CN103059037A - Method for enriching and purifying chelidonine in celandine - Google Patents
Method for enriching and purifying chelidonine in celandine Download PDFInfo
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- CN103059037A CN103059037A CN201210595639XA CN201210595639A CN103059037A CN 103059037 A CN103059037 A CN 103059037A CN 201210595639X A CN201210595639X A CN 201210595639XA CN 201210595639 A CN201210595639 A CN 201210595639A CN 103059037 A CN103059037 A CN 103059037A
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- stylophorine
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Abstract
The invention belongs to the field of natural organic chemistry, and relates to a method for enriching and purifying chelidonine in celandine by using the ultrasonic extraction-macroporous adsorption resin coupling method. The method has the following advantages: as the ultrasonic extraction is adopted, the extraction effect is better than that of the ethanol hot-dipping extraction; the macroporous adsorption resin is combined, so that the adsorption selectivity to the chelidonine is good, the adsorption is fast, the analysis is fast, and the adsorption capacity is higher; the extraction is convenient and quick; the production cost is low; the separation effect is remarkable; the chelidonine semi-finished product with content above 30% and the chelidonine finished product with content above 90% can be obtained; and the defects of lower conventional extraction rate and low extraction purity are overcome. The method chooses the macroporous adsorption resin with stable physicochemical property, larger surface area, higher exchange speed, high mechanical strength, strong pollution resistance and good heat stability, can selectively adsorb the chelidonine from the solution through the physical adsorption, and has the advantages of quick adsorption, quick analysis and larger adsorption capacity.
Description
Technical field:
The invention belongs to the natural organic chemistry field, relate to a kind of purification process of Stylophorine, particularly relate to a kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort.
Background technology:
Pilewort: [English name] Greater Celandine Herb, Herb of Greater Celandine[another name] Herba Munroniae henryi, Niu Jinhua, Root or stem of Wintergreen Barberry, eight steps are tight, Graceful Jessamine Herb, Shanxi melon, realgar grass, the mountain coptis, the false coptis, little Herba Stylophori Lasiocapi is careless, yellow stuff is sub, Rhizoma Picrorhizae, the little coptis.Medicinal material Ji Yuan: be the herb of bloodroot pilewort.Perennial herb, high 30-100cm contains safran milk.Main root is sturdy, taper shape, khaki color or dun, close living fibrous root.Stem is upright, and multi-branched has white powder, the elongated pubescence of tool white.Chemical Composition: Fresh Plants has dense orange-yellow emulsion, contains multiple alkaloid alkaloid 0.7% or 0.97～1.87% in the emulsion.Stylophorine 41%, protopine 22%, talatisamine 17%, allocryptopine 9%, Berberine 5%, white chelerythrine 3%, sanguinarine(e) 1.5%, sparteine 0.1% are wherein arranged, also having hydroxychelidonine is Oxychelidonine, methoxychelidonine, cryptopine, and chelilutine, chelidamine, homochelidonine, hydroxysanguinarine are Oxysanguinarine.Except alkaloid and, also contain jervasic acid, oxysuccinic acid, citric acid, succsinic acid, choline, methylamine, histamine, tyrasamine, saponin, flavonol, celidoniol, also contain cardiotonic glycoside, the highest at the content in flowering period.Root contains alkaloid and reaches 1.33% or 1.90～4.14%, contained identical of a part of alkaloid and over-ground part, and other contains coptisine, corysamine, chelirubine, chelidimerine, spinasterol, a small amount of ergosterol and rubber 0.118%.Leaf contains flavonoid 1.43%, volume vitamins C.Ascorbic content is the highest in flowering period, can reach 834 milligrams of %, and content is minimum when fruit maturation, is 231 milligrams of %.
Stylophorine is a kind of human health to be had the natural active matter of remarkable health-care effect, has caused the common concern of international community.Prove that through scientific research and clinical application Stylophorine is used for stomach and intestine angina, gastric and duodenal ulcer, renal colic, dysmenorrhoea and ascariasis of biliary tract pain relieving: be used local anaesthetics and can be used for women's sterilization operation; Compound Stylophorine analgesic activity is better, can be used for rheumatoid arthritis, injures cancer pain outward.In recent years, it is found that and contain a large amount of Stylophorines in the natural phant pilewort, because its raw material pilewort is distributed more widely, therefrom extracts Stylophorine and obtained encouraging progress.Name is called in the patent application of " a kind of Herba Chelidonii extract and preparation method thereof and use " and just discloses pilewort through ethanol, the technical process of chloroform extraction Herba Chelidonii extract.Herba Chelidonii extract extraction process in the above-mentioned patent documentation mainly is to extract through steps such as alcohol extracting, distillation, adjusting pH value, chloroform extraction techniques to form, and its separating effect is nothing like ultrasonic extraction-macroporous adsorbent resin coupled method and processes rear effect carrying out the silica gel column chromatography separation.
