CN103275012B - Preparation method and application of Maca alkaloid - Google Patents
Preparation method and application of Maca alkaloid Download PDFInfo
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Abstract
The invention discloses a preparation method and an application of Maca alkaloid. The method comprises the steps in sequence as follows: (1), Maca is smashed, a certain volume of an acid alcohol solution is added, supersonic extraction is performed for 1-3 times, the mixture is filtered, and an extracting solution is combined and concentrated, so that extractum is obtained; (2), the extractum is dissolved with acid water and filtered, the pH value is adjusted to be 9-12, the mixture is leached with chloroform or ethyl acetate, and an extraction liquid is subjected to pressure reduction concentration and dried, so that a Maca alkaloid crude extract is obtained; and (3), the Maca alkaloid crude extract is subjected to column chromatography with high-speed countercurrent and alkaline alumina, an eluent comprising a target compound is combined, and a solvent is recycled, so that a Maca alkaloid compound is obtained. According to the method, the technological operation is simple, the preparation amount is large, the sample loss is small, the separation effect is good, the energy consumption is low, the pollution is small the method is suitable for large-scale preparation, and a Maca alkaloid monomer obtained through separation is high in purity, has a better antineoplastic activity, is suitable for preparing antineoplastic drugs and neoplasm prevention health-care food, and has a better application and development prospect.
Description
Technical field
The effective ingredients in plant that the invention belongs in Separation of Natural Products technical field of purification extracts, the particularly alkaloidal preparation method of a kind of agate coffee and application.
Background technology
Agate coffee (Lepidium meyenii Walp.) is Cruciferae separate row Vegetable spp herbaceous plant, originate in the mountain area, Andean of Peru height above sea level 3500m-4450m, can comparatively large at day and night temperature, grow without under special conditions that is fertile, ice-bound and anoxic for a long time, have the good reputation of " Peru's ginseng ".High nutritive value, reasonably trophic structure and various active secondary metabolites make agate coffee have multiple nutrients nourishing function and pharmacological action, can be used for strengthening energy, raising fertility, improvement function, treatment climacteric syndrome, rheumatism, dysthymia disorders, anaemia etc., also have anticancer and leukemia effect.
Research shows, alkaloid is one of main active ingredient of agate coffee, and agate coffee alkaloid acts on pituitary gland and hypothalamus, endocrine regulation gland (as suprarenal gland, Tiroidina, pancreas, ovary etc.), balance hormone; Agate coffee alkaloid can be used to the deficiency of supplementary estradiol production, therefore can be used as exogenous hormone surrogate to alleviate hypothalamus and pituitary gland hyperfunction; Can sexual desire promoting containing components such as macamide and agate coffee alkene; In addition, the experiment of vitro inhibition cancer cells shows: maca imidazole Alkaloid lepidiline A has restraining effect (ED50 is 7.39 μ g/ml) to FDIGROV cell, lepidiline B then has restraining effect to multiple cancer cells, as UMUC3 cell, PACA2 cell, MDA231 cell and FDIGROV cell (ED50 is respectively 6.47,1.38,1.66 and 5.26 μ g/ml).
