CN1396161A - Process for purifying swainsonine in Feng grass - Google Patents
Process for purifying swainsonine in Feng grass Download PDFInfo
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- CN1396161A CN1396161A CN 02114590 CN02114590A CN1396161A CN 1396161 A CN1396161 A CN 1396161A CN 02114590 CN02114590 CN 02114590 CN 02114590 A CN02114590 A CN 02114590A CN 1396161 A CN1396161 A CN 1396161A
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- trihydroxyoctahydroindolizidine
- crude product
- chloroform
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- alkaloid
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- FXUAIOOAOAVCGD-FKSUSPILSA-N swainsonine Chemical compound C1CC[C@H](O)[C@H]2[C@H](O)[C@H](O)CN21 FXUAIOOAOAVCGD-FKSUSPILSA-N 0.000 title description 2
- 229960005566 swainsonine Drugs 0.000 title description 2
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- Medicines Containing Plant Substances (AREA)
Abstract
A process for purifying swainsonic contained in loco weed includes extracting macropolar alkaloid crude product by cationic exchange, further extracting with alkaline chloroform, silica gel chromatography to obtain swainsonic crude product and finally sublimating by steps. Its advantages are low cost and high purity.
Description
One, affiliated field
The invention belongs to the natural product chemistry research field, relate to natural product extraction, separation and authentication method, the purifying technique of trihydroxyoctahydroindolizidine in particularly a kind of loco weed.
Two, background technology
Loco weed (locoweed) is the general name that the pulse family whin belongs to (Oxytopis) and Astragalus (Astragalus) poisonous plants, is the most serious poisonous weeds of harm pasture animal husbandry in the world wide.In countries such as Russia, Canada, Morocco, Australia, Mexico, Spain, Brazil, Egypt, Iceland distribution is arranged all.In China, loco weed class plant has 44 kinds, mainly is distributed in provinces and regions such as the Inner Mongol, Ningxia, Gansu, Qinghai, Xinjiang, Tibet, Shaanxi, Sichuan, and distribution area surpasses 4,000,000 hectares, and the financial loss that caused because of loco weed poisons every year is just up to more than 1,200,000,000 yuan.Therefore, worldwide, the research of relevant loco weed and locoism has become numerous scholars' research focus.Abroad, about existing more than the 80 year history of the research of loco weed toxic ingredient, breakthrough starts from Australian scholar Colegate1979 and isolates indoles with the a-mannosidase first as toolenzyme from grey swainson pea and decide alkaloid-trihydroxyoctahydroindolizidine now.This is that the mankind obtain trihydroxyoctahydroindolizidine (swainsonine) first, and with its definite designation.Nineteen eighty-two American scholar Molyneux isolation identification from the spot pod Radix Astragali and silky crazyweed (Oxytropis sericea) goes out trihydroxyoctahydroindolizidine and nitrogen oxide trihydroxyoctahydroindolizidine (swainsonine N-oxide), and has determined that the toxic ingredient of loco weed is exactly a trihydroxyoctahydroindolizidine.
Domestic research to the loco weed toxic ingredient is started late.Phase early 1980s, domestic scholars has mainly been carried out the resource exploration of meadow loco weed kind, population regional distribution.The mid-80 has begun the Separation Research of loco weed toxic ingredient, Cao Guangrong in 1989 etc. first from China's yellowflower crazyweed herb isolation identification go out trihydroxyoctahydroindolizidine, and prove its main toxic ingredient.Between the more than ten years subsequently, the doctor, the Master degree candidate that mainly are Cao Guangrong seminar and cultivation thereof have systematically carried out research work around loco weeds such as China's Herba Oxytropis Kansuensis, yellowflower crazyweed herb, Astragalus strictus, the wide luxuriant whin variation Radixs Astragali, obtained some breakthroughs in succession, mainly contained: from multiple loco weed class plant, separated and identify trihydroxyoctahydroindolizidine; The quantitative analysis of trihydroxyoctahydroindolizidine in the multiple loco weed class plant; The extraction process and the parameter thereof of trihydroxyoctahydroindolizidine have been improved; This achievement in research obtained Ministry of Agriculture's scientific-technical progress third prize in 1996.
