CN103275012A - 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 the Separation of Natural Products purification technique field 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 be big at day and night temperature, do not have under the special conditions of fertile, long-term ice-bound and anoxic and grow, have the good reputation of " Peru's genseng ".High nutritive value, rational trophic structure and various active secondary metabolites make the agate coffee have multiple nutrients nourishing function and pharmacological action, can be used for strengthening energy, improve fertility, sexual function improving, treatment climacteric syndrome, rheumatism, dysthymia disorders, anaemia etc., also have anticancer and the leukemia effect.
Studies show that alkaloid is one of main active ingredient of agate coffee, 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 replenish the deficiency of estradiol secretion, alleviates hypothalamus and pituitary gland hyperfunction so can be used as external source estrogen replacement thing; Containing components such as agate coffee acid amides and agate coffee alkene can sexual desire promoting; In addition, the experiment of vitro inhibition cancer cells shows: the FDIGROV cell of maca imidazole Alkaloid lepidiline A has restraining effect (ED50 is 7.39 μ g/ml), lepidiline B then has restraining effect to multiple cancer cells, as the UMUC3 cell, the 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 the agate coffee is very low, only contain 0.2%-0.4%, this has brought difficulty for agate coffee alkaloid extraction and separation and purification, at present, agate coffee alkaloid extraction purifying process complexity in existing document and the patent, the column chromatography purification that needs multistep, as patent " indolizidine alkaloid of agate coffee Lepidium apetalum and uses thereof " (publication number: CN1684680A) earlier agate coffee root is obtained extract with the SDS extraction, then extract is dissolved in sour water-methanol solvate, obtains extract A E with dichloromethane extraction again; AE is gone up Diaion HP-20MG adsorb, carry out wash-out then, obtain four components, i.e. A1, A2, A3 and A4 detect by thin-layer chromatography again, find to contain alkaloid-positive spots in the A2 component; Then A2 is gone up silica gel column chromatography and carry out wash-out, obtain 7 components, namely B1-B7 obtains thick alkaloid fraction with the B5 component through the acidic alumina column chromatography again, this fraction is further carried out purifying by column chromatography and preparation type silica gel H PLC, obtains two kinds of alkaloid compounds.Patent " maca imidazole alkaloid is in the application of preparation in the cardiovascular agent " (publication number: CN101756965A) earlier with pueraria root powder after the sudden strain of a muscle formula is extracted, extracting solution concentrates after filtration, obtains extracting medicinal extract; Then medicinal extract is used 20% dissolve with methanol, last macroporous adsorbent resin is collected and is contained alkaloidal the 3rd elution fraction, concentrates, and obtains the maca imidazole alkaloid crude extract; Crude extract is gone up normal phase column chromatography (neutral alumina column chromatography or purification on normal-phase silica gel column chromatography) with chloroform dissolving back, collect and contain alkaloidal third and fourth elution fraction, concentrate, get medicinal extract; Medicinal extract is gone up C18 reverse phase silica gel post after with dissolve with methanol and is carried out chromatography again, collects second and third that contain maca imidazole alkaloid, four elution fractions, reclaims solvent, obtains the maca imidazole alkaloid composition; At last, the maca imidazole alkaloid composition is gone up solid-phase extraction column (be filler with C8 or C18 silica gel) after with dissolve with methanol be further purified, crystallization obtains the maca imidazole alkaloid monomer.Though above-mentioned patent can obtain highly purified alkaloid, but adopt repeatedly the method for chromatography to obtain monomer component, this method is complex procedures not only, wastes time and energy, and the alkaloidal rate of recovery is lower, and the consumption quantity of solvent is excessive, cost is too high, be not suitable for industrialized production.
Summary of the invention
In order to overcome existing deficiency and defective of separating 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 the agate coffee, to separate the method for preparing agate coffee alkaloid monomer with normal phase chromatography.