Total alkaloid content in agate coffee is very low, only containing 0.2%-0.4%, this brings difficulty to the alkaloidal Extraction and separation purifying of agate coffee, at present, in existing document and patent, the alkaloidal extraction and purification process of agate coffee is complicated, need the column chromatography purification of multistep, as agate coffee root SDS extraction is first extracted thing by patent " indolizidine alkaloid of agate coffee Lepidium apetalum and uses thereof " (publication number: CN1684680A), then extract is dissolved in sour water-methanol solvate, then obtains extract A E with dichloromethane extraction; Diaion HP-20MG on AE is adsorbed, then carries out wash-out, obtain four components, i.e. A1, A2, A3 and A4, then detected by thin-layer chromatography, find in A2 component containing alkaloid-positive spots; Then silica gel column chromatography on A2 is carried out wash-out, obtain 7 components, i.e. B1-B7, then B5 component is obtained thick alkaloid fraction through acidic alumina column chromatography, this fraction carries out purifying further by column chromatography and preparative silica gel H PLC, obtains two kinds of alkaloid compounds.Patent " maca imidazole alkaloid is preparing the application in cardiovascular agent " (publication number: CN101756965A) first by pueraria root powder after homogenate extraction, extracting solution is after filtration, concentrated, obtains extracting medicinal extract; Then medicinal extract is used 20% dissolve with methanol, upper macroporous adsorbent resin, collect containing alkaloidal 3rd elution fraction, concentrated, obtain maca imidazole alkaloid crude extract; Upper normal phase column chromatography (neutral alumina column chromatography or purification on normal-phase silica gel column chromatography) after crude extract being dissolved with chloroform, collects containing third and fourth elution fraction alkaloidal, concentrated, obtains medicinal extract; After medicinal extract dissolve with methanol, upper C18 reverse phase silica gel post carries out chromatography again, collect containing maca imidazole alkaloid second and third, four elution fractions, recycling design, obtains maca imidazole alkaloid composition; Finally, solid-phase extraction column (with C8 or C18 silica gel for filler) upper after maca imidazole alkaloid composition dissolve with methanol is further purified, crystallization, obtains maca imidazole alkaloid monomer.Although above-mentioned patent can obtain highly purified alkaloid, but adopt repeatedly the method for chromatography to obtain monomer component, the method is complex procedures not only, wastes time and energy, and the alkaloidal rate of recovery is lower, consumes excessive, the high cost of quantity of solvent, be not suitable for industrialized production.
Summary of the invention
In order to overcome deficiency and the defect of existing separation agate coffee alkaloid technology, the object of the present invention is to provide a kind of fast, the preparation method of high efficiency separation agate coffee alkaloid monomer, namely use high-speed countercurrent chromatography from agate coffee, to be separated with normal phase chromatography the method preparing agate coffee alkaloid monomer.
The technical solution adopted for the present invention to solve the technical problems is as follows:
(1) extract: pueraria root powder is broken, add 5-20 alcoholic solution doubly and extract 1-3 time, filter, united extraction liquid, concentrated, obtain medicinal extract;
(2) removal of impurities: dissolved by the medicinal extract acidic aqueous solution of step (1) gained, filters, filtrate again by lye pH adjustment value to 9-12, then with chloroform or extraction into ethyl acetate, extraction liquid concentrating under reduced pressure, dry, obtain agate coffee alkaloid crude extract;
(3) high-speed counter-current purifying: with alkane: fatty acid ester: fatty alcohol: water is solvent systems, and 4-10:1-13:2-13:1-10 mixes by volume, separates upper and lower phase in separating funnel after hold over night, degassed; Agate coffee alkaloid crude extract phase step (2) obtained, lower phase or the two isopyknic mixing solutions dissolve, and filter, obtain sample solution; Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph, adjust engine speed simultaneously, pump in post as moving phase down using what prepare, after the running balance of post internal solvent Establishing, extracting sample solution by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, obtain the agate coffee alkaloid of certain purity;
(4) column chromatography purification: the agate coffee alkaloid of certain purity step (3) obtained carries out silica gel column chromatography or alumina column chromatography, with chloroform-methanol or benzene-chloroform for eluting solvent carries out gradient elution, collect containing alkaloidal elutriant, recycling design, concentrated, drying, obtains agate coffee alkaloid compound.
Acidity alcohol solution in described step (1) is the mixing solutions of methanol solution, ethanolic soln or above-mentioned two kinds of alcohol, is preferably 70% ethanolic soln; Extracting method comprises supersound extraction, microwave extraction, lixiviate, refluxing extraction, is preferably supersound extraction.
Acidic aqueous solution in described step (2) is one or both in hydrochloric acid soln, acetic acid and sulphuric acid soln, and wherein, the massfraction of acid is 0.5-5%; Alkali lye is one or both in sodium hydroxide solution, ammoniacal liquor and sodium carbonate solution.
