CN103381200A - White mulberry root-bark total alkaloid extract and preparation and application thereof - Google Patents
White mulberry root-bark total alkaloid extract and preparation and application thereof Download PDFInfo
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- CN103381200A CN103381200A CN 201210137333 CN201210137333A CN103381200A CN 103381200 A CN103381200 A CN 103381200A CN 201210137333 CN201210137333 CN 201210137333 CN 201210137333 A CN201210137333 A CN 201210137333A CN 103381200 A CN103381200 A CN 103381200A
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Abstract
The invention relates to an extraction method and application of a white mulberry root-bark total alkaloid extract. According to the invention, a white mulberry root-bark is used as a raw material which successively undergoes normal temperature or heating extraction with water or water-containing alcohol, precipitation with alcohol or acid water for removal of impurities, decolouring with a macroporous resin, separation and purification with a cationic resin and an anion resin for obtainment of a crude extract and finally refining so as to obtain the total alkaloid extract with high purity, wherein the content of the total alkaloid extract can reach more than 50%.
Description
Technical field
The present invention relates to a kind of alkaloid activity composition and method of quality control of extracting from Cortex Mori, particularly a kind of have inhibiting Cortex Mori alkaloid of alpha-glucosidase and preparation method thereof and a method of quality control.
Technical background
In recent years, due to the change of growth in the living standard, dietary structure, rhythm of life and the few moving factors such as life style of sitting that day is becoming tight more, whole world onset diabetes rate rapid development, diabetes have become the chronic disease of the third-largest serious threat human health after tumor, cardiovascular pathological changes.According to IDF (IDF) statistics, in the whole world in 2000, diabetics 1.51 hundred million is arranged, and the whole world there is diabetics 2.85 hundred million at present, by present speed increment, estimate that the year two thousand thirty whole world will have nearly 500,000,000 people to suffer from diabetes.China is the maximum country of world population, the survey result demonstration of 2008, and in the adult more than 20 years old, the diabetes prevalence of age markization is 9.7%, and the ratio of prediabetes is more up to 15.5%.The treatment situation of diabetes is very severe.
The medicine that is used for the treatment of at present diabetes mainly contains sulphanylureas, biguanides and alpha-glucosidase inhibitor.But sulfonylurea drugs easily causes persistent hypoglycemia, biguanides can produce serious gastrointestinal reaction and lesions of liver and kidney, and alpha-glucosidase inhibitor gas in making abdomen increases, other untoward reaction are less, be widely used for reducing postprandial hyperglycemia, be considered to the ancillary drug of the insulinize of the drug of first choice of type 2 diabetes mellitus and type 1 diabetes, have broad application prospects.
Cortex Mori derives from the dry root bark that the moraceae plants Mulberry is removed cork, and sweet in the mouth is cold in nature, enters lung meridian.Be distributed in national most area, aboundresources.Theory of Chinese medical science is thought its tool eliminating pathogen from the lung for relieving asthma, the inducing diuresis to remove edema effect.The multiple alkaloids compositions such as the 1-DNJ that contains in Cortex Mori (DNJ) and derivant thereof, it is the inhibition active substance of the clear and definite alpha-glucosidase of effect, modern pharmacological research proof Cortex Mori can suppress blood sugar increasing, prevention and treatment diabetes.
By retrieval, still have following 4 pieces with Cortex Mori and have the research that alpha-glucosidase suppresses active Chinese medicine extract and report;
Application number is 200410021468.5: to relate to composition be total flavones to patent name for " production technology of Cortex Mori extract, " content, do not relate to alkaloid, and purposes is antiviral, antibiotic, antiinflammatory and acute and chronic bronchitis, and emphysema do not relate to diabetes.
Application number is 200610111644.3: patent name for " mulberry twigs alkaloid valid target is in the application of preparation in hypoglycemic drug; " wherein Chinese medicine extract is the total alkaloids that is got by the Ramulus Mori preparation, do not relate to Cortex Mori, and we carry out the DNJ detection to different regions Ramulus Mori and Cortex Mori respectively, find that in Cortex Mori, DNJ content is ten times of left and right of Ramulus Mori, in Ramulus Mori, DNJ content is too low, is more suitable for as crude drug with Cortex Mori.
Application number is 200610099049.2: patent name for " a kind of preparation method with Cortex Mori extract of glycosidase inhibiting function; " its extraction separation method is: the Cortex Mori crude drug is after cutting, with water or alcoholic solution as extracting solvent, centrifugal after the extracting solution concentrating under reduced pressure, activated-charcoal column on supernatant.This technique is too simple, active constituent content low (DNJ is 5%-9% approximately), and must take in a large number could onset.