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 concentrates, and obtains medicinal extract;
(2) removal of impurities: the medicinal extract of step (1) gained is dissolved with acidic aqueous solution, filters, filtrate again with the lye pH adjustment value to 9-12, then with chloroform or ethyl acetate extraction, the extraction liquid concentrating under reduced pressure, drying obtains agate coffee alkaloid crude extract;
(3) high-speed counter-current purifying: with alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): water is solvent systems, and 4-10:1-13:2-13:1-10 mixes by volume, tells upper and lower phase in separating funnel after the standing over night, the degassing; The agate coffee alkaloid crude extract that step (2) obtains is used phase, descended phase or the two isopyknic mixing solutions to dissolve, filter, obtain sample solution; With going up as stationary phase of preparing, pump in the spiral tube of high-speed counter-current chromatograph, adjust engine speed simultaneously, with pumping in the post as moving phase down of preparing, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, namely get the agate coffee alkaloid of certain purity;
(4) column chromatography purification: the agate coffee alkaloid of the certain purity that step (3) is obtained carries out silica gel column chromatography or alumina column chromatography, be that eluting solvent carries out gradient elution with chloroform-methanol or benzene-chloroform, collection contains alkaloidal elutriant, reclaim solvent, concentrate, drying obtains agate coffee alkaloid compound.
Acidity alcohol solution in the 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, and lixiviate, refluxing extraction is preferably supersound extraction.
Acidic aqueous solution in the described step (2) is one or both in hydrochloric acid soln, acetic acid and the 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 the sodium carbonate solution.
Solvent systems in the described step (3) comprises following two kinds: alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that water constitutes, or the polynary two phase solvent system of setting up on above-mentioned two-phase system basis.
Aluminum oxide in the described step (4) is neutral alumina or alkali alumina.
The application of the agate coffee alkaloid of the present invention's preparation in the preparation antitumor drug, 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 the antioxidant, amino acid, VITAMIN, carbohydrate and the plant milk extract that the present invention are not had obviously influence.Described preparation comprises tablet, capsule, granule, dripping pill and injection.
Method of the present invention is compared with the alkaloidal method of existing preparation agate coffee, has following technical superiority:
(1) the present invention earlier through the chloroform extraction removal of impurities, carries out purifying with high speed adverse current chromatogram with agate coffee alkaloid crude extract again, utilizes normal phase chromatography to be further purified at last and obtains agate coffee alkaloid monomer, and the technological process environmental protection does not have serious harm to environment.
(2) the present invention is simple to operate, and process cycle is short, saves reagent, and the recyclable utilization of eluent has reduced production cost, and the extraction efficiency height is suitable for suitability for industrialized production.
(3) agate coffee alkaloid preparation amount is big.According to required target sample amount, can select the high-speed counter-current chromatograph of different model to separate.Wherein, countercurrent chromatography instrument sample size can reach several grams even tens grams, and the amount of the agate coffee alkaloid monomer that flash liberation goes out is the reducible pilot scale level that reaches.
(4) separate the agate coffee alkaloid compound obtain in the external growth that can effectively suppress kinds of tumor cells, have notable antitumor activity.
Embodiment
Further describe the present invention with embodiment below, be conducive to the understanding to the present invention and advantage thereof, better effects if, but described embodiment only is used for explanation the present invention rather than 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 filters, and with residue supersound extraction twice as stated above again, the merging filtrate reclaim under reduced pressure gets medicinal extract to there not being the alcohol flavor.Dissolve above-mentioned medicinal extract with 1%HCl, filter, filtrate is used sodium hydroxide adjust pH to 9 again, uses chloroform extraction then, the extraction liquid concentrating under reduced pressure, and drying obtains agate coffee alkaloid crude extract.
With normal hexane: ethyl acetate: methyl alcohol: water is the 5:1:5:1 mixed by volume, isolate phase up and down after the standing over night in separating funnel, and ultrasonic degas 20min is standby.Get during 200mg agate coffee alkaloid crude extract is dissolved under the 20ml mutually, vibration makes it to dissolve fully, separates in order to high speed adverse current chromatogram.With going up as stationary phase of preparing, flow velocity with 20ml/min pumps in the spiral tube of high-speed counter-current chromatograph, ON cycle water-bath and to set thermostat temperature be 35 ℃, adjusting engine speed simultaneously is 700r/min, following to moving phase with what prepare, flow velocity with 2ml/min pumps in the post, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after treating sample introduction 3h, stop main frame and rotate, stationary phase is released, the stationary phase retention rate is 70%, gets purity and be 75% agate coffee alkaloid.