Solvent systems in described step (3) comprises following two kinds: alkane: fatty acid ester: fatty alcohol: the two phase solvent system that water is formed, or the polynary two phase solvent system set up on above-mentioned two-phase system basis.
Aluminum oxide in described step (4) is neutral alumina or alkali alumina.
The application in antitumor drug prepared by agate coffee alkaloid prepared by the present invention, the agate coffee alkaloid of above-mentioned preparation is mixed with medically acceptable auxiliary material, make corresponding preparation, described auxiliary material comprises carboxylic propyl methocel, polyvidone, stearyl alcohol, hexadecanol, Magnesium Stearate, Microcrystalline Cellulose, starch, N.F,USP MANNITOL, sodium bicarbonate, and to antioxidant, amino acid, VITAMIN, carbohydrate and plant milk extract that the present invention does not have a significant effect.Described preparation comprises tablet, capsule, granule, dripping pill and injection.
Method of the present invention is prepared compared with the alkaloidal method of agate coffee with existing, has following technical superiority:
(1) the present invention is by agate coffee alkaloid crude extract first through chloroform extraction removal of impurities, then carries out purifying with high speed adverse current chromatogram, and finally utilize normal phase chromatography to be further purified and obtain agate coffee alkaloid monomer, technological process environmental protection, to environment without serious harm.
(2) the present invention is simple to operate, and process cycle is short, and save reagent, eluent recoverable, reduces production cost, and extraction efficiency is high, is suitable for suitability for industrialized production.
(3) agate coffee alkaloid preparation amount is large.According to required target sample amount, the high-speed counter-current chromatograph of different model can be selected to be separated.Wherein, countercurrent chromatography instrument sample size can reach several grams even tens grams, the amount of the agate coffee alkaloid monomer that flash liberation goes out and reduciblely reach pilot scale level.
(4) be separated the growth that the agate coffee alkaloid compound obtained effectively can suppress kinds of tumor cells in vitro, there is significant anti-tumor activity.
Embodiment
Further describe the present invention by embodiment below, be conducive to the understanding to the present invention and advantage thereof, better effects if, but described embodiment is only for illustration of the present invention instead of restriction the present invention.
Embodiment 1: the alkaloidal preparation of agate coffee
Get pueraria root powder 1kg, add the ethanolic soln of 6L20%, mixing, supersound extraction 35min, filter, by residue supersound extraction twice as stated above again, merging filtrate reclaim under reduced pressure, to without alcohol taste, obtains medicinal extract.Dissolve above-mentioned medicinal extract with 1%HCl, filter, filtrate uses sodium hydroxide adjust pH to 9 again, then uses chloroform extraction, extraction liquid concentrating under reduced pressure, dry, obtains agate coffee alkaloid crude extract.
By normal hexane: ethyl acetate: methyl alcohol: water 5:1:5:1 ratio mixing by volume, in separating funnel, isolate phase up and down after hold over night, and ultrasonic degas 20min is for subsequent use.Get 200mg agate coffee alkaloid crude extract to be dissolved under 20ml mutually, vibration makes it to dissolve completely, is separated in order to high speed adverse current chromatogram.Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph with the flow velocity of 20ml/min, ON cycle water-bath to set thermostat temperature be 35 DEG C, adjust engine speed is 700r/min simultaneously, lower to moving phase using what prepare, pump in post with the flow velocity of 2ml/min, after the running balance of post internal solvent Establishing, extracting sample solution is by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after sample introduction 3h, main frame is stopped to rotate, stationary phase is released, stationary phase retention rate is 70%, obtain the agate coffee alkaloid that purity is 75%.
