CN102172366A - Method for separating total alkaloids from motherwort by using ion exchange resin - Google Patents
Method for separating total alkaloids from motherwort by using ion exchange resin Download PDFInfo
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Abstract
The invention discloses a method for separating total alkaloids from motherwort by using ion exchange resin, and belongs to the field of Chinese medicines. The method comprises the following steps of: adjusting the pH value of a motherwort extraction liquid to be 1-7, filtering, sieving by using a cation exchange resin column; washing with deionized water until the pH value is 5-7, eluting with a saline solution at appropriate concentration, and testing until no alkaloid is found; and collecting the eluted solution, adding alkali for neutralization, desalting and concentrating and drying to obtain motherwort total alkaloid. The method overcomes shortcomings of low extraction yield, high consumption of organic solvents, excessively high acid and alkali concentrations, complicated processes, inapplicability for large-scale production and the like in the prior art and can be used for high-efficient separation of high-purity motherwort total alkaloids from motherwort extraction liquid.
Description
Technical field
The present invention relates to a kind of in the extraction separation method of pharmaceutically active ingredient, particularly relate to a kind of from Chinese medicine Herba Leonuri extracting solution the method for extraction separation Herba Leonuri total alkaloids, belong to the field of Chinese medicines.
Background technology
Chinese traditional medicine biology alkali is the class important substance that occurring in nature exists, be separated to kind more than 700 at present, they are maximum class determined curative effects in the Chinese medicine, broad-spectrum effective ingredient, especially cancer, hepatopathy and cardiovascular and cerebrovascular vessel, rheumatism major disease etc. there is unique curative effect, be used for anti-inflammation as the berberine in the Rhizoma Coptidis, ephedrine in the Herba Ephedrae is used to relieving asthma, reserpine in the Radix Rauvolfiae is used for blood pressure lowering, galantamine in the Bulbus Lycoridis Radiatae is effective in cure to poliomyelitis sequela, and contained morphine base is famous analgesics in the Semen Papaveris peel; Quinine alkali is valuable antipyretic; Kurarinone class in the Radix Sophorae Flavescentis is effective in cure to hepatitis B; Capsaicin has many physiologically actives and strong and persistent anti-inflammatory analgesic action; Vincristine in the Herba Catharanthi Rosei is used for antitumor etc.Because the increase that modern disease threatens human existence, in recent decades, cardiovascular and cerebrovascular disease, tumor and hepatopathy become the chief reason that threatens the mankind day by day, so such preparation clinical demand amount is very big.
Because the content of alkaloid in Chinese crude drug (most content are 0.01%～1%) on the low side, guarantee the safe and effective of last preparation, must make with extra care purification to it, be prepared into effective site even effective ingredient is used, so the refining purification technique of effective site just seems very important.
" purification " technology of Chinese medicine preparation industry is modernization of Chinese medicine maximum " bottleneck ".Since the backwardness of " purification " technology, the state of Chinese medicine whole still " thick, big, black ".By the innovation purification technique, can reach the purpose of " discarding the dross and selecting the essential ", and the effective ingredient in Chinese that has been purified, then can satisfy various modern formulation requirements, prepare the modern Chinese medicine preparation of triple effect (efficient, quick-acting, long-acting), three little (dosage is little, toxicity is little, side effect little), five convenience (produce, transport, carry, store, use conveniently).
Present stage, contain the extraction of Chinese traditional medicine biology bases medical material, how according to alkaloidal physicochemical property, adopt The suitable solvent such as water, sour water, acid alcohol and alkali alcohol etc., utilize dipping, percolation, decoction, backflow, technology such as ultrasonic that the alkaloids composition is extracted, extraction process is more perfect, and the alkaloid extraction ratio can reach more than 90%.But because the relative amount of alkaloid in Chinese medicine be lower, certainly will introduce a large amount of impurity constituents during extraction, as a large amount of starch, natural gum, pectin, phlegmatic temperament, pigment etc., further be discarded the dross and selected the essential, purification refine brought very big difficulty.At present, the purification process of extensive use mainly is alkalization back organic solvent extractionprocess and macroporous adsorbent resin method (" Chemistry for Chinese Traditional Medicine ", Xiao Chonghou chief editor,, Shanghai science tech publishing house in 1997 in the production of alkaloid purifying process; " in the Chinese herbal medicine alkaloid extraction with separate ", Cai Yanhua, Sichuan chemical industry, 2005. (1) 39).
