CN104418852B - A kind of high-purity coptisine extracting method and application - Google Patents
A kind of high-purity coptisine extracting method and application Download PDFInfo
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- CN104418852B CN104418852B CN201310388306.4A CN201310388306A CN104418852B CN 104418852 B CN104418852 B CN 104418852B CN 201310388306 A CN201310388306 A CN 201310388306A CN 104418852 B CN104418852 B CN 104418852B
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- C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
- C07D455/03—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
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- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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Abstract
The present invention relates to extracting method and the application thereof of a kind of high-purity coptisine, its technological process is as follows: raw material (Rhizoma Coptidis or Rhizoma Coptidis fibrous root or Rhizoma Coptidis lime-ash) pulverize (as Rhizoma Coptidis fibrous root or Rhizoma Coptidis lime-ash be not required to pulverize) add 0.1～5% soak with sulphuric acid heat up extract four times filter calcium oxide (or calcium hydroxide) neutralize filter and wash precipitation pre-concentration removing impurity by means of precipitation precipitation coptisine coptisine recrystallization precipitation dry coptisine product.Rhizoma Coptidis content more than 90%.Product of the present invention has the effects such as good blood fat reducing, fat-reducing, blood sugar lowering, is treatment cardiovascular disease, obesity and the desirable feedstock of diabetes.
Description
Technical field
The invention belongs to biological extraction technology, be specifically related to a kind of high-purity coptisine extraction process technology.
Background technology
Rhizoma Coptidis begins to be loaded in Shennong's Herbal, is classified as top grade.Rhizoma Coptidis bitter in the mouth, cold in nature, there is the function of pathogenic fire purging, removing toxic substances, heat clearing away, dampness.For dysphoria with smothery sensation coma, vexed insomnia, damp and hot feeling of fullness, vomit, dysentery of suffering from abdominal pain, conjunctival congestion toxic swelling, aphtha of the mouth and tongue, eczema, scald, spit blood, epistaxis etc..Modern pharmacy result of study shows, Rhizoma Coptidis main active is Rhizoma Coptidis alkaloid, mainly includes (" Chinese Pharmacopoeias " 2010 editions) such as berberine (berberine), palmatine, coptisine and epiberberine；Rhizoma Coptidis (plain) has the pharmacologically active widely such as blood fat reducing and blood sugar lowering；Nearest research finds, in addition to berberine, Rhizoma Coptidis related alkaloids active component (coptisine, palmatine etc.) also has the effect (Hou Hong etc. such as blood fat reducing and blood sugar lowering.Rhizoma Coptidis alkaloid anti-hyperlipidemia and atherosclerosis experimentation.Time treasure's traditional Chinese medical science traditional Chinese medicines, 2011,22(10): 2462.)；The most also find, highly purified Rhizoma Coptidis alkaloid safety is the highest, and Rhizoma Coptidis alkaloid crude product will cause liver function to rise high safety issue (Jun Yi etc.Safety evaluation of main alkaloids from Rhizoma Coptidis.Journal of Ethnopharmacology, 2013.145:303-310).Extract highly purified Rhizoma Coptidis alkaloid and there is more preferable Development volue.
At present, the extraction process technology of berberine is a lot, and has been realized in industrialized production；The more existing patented technologies of extraction process technology and document report about coptisine.
Patent documentation (20091019620.7) reports a kind of coptisine extracting method, specifically including that raw material acid proposes acid extract neutralization and removing impurity by means of precipitation precipitation removes berberine solution alkali tune chromatographic column precipitation separation crystallographic product drying product, in prepared product, coptisine purity is up to more than 90%.This technology needs are by the feed liquid acid adjustment after neutralization remove impurity, upper chromatographic column separates, and goes up purification by macroporous resin the most again, finally uses ethanol elution；It is loaded down with trivial details that twice upper prop carries out chromatographic runs technique, utilizes ethanol elution cost the highest after macroporous resin adsorption.
Patent documentation (200910311352.8) reports the isolation and purification method of a kind of coptisine monomers, specifically include that extraction, concentrate, extract, the separation of dissolution filter, high performance preparative liquid chromatography, the step such as Product recycling.This technology utilizes high performance liquid chromatography, it is difficult to industrially apply.