The agate coffee alkaloid that the adverse current purifying is obtained concentrates upward silica gel column chromatography of back, with chloroform-methanol according to 10:1,7:3 carries out gradient elution, contain alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography collection, reclaim solvent, concentrate, drying obtains purity and is 98% agate coffee alkaloid compound.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column be Waters C18 pillar (5 μ m, 4.6mm * 250mm), column temperature is 30 ℃; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Be 30min detection time, and the detection 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 filters, and residue is extracted once more as stated above, and the merging filtrate reclaim under reduced pressure gets medicinal extract to there not being the alcohol flavor.With the above-mentioned medicinal extract of 2.5% acetate dissolution, to filter, filtrate is used ammoniacal liquor adjust pH to 10 again, uses ethyl acetate extraction then, the extraction liquid concentrating under reduced pressure, drying obtains agate coffee alkaloid crude extract.
With normal hexane: ethyl acetate: methyl alcohol: water is the 5:5:7:5 mixed by volume, isolates phase up and down in separating funnel after the standing over night, on be added to the triethylamine of 10mM, be added to the HCl of 10mM down, and ultrasonic degas 15min is standby.Get during 300mg agate coffee alkaloid crude extract is dissolved under the 20ml mutually, vibration makes it to dissolve fully, separates in order to high speed adverse current chromatogram.With going up as stationary phase of preparing, flow velocity with 10ml/min pumps in the spiral tube of high-speed counter-current chromatograph, ON cycle water-bath and to set thermostat temperature be 20 ℃, adjusting engine speed simultaneously is 800r/min, following to moving phase with what prepare, flow velocity with 3ml/min pumps in the post, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after treating sample introduction 4h, stop main frame and rotate, stationary phase is released, the stationary phase retention rate is 60%, gets purity and be 80% agate coffee alkaloid.
The agate coffee alkaloid that the adverse current purifying is obtained concentrates upward neutral alumina column chromatography of back, carries out wash-out with benzene-chloroform according to 1:1, contains alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography collection, reclaim solvent, concentrate, drying obtains purity and is 96% agate coffee alkaloid compound.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column be Waters C18 pillar (5 μ m, 4.6mm * 250mm), column temperature is 30 ℃; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Be 30min detection time, and the detection wavelength is 280nm.
Embodiment 3: the alkaloidal preparation of agate coffee
Get pueraria root powder 3kg, add 45L50% methyl alcohol, mixing, lixiviate 15min filters, and with residue lixiviate twice as stated above again, the merging filtrate reclaim under reduced pressure gets medicinal extract to there not being the alcohol flavor.Dissolve above-mentioned medicinal extract with 3%HCl, filter, filtrate is used yellow soda ash adjust pH to 11 again, uses ethyl acetate extraction then, the extraction liquid concentrating under reduced pressure, and drying obtains agate coffee alkaloid crude extract.
With normal hexane: ethyl acetate: ethanol: water is the 6:3:5:7 mixed by volume, isolate phase up and down after the standing over night in separating funnel, and ultrasonic degas 30min is standby.Get 400mg agate coffee alkaloid crude extract be dissolved in 20ml isopyknic up and down mutually in, vibration makes it to dissolve fully, separates in order to high speed adverse current chromatogram.With going up as stationary phase of preparing, flow velocity with 25ml/min pumps in the spiral tube of high-speed counter-current chromatograph, ON cycle water-bath and to set thermostat temperature be 25 ℃, adjusting engine speed simultaneously is 900r/min, following to moving phase with what prepare, flow velocity with 2.5ml/min pumps in the post, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after treating sample introduction 2.5h, stop main frame and rotate, stationary phase is released, the stationary phase retention rate is 65%, gets purity and be 85% agate coffee alkaloid.
The agate coffee alkaloid that the adverse current purifying is obtained concentrates upward alkali alumina column chromatography of back, with chloroform-methanol according to 10:0,8:2 carries out gradient elution, contain alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography collection, reclaim solvent, concentrate, drying obtains purity and is 99% agate coffee alkaloid compound.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column be Waters C18 pillar (5 μ m, 4.6mm * 250mm), column temperature is 30 ℃; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Be 30min detection time, and the detection wavelength is 280nm.
Embodiment 4: the alkaloidal preparation of agate coffee
Get pueraria root powder 2kg, add the 36L80% ethanolic soln, mixing, refluxing extraction 40min filters, with residue more as stated above refluxing extraction once, the merging filtrate reclaim under reduced pressure gets medicinal extract to there not being the alcohol flavor.With the above-mentioned medicinal extract of 0.5% sulfuric acid dissolution, to filter, filtrate is used ammoniacal liquor adjust pH to 12 again, uses chloroform extraction then, the extraction liquid concentrating under reduced pressure, drying obtains agate coffee alkaloid crude extract.