Organic solvent extractionprocess is to utilize the lipotropy alkaloid to be dissolved in the lipotropy organic solvent, and the water-soluble character of its salt, by the alkaloid soda acid is transformed, utilize organic solvent (commonly used as benzene, chloroform, ether etc.) extraction, remove a large amount of impurity, obtain Chinese medicine total alkaloids effective site at last.Organic solvent extractionprocess is used comparatively extensive, but easily bring impurity such as a large amount of neutrality and nonpolar pigment into after the alkalization, therefore separation efficiency and purity are lower, and have use a large amount of organic solvents (generally extracting 5 times), (extractor efficient is not high in operation inconvenience, and easily emulsifying), not good (organic solvent toxicity is big in safety, and inflammable and explosive) etc. shortcoming, belong to laboratory process, be not suitable for that industry is big produces.
During by macroporous adsorbent resin isolation technics purification Chinese medicine extraction liquid, can selective absorption effective ingredient wherein, remove impurity simultaneously.Compare with technology with traditional impurity-removing method, this law can be dwindled medication dose, improves the quality of preparation.This method to water solublity preferably flavone, saponin etc. good separation and purification effect is arranged, as in Semen Ginkgo class preparation and ginsenoside's preparation, often using at present.But for alkaloidal refining, the deficiency of this law maximum is: at first be the absorption difficulty, last sample medicinal liquid must be a weak solution, and basicity will suit: the low then alkaloid of basicity still is ionization state, and the adsorption rate of resin is low; The basicity height, alkaloid is free type, macromole alkaloid poorly water-soluble and separating out still can't upper prop.So the alkaloid scope that this method is suitable for is very little, and kind very little; Secondly, the nonpolar electrostatic absorption principle of macroporous adsorbent resin is too poor to the separation selectivity of alkaloid effective ingredient and oil-soluble impurities, and in the solvent elution process owing to there is not a specific eluting solvent, alkaloid and a large amount of oil-soluble impurities often are washed down together, and the total alkaloids purity that obtains at last is lower.
And emerging ion exchange resin isolation technics then is the purpose that reaches separation and purification by ion-exchange reactions.At present be: after will containing alkaloidal solution and crossing post,, collect reuse organic solvent extraction total alkaloids behind the eluent with ammoniacal liquor or NaOH eluant solution in the general technology that adopts of the field of Chinese medicines; Perhaps will adsorb resin behind the alkaloid with dipping by lye, reuse organic solvent reflux, extract, total alkaloids.But the said method complex steps is carried out technology such as organic solvent reflux, extract, industrial difficult the realization with organic solvent extraction or with resin, and very low with the alkaloidal efficient of alkali liquor eluting.This method only stays in the level of prepared in laboratory small sample at present, also can't realize big production.
In sum, above method ubiquity cost height, organic solvent consumption are big, poor selectivity, total alkaloids purity is low and can't realize defective such as big production, and therefore, purification technique is still " bottleneck " problem of separating total alkaloids from Chinese medicine.
Summary of the invention
The method that the purpose of this invention is to provide a kind of safe, low-cost, high efficiency extraction separation Chinese medicine Herba Leonuri total alkaloids.
This goal of the invention realizes by following technology: get the extracting solution that contains the Chinese medicine Herba Leonuri, adjust pH value 1～7, filter, getting filtrate is added on the cation exchange resin column, earlier be washed till pH value 5～7,, collect eluent again with 0.5%～20% saline solution eluting with deionized water, through desalting processing, promptly get required Herba Leonuri total alkaloids.