Application number is 200710098987.5: patent name for " a kind of Cortex Mori extract with glycosidase inhibiting function and preparation method thereof; " this patent has been used active carbon when decolouring, and active carbon can not be reused, we have used reusable macroporous resin, greatly saved cost, the while with the acid adjustment of macroporous resin eluent, not only can be removed a large amount of non-alkaloids impurity before upper resin anion (R.A.) post, increase simultaneously the adsorbance of resin, reduced resin stain.
Summary of the invention
The preparation method that the purpose of this invention is to provide a kind of Cortex Mori total alkaloids, the method for taking is column chromatography.
Another object of the present invention is to provide the preparation method of Cortex Mori total alkaloids extract.
Another object of the present invention is to provide the Chinese medicine preparation of described extract.
The object of the invention also is to provide the application of this Cortex Mori total alkaloids extract in the treatment diabetes.
The present invention seeks to be achieved through the following technical solutions: it is characterized in that described extract total alkaloid content by weight percentage is 50-90%; Contained alkaloid wherein: 1-deoxynojirimycin 10-90%, 3-epi-fagmine 0.1-10%, 1,4-dideoxy-1,4-miino-D-arabinitol 0.1-10%, fagmine 0.1-10%, 2-O-α-D-galactopyranosl-1-deoxynojirimycin 0.1-10%, related alkaloids composition surplus.
The object of the invention Cortex Mori total alkaloids extract preparation method operates as follows:
A. the raw medicinal material Cortex Mori is through after cutting, and as solvent extraction 1～5 time, each 0.5～3 hour, the extraction temperature was room temperature～100 ℃ with the alcoholic solution of water or 5%～50%, and the total solvent amount is 18～36 times of Cortex Mori raw medicinal material weight;
B. to be concentrated into relative density be 1.0～1.2 to extracting solution boiling lower than 80 ℃ of lower concentrating under reduced pressure or 100 ℃, adds 90%～100% ethanol, regulates concentration of alcohol to 40%～80%, and cold preservation is spent the night, and filters or centrifugal;
C. the gained clear liquid is removed ethanol through reduction vaporization, gets clear paste after precipitate with ethanol, adds deionized water and is diluted to every 1ml and contains Cortex Mori medical material 0.5～1.0g, and acid adding is transferred pH value to 2 ~ 6, and the standing cold preservation rear high speed centrifugation that spends the night separates, and gets acid clear paste;
D. get acid clear paste, remove pigment or directly go up the strongly acidic cation-exchange post by macroporous adsorptive resins, be eluted to colourlessly with deionized water, and pH value is 5 ~ 6, then uses the aqueous alkali eluting of 2 ~ 8 times of column volumes, with aqueous alkali eluent concentrating under reduced pressure, add the clear paste that deionized water makes into 0.25 ~ 1.0g medical material/ml, upper strong-base anion-exchange resin post, and be washed to neutrality, merge upper prop effluent and water lotion, concentrating under reduced pressure gets the Cortex Mori crude extract.
E. get crude extract, further make with extra care to get the high-purity total alkaloids extract, faint yellowly to the brown color powder dry, its total alkaloid content can reach more than 50%.
In above-mentioned technical process, after the extracting solution that will contain total alkaloids transfers to suitable acidity, alkaloid ionization is cation, nonbasic substances is not ionized, after extracting solution was crossed cation exchange resin column, the alkaloid organic ion of ionization and the hydrion on resin column exchanged and are adsorbed on resin securely.In extracting solution acid adjustment process, we find that pH value is unsuitable too high or too low, found through experiments pH value proper between 2-6.
The Chinese medicine preparation of Cortex Mori total alkaloids of the present invention is characterized in that described Cortex Mori total alkaloids extract is effective ingredient, according to different medication demands, can be made into tablet, drop pill, granule, capsule, oral liquid etc. after adding pharmaceutic adjuvant.
Compared with the prior art, benefit of the present invention is embodied in:
1, Cortex Mori extract of the present invention is with total alkaloid content, 1-deoxynojirimycin, 3-epi-fagmine, 1,4-dideoxy-1,4-miino-D-arabinitol, fagmine, the content of 2-O-α-D-galactopyranosl-1-deoxynojirimycin carries out definite sign to it.
