CN101697986B - Method for separating catharanthus roseus total alkaloids by ion exchange resin - Google Patents
Method for separating catharanthus roseus total alkaloids by ion exchange resin Download PDFInfo
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Abstract
The invention discloses a method for separating traditional Chinese medicine catharanthus roseus total alkaloids, in particular to a method for separating total alkaloids from traditional Chinese medicine catharanthus roseus extract, belonging to the traditional Chinese medicine field. The method includes the following technical scheme: catharanthus roseus extract is taken, pH value is adjusted to be 1-7, filtering is carried out, filtrate is taken and added onto cation exchange resin, eluting by water is firstly carried out to remove impurities, and then 0.5-20% saline solution is used for socking the resin, and then 0.5-20% saline solution containing acid or alcohol is used for eluting until no alkaloid is detected, eluant is collected and added with alkali to be natural, and desalinization treatment and concentrated drying are carried out to obtain the traditional Chinese medicine catharanthus roseus total alkaloids. The method overcomes the defects of low extraction rate, high organic solvent dosage, excessive high acid-base concentration, complex technology and impossible for large production and the like of the prior art and can efficiently separate high purity catharanthus roseus total alkaloids in traditional Chinese medicine catharanthus roseus extract.
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 Catharanthi Rosei extracting solution the method for extraction separation catharanthus roseus total alkaloids, belong to the field of Chinese medicines.
Background technology
Chinese traditional medicine biology alkali is one type of important substance that occurring in nature exists; Be separated to kind more than 700 at present, they are maximum one type of 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; Berberine as in the Rhizoma Coptidis is used for anti-inflammation, and the ephedrine in the Herba Ephedrae is used to relieving asthma, and the 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; The total alkaloids for treating senile dementia; Capsaicin has many physiologically actives and strong and persistent anti-inflammatory analgesic action; Vincaleucoblastine in the Herba Catharanthi Rosei and the vincristine in the Herba Catharanthi Rosei are 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 the modernization of Chinese medicine maximum " bottleneck ".Since the backwardness of " purification " technology, the state of Chinese medicine whole still " thick, big, black ".Through the innovation purification technique; Can reach the purpose of " discarding the dross and selecting the essential "; And by sublimed effective ingredient in Chinese; 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, storage, convenient in application).
Present stage; Contain the extraction of Chinese traditional medicine biology bases medical material; How, 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 according to alkaloidal physicochemical property; 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, like a large amount of starch, natural gum, pectin, phlegmatic temperament, pigment etc., further 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; Through the alkaloid soda acid is transformed; Utilize organic solvent (commonly used like 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 be prone to bring into impurity such as a large amount of neutrality and nonpolar pigment after the alkalization, so separation efficiency and purity are lower, and having a large amount of organic solvents (generally extracting 5 times) of use, operation, inconvenient (extractor efficient is not high; And easy 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 through 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 maximum deficiency of this law is: at first be the absorption difficulty, last appearance 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; Basicity is high, and alkaloid is free type, and 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 often is washed down with a large amount of oil-soluble impuritieses 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 through 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 difficult realizations in industry such as organic solvent reflux, extract, 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 is high, organic solvent consumption is 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 catharanthus roseus total alkaloids.
This goal of the invention realizes through following technology: get the extracting solution that contains the Chinese medicine Herba Catharanthi Rosei, adjustment pH value 1 to 7 filters; Getting filtrating and being added on the cation exchange resin column,, soaking resin column with 0.5%～20% saline solution more earlier with the water elution remove impurity; Again to contain acid or alcoholic acid saline solution eluting; Collect eluent,, promptly get required catharanthus roseus total alkaloids through desalting processing.