In above-mentioned technical process, the Herba Leonuri extracting solution transferred to proper acidity after, alkaloid is ionized to and is cation, nonbasic substances is not ionized.After extracting solution was crossed cation exchange resin column, the hydrion on ionized alkaloid ion and the resin column exchanged and is adsorbed on the resin column securely.Here the pH value of extracting solution is unsuitable too high or too low: if too high, it is neutral or alkaline that extracting solution is, and alkaloid can not be ionized, and also just can't finish the exchange process on the resin; If cross lowly, the hydrogen ion concentration in the extracting solution is too high, has hindered the hydrion on the resin column and has dissociated, and can not successfully finish the resins exchange process equally, thereby finds in the experiment that the pH value of adjusting extracting solution is 1～7th, and is proper.During with the water elution resin column, select to use deionized water, to guarantee not introduce new ion interference.In elution process, those are not easy to be got off by water elution by the nonbasic substances of resin absorption, thereby come with the alkaloid ion isolation that is attracted on the resin column.After this add saline solution earlier and soak, the cation of salt and alkaloidal cation are fully exchanged, and then with the saline solution eluting.At last, adopt suitable method that the alkaloid in the eluent is separated with salt, obtain the solution that salt then can be made into suitable concentration and reuse as eluent and separate, the while can obtain the higher Herba Leonuri total alkaloids of purity.
This technical process compared with prior art, not only technological process is simple, and its important contribution also is: that has broken that traditional theory thinks can only adopt alkali liquor or ammoniacal liquor eluting alkaloid, do not use simultaneously the organic solvent of high concentration, the acid solution of high concentration, the alkali liquor of high concentration, but adopt saline solution as eluent, to the basic non-corrosiveness of equipment, the equipment of industrial product is required to have reduced, cost reduces greatly, the purpose of the big production of easier realization.
It all is that alkali liquor or ammoniacal liquor carry out eluting that traditional theory is thought in the ionic exchange resin of total alkaloids, perhaps earlier with in the alkali liquor and the free in theory alkaloid of resin column reuse organic solvent reflux, extract,, and the effect in the practical application is very undesirable and be unsuitable for big production.In technical scheme of the present invention, adopt the strong Ca of ion-exchange capacity
2+, NH
4 +, Na
+Saline solution as eluant, thereby the shortcoming of having avoided strong acid-base solution to be inconvenient to operate can make the alkaloid ion that exchanges be dissolved in the eluent rapidly, and be rushed out pillar, has guaranteed the ionic elute effect of alkaloid thus.And the recyclable recycling of the salt in the eluent reduces cost.
Based on above-mentioned technical process, the inventor has carried out further research again, and the parameter of each step of refinement filters out optimal case, and particular content is as follows:
1, the pH value scope of adjustment medicinal liquid is preferably 2～5 before the upper prop, and effect is more obvious.
2, used cation exchange resin can be macroporous type or gel-type strong acid ion exchange resin, can according to circumstances be processed in Hydrogen or the salt type any when reality is used, and is preferably the salt type, as the sodium type.
3, the post of used cation exchange resin column footpath there is no a fixed pattern with the ratio of post height, but considers to produce reality, post footpath: post height=1: 5～can obtain best effect at 1: 10 o'clock.
4, the liquor strength to sample on the resin column is that every ml is equivalent to crude drug 0.1～3g, and concrete condition can determine that if adopt modes such as dipping, percolation, the medicinal liquid that obtains will be rarer according to the pre-treatment mode of medicinal liquid; And if modes such as reflux, extract,, decoction are especially passed through concentration, the concentration of medicinal liquid will be bigger, but so long as in this scope, can both realize sample.Simultaneously, last sample speed is also adjusted according to the concentration of medicinal liquid in good time, satisfies substantially at 0.5～5 times of column volume/hour get final product.
5, go up sample loading mode and be typically chosen in from the post upper end upward sample, medicinal liquid flows through whole pillar by gravity flow power; The inventor finds, if sample on the adverse current, be about to medicinal liquid sample from the pillar bottom, by certain pressure, make medicinal liquid fill with whole pillar, thisly go up the maximum exchange efficient that sample loading mode can be brought into play pillar, when also having avoided from the upper end sample since the alkaloid exchange not exclusively take place shift to an earlier date penetration problem.This point also is the creationary discovery of inventor.