Upper silica gel column chromatography after the agate coffee alkaloid obtained by countercurrent purification is concentrated, with chloroform-methanol according to 10:1,7:3 carries out gradient elution, collect containing alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography, recycling design, concentrated, dry, obtain the agate coffee alkaloid compound that purity is 98%.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column is Waters C18 pillar (5 μm, 4.6mm × 250mm), and column temperature is 30 DEG C; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Detection time is 30min, and determined wavelength is 280nm.
Embodiment 2: the alkaloidal preparation of agate coffee
Get pueraria root powder 5kg, add the methanol solution of 50L60%, mixing, microwave extraction 20min, filter, extracted as stated above once by residue again, merging filtrate reclaim under reduced pressure, to without alcohol taste, obtains medicinal extract.With the above-mentioned medicinal extract of 2.5% acetate dissolution, filter, filtrate uses ammoniacal liquor adjust pH to 10 again, is then extracted with ethyl acetate, extraction liquid concentrating under reduced pressure, dry, obtains agate coffee alkaloid crude extract.
By normal hexane: ethyl acetate: methyl alcohol: water 5:5:7:5 ratio mixing by volume, in separating funnel, isolate phase up and down after hold over night, be above added to the triethylamine of 10mM, under be added to the HCl of 10mM, and ultrasonic degas 15min is for subsequent use.Get 300mg agate coffee alkaloid crude extract to be dissolved under 20ml mutually, vibration makes it to dissolve completely, is separated in order to high speed adverse current chromatogram.Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph with the flow velocity of 10ml/min, ON cycle water-bath to set thermostat temperature be 20 DEG C, adjust engine speed is 800r/min simultaneously, lower to moving phase using what prepare, pump in post with the flow velocity of 3ml/min, after the running balance of post internal solvent Establishing, extracting sample solution is by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after sample introduction 4h, main frame is stopped to rotate, stationary phase is released, stationary phase retention rate is 60%, obtain the agate coffee alkaloid that purity is 80%.
After the agate coffee alkaloid obtained by countercurrent purification is concentrated, upper neutral alumina column chromatography, carries out wash-out with benzene-chloroform according to 1:1, collects containing alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography, recycling design, concentrated, dry, obtain the agate coffee alkaloid compound that purity is 96%.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column is Waters C18 pillar (5 μm, 4.6mm × 250mm), and column temperature is 30 DEG C; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Detection time is 30min, and determined wavelength is 280nm.
Embodiment 3: the alkaloidal preparation of agate coffee
Get pueraria root powder 3kg, add 45L50% methyl alcohol, mixing, lixiviate 15min, filter, by residue lixiviate twice as stated above again, merging filtrate reclaim under reduced pressure, to without alcohol taste, obtains medicinal extract.Dissolve above-mentioned medicinal extract with 3%HCl, filter, filtrate uses sodium carbonate adjust pH to 11 again, is then extracted with ethyl acetate, extraction liquid concentrating under reduced pressure, dry, obtains agate coffee alkaloid crude extract.
By normal hexane: ethyl acetate: ethanol: water 6:3:5:7 ratio mixing by volume, in separating funnel, isolate phase up and down after hold over night, and ultrasonic degas 30min is for subsequent use.Get 400mg agate coffee alkaloid crude extract be dissolved in 20ml isopyknic up and down mutually in, vibration make it to dissolve completely, be separated in order to high speed adverse current chromatogram.Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph with the flow velocity of 25ml/min, ON cycle water-bath to set thermostat temperature be 25 DEG C, adjust engine speed is 900r/min simultaneously, lower to moving phase using what prepare, pump in post with the flow velocity of 2.5ml/min, after the running balance of post internal solvent Establishing, extracting sample solution is by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after sample introduction 2.5h, main frame is stopped to rotate, stationary phase is released, stationary phase retention rate is 65%, obtain the agate coffee alkaloid that purity is 85%.
Upper alkali alumina column chromatography after the agate coffee alkaloid obtained by countercurrent purification is concentrated, with chloroform-methanol according to 10:0,8:2 carries out gradient elution, collect containing alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography, recycling design, concentrated, dry, obtain the agate coffee alkaloid compound that purity is 99%.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column is Waters C18 pillar (5 μm, 4.6mm × 250mm), and column temperature is 30 DEG C; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Detection time is 30min, and determined wavelength is 280nm.