With normal hexane: ethyl acetate: ethanol: water is the 6:2:3:7 mixed by volume, isolates phase up and down in separating funnel after the standing over night, on be added to the triethylamine of 5mM, be added to the HCl of 10mM down, and ultrasonic degas 20min is standby.Get 300mg agate coffee alkaloid crude extract be dissolved in 15ml isopyknic up and down mutually in, vibration makes it to dissolve fully, separates in order to high speed adverse current chromatogram.With going up as stationary phase of preparing, flow velocity with 15ml/min pumps in the spiral tube of high-speed counter-current chromatograph, ON cycle water-bath and to set thermostat temperature be 18 ℃, adjusting engine speed simultaneously is 750r/min, following to moving phase with what prepare, flow velocity with 2ml/min pumps in the post, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, after treating sample introduction 6h, stop main frame and rotate, stationary phase is released, the stationary phase retention rate is 55%, gets purity and be 70% agate coffee alkaloid.
The agate coffee alkaloid that the adverse current purifying is obtained concentrates upward silica gel column chromatography of back, carries out wash-out with benzene-chloroform according to 1:2, contains alkaloidal elutriant in conjunction with liquid chromatography and thin layer chromatography collection, reclaim solvent, concentrate, drying obtains purity and is 98.6% agate coffee alkaloid compound.The alkaloidal liquid-phase chromatography method of agate coffee is as follows: chromatographic column be Waters C18 pillar (5 μ m, 4.6mm * 250mm), column temperature is 30 ℃; Moving phase is acetonitrile: water=90:10, and flow velocity is 0.8ml/min; Be 30min detection time, and the detection wavelength is 280nm.
Embodiment 5: the preparation of agate coffee alkaloid tablet
Get the agate coffee alkaloid 120g of preparation among the embodiment 1, add Vltra tears 160g, 30 POVIDONE K 30 BP/USP 3055g, stearyl alcohol 160g, Magnesium Stearate 5g mixes, dry granulation, compressing tablet is made respective tablets (1000), and every contains agate coffee alkaloid 100mg.
Embodiment 6: the preparation of agate coffee alkaloid capsule
Get the agate coffee alkaloid 100g of preparation among the embodiment 2,50 ℃ of dryings grind, cross 80 mesh sieves, added medical starch 97g and the Magnesium Stearate 3g of 80 mesh sieves, mixing, make softwood in right amount with 80% ethanol, cross 30 mesh sieves particle processed, oven dry, make moisture less than 5%, cross the whole grain of 40 mesh sieves, packing contains agate coffee alkaloid 0.1g in every capsules, use aluminium plastic composite packaging, namely.
Embodiment 7: the alkaloidal antitumor action of agate coffee
The agate coffee alkaloid of getting embodiment 3 preparations is made into two fun gi polysaccharides for test usefulness, and the cancer cells of test usefulness comprises human cervical carcinoma cell strain Hela, human lung adenocarcinoma epithelial cell strain A549, the former leukemia cell line K562 of the chronic marrow of people, hepatoma cell strain HUH7, breast cancer cell line mcf-7.
The cell in vegetative period of taking the logarithm, cell count to 1 * 105/ml is adjusted in trysinization; Cell suspension is joined in 96 orifice plates, and each hole adds cell 100 μ l, 5%CO
2, cultivate 24h for 37 ℃.The agate coffee alkaloid solution 100 μ l that add different concns, each concentration establish 5 parallel, contrast adds full nutrient solution 100 μ l, continues to 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, after the vibration, uses the enzyme linked immunological instrument to measure the OD value in every hole under the 492nm gently.
The average OD value that the average OD value that the inhibiting rate %=[(control group of tumour cell is measured-dosing group is measured)/the average OD value of control group mensuration] * 100%
Table 1 agate coffee alkaloid is to half inhibiting rate of different tumor cell proliferations
| Tumor cell line | IC50（ug/ml） |
| Human cervical carcinoma cell strain Hela | 3.5 |
| Human lung adenocarcinoma epithelial cell strain A549 | 6.1 |
| The former leukemia cell line 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, the agate coffee alkaloid of the present invention's preparation has tangible tumor cell proliferation restraining effect, can be used as inhibition of cell proliferation or antineoplastic agent and is used for antineoplastic research.