6, the concentration of the saline solution of Jin Paoing is preferably 10%～15%, and the concentration of salt solution that eluting is used is preferably 5%～15%.
7, the time with saline solution immersion resin should be more than 1 hour, generally need not surpass 5 hours, carry out eluting again.Through after leaving standstill, a large amount of alkaloid ions are exchanged, perhaps are in half absorption, half dissociated state, and carry out eluting this moment, and the alkaloid concentration effect is obvious, and elution efficiency obviously improves.
8, eluting can be any in the inorganic salt of sodium, potassium, calcium, ammonium with the salt in the saline solution, is preferably sodium salt, preferably NaCl.
9, the flow velocity of eluent is unsuitable too fast or slow excessively during eluting, if too fast, then the ionic exchange of salt ion and alkaloid is insufficient, and eluent is wasted more; If cross slowly, the alkaloid ion that then disintegrates down may adsorb back on the resin column again, influences elution efficiency.Experiment finds, elution speed remains on 2～6 times of column volumes/hour be advisable.Further preferred, elution speed remains on 4～6 times of column volumes/hour best.
10, said desalination process can be the desalination method that condensing crystallizing desalination, dialysis desalination, membrane filtration are crossed desalination and other various medicine biological field routines in the technology, these methods comparative maturity all at present, and can channelization production.
11, in this method said Herba Leonuri extracting solution can be by dipping, percolation, ultrasonic, microwave or any mode in decocting extract and make, and the solvent that extracts can be in water, sour water, alcoholic solution, the acid ethanol solution any, but all belongs to the conventional extracting method of the present field of Chinese medicines.Distinctive points is, if make with dipping or percolation method, can directly go up sample; Obtain if extract, also will pass through steps such as precipitate with ethanol, filtration or centrifugal remove impurity, to guarantee the smooth of sample with additive method.
Need to prove that above-mentioned preferred technical parameter can carry out combination in any with basic technology, and all can obtain good effect according to the needs of practical situation, the technical solution problem reaches purpose of the present invention.
Below by the contrast experiment advantage of the present invention is described.
One, experimental program:
Get Herba Leonuri medical material 4kg, respectively add 12 times of water gagings and decoct, extract each 2 hours 2 times, the condensed water extract is to 4000ml, add ethanol and make determining alcohol reach 80% back cold preservation, standing over night is filtered, reclaim ethanol, medicinal liquid is concentrated into 1: 1, and this medicinal liquid is four parts, carries out alkaloidal separation by following four kinds of methods respectively and purifies:
(1) ion exchange resin+saline solution eluting: medicinal liquid is regulated pH=3, on handled strong cation-exchanging resin 001 * 2.5 (sodium type well, blade diameter length ratio is 1: 7), last sample speed be 3 times of column volumes/hour, have precipitation to stop to go up sample with the silico-tungstic acid inspection to effluent, water is not eluted to when eluent has color substantially and stops.Reuse 10% sodium chloride salt acid solution eluting, elution speed be 4 times of column volumes/hour, eluent begins to receive when with the silico-tungstic acid inspection precipitation being arranged, receive eluent to do not have with the silico-tungstic acid inspection precipitation up to, merge eluent, be neutralized to about pH=7 desalination (standby) with ammonia, concentrate drying gets Herba Leonuri total alkaloids.
(2) ion exchange resin+reflux, extract: medicinal liquid is regulated pH=3, on handled strong cation-exchanging resin 001 * 2.5 (sodium type, blade diameter length ratio are 1: 7) well, last sample speed be 3 times of column volumes/hour, there is precipitation to stop to go up sample with the silico-tungstic acid inspection to effluent, water is not eluted to when eluent has color substantially and stops eluting, and resin by pouring out in the post, is put in the porcelain dish and dried, it is an amount of to add 10% ammonia, stir, touched damp, be not advisable but do not touch with one's hand with hands.This resin is put in the apparatus,Soxhlet's,, chloroform solution is added anhydrous Na with chloroform reflux, extract, 2 hours
2SO
4After the dehydration, reclaim chloroform, the dry Herba Leonuri total alkaloids that gets to the dried molassed pulpous state.