2, the present invention adopts high performance liquid chromatography, use Derivative, can be to 1-deoxynojirimycin, 3-epi-fagmine, Isosorbide-5-Nitrae-dideoxy-1,4-miino-D-arabinitol, fagmine, the alkaloid components such as 2-O-α-D-galactopyranosl-1-deoxynojirimycin carry out clear and definite quantitative and qualitative analysis.
3, the present invention has all adopted reusable resin in decolouring and purge process, has greatly saved cost.
4, the present invention has regulated pH value before upper prop, after extracting solution transfers to suitable acidity, alkaloid ionization is cation, simultaneously, the part composition is met Acid precipitation, after filtration, after acidic extraction liquid was crossed cationic resin, the alkaloid organic ion of ionization and the hydrion on resin column exchanged and firmly are adsorbed on resin column.By regulating pH value, removed more impurity, increased the adsorbance of resin column, improved the quality of product.
5, Cortex Mori extract of the present invention is to obtain with the extraction from Cortex Mori of new extracting method, for diabetes, reliable curative effect is arranged.
6, the present invention can satisfy vast diabetics to the requirement of different dosage form.
The specific embodiment
Below will further describe the present invention by embodiment, but these embodiment only illustrate the present invention, and should not be construed as any limitation of the invention.
Embodiment 1:
Cortex Mori is through after cutting, with water as solvent extraction three times, each 1h, extracting temperature is 100 ℃, quantity of solvent is respectively 10 times of Cortex Mori raw medicinal material weight, 8 times, 8 times, extracting solution merges, being concentrated into relative density is 1.1, be sink to 50% with 95% ethanol alcohol, cross leaching filtrate, pressure reducing and steaming ethanol, it is 4 that gained solution is transferred pH value again, the standing rear high speed centrifugation of cold preservation, get supernatant, upper AB-8 macroporous adsorptive resins, be eluted to clarification color light (ninhydrin reaction is negative) with deionized water, merge loading effluent and eluent, upper 001*7 strongly acidic cation-exchange post, be eluted to pH value with deionized water closely neutral, use again the ammonia eluting of 1% volumetric concentration, when being positive, ninhydrin reaction begins to collect, effluent volume is about 3 times of column volumes, ammonia eluent concentrating under reduced pressure is waved most ammonia, pH value is 8 ~ 10, 201*7 gel strong basic type anion-exchange resin post on concentrated solution, and be washed to neutrality (ninhydrin reaction is negative), collect loading effluent and water lotion, merge the concentrating under reduced pressure drying and obtain Cortex Mori extract.
Embodiment 2
the raw medicinal material Cortex Mori is through after cutting, with water as solvent extraction 3 times, each 1h, extracting temperature is 100 ℃, quantity of solvent is respectively 10 times, 8 times, 8 times of Cortex Mori raw medicinal material weight, merge extractive liquid, at 70 ℃ of lower concentrating under reduced pressure, centrifugal after cold preservation, get supernatant, transfer pH value 5, centrifugal after cold preservation, get supernatant, by the D-101 macroporous adsorptive resins, use the deionized water eluting, collect loading effluent and eluent, 001*7 type cation exchange resin column on loading effluent and water elution liquid, the pH value of upper prop liquid is 5, the effluent pH value drops to 2, be washed till colourless with deionized water, and pH value is closely neutral, use again the diethylamine eluting of 2% volumetric concentration, when being positive, ninhydrin reaction begins to collect, effluent volume is about 3 times of column volumes, ammonia eluent concentrating under reduced pressure is waved most ammonia, pH value is 8 ~ 10, 201*7 gel strong basic type anion-exchange resin post on concentrated solution, and be washed to neutrality (ninhydrin reaction is negative), collect loading effluent and water lotion, concentrate to get thick paste, refining through alkali alumina again, lyophilization obtains Cortex Mori extract.
Embodiment 3: Cortex Mori total alkaloids chewable tablet
Preparation process: citric acid is added water 30ml, ultrasonicly make dissolving, standby.Cortex Mori total alkaloids and mannitol, steviosin and citric acid mixed grinding make evenly, add aqueous solution of citric acid, grind well, then add 70% alcoholic solution, grind well, and soft material processed, 20 mesh sieve granules processed, drying is pressed 1000 after granulate
Embodiment 4: Cortex Mori total alkaloids ordinary tablet
Preparation process: Cortex Mori total alkaloids extract, dextrin, microcrystalline Cellulose, low-substituted hydroxypropyl methylcellulose are crossed respectively 80 mesh sieves; With Cortex Mori total alkaloids and dextrin, the low-substituted hydroxypropyl methylcellulose of 2/3 amount and 70% alcoholic solution mixed grinding, make evenly soft material processed; 20 mesh sieve granules processed again, drying adds remaining low-substituted hydroxypropyl methylcellulose after granulate, mixing, then add the magnesium stearate mixing, press 1000.