Patent documentation (200810233362.X) reports a kind of with Rhizoma Coptidis fibrous root for raw material coproduction Rhizoma Coptidis alkaloid extracting method, specifically include that Rhizoma Coptidis fibrous root add appropriate 0.05～1% soak with sulphuric acid neutralization precipitation remove impurity on the packet point decolouring recycling design fractional precipitation crystallization of cation separation post stepwise elution dry jateorhizine monomer, berberine monomer, a Rhizoma Coptidis compound bio alkali.The patent proposes berberine and Technology that two monomers of jateorhizine extract simultaneously, but the extraction not mentioning coptisine separates.
Patent documentation (201010519053.6) reports a kind of method that Rhizoma Coptidis mixed biologic alkali based on structural features prepares coptisine, specifically includes that Rhizoma Coptidis total alkaloids is converted into coptisine by utilization chemistry reflection.This technology is not related to extract coptisine.
Patent documentation (200510044345.8) reports a kind of Rhizoma Coptidis total alkaloids extractive technique, but is not related to the extractive technique that coptisine monomers is.
Patent documentation (200810069671.8) reports a kind of coptis chinensis total alkaloid extracting technique technology, but is not related to the extraction of coptisine monomers.
Patent documentation (201110053453.7) reports a kind of method utilizing the isolated and purified Rhizoma Coptidis alkaloid of polyamide, specifically include that Rhizoma Coptidis crude extract polyamide goes up polyamide column after mixing sample, with dichloromethane eluent or with the mixed solvent gradient elution of dichloromethane and methanol, with thin layer chromatography inspection stream part, merge same stream part, it is dried, gets final product seven alkaloids such as isolated berberine, palmatine, coptisine, Columbamine, jateorhizine, magnoflorine, Groenlandcine.What this Technology separated is Rhizoma Coptidis alkaloid monomer, relates to, during isolated and purified, the dichloromethane equal solvent that toxicity is the biggest, and safety is difficult to ensure.
Li Feng etc. provide a kind of method extracting coptisine from Rhizoma Coptidis: Rhizoma Coptidis with 1% hydrochloric acid extraction, acid adjustment degree removing impurity by means of precipitation after acid extract concentrating under reduced pressure；The potassium iodide precipitation coptisine crude product that supernatant adds 5%, coptisine crude product chloroform and methanol (1:1) heating for dissolving, crystallisation by cooling, obtain coptisine secondary crude product；Alumina column chromatography on secondary crude product, finally obtains coptisine sterling (Li Feng, the chemical composition of Rhizoma Coptidis and quality standard research.Sichuan University's master thesis, 2007)；This Technology needs to use valuable reagent potassium iodide, simultaneously need to carry out upper prop separation, technique is loaded down with trivial details；And Gossypol recrystallized from chloroform safety is the most bad, it is difficult to meet the needs of extensive preparation.
The method of " Separation of dehydrocavidine from Corydalis saxicola by preparative chromatography " of Jiang Weizhe report reports and utilizes preparative liquid chromatography isolated and purified coptisine method (Jiang Weizhe etc., Separation of dehydrocavidine from Corydalis saxicola by preparative chromatography.Chinese herbal medicine, 2006,37(7): 1017)；Owing to using preparative liquid chromatography, apparatus expensive, investment is big, is unfavorable for equally realizing industrialized production.
In sum, although the Technology extracting coptisine has some to report, but, or Technology is complicated, or need expensive reagent, or relate to the deficiencies such as poisonous raw material.
Summary of the invention
The present invention is directed to these not enough, purpose is to provide a kind of high-purity coptisine extraction process, it with Rhizoma Coptidis and by-product (including other raw materials containing coptisine) thereof as raw material, extract the coptisine in Rhizoma Coptidis and Rhizoma Coptidis fibrous root and Rhizoma Coptidis lime-ash, Technology is simple, it is not necessary to expensive reagent and toxic raw materials, it is not required that carry out chromatographic isolation, product purity is high, and there is not been reported for the Technology of the present invention.
The present invention, with Rhizoma Coptidis and coptis plant as raw material, uses sulfuric acid solution soak extraction to produce coptisine, and described method includes:
1), raw material prepares
Rhizoma Coptidis or Rhizoma Coptidis fibrous root or Rhizoma Coptidis lime-ash are removed silt, pulverizes or shredding (as being not required to pulverize or shredding for Rhizoma Coptidis fibrous root or Rhizoma Coptidis lime-ash), as the raw material of processing.