Embodiment 4: the alkaloidal preparation of agate coffee
Get pueraria root powder 2kg, add 36L80% ethanolic soln, mixing, refluxing extraction 40min, filter, by residue more as stated above refluxing extraction once, merging filtrate reclaim under reduced pressure to without alcohol taste, obtains medicinal extract.With the above-mentioned medicinal extract of 0.5% sulfuric acid dissolution, filter, filtrate uses ammoniacal liquor adjust pH to 12 again, then uses chloroform extraction, extraction liquid concentrating under reduced pressure, dry, obtains agate coffee alkaloid crude extract.
By normal hexane: ethyl acetate: ethanol: water 6:2:3:7 ratio mixing by volume, in separating funnel, isolate phase up and down after hold over night, be above added to the triethylamine of 5mM, under be added to the HCl of 10mM, and ultrasonic degas 20min is for subsequent use.Get 300mg agate coffee alkaloid crude extract be dissolved in 15ml isopyknic up and down mutually in, vibration make it to dissolve completely, be separated in order to high speed adverse current chromatogram.Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph with the flow velocity of 15ml/min, ON cycle water-bath to set thermostat temperature be 18 DEG C, adjust engine speed is 750r/min simultaneously, lower to moving phase using what prepare, pump in post with the flow velocity of 2ml/min, after the running balance of post internal solvent Establishing, extracting sample solution is by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after sample introduction 6h, main frame is stopped to rotate, stationary phase is released, stationary phase retention rate is 55%, obtain the agate coffee alkaloid that purity is 70%.
After the agate coffee alkaloid obtained by countercurrent purification is concentrated, upper silica gel column chromatography, carries out wash-out with benzene-chloroform according to 1:2, collects containing alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography, recycling design, concentrated, dry, obtain the agate coffee alkaloid compound that purity is 98.6%.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column is Waters C18 pillar (5 μm, 4.6mm × 250mm), and column temperature is 30 DEG C; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Detection time is 30min, and determined wavelength is 280nm.
Embodiment 5: the preparation of agate coffee alkaloid tablet
The agate coffee alkaloid 120g of preparation in Example 1, adds Vltra tears 160g, PVP K30 55g, stearyl alcohol 160g, Magnesium Stearate 5g, mixes, dry granulation, compressing tablet, makes respective tablets (1000), and every sheet is containing agate coffee alkaloid 100mg.
Embodiment 6: the preparation of agate coffee alkaloid capsule
The agate coffee alkaloid 100g of preparation in Example 2,50 DEG C of dryings, grind, cross 80 mesh sieves, added medical starch 97g and the Magnesium Stearate 3g of 80 mesh sieves, mixing, make softwood with 80% appropriate amount of ethanol, cross 30 mesh sieve particles, dry, moisture is made to be less than 5%, cross the whole grain of 40 mesh sieve, packing, containing agate coffee alkaloid 0.1g in every capsules, with aluminium plastic composite packaging, to obtain final product.
Embodiment 7: the alkaloidal antitumor action of agate coffee
Agate coffee alkaloid prepared by Example 3 is made into certain density medicine for test, and the cancer cells of test comprises human cervical carcinoma cell lines Hela, human lung adenocarcinoma epithelial cell strain A549, the former malignant myeloid cell lines K562 of the chronic marrow of people, hepatoma cell strain HUH7, breast cancer cell line mcf-7.
To take the logarithm cell in vegetative period, trysinization, adjustment cell count to 1 × 105/ml; Joined by cell suspension in 96 orifice plates, each hole adds cell 100 μ l, 5%CO
2, cultivate 24h for 37 DEG C.Add the agate coffee alkaloid solution 100 μ l of different concns, each concentration establish 5 parallel, contrast add full nutrient solution 100 μ l, continue cultivate 48hr; Every hole adds each 20 μ l(5mg/ml of MTT solution), continue to cultivate 4h; Abandon nutrient solution, every hole adds dimethyl sulfoxide (DMSO) 150 μ l, gently after vibration, and the OD value in every hole under using enzyme linked immunological instrument to measure 492nm.