External major technique technology of separating purifying spherosinin is earlier with plant sample lixiviate in organic solvent, extractive substance is crossed cationic exchange coloum and sepharose post again, and last recrystallization obtains trihydroxyoctahydroindolizidine.Its deficiency is that extraction yield is low, not can manufacture.Domestic major technique process aspect of purifying at trihydroxyoctahydroindolizidine still belongs to vacancy.
Three, summary of the invention
At defective that exists in the above-mentioned prior art and deficiency, the objective of the invention is to, the purifying technique of trihydroxyoctahydroindolizidine in a kind of loco weed is provided, this purifying technique adopts cation exchange column chromatography, silica gel column chromatography, technology such as segmentation distillation, obtain trihydroxyoctahydroindolizidine at last, the extraction yield of trihydroxyoctahydroindolizidine is increased, product purity improves.Strengthen the power of striving unexpectedly in market, promote the industrialization of trihydroxyoctahydroindolizidine as exploitations such as antitumor drug and immunostimulants.
To achieve these goals, the technical solution adopted in the present invention is, the purifying technique of trihydroxyoctahydroindolizidine in the loco weed, be characterized in, according to the basic physicochemical property of trihydroxyoctahydroindolizidine, at first adopt base exchange method to extract high polarity alkaloid crude product, use alkaline chloroform extracting again, the extracting part obtains the trihydroxyoctahydroindolizidine crude product through the silica gel column chromatography separation, and last fractional sublimation obtains the pure product of trihydroxyoctahydroindolizidine.
Concrete purifying technique of the present invention divides following five stages to finish:
The first step, medicinal extract extracts the stage: extract the total medicinal extract of organic composition with industrial methanol from loco weed class plant.
Second step, the coarse biometric alkaline extraction stage: adopt base exchange method to extract coarse biometric alkali.
In the 3rd step, the trihydroxyoctahydroindolizidine crude product extracts the stage: adopt solvent extraction process to extract the trihydroxyoctahydroindolizidine crude product.
In the 4th step, trihydroxyoctahydroindolizidine extracts the stage: adopt silica gel column chromatography to extract trihydroxyoctahydroindolizidine.
The 5th step, trihydroxyoctahydroindolizidine purification phase: adopt the fractional sublimation method to extract the pure product of trihydroxyoctahydroindolizidine.
Purifying technique extraction cost of the present invention is low, the product purity height, and the trihydroxyoctahydroindolizidine extraction yield obviously is better than foreign technology.
Four, description of drawings
Fig. 1 is a trihydroxyoctahydroindolizidine purifying technique schema of the present invention;
Fig. 2 is a trihydroxyoctahydroindolizidine spectroscopic data graphic representation of the present invention; Spectroscopic data: UV λ
Max: 289
(log4.67)，249(log3.83)；
Fig. 3 is a trihydroxyoctahydroindolizidine IR spectrogram of the present invention; Spectroscopic data: IR
Max(KBr) cm
-1: 3431,3370 (OH), 2946,2806 (CH), 2806～2725 (Bohlman band), 1074 absorption peaks such as (C-O);
Fig. 4 is a trihydroxyoctahydroindolizidine MS spectrogram; Spectroscopic data: EIMSm/e:173 (M
+), 155 (M-H
2O), 138 (M-H
2O-OH), 116,115,96,84,72,43 (parent nucleus cleaved fragment peaks).
Five, embodiment
For a more clear understanding of the present invention, the present invention is described in further detail below in conjunction with drawings and Examples.
The present invention is a material with loco weed class plant-yellowflower crazyweed herb abundant on the NORTHWEST CHINA grassland, Herba Oxytropis Kansuensis, glabrous crazyweed, Astragalus strictus, the variation Radix Astragali etc., basic physicochemical property according to trihydroxyoctahydroindolizidine, adopt base exchange method to extract high polarity alkaloid crude product, use alkaline chloroform extracting again, the extracting part obtains the trihydroxyoctahydroindolizidine crude product through the silica gel column chromatography separation, and last fractional sublimation obtains the pure product of trihydroxyoctahydroindolizidine.