2), soak
Add the raw material into fermentation vat or extract in still, add appropriate (3～15 times of volumes), concentration be 0.1～5% aqueous sulfuric acid (W/V) soak 1～48 hour.
3), extract
After immersion, being warmed up to 20～100 DEG C, insulation is extracted 0.1～5 hour, extracts four times；It is used for producing product with second time extracting solution for the first time；Extracting solution is for the soak of next group raw material for the third time, and soak is directly as primary extracting solution；4th extracting solution is used for the second time extracting solution of next group raw material.
4), feed liquid neutralizes
To merge with second time extracting solution for the first time, filter；Filtrate neutralizes the pH=2～12 of solution with Calx (or calcium hydroxide), filters；Residue washs 1～3 time with appropriate (0.1～3 times of level of residue) water again, reclaims the alkaloid that residue is taken away.
5), pre-concentration
Neutralize latter incorporated filtrate and washing liquid, concentrating under reduced pressure 0～20 times (0 times is directly used in subsequent treatment for not concentrating).
6), remove impurity
Concentrated solution adds 0.1～the concentrated hydrochloric acid (V/V) of 10% and liquor capacity 0～the sodium chloride (W/V) of 10%, and low temperature (0～30 DEG C) is placed 1～100 hour；Filter, a small amount of water (precipitation capacity 0.1～1 times of volume) washing precipitation.
7), Rhizoma Coptidis crude product produces
Filtrate and washing liquid after remove impurity merge, interpolation 0.1～the concentrated sulphuric acid (V/V) of 10% and 1～the sulfate (W/V) of 20%, and after being sufficiently stirred for, low temperature (0～30 DEG C) is placed 1～100 hour；Filter, 0.1～1 times of precipitation capacity water washing precipitation (coptisine), obtain coptisine crude product.This step is the key of the present invention, only uses this mixed precipitant system, could selective extraction coptisine.
8), coptisine recrystallization
Coptisine crude product is dissolved in 1～10 times of volume of solvent (water or methanol or ethanol), rising temperature for dissolving, and low temperature (0～30 DEG C) places crystallization 1～100 hour；Filter, 0.1～1 times of precipitation capacity solvent washing precipitation (coptisine), obtain coptisine finished product.
9), dry
Coptisine is dried at 60 DEG C, is high-purity coptisine product.
The content more than 50% of the coptisine that above method obtains, preferably up to more than 90%, can be used in the medicine of blood fat reducing or treatment obesity.
The following is about coptisine safety and treatment relevant disease in terms of contrast experiment:
1., subchronic toxicity testing
Test material be according to the inventive method prepare coptisine 1(content 90%), coptisine 2(content 50%), coptisine crude product (content 40%).
Subchronic toxicity testing method is carried out with reference to " natural drug (Chinese medicine) new drug research guideline ": SD rat is divided into four groups, be respectively Normal group, coptisine 1(content 90%), coptisine 2(content 50%), coptisine crude product (content 40%), often group 10, dosage is 1g/Kg；Fill continuously and feed 3 months, then adopt the biochemical indicators such as fasting blood monitoring liver function.
Table 1, the impact on rat liver function of the coptisine purity
From table it will be seen that compared with Normal group, coptisine 1 and coptisine 2 do not have any impact to liver function；The liver function of coptisine crude product group mouse is significantly raised, shows that the liver of animal is adversely affected by the coptisine of low-purity.The result that this result is reported with document is similar (Jun Yi etc.Safety evaluation of main alkaloids from Rhizoma Coptidis.Journal of Ethnopharmacology, 2013.145:303-310).
2., hypolipidemic activity experiment
Test material be the inventive method prepare coptisine 1(content 90%), coptisine 2(content 50%), coptisine crude product (content 40%).