Embodiment 5: Cortex Mori total alkaloids drop pill
Preparation process: with Macrogol 4000, polyethylene glycol 6000 heating and melting, then Cortex Mori total alkaloids extract powder is added wherein, fully stir, medicine is well-dispersed in substrate; Said mixture is transferred on pill dripping machine, and 70 ℃ of-80 ℃ of sealed thermal insulatings are made condensing agent with anhydrous methyl-silicone oil, and 10 ℃ of-20 ℃ of gradients are cooling, with 30-60 drip/minute carry out dripping, dripping becomes 1000 balls.Collect drop pill, absorb condensed fluid, dry, packing gets final product.
Embodiment 6: Cortex Mori total alkaloids granule
Preparation process: Cortex Mori total alkaloids extract, starch, dextrin are crossed respectively 100 mesh sieves, and mixing is crossed 15 mesh sieves, adds 70% appropriate ethanol soft material processed, crosses 14 mesh sieve granules processed, 60 ℃ of dryings, and granulate, pack is made altogether 1000 bags, and be get final product
Embodiment 7: Cortex Mori total alkaloids capsule
Preparation process: with starch, Cortex Mori total alkaloids extract mixing, cross 60 mesh sieves, add 70% appropriate ethanol soft material processed, cross 24 mesh sieve granules processed, 60 ℃ of dryings, 20 mesh sieve granulate, encapsulated, make altogether 1000, and get final product.
Embodiment 8: Cortex Mori total alkaloids oral liquid
Preparation process: the Cortex Mori total alkaloids extract is soluble in water, add steviosin, sodium benzoate, mannitol, citric acid to be stirred to dissolve, make 10000ml, sterilization is distributed into 1000, and get final product.
Claims (11)
1. Cortex Mori total alkaloids extract, it is characterized in that described total alkaloids extract by weight percentage content be 50-90%, wherein contain alkaloid: 1-deoxynojirimycin 10-90%, 3-epi-fagmine 0.1-10%, Isosorbide-5-Nitrae-dideoxy-1,4-miino-D-arabinitol 0.1-10%, fagmine 0.1-10%, 2-O-α-D-galactopyranosl-1-deoxynojirimycin 0.1-10% etc., related alkaloids composition surplus.
2. according to claim 1 alkaloid valid target, is characterized in that, in described total alkaloids compositions, the percentage by weight of 1-deoxynojirimycin must not be lower than 50% of total alkaloids.
3. the preparation method of Cortex Mori total alkaloids extract according to claim 1 is characterized in that operating as follows:
A. the raw medicinal material Cortex Mori is through after cutting, and as solvent extraction 1～5 time, each 0.5～3 hour, the extraction temperature was room temperature～100 ℃ with the alcoholic solution of water or 5%～50%, and the total solvent amount is 18～36 times of Cortex Mori raw medicinal material weight;
B. to be concentrated into relative density be 1.0～1.2 to extracting solution boiling lower than 80 ℃ of lower concentrating under reduced pressure or 100 ℃, adds 90%～100% ethanol, regulates concentration of alcohol to 40%～80%, and cold preservation is spent the night, and filters or centrifugal;
C. the gained clear liquid is removed ethanol through reduction vaporization, gets clear paste after precipitate with ethanol, adds deionized water and is diluted to every 1ml and contains Cortex Mori medical material 0.5～1.0g, and acid adding is transferred pH value to 2～6, and the standing cold preservation rear high speed centrifugation that spends the night separates, and gets acid clear paste;
D. get acid clear paste, remove pigment or directly go up the strongly acidic cation-exchange post by macroporous adsorptive resins, be eluted to colourlessly with deionized water, and pH value is 5～6, then uses the aqueous alkali eluting of 2～8 times of column volumes, with aqueous alkali eluent concentrating under reduced pressure, add the clear paste that deionized water makes into 0.25～1.0g medical material/ml, upper strong-base anion-exchange resin post, and be washed to neutrality, merge upper prop effluent and water lotion, concentrating under reduced pressure gets the Cortex Mori crude extract;
E. get crude extract, get the high-purity total alkaloids extract through an one-step refining, faint yellowly to the brown color powder dry, its total alkaloid content can reach more than 50%.
4. method according to claim 3, is characterized in that the concentration of alcohol for precipitate with ethanol is 50-70% in described step b.