(3) macroporous adsorbent resin method: medicinal liquid is regulated pH=10, on the DF01 type macroporous adsorbent resin handled well, last sample speed be 4 times of column volumes/hour, have precipitation to stop to go up sample with the silico-tungstic acid inspection to effluent, water is eluted to and stops eluting when eluent does not have color substantially.With alcohol-water (70: 30) is eluant, with 2.5 times of column volumes/hour speed carry out eluting, eluent has precipitation the time to begin to receive with the silico-tungstic acid inspection, receive eluent to do not have with the silico-tungstic acid inspection precipitation up to, merge eluent, reclaim ethanol, drying gets Herba Leonuri total alkaloids.
(4) organic solvent extractionprocess: medicinal liquid is regulated pH=10, divide three extractions with 10 times of amount chloroforms earlier, 10 times of amounts of reuse ethyl acetate extraction three times merges all extracts, and evaporate to dryness gets Herba Leonuri total alkaloids.
Two, the result calculates: distinguish the weight of four kinds of technology gained of weighing total alkaloids, and be divided by with the medical material amount, get the yield of total alkaloids; With acid-base titrations four kinds of method gained total alkaloidss are carried out assay respectively, calculate the purity of total alkaloids.
Three, result's contrast, see the following form:
The effect assessment table of four kinds of technologies
By above comparing result as seen, technical scheme of the present invention can solve deficiency of the prior art, and improve significantly product quality, reduce cost, improve safety and producing feasibility, the present invention has reached goal of the invention.
The specific embodiment
Embodiment 1:
Get Herba Leonuri medical material 1kg, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, centrifugal, regulate pH=3, centrifugal, supernatant is so that 4 times of column volumes/hour by cation exchange resin (001 * 2.5, sodium type, blade diameter length ratio 1: 8), being washed to effluent does not have color.Reuse 15% sodium chloride solution eluting, elution speed be 2 times of column volumes/hour, check negative (effluent drips 10% silico-tungstic acid does not have precipitation) to alkaloid, collect eluent, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets Herba Leonuri total alkaloids.
Embodiment 2:
Get Herba Leonuri medical material 10kg, decoct with water 2 times, each 10 times of amounts, decocted 2 hours, and merged decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 80%, leaves standstill, supernatant reclaims ethanol, adds water to 10000ml, regulates pH=3, centrifugal, supernatant with 3 times of column volumes/hour by last cation exchange resin (001 * 4, the sodium type, blade diameter length ratio 1: 8), being washed to effluent does not have color, and reuse 10% sodium chloride solution soaked resin 3 hours.Reuse 8% sodium chloride eluting, elution speed be 6 times of column volumes/hour, check negative (effluent drips 10% silico-tungstic acid does not have precipitation) to alkaloid, the collection eluent is neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets Herba Leonuri total alkaloids.
Embodiment 3:
Get Herba Leonuri medical material 5kg, add 80% ethanol ultrasonic extraction twice, add 5 times of amounts at every turn, ultrasonic 0.5 hour, merge extractive liquid,, centrifugal, reclaim ethanol, add water to 5000ml, regulate pH=2, centrifugal, supernatant is with 2 times of column volumes/hour by last cation exchange resin (001 * 10, sodium type, blade diameter length ratio 1: 7), being washed to effluent does not have color, and 0.5% hydrochloric acid solution of reuse 20% sodium chloride soaked resin 1 hour.Reuse 8% sodium chloride solution eluting, elution speed be 5 times of column volumes/hour, check negative (effluent drips 10% silico-tungstic acid does not have precipitation) to alkaloid, collect eluent, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets Herba Leonuri total alkaloids.
Embodiment 4:
Get Herba Leonuri medical material 2kg, the hydrochloric acid solution that adds pH=3, microwave extraction 2 times adds 5 times of volume microwave extractioies 0.5 hour at every turn, filter, the filtrate thin up is regulated pH=6 to 1600ml, and is centrifugal, supernatant is with 2 times of column volumes/hour by last cation exchange resin (001 * 15, Hydrogen, blade diameter length ratio 1: 6), being washed to effluent does not have color.Reuse 10% sodium chloride solution eluting, elution speed be 5 times of column volumes/hour, check negative (effluent drips 10% silico-tungstic acid does not have precipitation) to alkaloid, collect eluent, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets Herba Leonuri total alkaloids.