In above-mentioned technical process, the Herba Catharanthi Rosei extracting solution transferred to proper acidity after, alkaloid is ionized to and is cation, nonbasic substances is not by ionization.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 also just can't be accomplished the exchange process on the resin by ionization; 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 accomplish the resins exchange process equally, thereby the pH value of finding the adjustment extracting solution in the experiment is 1 to 7 to be proper.During with the water elution resin column, select to use deionized water, to guarantee not introduce new ion interference.Loyal in elution process, those are not easy to 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 again and add a spot of acid solution in saline solution or the saline solution and soak, the cation of salt and alkaloidal cation are fully exchanged, and then to contain the saline solution eluting of acid or alcohol.Because the cation concn in the eluent is very high at this moment, it will combine with resin on competitive ground, thereby the alkaloid ion exchange that will be adsorbed on the resin column is got off.Consider that the alkaloid of replacing can saltout in the saline solution of high concentration, so add an amount of acid or pure in the eluent, make the alkaloid dissolving that exchange, and, accomplish alkaloidal enrichment process with eluent outflow resin column.At last, adopt suitable method that the alkaloid in the eluent is separated with salt, and separation obtains the solution that salt then can be made into suitable concentration and reuses as eluent, the while can obtain the higher catharanthus roseus 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 (containing diluted acid or rare alcohol) as eluent, to the basic non-corrosiveness of equipment, the equipment of industrial product is required to have reduced; Cost reduces greatly, more is prone to realize big purpose of producing.
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 inappropriate 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 adds simultaneously diluted acid (as with containing Ca in eluent
2+Salt, hydrochloric acid is selected in acid) or rare alcohol, the alkaloid ion that exchanges is dissolved in the acid solution rapidly, and is rushed out pillar, 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 following:
1, the pH value scope of adjustment medicinal liquid is preferably 2 to 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, when reality is used, can according to circumstances be processed in Hydrogen or the salt type any, is preferably salt type such as sodium type or ammonium type.
3, the post of used cation exchange resin column footpath does not have a fixed pattern with the ratio of post height, but consider to produce actual, post directly: can obtain best effect during post height=1:5～1:10.
4, the liquor strength to appearance 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 appearance.Simultaneously, last appearance speed is also adjusted according to the concentration of medicinal liquid in good time, satisfies basically 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 appearance, medicinal liquid leans on gravity flow power and flows through whole pillar; The inventor finds; If appearance on the adverse current; Be about to medicinal liquid appearance from the pillar bottom,, make medicinal liquid fill with whole pillar through certain pressure; 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 appearance 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, can contain hydrochloric acid or the sulphuric acid of 0-0.5% in the saline solution that the immersion resin is used, promptly the solvent of this saline solution is hydrochloric acid or the sulphuric acid below the water or 0.5% (containing), can obtain better exchange effect like this.
The concentration of the saline solution that 7, soaks is preferably 1%～5%, and the concentration of salt solution that eluting is used is preferably 3%～8%.
8, 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.
9, in order to strengthen the eluting power of eluent, in the saline solution that eluting is used, can also add small amount of acid or ethanol, the acid of adding can be hydrochloric acid or sulphuric acid, makes the concentration of acid in the saline solution remain on 0.05%-0.5%; When adding acid or add ethanol separately, make the concentration of alcohol in the saline solution remain on 5%-30%.Above parameter preferably adds hydrochloric acid through after the repetition test, makes solution acid concentration reach 0.1%-0.5%; Or adding ethanol, make the solution determining alcohol reach 10%-20%.
10, eluting can be CaCl with the salt in the saline solution
2, NaCl, NH
4Among Cl, the KCl any is preferably NaCl, NH
4Cl.
11, 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 possibly 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.
12, said desalination process can be that condensing crystallizing desalination, dialysis desalination, membrane filtration are crossed desalination and the conventional desalination method of other various medicine biological fields in the technology, these methods comparative maturity all at present, and can channelization production.
13, in this method said Herba Catharanthi Rosei extracting solution can be through 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 method for distilling of the present field of Chinese medicines.Distinctive points is, if make with dipping or percolation method, can directly go up appearance; Obtain if extract, also will pass through steps such as precipitate with ethanol, filtration or centrifugal remove impurity, to guarantee the smooth of appearance 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 the object of the invention.
Through the contrast experiment advantage of the present invention is described below.