Lipid-lowering test method is carried out with reference to " natural drug (Chinese medicine) new drug research guideline ": Golden Hamster is divided into 5 groups, and one group (10) give chow diet, and one group (40) give high lipid food；After 4 weeks, detect blood lipids index, more successful for modeling Golden Hamster be divided into 3 groups (often group 8～10): high fat matched group, coptisine 1(content 90%), coptisine 2(content 50%), coptisine crude product (content 40%).Medicine group gives 500mg/Kg medicine；Filling continuously and feed 4 weeks, blood lipids index is surveyed in the most mined out abdomen blood examination.
Table 2, Rhizoma Coptidis total alkaloids lipid-lowering effect compare
From table it will be seen that compared with Normal group, each test group blood fat all has rising, shows modeling success；Compared with high fat group, each medicine group cholesterol (TC) has declined, and shows all to have hypolipidemic activity；Wherein the hypolipidemic activity of coptisine 1 is best, is better than coptisine 2 and coptisine crude product；The hypolipidemic activity of coptisine 2 is better than coptisine crude product.
3., weight-reducing experiment
Test material be the inventive method prepare coptisine 1(content 90%), coptisine 2(content 50%), coptisine crude product (content 40%).
Weight-reducing experiment method is carried out with reference to " natural drug (Chinese medicine) new drug research guideline ": Golden Hamster is divided into 5 groups, one group (10) give chow diet, one group gives high lipid food (positive control), gives high lipid food respectively and give coptisine 1(content 90% simultaneously for other three groups), coptisine 2(content 50%), coptisine crude product (content 40%)；After 1 month, monitor the body weight of four groups of mouse, after 2 months, monitor the body weight of a mouse again.
Table 3, the impact of Rhizoma Coptidis total alkaloids team Golden Hamster body weight
From table it will be seen that compared with Normal group, high lipid food group body weight substantially increases；Compared with Normal group, the mouse body weight giving Rhizoma Coptidis medicine group is slightly increased；Compared with high lipid food matched group, coptisine 1 medicine group mouse weight loss fairly obvious (close with normal group), the weight loss of coptisine 2 is it is also obvious that and coptisine crude product medicine group mouse body weight has declined but inconspicuous.Relevant result shows, coptisine has the adverse effect that significantly mouse body weight is increased by control high lipid food, has preferable antiobesity action.
The studies above result shows, no matter from safety or from the perspective of drug effect (blood fat reducing and fat-reducing), highly purified coptisine value of exploiting and utilizing is substantially better than the coptisine of low-purity.
The invention have the advantages that
1., the Technology of the present invention with Rhizoma Coptidis and by-product thereof as raw material, especially with by-products such as Rhizoma Coptidis fibrous root and Rhizoma Coptidis lime-ash as raw material, belong to comprehensive utilization of resources scope, low cost product.
2., extracting solution by Calx (or calcium hydroxide) neutralisation of sulphuric acid, utilize calcium sulfate absorption to remove the impurity in extracting solution, compared with other impurity and purification techniques, this technological operation is simple, with low cost.
3., utilizing mixed precipitant (sulphuric acid and sulfate) selective precipitation coptisine, then carry out being recrystallized to give coptisine sterling, Technology is simple, low cost, and coptisine yield is high, and purity can reach more than 90%, and safety is fabulous.
4., whole technological operation simple, practical, industrial cost is substantially reduced, and productivity effect significantly improves.
5., the coptisine that obtains of the present invention have the effects such as preferable blood fat reducing, fat-reducing and blood sugar lowering, the product coptisine of the present invention may be used for treating hyperlipidemia, obesity and cardiovascular disease, diabetes etc..
Detailed description of the invention
By the following examples the present invention is specifically described; be necessary it is pointed out here that be that the present embodiment is served only for being further described the present invention; it is not intended that limiting the scope of the invention, the person skilled in the art in this field can make some nonessential improvement and adjustment according to the content of the invention described above.
Embodiment 1:
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 3 liter 5%, after soaking 1 hour, is warmed up to 100 DEG C of reflux, extract, 0.1 hour, filters；Residue is warmed up to 100 DEG C with the sulfuric acid solution of 3 liter 5% again and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=2, filters, and precipitation utilizes the water of 0.1 times of precipitation capacity to wash precipitation 3 times；Merging filtrate and washing liquid, add 10% concentrated hydrochloric acid, and low temperature (0 DEG C) is placed 100 hours, filtering and impurity removing, the precipitation water washing of 0.1 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated sulphuric acid (V/V) and the ammonium sulfate (W/V) of 20% of 0.1%, after being sufficiently stirred for, places 1 hour at 0 DEG C, filters, and precipitates and washs with the water of 1 times of volume；Precipitation is dissolved in the water of 10 times of volumes, and rising temperature for dissolving is placed 100 hours at 30 DEG C, filters, and precipitates with 0.1 times of water washing；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 90% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 91%.