5. method according to claim 3, is characterized in that acid used in described step c is concentrated hydrochloric acid or concentrated sulphuric acid, regulates pH value to 4～6.
6. method according to claim 3 is characterized in that the resin that is used for chromatographic column in described steps d is: wherein macroporous adsorbent resin is selected and is comprised nonpolar, low pole and intermediate-polarity macroporous adsorption resin; In steps d, strongly acidic cation-exchange is selected 001*2,001*3,001*7, WA-2 gel type cation exchanger resin or D072, D101, HD-8 large pores cation exchange resin; In steps d, strong basic type anion-exchange resin is selected 201*7 gel-type anion exchange resin or D293 macroporous type anion exchange resin.
7. method according to claim 3, the aqueous alkali that it is characterized in that being used for the chromatographic column eluting in described steps d is one or more mixed solutions wherein such as the ammonia of 0.1%-5.0%, diethylamine, triethylamine, pyridine.
8. Cortex Mori total alkali method for preparing extractive according to claim 3, it is characterized in that: in step e, refining used carrier is one or more mixture wherein such as polyamide, active carbon, aluminium oxide, silica gel, pass through column chromatography for separation, after the organic solvents such as chloroform, ethyl acetate, acetone, methanol, ethanol or water elution, concentrating under reduced pressure, drying, and get final product.
9. method according to claim 7, the concentration of its acetone, methanol, ethanol is 5%-100%.
10. according to claim 1-8 described Cortex Mori total alkaloids extract, it is characterized in that: described Cortex Mori total alkaloids extract can be made different dosage forms according to different medication demands, comprises tablet, drop pill, capsule, granule, oral liquid etc.
11. pharmaceutical composition according to claim 9 is characterized in that, the alkaloid valid target that contains is counted the 20-500mg/ sheet by effective part extract.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104402801A (en) * | 2014-10-31 | 2015-03-11 | 西南大学 | Methods for separating DNJ (1-deoxynojirimycin) and preparing DNJ nanometer suspension |
| CN105153015A (en) * | 2015-08-29 | 2015-12-16 | 合肥华方医药科技有限公司 | Method for separating trace alkaloid monomers from white mulberry root-bark total alkaloid |
| CN106420933A (en) * | 2016-09-30 | 2017-02-22 | 毛华养 | Operation method of white mulberry root bark extracts |
| CN113292482A (en) * | 2021-05-27 | 2021-08-24 | 湖南德诺贝莱健康产业有限公司 | Method for extracting high-content deoxynojirimycin from cortex mori |
-
2012
- 2012-05-04 CN CN 201210137333 patent/CN103381200A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104402801A (en) * | 2014-10-31 | 2015-03-11 | 西南大学 | Methods for separating DNJ (1-deoxynojirimycin) and preparing DNJ nanometer suspension |
| CN104402801B (en) * | 2014-10-31 | 2016-06-15 | 西南大学 | Separate DNJ and the method preparing DNJ nano suspension |
| CN105153015A (en) * | 2015-08-29 | 2015-12-16 | 合肥华方医药科技有限公司 | Method for separating trace alkaloid monomers from white mulberry root-bark total alkaloid |
| CN105153015B (en) * | 2015-08-29 | 2018-02-27 | 合肥华方医药科技有限公司 | A kind of method that Alkaloid monomer is separated in the biology total alkali from the root bark of white mulberry |
| CN106420933A (en) * | 2016-09-30 | 2017-02-22 | 毛华养 | Operation method of white mulberry root bark extracts |
| CN113292482A (en) * | 2021-05-27 | 2021-08-24 | 湖南德诺贝莱健康产业有限公司 | Method for extracting high-content deoxynojirimycin from cortex mori |
| CN113292482B (en) * | 2021-05-27 | 2022-11-01 | 湖南德诺贝莱健康产业有限公司 | Method for extracting high-content deoxynojirimycin from cortex mori |
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Address after: Science and Technology Industrial Park D-5 No. 168 high tech Zone camphor road in Hefei city of Anhui Province in 230088 Applicant after: Hefei Huafang Pharmaceutical Sciences & Technology Co., Ltd. Applicant after: Anhui Huatuo Traditional Chinese Medicine Co., Ltd. Address before: Tianda high tech Zone 230088 Hefei Road, Anhui province No. 71 Huayi Science Park G block four building Applicant before: Hefei Huafang Pharmaceutical Sciences & Technology Co., Ltd. Applicant before: Anhui Huatuo Traditional Chinese Medicine Co., Ltd. |
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Application publication date: 20131106 |