Summary of the invention:
The object of the invention is to overcome the shortcomings such as the routine techniques extraction yield is relatively low, dna purity is low, a kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort is provided.
For achieving the above object, the technical solution used in the present invention is:
A kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort, its step is as follows:
(1) dried pilewort being pulverized, is that 65%～80% ethanolic soln carries out ultrasonic extraction with concentration, and extracting temperature is 40～60 ℃, and extraction frequency 60～100Hz, extracts 3 times at 1～2 hour extraction time;
(2) the pilewort extracting liquid filtering of (1) being processed merges No. three times extracting solution, concentrating under reduced pressure, and Recycled ethanol is to relative density;
(3) regulate pH value to 3.4～3.5 with the acid of 1mol/L, suction filtration behind 5 ℃ of refrigeration 48h; Filtrate is regulated pH value to 11 with 30% sodium hydroxide, use and the chloroform extraction of filtrate equivalent 4 times, combined chloroform liquid, the recovery chloroform gets the Stylophorine crude extract;
(4) gained Stylophorine crude extract is adsorbed with macroporous adsorbent resin;
(5) ethanol with 50%～80% different concns carries out wash-out to macroporous adsorbent resin, follows the tracks of with tlc and detects, and collects each stepwise elution liquid of Stylophorine;
(6) with each stepwise elution liquid concentration and recovery ethanol of Stylophorine, get the Stylophorine work in-process, its content is more than 30%;
(7) with silica gel column chromatography on the gained Stylophorine work in-process, and carry out wash-out with chloroform with 5: 1 volume ratio, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of Stylophorine;
(8) with collected Stylophorine elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care to get Stylophorine, its content is more than 90%.
In the described step (4), used macroporous adsorbent resin is a kind of in Tianjin Nankai university X-5, the NKA-9, S-8, D 3520, D4006, H103, D4020, AB-8H and the NKA-II type resin that produce, is adsorbed as Static Adsorption or dynamic adsorption.
Using ultrasound ripple extraction-macroporous adsorbent resin coupled method separation and purification Stylophorine of the present invention, good to the adsorption selectivity of Stylophorine, the fast parsing of absorption is also fast, and loading capacity is larger; Extract convenient and swiftly, raw material sources are abundant, low production cost, separating effect is obvious, dna purity is high, can obtain Stylophorine work in-process and content the Stylophorine finished product 90% or more of content more than 30%, has overcome the shortcoming that conventional extraction yield is relatively low, dna purity is low.
Embodiment:
Case study on implementation 1: a kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort, its step is as follows:
(1) the dried pilewort of 500g is pulverized rear 60 mesh sieves of crossing, extract with ultrasonic extraction, its concentration is 65% ethanolic soln, and solid-liquid ratio is 8 times of the pulverizing volume, and extracting temperature is 40 ℃, and extraction frequency 60Hz, extracts 3 times at 1 hour extraction time;
(2) the pilewort extracting solution of (1) being processed merges, and decompression recycling ethanol is concentrated into relative density 1.08;
(3) with 1mol/L phosphoric acid regulating ph value to 3.4; Suction filtration behind 5 ℃ of refrigeration 48h.Filtrate is regulated pH value to 11 with 30% sodium hydroxide, use and the chloroform extraction of filtrate equivalent 4 times, combined chloroform liquid, the recovery chloroform gets the Stylophorine crude extract;
(4) the gained Stylophorine is carried thing with adsorbing on the macroporous adsorption resin chromatography post, used macroporous resin is the NKA-II type resin that Tianjin Nankai university produces;
(5) in macroporous adsorbent resin layer post, carry out successively gradient elution with 50%, 60%, 70%, 80% ethanol, follow the tracks of with tlc and detect, collect each stepwise elution liquid of Stylophorine;
(6) with each stepwise elution liquid concentration and recovery ethanol of Stylophorine, obtain content and be 30% Stylophorine work in-process;
(7) carry out gradient elution with silica gel column chromatography on the gained Stylophorine work in-process, and with chloroform with 5: 1 volume ratio, tlc is followed the tracks of and is detected, and collects the Stylophorine elutriant;
(8) with collected Stylophorine elutriant, adopt activated carbon decolorizing, refining content 902% Stylophorine that obtains of recrystallization method.