1. the purification principle of trihydroxyoctahydroindolizidine
Trihydroxyoctahydroindolizidine belongs to western pyridine Alkaloid in the poly-hydroxy indoles, and molecular weight is little, and polarity is big, soluble in water, methyl alcohol, ethanol, alkaline chloroform etc., and the easy moisture absorption, and have the distillation characteristic.Present technique technology is according to the basic physicochemical property of trihydroxyoctahydroindolizidine, at first adopt base exchange method to extract high polarity alkaloid crude product, use alkaline chloroform extracting again, the extracting part obtains the trihydroxyoctahydroindolizidine crude product through the silica gel column chromatography separation, and last fractional sublimation obtains the pure product of trihydroxyoctahydroindolizidine.2. the trihydroxyoctahydroindolizidine operational path of purifying
The first step: organic total composition extracts in the plant sample (loco weed grass meal).Take by weighing a certain amount of loco weed grass meal, with 5-10 doubly measure industrial methanol repeatedly cold soaking extract 4-6 time, each cold soaking 7 days filters, the merging methanol solution, reclaim under reduced pressure methyl alcohol gets medicinal extract.
Second step: the alkaloid crude product extracts.Reclaim the medicinal extract of methyl alcohol gained, with doubly (weight/volume) amount 1N hydrochloric acid repeated multiple times dissolving of 10-50, filter, to the last an acid liquid alkaloid is checked till the feminine gender.Merging filtrate is transferred pH4-5 with strong aqua, and by the hydrogen type cation exchange resin post, it is colourless earlier being eluted to effluent liquid with deionized water, uses 1N ammoniacal liquor wash-out again, collects alkaline eluant, reclaims solvent, drain the alkaloid crude product.
The 3rd step: the trihydroxyoctahydroindolizidine crude product extracts.The alkaloid crude product that second step obtained is used earlier the analytical pure dissolve with methanol, filters, and residue is abandoned or adopted, and methanol solution decompression and solvent recovery, gained medicinal extract are used alkaline chloroform extracting again, and combined chloroform liquid reclaims chloroform at last and gets the trihydroxyoctahydroindolizidine crude product.
The 4th step: trihydroxyoctahydroindolizidine separates.Adopt silica gel column chromatography, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water gradient elution, TLC detects, and merges the trihydroxyoctahydroindolizidine part, and decompressing and extracting gets buff powder sample trihydroxyoctahydroindolizidine.
The 5th step: trihydroxyoctahydroindolizidine purifying.The buff powder sample trihydroxyoctahydroindolizidine that the 4th step obtained adopts the fractional sublimation method to obtain the pure product of trihydroxyoctahydroindolizidine.
The contriver has provided following embodiment, but the invention is not restricted to these embodiment.
Embodiment 1: the extraction of trihydroxyoctahydroindolizidine in the Herba Oxytropis Kansuensis
The first step, medicinal extract extracts the stage: Herba Oxytropis Kansuensis hay powder 5kg, extract 4 times with 30 liters of industrial methanol cold soakings, each cold soaking 7 days filters, and merges methanol solution, and reclaim under reduced pressure methyl alcohol obtains total medicinal extract 680g, and paste-forming rate is 13.6%.
In second step, the total medicinal extract of coarse biometric alkaline extraction stage: 680g grinds molten with 6800mL1N hydrochloric acid repeated multiple times, filters, and acid liquid improvement bismuth potassium iodide inspection to the last merges acid liquid till being negative and reacting.The gained acid liquid is transferred pH to 4-5 with strong aqua, and by the hydrogen type cation exchange resin post, 100 times of amount deionized waters with applied sample amount are eluted to colourless earlier, again with 20 times of amount 1N ammoniacal liquor wash-outs, collect the basic stream fluid, decompression and solvent recovery gets 62.56g alkaloid crude product, and going out alkali content is 1.3%.