The inhibiting rate %=[mean OD value that (mean OD value that the mean OD value that control group measures-dosing group measures)/control group measures] × 100% of tumour cell
Table 1 agate coffee alkaloid is to half inhibiting rate of different tumor cell proliferation
| Tumor cell line | IC50（ug/ml） |
| Human cervical carcinoma cell lines Hela | 3.5 |
| Human lung adenocarcinoma epithelial cell strain A549 | 6.1 |
| The former malignant myeloid cell lines K562 of the chronic marrow of people | 4.2 |
| Hepatoma cell strain HUH7 | 2.7 |
| Breast cancer cell line mcf-7 | 5.3 |
As shown in Table 1, agate coffee alkaloid prepared by the present invention has obvious Cytostatic to tumor cell effect, can be used as inhibition of cell proliferation or antineoplastic agent for antineoplastic research.
Claims (12)
1. a preparation method for agate coffee alkaloid monomer, is characterized in that comprising the following steps:
(1) extract: pueraria root powder is broken, add 5-20 alcoholic solution doubly and extract 1-3 time, filter, united extraction liquid, concentrated, obtain medicinal extract;
(2) removal of impurities: dissolved by the medicinal extract acidic aqueous solution of step (1) gained, filters, filtrate again by lye pH adjustment value to 9-12, then with chloroform or extraction into ethyl acetate, extraction liquid concentrating under reduced pressure, dry, obtain agate coffee alkaloid crude extract;
(3) high-speed counter-current purifying: with alkane: fatty acid ester: fatty alcohol: water is solvent systems, and 4-10:1-13:2-13:1-10 mixes by volume, separates upper and lower phase in separating funnel after hold over night, degassed; Agate coffee alkaloid crude extract phase step (2) obtained, lower phase or the two isopyknic mixing solutions dissolve, and filter, obtain sample solution; Upper as stationary phase using what prepare, pump in the spiral tube of high-speed counter-current chromatograph, adjust engine speed simultaneously, pump in post as moving phase down using what prepare, after the running balance of post internal solvent Establishing, extracting sample solution by sampling valve sample introduction, according to detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, obtain the agate coffee alkaloid of certain purity;
(4) column chromatography purification: the agate coffee alkaloid of certain purity step (3) obtained carries out silica gel column chromatography or alumina column chromatography, with chloroform-methanol or benzene-chloroform for eluting solvent carries out gradient elution, collect containing alkaloidal elutriant, recycling design, concentrated, drying, obtains agate coffee alkaloid compound.
2. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, the alcoholic solution that it is characterized in that in described step (1) is the mixing solutions of methanol solution, ethanolic soln or above-mentioned two kinds of alcohol.
3. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, is characterized in that in described step (1), extracting method is supersound extraction, microwave extraction, lixiviate, refluxing extraction.
4. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, the acidic aqueous solution that it is characterized in that in described step (2) is one or both in hydrochloric acid soln, acetic acid and sulphuric acid soln, wherein, the massfraction of acid is 0.5-5%.
5. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, the alkali lye that it is characterized in that in described step (2) is one or both in sodium hydroxide solution, ammoniacal liquor and sodium carbonate solution.
6. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, is characterized in that the solvent systems in described step (3) is normal hexane: ethyl acetate: ethanol: water=6:3:5:7.
7. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1, is characterized in that the aluminum oxide in described step (4) is neutral alumina or alkali alumina.
8. the preparation method of a kind of agate coffee alkaloid monomer according to claim 1-7 any one, it is characterized in that the liquid-phase chromatography method of the agate coffee alkaloid compound that step (4) obtains is as follows: chromatographic column is 5 μm, the WatersC18 pillar of 4.6mm × 250mm, column temperature is 30 DEG C; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Detection time is 30min, and determined wavelength is 280nm.