Embodiment 2
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 15 liter 0.1%, after soaking 48 hours, is warmed up to 20 DEG C of insulations and extracts 5 hours, filter；Residue is incubated with the sulfuric acid solution of 15 liter 0.1% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=12, filters, and precipitation utilizes the water of 3 times of precipitation capacities to wash precipitation 1 time；Merging filtrate and washing liquid, concentrating under reduced pressure 20 times；Adding the sodium chloride of 0.1% concentrated hydrochloric acid and 10%, low temperature (30 DEG C) is placed 1 hour, filtering and impurity removing, the precipitation water washing of 1 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated sulphuric acid (V/V) and the sodium sulfate (W/V) of 1% of 10%, after being sufficiently stirred for, places 100 hours at 30 DEG C, filters, and precipitates and washs with the water of 0.1 times of volume；Precipitation is dissolved in the methanol of 1 times of volume, and rising temperature for dissolving is placed 1 hour at 0 DEG C, filters, and precipitates with 1 times of methanol washing；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 91% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 90%.
Embodiment 3
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated sulphuric acid (V/V) and the potassium sulfate (W/V) of 16% of 2%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 96% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 95%.
Embodiment 4
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated sulphuric acid (V/V) and the zinc sulfate (W/V) of 4% of 5%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 96% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 96%.
Embodiment 5
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated sulphuric acid (V/V) and the ammonium sulfate (W/V) of 12% of 3%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 98% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 96%.
Comparative example 6
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated hydrochloric acid (V/V) and the ammonium sulfate (W/V) of 12% of 3%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 45% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 91%.
Comparative example 7
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add strong phosphoric acid (V/V) and the ammonium sulfate (W/V) of 12% of 3%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 43% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 85%.
Comparative example 8
1 kilogram of Rhizoma Coptidis is pulverized or section, adds the sulfuric acid solution of 8 liter 3%, after soaking 12 hours, is warmed up to 70 DEG C of insulations and extracts 2 hours, filter；Residue is incubated with the sulfuric acid solution of 8 liter 3% and extracts four times；Merge first time and the second extracting solution (extract for three times and extract the process being used for next group raw material four times), after cooling, supernatant is separated (or being filtrated to get supernatant)；Supernatant utilizes Calx (calcium oxide) to neutralize pH=6, filters, and precipitation utilizes the water of 1 times of precipitation capacity to wash precipitation 2 times；Merging filtrate and washing liquid, concentrating under reduced pressure 6 times；Adding the sodium chloride of 5% concentrated hydrochloric acid and 5%, low temperature (10 DEG C) is placed 24 hours, filtering and impurity removing, the precipitation water washing of 0.5 times of volume of precipitation；Merging filtrate and washing liquid, add concentrated nitric acid (V/V) and the ammonium sulfate (W/V) of 12% of 3%, after being sufficiently stirred for, places 24 hours at 10 DEG C, filters, and precipitates and washs with the water of 0.5 times of volume；Precipitation is dissolved in the ethanol of 4 times of volumes, and rising temperature for dissolving is placed 24 hours at 10 DEG C, filters, and precipitates by 0.5 times of washing with alcohol；It is deposited at 50～70 DEG C drying, obtains product.Coptisine content 32% in high performance liquid chromatography (HPLC) research and application product, coptisine yield 68%.
Can be seen that from above comparative example, on the premise of other process conditions are the most constant, the only mixed solution of concentrated hydrochloric acid and sulfate can carry out selective precipitation to the filtrate after remove impurity and washing liquid, makes the coptisine content of the product finally obtained and coptisine yield be much higher than and carries out, with other mixed solution, the product that processes.