The 3rd step, the trihydroxyoctahydroindolizidine crude product extracts the stage: 62.56g alkaloid crude product is used earlier the analytical pure dissolve with methanol, filter, reclaim solvent, medicinal extract is used alkaline chloroform extracting again, till extract can't check trihydroxyoctahydroindolizidine, merges extract, the reclaim under reduced pressure chloroform extraction goes out trihydroxyoctahydroindolizidine crude product 4g, and extraction yield is 0.08%.
In the 4th step, trihydroxyoctahydroindolizidine extracts the stage: the column chromatography silica gel mixed sample of 4g trihydroxyoctahydroindolizidine crude product and equivalent, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water gradient elution, every 20mL collects 1 part, collects 300 components altogether, TLC checks that merging is similar, extracts trihydroxyoctahydroindolizidine 0.5g.
The 5th step trihydroxyoctahydroindolizidine purification phase: the 0.5g trihydroxyoctahydroindolizidine vacuumizes in special sublimation pipe and draws driedly, puts oil bath, adopts the fractional sublimation method to obtain the pure product 75mg of trihydroxyoctahydroindolizidine, and extraction yield is 0.0015%.
Embodiment 2: the extraction of trihydroxyoctahydroindolizidine in the yellowflower crazyweed herb
The first step medicinal extract extracts the stage: yellowflower crazyweed herb hay powder 11.5kg, extract 6 times with 70 liters of industrial methanol cold soakings, and each cold soaking 7 days filters, and merges methanol solution, and reclaim under reduced pressure methyl alcohol obtains total medicinal extract 1600g, and paste-forming rate is 13.9%.
The second coarse biometric alkaline extraction stage in step: the total medicinal extract of 1600g grinds molten with 16000mL1N hydrochloric acid repeated multiple times, filters, and acid liquid improvement bismuth potassium iodide inspection to the last merges acid liquid till being negative and reacting.The gained acid liquid is transferred pH to 4-5 with strong aqua, and by the hydrogen type cation exchange resin post, 100 times of amount deionized waters with applied sample amount are eluted to colourless earlier, again with 20 times of amount 1N ammoniacal liquor wash-outs, collect the basic stream fluid, decompression and solvent recovery gets 127g alkaloid crude product, and going out alkali content is 1.1%.
The 3rd step trihydroxyoctahydroindolizidine crude product extracts the stage: 127g alkaloid crude product is used earlier the analytical pure dissolve with methanol, filter, reclaim solvent, medicinal extract is used alkaline chloroform extracting again, till extract can't check trihydroxyoctahydroindolizidine, merge extract, the reclaim under reduced pressure chloroform extraction goes out trihydroxyoctahydroindolizidine crude product 5g, and extraction yield is 0.0043%.
The 4th step trihydroxyoctahydroindolizidine extracts the stage: the column chromatography silica gel mixed sample of g trihydroxyoctahydroindolizidine crude product and equivalent, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water gradient elution, every 30mL collects 1 part, collect 400 components altogether, TLC checks that merging is similar, extracts trihydroxyoctahydroindolizidine 0.58g.
The 5th step trihydroxyoctahydroindolizidine purification phase: the 0.58g trihydroxyoctahydroindolizidine vacuumizes in special sublimation pipe and draws driedly, puts oil bath, adopts the fractional sublimation method to obtain the pure product 105mg of trihydroxyoctahydroindolizidine, and extraction yield is 0.0009%.
Embodiment 3: the extraction of trihydroxyoctahydroindolizidine in the Herba Oxytropis Kansuensis
The first step medicinal extract extracts the stage: Herba Oxytropis Kansuensis hay powder 20kg, extract 4 times with 100 liters of industrial methanol cold soakings, and each cold soaking 5 days filters, and merges methanol solution, and reclaim under reduced pressure methyl alcohol obtains total medicinal extract 2230g, and paste-forming rate is 11.2%.