9. the agate coffee alkaloid monomer that according to claim 8 prepared by preparation method is preparing the application in antitumor drug.
10. the application of agate coffee alkaloid monomer according to claim 9, is characterized in that described medicine is with agate coffee alkaloid for main raw material, the different formulation made from pharmaceutically acceptable auxiliary material combination.
The application of 11. agate coffee alkaloid monomers according to claim 10, is characterized in that pharmaceutically acceptable auxiliary material is carboxylic propyl methocel, polyvidone, stearyl alcohol, hexadecanol, Magnesium Stearate, Microcrystalline Cellulose, starch, N.F,USP MANNITOL, sodium bicarbonate, amino acid and VITAMIN.
The application of 12. agate coffee alkaloid monomers according to claim 10, is characterized in that formulation comprises tablet, capsule, granule, dripping pill and injection.
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Families Citing this family (16)
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|---|---|---|---|---|
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| CN104513173B (en) * | 2013-09-30 | 2018-03-06 | 中国科学院过程工程研究所 | A kind of Maca extract, Its Preparation Method And Use |
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| CN106666736A (en) * | 2016-07-11 | 2017-05-17 | 周晗生 | Extraction method for maca extract |
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| CN107459474A (en) * | 2017-09-15 | 2017-12-12 | 陕西科技大学 | A kind of alkaloid and its extracting method and application |
| CN110632238B (en) * | 2019-09-26 | 2021-10-08 | 吕梁学院 | Method for evaluating oxidation resistance of alkaloid in rice bran by TLC-CMS technology |
| CN114621224B (en) * | 2022-03-23 | 2023-08-01 | 云南省农业科学院质量标准与检测技术研究所 | Maca alkaloid and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6267995B1 (en) * | 1999-03-03 | 2001-07-31 | Pure World Botanicals, Inc. | Extract of Lepidium meyenii roots for pharmaceutical applications |
| CN1473594A (en) * | 2003-07-01 | 2004-02-11 | 华中科技大学 | Extract of Maka root |
| CN1684680A (en) * | 2002-08-14 | 2005-10-19 | 纯净世界植物学股份有限公司 | Imidazole alkaloids from lepidium meyenii and methods of usage |
| CN101756965A (en) * | 2010-01-29 | 2010-06-30 | 华中科技大学 | Application of maca imidazole alkaloid in preparation of cardiovascular drugs |
Family Cites Families (1)
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6267995B1 (en) * | 1999-03-03 | 2001-07-31 | Pure World Botanicals, Inc. | Extract of Lepidium meyenii roots for pharmaceutical applications |
| CN1684680A (en) * | 2002-08-14 | 2005-10-19 | 纯净世界植物学股份有限公司 | Imidazole alkaloids from lepidium meyenii and methods of usage |
| CN1473594A (en) * | 2003-07-01 | 2004-02-11 | 华中科技大学 | Extract of Maka root |
| CN101756965A (en) * | 2010-01-29 | 2010-06-30 | 华中科技大学 | Application of maca imidazole alkaloid in preparation of cardiovascular drugs |
Non-Patent Citations (4)
| Title |
|---|
| Imidazole Alkaloids from Lepidium meyenii;Baoliang Cui等;《Journal of Natural Product》;20030715;第66卷(第8期);第1101-1103页 * |
| New Alkamides from Maca (Lepidium meyenii);Jianping Zhao等;《Journal of Agricultural and Food Chemistry》;20050119;第53卷(第3期);第690-693页 * |
| 超声波提取玛咖生物碱的工艺研究;杜广香等;《广东农业科学》;20110214(第3期);第74-76页 * |
| 金文闻.药食两用植物玛咖(Lepidium meyenii)的功效物质研究.《中国博士学位论文全文数据库》.2011, * |
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