Case study on implementation 2: a kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort, its step is as follows:
(1) the dried pilewort of 500g is pulverized rear 70 mesh sieves of crossing, extract with ultrasonic extraction, its concentration is 70% ethanolic soln, and solid-liquid ratio is 9 times of the pulverizing volume, and extracting temperature is 50 ℃, and extraction frequency 80Hz, extracts 3 times at 1.5 hours extraction times;
(2) the pilewort extracting solution of (1) being processed merges, and decompression recycling ethanol is concentrated into relative density 1.10;
(3) with 1mol/L phosphoric acid regulating ph value to 3.4, suction filtration behind 5 ℃ of refrigeration 48h; Filtrate is regulated pH value to 11 with 30% sodium hydroxide, use and the chloroform extraction of filtrate equivalent 4 times, combined chloroform liquid, the recovery chloroform gets the Stylophorine crude extract;
(4) gained Stylophorine crude extract is adsorbed with macroporous adsorbent resin, used macroporous resin is the D4006 type resin that Tianjin Nankai university produces;
(5) carry out wash-out with 50%, 60%, 70%, 80% ethanol at macroporous adsorbent resin, follow the tracks of with tlc and detect, collect each stepwise elution liquid of Stylophorine;
(6) with each stepwise elution liquid concentration and recovery ethanol of Stylophorine, obtain the Stylophorine work in-process of content 33%;
(7) carry out gradient elution with silica gel column chromatography on the gained Stylophorine work in-process, and with chloroform with 5: 1 volume ratio, tlc is followed the tracks of and is detected, and collects the Stylophorine elutriant;
(8) with collected Stylophorine elutriant, adopt activated carbon decolorizing, refining content 91.7% Stylophorine that obtains of recrystallization method.
Case study on implementation 3: a kind of method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort, its step is as follows:
(1) the dried pilewort of 500g is pulverized rear 80 mesh sieves of crossing, extract with ultrasonic extraction, its concentration is 75% ethanolic soln, and solid-liquid ratio is 10 times of the pulverizing volume, and extracting temperature is 60 ℃, and extraction frequency 100Hz, extracts 3 times at 2 hours extraction times;
(2) the pilewort extracting solution of (1) being processed merges, and decompression recycling ethanol is concentrated into relative density 1.12;
(3) with 1mol/L phosphoric acid regulating ph value to 3.5, suction filtration behind 5 ℃ of refrigeration 48h; Filtrate is regulated pH value to 11 with 30% sodium hydroxide, use and the chloroform extraction of filtrate equivalent 4 times, combined chloroform liquid, the recovery chloroform gets the Stylophorine crude extract;
(4) with gained Stylophorine crude extract with the macroporous adsorption resin chromatography pillar, on adsorb, used macroporous resin is the NKA-9 type resin that Tianjin Nankai university produces;
(5) in the macroporous adsorption resin chromatography pillar, carry out successively gradient elution with 50%, 60%, 70%, 80% ethanol, follow the tracks of with tlc and detect, collect each stepwise elution liquid of Stylophorine;
(6) with each stepwise elution liquid concentration and recovery ethanol of Stylophorine, obtain content 32% Stylophorine work in-process;
(7) carry out gradient elution with silica gel column chromatography on the gained Stylophorine work in-process, and with chloroform with 5: 1 volume ratio, tlc is followed the tracks of and is detected, and collects the Stylophorine elutriant;
(8) with collected Stylophorine elutriant, adopt activated carbon decolorizing, refining content 923% Stylophorine that obtains of recrystallization method.