One, experimental program:
Get Herba Catharanthi Rosei medical material 5000g, add 8 times of amounts of sour water of pH=3, soak a night, percolation is collected percolate extremely with till the silico-tungstic acid inspection nothing deposition, merges percolate, and is centrifugal, gets supernatant; Supernatant is five parts, carries out alkaloidal separation by following five kinds of methods respectively and purify:
(1) the acid solution eluting of ion exchange resin+salt: supernatant is regulated pH=3; On handled strong cation-exchanging resin 001 * 2.5 (ammonium type well; Blade diameter length ratio is 1:7), last appearance speed be 3 times of column volumes/hour, have deposition to stop to go up appearance with the silico-tungstic acid inspection to effluent; Water is not eluted to when eluent has color basically and stops, and 0.5% hydrochloric acid solution of reuse 2% ammonium chloride soaks resin.0.5% hydrochloric acid solution eluting of reuse 2% ammonium chloride, elution speed be 4 times of column volumes/hour, eluent begins to receive when with the silico-tungstic acid inspection deposition being arranged; Receive eluent to do not have with the silico-tungstic acid inspection deposition up to; Merge eluent, be neutralized to about pH=7 desalination (subsequent use) with ammonia; Concentrate drying gets catharanthus roseus total alkaloids.
(2) the alcoholic solution eluting of ion exchange resin+salt: supernatant is regulated pH=3; On handled strong cation-exchanging resin 001 * 2.5 (ammonium type well; Blade diameter length ratio is 1:7), last appearance speed be 3 times of column volumes/hour, have deposition to stop to go up appearance with the silico-tungstic acid inspection to effluent; Water is eluted to and stops eluting when eluent does not have color basically, and reuse 2% ammonium chloride solution soaks resin.10% alcoholic solution eluting of reuse 2% ammonium chloride, again with 4 times of column volumes/hour the speed eluting, eluent begins to receive when with silico-tungstic acid inspection deposition being arranged; Receive eluent to do not have with the silico-tungstic acid inspection deposition up to, merge eluent, desalination (subsequent use); Drying gets catharanthus roseus total alkaloids.
(3) ion exchange resin+reflux, extract: supernatant is regulated pH=3, on handled strong cation-exchanging resin 001 * 2.5 (Hydrogen, blade diameter length ratio are 1:7) well; Last appearance speed be 3 times of column volumes/hour, have deposition to stop to go up appearance with the silico-tungstic acid inspection to effluent, water is eluted to and stops eluting when eluent does not have color basically; Resin by pouring out in the post, is put in the porcelain dish and dried, and 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 to dried molassed pulpous state, the dry catharanthus roseus total alkaloids that gets.
(4) macroporous adsorbent resin method: supernatant is regulated pH=10; On the DF01 type macroporous adsorbent resin handled well; Last appearance speed be 4 times of column volumes/hour, have deposition to stop to go up appearance with the silico-tungstic acid inspection to effluent, water is eluted to and stops eluting when eluent does not have color basically.With ethanol-water (70:30) is eluant, with 2.5 times of column volumes/hour speed carry out eluting, eluent begins to receive when with silico-tungstic acid inspection deposition being arranged; Receive eluent to do not have with the silico-tungstic acid inspection deposition up to, merge eluent, reclaim ethanol; Drying gets catharanthus roseus total alkaloids.
(5) organic solvent extractionprocess: supernatant 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 catharanthus roseus total alkaloids.