The second coarse biometric alkaline extraction stage in step: the total medicinal extract of 2230g grinds molten with 15000mL1N hydrochloric acid repeated multiple times, filters, and acid liquid improvement bismuth potassium iodide inspection to the last merges acid liquid till being negative and reacting.The gained acid liquid is transferred pH to 4-5 with strong aqua, and by the hydrogen type cation exchange resin post, 100 times of amount deionized waters with applied sample amount are eluted to colourless earlier, again with 30 times of amount 1N ammoniacal liquor wash-outs, collect the basic stream fluid, decompression and solvent recovery gets 219.1g alkaloid crude product, and going out alkali content is 1.095%.
The 3rd step trihydroxyoctahydroindolizidine crude product extracts the stage: 219.1g alkaloid crude product is used earlier the analytical pure dissolve with methanol, filter, reclaim solvent, get medicinal extract 155.4g, use alkaline chloroform extracting again, till extract can't check trihydroxyoctahydroindolizidine, merge extract, the reclaim under reduced pressure chloroform extraction goes out trihydroxyoctahydroindolizidine crude product 15g, and extraction yield is 0.075%.
The 4th step trihydroxyoctahydroindolizidine extracts the stage: the column chromatography silica gel mixed sample of 15g trihydroxyoctahydroindolizidine crude product and equivalent, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water gradient elution, every 50mL collects 1 part, collect 350 components altogether, TLC checks that merging is similar, extracts trihydroxyoctahydroindolizidine 2.1g.
The 5th step trihydroxyoctahydroindolizidine purification phase: the 2.1g trihydroxyoctahydroindolizidine vacuumizes in special sublimation pipe and draws driedly, puts oil bath, adopts the fractional sublimation method to obtain the pure product 236mg of trihydroxyoctahydroindolizidine, and extraction yield is 0.0012%.
Embodiment 4: the extraction of trihydroxyoctahydroindolizidine in the variation Radix Astragali
The first step medicinal extract extracts the stage: variation Radix Astragali hay powder 2kg, extract 6 times with 10 liters of industrial methanol cold soakings, and each cold soaking 7 days filters, and merges methanol solution, and reclaim under reduced pressure methyl alcohol obtains total medicinal extract 219g, and paste-forming rate is 10.95%.
The second coarse biometric alkaline extraction stage in step: the total medicinal extract of 219g grinds molten with 2500mL1N hydrochloric acid repeated multiple times, filters, and acid liquid improvement bismuth potassium iodide inspection to the last merges acid liquid till being negative and reacting.The gained acid liquid is transferred pH to 4-5 with strong aqua, and by the hydrogen type cation exchange resin post, 100 times of amount deionized waters with applied sample amount are eluted to colourless earlier, again with 30 times of amount 1N ammoniacal liquor wash-outs, collect the basic stream fluid, decompression and solvent recovery gets 22.797g alkaloid crude product, and going out alkali content is 1.14%.
The 3rd step trihydroxyoctahydroindolizidine crude product extracts the stage: 22.797g alkaloid crude product is used earlier the analytical pure dissolve with methanol, filter, reclaim solvent, medicinal extract is used alkaline chloroform extracting again, till extract can't check trihydroxyoctahydroindolizidine, merge extract, the reclaim under reduced pressure chloroform extraction goes out trihydroxyoctahydroindolizidine crude product 2.6g, and extraction yield is 0.13%.
The 4th step trihydroxyoctahydroindolizidine extracts the stage: the column chromatography silica gel mixed sample of 2.6g trihydroxyoctahydroindolizidine crude product and equivalent, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water (70: 26: 2: 2) wash-out, every 10mL collects 1 part, collect 200 components altogether, TLC checks that merging is similar, extracts trihydroxyoctahydroindolizidine 0.3g.
The 5th step trihydroxyoctahydroindolizidine purification phase: the 0.3g trihydroxyoctahydroindolizidine is placed special sublimation pipe, vacuumize and draw driedly, adopt the fractional sublimation method to obtain the pure product 35mg of trihydroxyoctahydroindolizidine, extraction yield is 0.0018%.