Claims (6)
1. an extracting method for high-purity coptisine, with Rhizoma Coptidis and coptis plant as raw material, uses sulfuric acid solution soak extraction to produce high-purity coptisine, and described method includes:
1) raw material prepares
Rhizoma Coptidis or Rhizoma Coptidis fibrous root or Rhizoma Coptidis lime-ash are removed silt, pulverizes or shredding；
2) soak
Add aqueous sulfuric acid in the feed to soak；
3) extract
After immersion, it is extracted as four times；It is used for producing product with second time extracting solution for the first time；Extracting solution is for the soak of next group raw material for the third time, and soak is directly as primary extracting solution；4th extracting solution is used for the second time extracting solution of next group raw material；
4) feed liquid neutralizes
To merge with second time extracting solution for the first time, filter；Filtrate Calx or calcium hydroxide neutralize the pH=2～12 of solution, filter；Residue is again with appropriate water washing；Reclaim the alkaloid that residue is taken away, obtain washing liquid；Merge washing liquid and filtrate, for subsequent treatment；
5) remove impurity
Concentrated hydrochloric acid and liquor capacity the ratio 0～sodium chloride of 10% is added, placement 1～100 hour at low temperature 0～30 DEG C according to the 0.1～10% of filtrate volume；Filter, then wash with appropriate water；Merge washing liquid and filtrate, for subsequent treatment；
6) coptisine crude product produces
Adding 0.1～the concentrated sulphuric acid of 10% and 1～the sulfate of 20% (W/V) according to liquor capacity, after being sufficiently stirred for, low temperature 0～30 DEG C are placed 1～100 hour；Filter, wash precipitation with water, obtain coptisine crude product；
7) coptisine recrystallization
Coptisine crude product is dissolved in 1～10 times of volume of solvent, rising temperature for dissolving, and low temperature 0～30 DEG C of placements crystallize 1～100 hour；Filter, 0.1～1 times of precipitation capacity solvent washing precipitation, obtain coptisine finished product；
8) dry
Coptisine finished product is dried at 60 DEG C, is high-purity coptisine product, the content more than 90% of coptisine.
2. according to the extracting method of the high-purity coptisine described in claim 1, it is characterised in that the ratio adding concentrated sulphuric acid and sulfate is 2～5%(V/V) concentrated sulphuric acid and 4～the sulfate of 16% (W/V).
3. according to the extracting method of the high-purity coptisine described in claim 1 or 2, it is characterised in that described sulfate refers to ammonium sulfate, sodium sulfate, potassium sulfate, magnesium sulfate, zinc sulfate.
4. according to the extracting method of the high-purity coptisine described in claim 1 or 2, it is characterised in that described step 7) coptisine crude product recrystallization solvent is water or methanol or ethanol；Step 2) soak consumption is raw material the 3～15 times of volumes of aqueous sulfuric acid used, the concentration of aqueous sulfuric acid is 0.1～5%, and soak time is 1～48 hour；The extraction of step 3) is that after being warmed up to 20～100 DEG C, insulation is extracted 0.1～5 hour.
5. according to the extracting method of the high-purity coptisine described in claim 1 or 2, it is characterised in that can also merge the filtrate of step 4) and washing liquid and then carry out pre-concentration, concentrating under reduced pressure 1～20 times, concentrated solution carries out the 5th again) process of step.
6. according to the extracting method of the high-purity coptisine described in claim 1 or 2, it is characterised in that described high-purity coptisine is applied in the medicine that preparation blood fat reducing or treatment are fat.
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| CN1069493A (en) * | 1991-08-09 | 1993-03-03 | 廖自基 | Extract refining berberine technology from " Mahonia fortunei " |
| CN1165822A (en) * | 1996-05-22 | 1997-11-26 | 湖南省桑植县药材公司 | Process for extracting berberine hydrochloride |
| CN101269132A (en) * | 2008-05-14 | 2008-09-24 | 西南大学 | Coptis chinensis total alkaloid extracting technique |
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| CN1069493A (en) * | 1991-08-09 | 1993-03-03 | 廖自基 | Extract refining berberine technology from " Mahonia fortunei " |
| CN1165822A (en) * | 1996-05-22 | 1997-11-26 | 湖南省桑植县药材公司 | Process for extracting berberine hydrochloride |
| CN101269132A (en) * | 2008-05-14 | 2008-09-24 | 西南大学 | Coptis chinensis total alkaloid extracting technique |
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