column chromatography on the liquid concentrator,, use 75-90% low-carbon alcohol or low-carbon (LC) ketone wash-out again, collect the elutriant of 75-90% part, concentrated coarse-grain earlier with 20-40% low-carbon alcohol or low-carbon (LC) ketone wash-out;
(5) recrystallization: with acetone backflow dissolving, cooling crystallization filters with coarse-grain, and drying obtains highly purified finished product.
2. the plain working method of a kind of Si Pinuo as claimed in claim 1; It is characterized in that: the said hot water temperature of step (1) is 60-100 ℃; Solid-liquid ratio is 1: 5-10 (L/kg); Process for extracting is decocting method, extracting at constant temperature or ultrasonic extraction, and extraction time 0.5-3 hour, extraction time was 2-3 time.
3. the working method that a kind of Si Pinuo as claimed in claim 1 is plain is characterized in that: the described macroporous resin of step (2) is a kind of in X-5 type macroporous resin, AB-8 type macroporous resin, the S-8 type macroporous resin.
4. the plain working method of a kind of Si Pinuo as claimed in claim 1, it is characterized in that: the said cold water elution volume of step (2) consumption is a 1-2 times of column volume; Said 50-95% ethanol elution volumetric usage is a 2-5 times of column volume.
5. the plain working method of a kind of Si Pinuo as claimed in claim 1, it is characterized in that: the said polymeric amide of step (4) is the 50-100 order, and said eluent low-carbon alcohol is an ethanolic soln, and said low-carbon (LC) ketone is acetone soln, the eluent consumption is a 2-5 times of column volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100262068A CN102603832A (en) | 2011-01-25 | 2011-01-25 | Production method of spinosin |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011100262068A CN102603832A (en) | 2011-01-25 | 2011-01-25 | Production method of spinosin |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103877110A (en) * | 2014-03-25 | 2014-06-25 | 郭贤卿 | Pharmaceutical composition for preventing and treating viral hepatitis B and application of pharmaceutical composition |
| CN106473124A (en) * | 2015-08-31 | 2017-03-08 | 广州棒棒聪生物科技有限公司 | A kind of compositionss promoting infant intelligent development |
| CN109879919A (en) * | 2019-02-20 | 2019-06-14 | 滨州医学院 | A method of three kinds of flavonoid glycosides of separation preparation from semen ziziphi spinosae |
| CN110229205A (en) * | 2019-07-09 | 2019-09-13 | 山西大学 | The preparation method and application of semen ziziphi spinosae monomer component spinosin sodium |
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| CN1401643A (en) * | 2002-09-13 | 2003-03-12 | 中国药科大学 | Process for preparing total jujuboflavone with calmative and hypnosis action |
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2011
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Patent Citations (2)
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| WO2002068644A1 (en) * | 2001-02-24 | 2002-09-06 | Scigen Harvest Co., Ltd | Promoter of s-adenosyl-l-methionine: jasmonic acid carboxyl methyl transferase gene and method for producing target protein in plants using the same |
| CN1401643A (en) * | 2002-09-13 | 2003-03-12 | 中国药科大学 | Process for preparing total jujuboflavone with calmative and hypnosis action |
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| 《首都师范大学硕士学位论文》 20050815 姚煜 "酸枣仁提取工艺及成份研究" 第11-24页 1-5 , * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103877110A (en) * | 2014-03-25 | 2014-06-25 | 郭贤卿 | Pharmaceutical composition for preventing and treating viral hepatitis B and application of pharmaceutical composition |
| CN103877110B (en) * | 2014-03-25 | 2016-03-23 | 郭贤卿 | A kind of pharmaceutical composition and application thereof preventing and treating hepatitis B |
| CN106473124A (en) * | 2015-08-31 | 2017-03-08 | 广州棒棒聪生物科技有限公司 | A kind of compositionss promoting infant intelligent development |
| CN109879919A (en) * | 2019-02-20 | 2019-06-14 | 滨州医学院 | A method of three kinds of flavonoid glycosides of separation preparation from semen ziziphi spinosae |
| CN109879919B (en) * | 2019-02-20 | 2022-03-15 | 滨州医学院 | Method for separating and preparing three flavonoid glycosides from spina date seeds |
| CN110229205A (en) * | 2019-07-09 | 2019-09-13 | 山西大学 | The preparation method and application of semen ziziphi spinosae monomer component spinosin sodium |
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Application publication date: 20120725 |