Institute of the present invention produce an effect is:
1. spherosin recovery rate
China loco weed class plant has 44 kinds, and distribution area surpasses 4,000,000 hectares, because its ecoenvironment of growth difference, so that the contained spherosin amount of loco weed class plant not of the same race is also variant. Adopt the technology of the present invention that spherosin in the following loco weed is extracted, extract and the results are shown in Table 1.
Spherosin extracts the result in table 1 China main loco weed class plant
Paste-forming rate goes out alkali content spherosin recovery rate
Botanical name sampling position
(%) (%) (%) kansu crazyweed herb (Qinghai) herb 13.6 1.3 0.0015 yellowflower crazyweed herbs (Ningxia) 13.9 1.1 0.0009 maos of lobe whins of herb (Tibet) herb 8.95 0.78 0.0008 wide luxuriant whins (Qinghai) herb 9.95 0.7 0.0012 on the ground on the ground on the ground on the ground
The ground herb
12.6 0.95 0.007 sharp turn whin (Qinghai) glacier whin (Tibet) is herb 8.96 0.85 0.0007 Astragalus strictus (Tibet) the ground herb 10.2 1.0 0.0006 variation Radixs Astragali (Ningxia) ground herbs 10.95 1.14 0.0018 on the ground
2. spherosin technical indicator
Table 2 has been listed the technical performance index of the trihydroxyoctahydroindolizidine of the present invention's extraction.
Trihydroxyoctahydroindolizidine white, needle-shaped crystals, molecular weight are 173,144～145 ℃ of fusing points, purity 〉=98%.UV, IR, GC, MS, NMR spectroscopic data and trihydroxyoctahydroindolizidine standard spectrum data fit like a glove.
Trihydroxyoctahydroindolizidine UV spectroscopic data: UV λ
Max: 289 (log4.67), 249 (log3.83) (referring to Fig. 2).
Trihydroxyoctahydroindolizidine IR spectroscopic data: IR
Max(KBr) cm
-1: 3431,3370 (OH), 2946,2806 (CH), 2806～2725 (Bohlman band), 1074 absorption peaks (referring to Fig. 3) such as (C-O).
Trihydroxyoctahydroindolizidine GC spectroscopic data: contain hydroxyl in the trihydroxyoctahydroindolizidine molecular structure, with the monose structural similitude, so the GC direct injection do not go out the peak, analyzes and must at first be prepared into the silicon ether derivant so will carry out GC.Trihydroxyoctahydroindolizidine dissolves with 100 μ l pyridines, with 100 μ l BSTFA room temperature treatment 30min, forms the TMS derivative.Carry out gas-chromatography at 0.32mm * 30m SE-30 capillary column (tool injection post).Column temperature is increased to 300 ℃ gradually from 120 ℃, and 5 ℃/min, 13.77min goes out the peak.
Trihydroxyoctahydroindolizidine MS spectroscopic data: EIMSm/e:173 (M
+), 155 (M-H
2O), 138 (M-H
2O-OH), 116,115,96,84,72,43 (parent nucleus cleaved fragment peak) (referring to Fig. 4).
3. purifying technique extraction cost of the present invention is low, the product purity height, and the trihydroxyoctahydroindolizidine extraction yield obviously is better than foreign technology.
The technical performance index of table 2 trihydroxyoctahydroindolizidine
| Index system | Technical feature |
| Outward appearance | The white fine needle crystal |
| Fusing point | ????144～145℃ |
| Thin-layer chromatography | R on silica-gel plate fBe worth 0.44 place one red-purple spot is arranged |
| Infrared spectra (IR) | There is typical B ohlman band to absorb |
| Mass spectrum (MS) | Molecular ion peak is 173 |
| Gas-chromatography (GC) | TMS derivative retention time 13.77min |
| Quality guaranteed period | Asepticly vacuumize packing, sealing is stored in-4 ℃ of refrigerators, 3 years quality guaranteed perioves |
| Biochemical characteristic | Efficient, the single-minded inhibitor of a-mannosidase |
| Molecular weight | ????173 |
| Acid dissociation constant | ????Pka?7.4 |
Claims (2)
1. the purifying technique of trihydroxyoctahydroindolizidine in the loco weed, it is characterized in that, basic physicochemical property according to trihydroxyoctahydroindolizidine, at first adopt base exchange method to extract high polarity alkaloid crude product, use alkaline chloroform extracting again, the extracting part obtains the trihydroxyoctahydroindolizidine crude product through the silica gel column chromatography separation, and last fractional sublimation obtains the pure product of trihydroxyoctahydroindolizidine.