Claims (11)
1. alkaloidal preparation method of agate coffee is characterized in that may further comprise the steps:
(1) extract: pueraria root powder is broken, add 5-20 alcoholic solution doubly and extract 1-3 time, filter, united extraction liquid concentrates, and obtains medicinal extract;
(2) removal of impurities: the medicinal extract of step (1) gained is dissolved with acidic aqueous solution, filters, filtrate again with the lye pH adjustment value to 9-12, then with chloroform or ethyl acetate extraction, the extraction liquid concentrating under reduced pressure, drying obtains agate coffee alkaloid crude extract;
(3) high-speed counter-current purifying: with alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): water is solvent systems, and 4-10:1-13:2-13:1-10 mixes by volume, tells upper and lower phase in separating funnel after the standing over night, the degassing; The agate coffee alkaloid crude extract that step (2) obtains is used phase, descended phase or the two isopyknic mixing solutions to dissolve, filter, obtain sample solution; With going up as stationary phase of preparing, pump in the spiral tube of high-speed counter-current chromatograph, adjust engine speed simultaneously, with pumping in the post as moving phase down of preparing, after treating that post internal solvent system is set up running balance, extracting sample solution is by the sampling valve sample introduction, according to the detector spectrogram, collect in conjunction with liquid chromatography and thin layer chromatography, namely get the agate coffee alkaloid of certain purity;
(4) column chromatography purification: the agate coffee alkaloid of the certain purity that step (3) is obtained carries out silica gel column chromatography or alumina column chromatography, be that eluting solvent carries out gradient elution with chloroform-methanol or benzene-chloroform, collection contains alkaloidal elutriant, reclaim solvent, concentrate, drying obtains agate coffee alkaloid compound.
2. the alkaloidal preparation method of a kind of agate coffee according to claim 1 is characterized in that the alcoholic solution in the described step (1) is the mixing solutions of methanol solution, ethanolic soln or above-mentioned two kinds of alcohol.
3. the alkaloidal preparation method of a kind of agate coffee according to claim 1 is characterized in that extracting method comprises supersound extraction, microwave extraction, lixiviate, refluxing extraction in the described step (1).
4. the alkaloidal preparation method of a kind of agate coffee according to claim 1 is characterized in that acidic aqueous solution in the described step (2) is one or both in hydrochloric acid soln, acetic acid and the sulphuric acid soln, and wherein, the massfraction of acid is 0.5-5%.
5. the alkaloidal preparation method of a kind of agate coffee according to claim 1 is characterized in that alkali lye in the described step (2) is one or both in sodium hydroxide solution, ammoniacal liquor and the sodium carbonate solution.
6. the alkaloidal preparation method of a kind of agate coffee according to claim 1, it is characterized in that the solvent systems in the described step (3) comprises following two kinds: alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that water constitutes, or the polynary two phase solvent system of setting up on above-mentioned two-phase system basis.
7. the alkaloidal preparation method of a kind of agate coffee according to claim 1 is characterized in that the aluminum oxide in the described step (4) is neutral alumina or alkali alumina.
8. according to the application of any described agate coffee alkaloid among the claim 1-7 in the preparation antitumor drug.
9. the alkaloidal application of agate coffee according to claim 8 is characterized in that described medicine is is main raw material with agate coffee alkaloid, the different formulation made from the acceptable accessories combination.
10. the alkaloidal application of agate coffee according to claim 9, it is characterized in that acceptable accessories is carboxylic propyl methocel, polyvidone, stearyl alcohol, hexadecanol, Magnesium Stearate, Microcrystalline Cellulose, starch, N.F,USP MANNITOL, sodium bicarbonate, and the antioxidant, amino acid, VITAMIN, carbohydrate and the plant milk extract that the present invention are not had obviously influence.
11. the alkaloidal application of agate coffee according to claim 9 is characterized in that formulation comprises tablet, capsule, granule, dripping pill and injection.
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| CN110632238A (en) * | 2019-09-26 | 2019-12-31 | 吕梁学院 | Method for evaluating oxidation resistance of alkaloid in rice bran by TLC-CMS technology |
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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 |
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