Claims (5)
1. method of utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort, its step is as follows:
(1) dried pilewort being pulverized, is that 65%～80% ethanolic soln carries out ultrasonic extraction with concentration, and extracting temperature is 40～60 ℃, and extraction frequency 60～100Hz, extracts 3 times at 1～2 hour extraction time;
(2) the pilewort extracting liquid filtering of (1) being processed merges No. three times extracting solution, concentrating under reduced pressure, and Recycled ethanol is to relative density;
(3) regulate pH value to 3.4～3.5 with the acid of 1mol/L, suction filtration behind 5 ℃ of refrigeration 48h; Filtrate is regulated pH value to 11 with 30% sodium hydroxide, use and the chloroform extraction of filtrate equivalent 4 times, combined chloroform liquid, the recovery chloroform gets the Stylophorine crude extract;
(4) gained Stylophorine crude extract is adsorbed with macroporous adsorbent resin;
(5) ethanol with 50%～80% different concns carries out wash-out to macroporous adsorbent resin, follows the tracks of with tlc and detects, and collects each stepwise elution liquid of Stylophorine;
(6) with each stepwise elution liquid concentration and recovery ethanol of Stylophorine, get the Stylophorine work in-process, its content is more than 30%:
(7) with silica gel column chromatography on the gained Stylophorine work in-process, and carry out wash-out with chloroform with 5: 1 volume ratio, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of Stylophorine;
(8) with collected Stylophorine elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care to get Stylophorine, its content is more than 90%.
2. the method for utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort according to claim 1 is characterized in that: in the described step (1), cross 60～80 mesh sieves after pilewort is pulverized.
3. the method for utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort according to claim 2 is characterized in that: in the described step (1), the volume that adds ethanol is for pulverizing 8～10 times of volume.
4. the method for utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort according to claim 3, it is characterized in that: in the described step (2), the relative density of concentrated solution is controlled between 1.02～1.12.
5. the method for utilizing Stylophorine in ultrasonic extraction-macroporous adsorbent resin coupled method enriching and purifying pilewort according to claim 4 is characterized in that: in the described step (3), the acid of regulating the used 1mol/L of pH value is phosphoric acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201210595639.XA CN103059037B (en) | 2012-12-13 | 2012-12-13 | A kind of method of Chelidonine in enriching and purifying greater celandine |
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| CN201210595639.XA CN103059037B (en) | 2012-12-13 | 2012-12-13 | A kind of method of Chelidonine in enriching and purifying greater celandine |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106138054A (en) * | 2015-04-28 | 2016-11-23 | 沈阳伟嘉牧业技术有限公司 | Chelerythrine application in treatment fowl drug resistance colibacillosis |
| CN109999014A (en) * | 2019-03-28 | 2019-07-12 | 佛山市正典生物技术有限公司 | It is a kind of can the new merit chigger disease of prevention and control plants essential oil and preparation method thereof |
| CN111297950A (en) * | 2018-12-11 | 2020-06-19 | 泰州医药城国科化物生物医药科技有限公司 | Preparation method and application of chelidonine |
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| DE2755577A1 (en) * | 1977-12-09 | 1979-06-13 | Steiner & Co Arznei | Isolation of spasmolytic, antibacterial and antimycotic alkaloid(s) - from e.g. Cheledonius root by moistening with ammonia soln., extracting with chloroform and working up extract |
| CN1429611A (en) * | 2002-09-13 | 2003-07-16 | 张平 | Greater celandine extract and its preparation method and application |
| CN101955485A (en) * | 2010-05-26 | 2011-01-26 | 南京泽朗医药科技有限公司 | Preparation method of chelidonine |
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2012
- 2012-12-13 CN CN201210595639.XA patent/CN103059037B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2755577A1 (en) * | 1977-12-09 | 1979-06-13 | Steiner & Co Arznei | Isolation of spasmolytic, antibacterial and antimycotic alkaloid(s) - from e.g. Cheledonius root by moistening with ammonia soln., extracting with chloroform and working up extract |
| CN1429611A (en) * | 2002-09-13 | 2003-07-16 | 张平 | Greater celandine extract and its preparation method and application |
| CN101955485A (en) * | 2010-05-26 | 2011-01-26 | 南京泽朗医药科技有限公司 | Preparation method of chelidonine |
Non-Patent Citations (1)
| Title |
|---|
| 石广亮: "白屈菜生物碱类成分提取、分离、纯化及体外抑菌作用研究", 《吉林农业大学硕士学位论文》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106138054A (en) * | 2015-04-28 | 2016-11-23 | 沈阳伟嘉牧业技术有限公司 | Chelerythrine application in treatment fowl drug resistance colibacillosis |
| CN111297950A (en) * | 2018-12-11 | 2020-06-19 | 泰州医药城国科化物生物医药科技有限公司 | Preparation method and application of chelidonine |
| CN109999014A (en) * | 2019-03-28 | 2019-07-12 | 佛山市正典生物技术有限公司 | It is a kind of can the new merit chigger disease of prevention and control plants essential oil and preparation method thereof |
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