2. the purifying technique of trihydroxyoctahydroindolizidine is characterized in that in the loco weed according to claim 1, and its concrete purifying technique carries out according to the following steps:
The first step: organic total composition extracts in the plant sample (loco weed grass meal)
Take by weighing a certain amount of loco weed grass meal, with (5-10) doubly measure industrial methanol repeatedly cold soaking extract 4 times-6 times, each cold soaking 7 days filters, and merges methanol solution, reclaim under reduced pressure methyl alcohol gets medicinal extract;
Second step: the alkaloid crude product extracts
To reclaim the medicinal extract of methyl alcohol gained, with (10-50) doubly (weight/volume) amount 1N hydrochloric acid repeated multiple times dissolving, filter, to the last an acid liquid alkaloid is checked till the feminine gender; Merging filtrate is transferred pH4-pH5 with strong aqua, and by the hydrogen type cation exchange resin post, it is colourless earlier being eluted to effluent liquid with deionized water, uses 1N ammoniacal liquor wash-out again, collects alkaline eluant, reclaims solvent, drain the alkaloid crude product;
The 3rd step: the trihydroxyoctahydroindolizidine crude product extracts
With the alkaloid crude product that second step obtained, use earlier the analytical pure dissolve with methanol, filter, residue is abandoned or adopted, and methanol solution decompression and solvent recovery, gained medicinal extract are used alkaline chloroform extracting again, and combined chloroform liquid reclaims chloroform at last and gets the trihydroxyoctahydroindolizidine crude product;
The 4th step: trihydroxyoctahydroindolizidine separates
Adopt silica gel column chromatography, sample on the dry method, chloroform-methyl alcohol-ammoniacal liquor-water gradient elution, TLC detects, and merges the trihydroxyoctahydroindolizidine part, and decompressing and extracting gets buff powder sample trihydroxyoctahydroindolizidine;
The 5th step: trihydroxyoctahydroindolizidine purifying
With the buff powder sample trihydroxyoctahydroindolizidine that the 4th step obtained, adopt the fractional sublimation method to obtain the pure product of trihydroxyoctahydroindolizidine.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331468C (en) * | 2003-05-21 | 2007-08-15 | 杨凌大农生物技术有限公司 | Plant extractive for treating cancer |
| CN102432609A (en) * | 2011-12-15 | 2012-05-02 | 西藏自治区农牧科学院 | Microwave-assisted extraction process for swainsonine in astragalus strictus |
| CN101270118B (en) * | 2007-03-22 | 2012-05-23 | 贺武利 | Technique for spherosinin purification with continuous column chromatography |
| CN103319481A (en) * | 2013-07-05 | 2013-09-25 | 西藏自治区农牧科学院 | Extracting process of swainsonine in astragalus strictus |
-
2002
- 2002-05-24 CN CNB021145903A patent/CN1182130C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331468C (en) * | 2003-05-21 | 2007-08-15 | 杨凌大农生物技术有限公司 | Plant extractive for treating cancer |
| CN101270118B (en) * | 2007-03-22 | 2012-05-23 | 贺武利 | Technique for spherosinin purification with continuous column chromatography |
| CN102432609A (en) * | 2011-12-15 | 2012-05-02 | 西藏自治区农牧科学院 | Microwave-assisted extraction process for swainsonine in astragalus strictus |
| CN103319481A (en) * | 2013-07-05 | 2013-09-25 | 西藏自治区农牧科学院 | Extracting process of swainsonine in astragalus strictus |
| CN103319481B (en) * | 2013-07-05 | 2014-12-17 | 西藏自治区农牧科学院 | Extracting process of swainsonine in astragalus strictus |
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