Embodiment 5:
Get Herba Leonuri medical material 1kg, the sour water percolation that adds pH=5, merge the percolate thin up to 10000ml, centrifugal, regulate pH=1, centrifugal, supernatant is with 5 times of column volumes/hour by cation exchange resin (001 * 7, sodium type, blade diameter length ratio 1: 5), being washed to effluent does not have color, and reuse 15% sodium chloride soaked resin 2 hours.Reuse 5% sodium chloride solution eluting, elution speed be 4 times of column volumes/hour, soaked 5 hours behind 2 times of column volumes of eluting, be eluted to alkaloid again and check negative (effluent drips 10% silico-tungstic acid does not have precipitation), collect eluent, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets Herba Leonuri total alkaloids.
Claims (9)
1. method of from the Herba Leonuri extracting solution, separating Herba Leonuri total alkaloids, it is characterized in that this method is: get the Herba Leonuri extracting solution, regulate pH value 1～7, filter, cross cation exchange resin column, be washed till pH value 5～7 with deionized water earlier, again with 0.5%～20% saline solution eluting, collect eluent, through desalting processing, concentrate drying gets Herba Leonuri total alkaloids.
2. the method for separation Herba Leonuri total alkaloids according to claim 1, the pH value scope that it is characterized in that Herba Leonuri extract solution is 2～5.
3. the method for separation Herba Leonuri total alkaloids according to claim 1 is characterized in that sample loading mode is that adverse current is crossed the post method on the medicinal liquid, and saline solution that can 0.5%～25% before with the saline solution eluting soaks resin column.
4. according to the method for claim 1,3 described separation Herba Leonuri total alkaloidss, the concentration of salt solution that it is characterized in that used immersion resin is 10%～15%, and the eluting concentration of salt solution is 5%～15%.
5. the method for separation Herba Leonuri total alkaloids according to claim 1 is characterized in that used salt is any in the inorganic salt of sodium, potassium, calcium, ammonium.
6. the method for separation Herba Leonuri total alkaloids according to claim 5 is characterized in that used salt is sodium salt.
7. the method for separation Herba Leonuri total alkaloids according to claim 1, the time that it is characterized in that used salt solution soaking resin is 1～5 hour, elution speed is 4～6 times of column volumes per hour.
8. the method for separation Herba Leonuri total alkaloids according to claim 1 is characterized in that used resin is Hydrogen or salt type.
9. the Herba Leonuri total alkaloids of each described method preparation in the claim 1～8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106265895A (en) * | 2016-08-08 | 2017-01-04 | 桂林市晟博科技服务有限公司 | A kind of preparation method of Herba Leonuri extract |
| CN109180534A (en) * | 2018-09-10 | 2019-01-11 | 合肥科佳高分子材料科技有限公司 | A kind of method of Total alkaloids eluent desalination |
| CN109806293A (en) * | 2017-11-20 | 2019-05-28 | 成都中医药大学 | Purposes of the motherwort total alkaloid in the drug that preparation has angiogenesispromoting effect |
-
2008
- 2008-08-13 CN CN2008101183150A patent/CN102172366A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106265895A (en) * | 2016-08-08 | 2017-01-04 | 桂林市晟博科技服务有限公司 | A kind of preparation method of Herba Leonuri extract |
| CN109806293A (en) * | 2017-11-20 | 2019-05-28 | 成都中医药大学 | Purposes of the motherwort total alkaloid in the drug that preparation has angiogenesispromoting effect |
| CN109180534A (en) * | 2018-09-10 | 2019-01-11 | 合肥科佳高分子材料科技有限公司 | A kind of method of Total alkaloids eluent desalination |
| CN109180534B (en) * | 2018-09-10 | 2021-04-23 | 合肥科佳高分子材料科技有限公司 | Method for desalting motherwort total alkali eluent |
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Open date: 20110907 |