Two, the result calculates: distinguish the weight of five 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 five 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 five kinds of technologies
Visible by above comparing result, 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 Catharanthi Rosei decoction pieces 1000g, decocte with water 2 times, each 10 times of amounts decocted 2 hours; Merge decoction liquor, centrifugal, regulate pH=3; Centrifugal, supernatant with 4 times of column volumes/hour through cation exchange resin (001 * 2.5, the ammonium type; Blade diameter length ratio 1:8), wash to effluent and do not have color, 0.5% hydrochloric acid solution of reuse 2% ammonium chloride soaked resin 2 hours.0.1% hydrochloric acid solution eluting of reuse 2% ammonium chloride, elution speed be 2 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 2:
Get Herba Catharanthi Rosei decoction pieces 1000g, add the sour water percolation of pH=5, merge the percolate thin up to 10000ml; Centrifugal, regulate pH=1, centrifugal; Supernatant is with 5 times of column volumes/hour through cation exchange resin (001 * 7, sodium type, blade diameter length ratio 1:5); Washing to effluent does not have color, and 0.5% hydrochloric acid solution of reuse 1% sodium chloride soaked resin 2 hours.0.3% hydrochloric acid solution eluting of reuse 5% sodium chloride; Elution speed be 4 times of column volumes/hour, soaked behind 2 times of column volumes of eluting 5 hours, be eluted to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition) again; Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 3:
Get Herba Catharanthi Rosei decoction pieces 5kg, add 70% 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 with 2 times of column volumes/hour through last cation exchange resin (001 * 10, the sodium type; Blade diameter length ratio 1:7), wash to effluent and do not have color, 0.5% hydrochloric acid solution of reuse 20% sodium chloride soaked resin 1 hour.0.2% hydrochloric acid solution eluting of reuse 8% sodium chloride, elution speed be 5 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 4:
Get Herba Catharanthi Rosei decoction pieces 10kg, decocte with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor; Filter, it is 1.1 (60 ℃) that filtrating is concentrated into relative density, and adding ethanol to concentration is 70%, leaves standstill, and supernatant reclaims ethanol; Add water to 3000ml, regulate pH=4, centrifugal, supernatant is with 3 times of column volumes/hour through last cation exchange resin (011 * 6; Ammonium type, blade diameter length ratio 1:9), wash to effluent and do not have color, 0.5% sulfuric acid solution of reuse 5% ammonium chloride soaked resin 5 hours.0.1% sulfuric acid solution eluting of reuse 5% sodium chloride, elution speed be 6 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 5:
Get Herba Catharanthi Rosei decoction pieces 10kg, decocte with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor; Filter, it is 1.1 (60 ℃) that filtrating is concentrated into relative density, and adding ethanol to concentration is 70%, leaves standstill, and supernatant reclaims ethanol; Add water to 5000ml, regulate pH=5, centrifugal, supernatant is with 3 times of column volumes/hour through last cation exchange resin (D001 * 4; Hydrogen, blade diameter length ratio 1:10), wash to effluent and do not have color, 0.4% sulfuric acid solution of reuse 6% ammonium chloride soaked resin 5 hours.5% alcoholic solution eluting of reuse 6% sodium chloride, elution speed be 3 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 6:
Get Herba Catharanthi Rosei decoction pieces 2000g, add the hydrochloric acid solution of pH=3, microwave extraction 2 times adds 5 times of volume microwave extractioies 0.5 hour at every turn; Filter, the filtrating thin up is regulated pH=6 to 1600ml; Centrifugal, supernatant with 2 times of column volumes/hour through last cation exchange resin (001 * 15, Hydrogen; Blade diameter length ratio 1:6), wash to effluent and do not have color, 0.1% hydrochloric acid solution of reuse 10% sodium chloride soaked resin 4 hours.30% alcoholic solution eluting of reuse 10% sodium chloride, elution speed be 5 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 7:
Get Herba Catharanthi Rosei decoction pieces 10kg, decocte with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor; Filter, it is 1.1 (60 ℃) that filtrating is concentrated into relative density, and adding ethanol to concentration is 70%, leaves standstill, and supernatant reclaims ethanol; Add water to 4000ml, regulate pH=4, centrifugal, supernatant is with 0.5 times of column volume/hour through last cation exchange resin (001 * 18; Hydrogen, blade diameter length ratio 1:6), wash to effluent and do not have color, 0.1% hydrochloric acid solution of reuse 0.5% calcium chloride soaked resin 2 hours.20% alcoholic solution eluting of reuse 4% calcium chloride, elution speed be 5 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 8:
Get Herba Catharanthi Rosei decoction pieces 10kg, decocte with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor; Filter, it is 1.1 (60 ℃) that filtrating is concentrated into relative density, and adding ethanol to concentration is 70%, leaves standstill, and supernatant reclaims ethanol; Add water to 10000ml, regulate pH=3, centrifugal, supernatant is with 3 times of column volumes/hour through last cation exchange resin (001 * 4; Potassium type, blade diameter length ratio 1:8), wash to effluent and do not have color, 0.5% sulfuric acid solution of reuse 4% ammonium chloride soaked resin 3 hours.0.05% sulfuric acid solution eluting of reuse 4% ammonium chloride, elution speed be 6 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Embodiment 9:
Get Herba Catharanthi Rosei decoction pieces 1000g, decocte with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor; Filter, it is 1.1 (60 ℃) that filtrating is concentrated into relative density, and adding ethanol to concentration is 70%, leaves standstill, and supernatant reclaims ethanol; Add water to 1000ml, regulate pH=3, centrifugal, supernatant is with 3 times of column volumes/hour through last cation exchange resin (001 * 7; Hydrogen, blade diameter length ratio 1:8), wash to effluent and do not have color, 0.3% hydrochloric acid solution of reuse 3% calcium chloride soaked resin 2 hours.0.3% hydrochloric acid solution eluting of reuse 3% calcium chloride, elution speed be 4 times of column volumes/hour, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have deposition); Collect eluent; Be neutralized to neutrality with sodium hydroxide, desalination, the filtrating concentrate drying gets catharanthus roseus total alkaloids.
Claims (10)
1. the method for a catharanthus roseus total alkaloids separation from Chinese medicine Herba Catharanthi Rosei extracting solution is characterized in that this method is: get the Herba Catharanthi Rosei extracting solution, regulate pH value 1 to 7; Filter, cross cation exchange resin column, earlier with the washing remove impurity; Soak resin column with 0.5%～20% saline solution again, to contain acid or alcoholic acid 0.5%～20% saline solution eluting, collect eluent again; Through desalting processing, concentrate drying gets catharanthus roseus total alkaloids.
2. the method for catharanthus roseus total alkaloids separation according to claim 1, the pH value scope that it is characterized in that Chinese medicine extract solution is 2 to 5.
3. the method for catharanthus roseus total alkaloids separation according to claim 1 is characterized in that sample loading mode is that adverse current is crossed the post method on the medicinal liquid.
4. the method for catharanthus roseus total alkaloids separation according to claim 1, the concentration of salt solution that it is characterized in that used immersion resin is 1%-5%, eluting concentration is 3%-8%.
5. the method for catharanthus roseus total alkaloids separation according to claim 1 is characterized in that used salt is CaCl
2, KCl, NaCl, NH
4Among the Cl any.
6. the method for catharanthus roseus total alkaloids separation according to claim 1, the time that it is characterized in that used salt solution soaking resin is 1-5 hour.
7. the method for catharanthus roseus total alkaloids separation according to claim 1 is characterized in that eluting contains the hydrochloric acid of 0-0.5% or contains the sulphuric acid of 0-0.5% and/or the ethanol of 5%-30% with saline solution.
8. the method for catharanthus roseus total alkaloids separation according to claim 1 is characterized in that used resin is Hydrogen and salt type.
9. the method for catharanthus roseus total alkaloids separation according to claim 8 is characterized in that used resin is sodium type or the ammonium type in the salt type.
10. the method for catharanthus roseus total alkaloids separation according to claim 1 is characterized in that elution speed is 4～6 times of column volumes per hour.
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| CN1467204A (en) * | 2002-07-10 | 2004-01-14 | 江苏康缘药业股份有限公司 | Method for preparing motherwort total alkaloid and new medicine uses |
| CN1500778A (en) * | 2002-11-15 | 2004-06-02 | 北京天衡药物研究院 | Preparing method for motherwort general alkaloid |
| CN1800189A (en) * | 2005-12-19 | 2006-07-12 | 东北林业大学 | Method for separating vindoline , catharanthine, vinblastine and vincristine from Vinca rosea |
| CN1951432A (en) * | 2005-09-13 | 2007-04-25 | 山东绿叶天然药物研究开发有限公司 | Gingseng and aconite composition, its preparation, preparation process and usage |
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| CN1467204A (en) * | 2002-07-10 | 2004-01-14 | 江苏康缘药业股份有限公司 | Method for preparing motherwort total alkaloid and new medicine uses |
| CN1500778A (en) * | 2002-11-15 | 2004-06-02 | 北京天衡药物研究院 | Preparing method for motherwort general alkaloid |
| CN1951432A (en) * | 2005-09-13 | 2007-04-25 | 山东绿叶天然药物研究开发有限公司 | Gingseng and aconite composition, its preparation, preparation process and usage |
| CN1800189A (en) * | 2005-12-19 | 2006-07-12 | 东北林业大学 | Method for separating vindoline , catharanthine, vinblastine and vincristine from Vinca rosea |
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