CN101491555A - Camptotheca acuminata total alkaloids separation method in Camptotheca acuminata extract - Google Patents
Camptotheca acuminata total alkaloids separation method in Camptotheca acuminata extract Download PDFInfo
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Abstract
The invention discloses a method for separating total alkaloid from traditional Chinese medicine, in particular a method for separating total alkaloid from an extracting solution of the traditional Chinese medicine, namely camplotheca acuminata, and belongs to the field of traditional Chinese medicines. The method comprises the following technical proposal: taking the extracting solution of the camplotheca acuminata; adjusting the pH value of the extracting solution to between 1 and 7; filtering the extracting solution; taking and adding the filtrate to a cation exchange resin column; rinsing the filtrate and removing the impurities; eluting the filtrate with 0.5 to 15 percent acid liquor; checking eluent until no alkaloid exists; collecting the eluent; adding alkali to the eluent for neutralization; performing desalting treatment on the mixed solution; and performing concentrated drying to obtain the camplotheca acuminata total alkaloid. The method overcomes the defects of low extracting rate, large using level of organic solvent, complicated process, inability of large-scale production, and the like in the prior art and can efficiently separate high purity of the camplotheca acuminata total alkaloid from the extracting solution of the camplotheca acuminata.
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 Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) extracting solution the method for extraction separation common camptotheca fruit total alkaloids, belong to the field of Chinese medicines.
Background technology
Alkaloid is the big class basic nitrogen compound that occurring in nature extensively exists, and is many medium-height grass the effective elements of the medicines, is the class important substance that occurring in nature exists.Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) is China's endemic plant, originate to be Nyssaceae Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) platymiscium Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) Camptotheca acuminata Decne. because root, really, bark, branch, leaf etc. can extract multiple alkaloid, can treat multiple cancer, therefore have very high exploitation and be worth.Cool in nature, the bitter and puckery flavor of Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) has effects such as anticancer, heat clearing away, parasite killing.Studies show that, common camptotheca fruit total alkaloids is its main effective site, comprises camptothecine (Camptothecine), 10-hydroxycamptothecine (10-Hydroxy camptothecine), methoxycamptothecine (Methoxy camptothecine), venoterpine (venoterpine) etc.Clinical preparation is mainly used in treatment gastric cancer, colon cancer, rectal cancer, bladder cancer, chronic myelocytic leukemia, acute lymphoblastic leukemia etc.Therefore, common camptotheca fruit total alkaloids effective parts formulation clinical demand amount is very big.
Present stage, the extraction process of Chinese traditional medicine biology alkali is ripe, according to alkaloidal physicochemical property, adopt The suitable solvent such as water, sour water, acid alcohol and alkali alcohol etc., utilize technology such as dipping, percolation, decoction, backflow, ultrasonic, enzymolysis that the alkaloids composition is extracted, extraction ratio can reach more than 90%.But because the relative amount of alkaloid in Chinese medicine lower (most content are 0.01%-1%), certainly will introduce a large amount of other drug compositions and impurity constituents during extraction, 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 alkaloid valid target just seems very important.But since other multiple compositions and deposit and the interference of impurity such as starch, natural gum, pectin, phlegmatic temperament, pigment, the purification techniques of alkaloid valid target is the modernization of Chinese medicine " bottleneck " always, has had a strong impact on the requirement of modern Chinese medicine preparation to 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).Now, the purification techniques of alkaloid valid target mainly is organic solvent extractionprocess, Amberlyst process and ion exchange resin method etc.
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, and the alkaloid soda acid is transformed, and utilizes organic solvent (commonly used as benzene, chloroform, ether etc.) extraction, removes a large amount of impurity.Organic solvent extractionprocess is used comparatively extensive in the refining purification of common camptotheca fruit total alkaloids, but there is a lot of problems in this method: impurity such as a large amount of neutrality in alkaloids extract alkalization back and nonpolar pigment easily are brought into, therefore separation efficiency and purity are lower, and use a large amount of organic solvents (generally extracting 5 times), operation inconvenience (extractor efficient is not high, and easily emulsifying), safety is not good, and (organic solvent toxicity is big, and inflammable and explosive), be laboratory process, be not suitable for the big production of industry.
The macroporous adsorbent resin isolation technics, effective ingredient in the selective absorption Chinese medicine extraction liquid is removed impurity.Compare with technology with traditional impurity-removing method, can dwindle medication dose, improved the quality of preparation, this method to water solublity preferably flavone, saponin etc. good separation and purification effect is arranged, as Semen Ginkgo class preparation at present commonly used and ginsenoside's preparation etc.But make with extra care for alkaloidal, 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 an ionization state, the adsorption rate of resin is low, the basicity height, and alkaloid is free type, macromole alkaloid poorly water-soluble and separating out, still can't upper prop, suitable alkaloid scope is very little, and kind is very little; Secondly, the nonpolar electrostatic absorption principle of macroporous adsorbent resin is too poor to the separation selectivity of alkaloid active 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 final refining obtains is not high.
Also having emerging ion exchange resin isolation technics, is the purpose that reaches separation and purification by ion-exchange reactions.At present the technology that generally adopts in field of Chinese medicines institute is, after will containing alkaloidal solution and crossing post, with ammoniacal liquor or NaOH eluant solution, behind the collection eluent again with the organic solvent extraction alkaloid; The resin that perhaps will go up sample is with dipping by lye, and then with organic solvent reflux, extract, alkaloid.But this method step is loaded down with trivial details, organic solvent extraction, resin column is carried out steps such as organic solvent reflux, extract, in industrial very difficult realization, and very low with the efficient of alkali liquor eluting, only stays in the level of prepared in laboratory small sample, also can't realize big production.
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 common camptotheca fruit total alkaloids.
This goal of the invention realizes by following technology: get the Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) extracting solution, adjust pH value 1 to 7, filter, get filtrate and be added on the cation exchange resin column, earlier with the water elution remove impurity, reuse 0.5%～15% acid solution eluting, collect eluent, add the alkali neutralization,, promptly get common camptotheca fruit total alkaloids through desalting processing.
In above-mentioned technical process, after the extracting solution that will contain total alkaloids transfers to proper acidity, alkaloid ionization becomes cation, nonbasic substances is not ionized, after extracting solution was crossed cation exchange resin column, the hydrion on ionized alkaloid organic 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 ionization, also just can not 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 finish the resins exchange process well equally, thereby the pH value of finding to adjust extracting solution in the experiment is 1 to 7 to be proper.During with water elution, use be deionized water, to guarantee not introducing new ion interference, those just are not easy to be got off by water elution by the nonbasic substances of resin absorption during eluting, thus be attracted to resin column on the alkaloid organic ion separated.After this acid solution that adds higher concentration is again carried out eluting, because the hydrogen ion concentration in the acid solution is very high at this moment, will combine with resin competitively, thereby the alkaloid organic ion that will be adsorbed on the resin column exchanges, be dissolved in the sour eluent, flow out resin column, finish alkaloidal enrichment process.Afterwards, in eluent, add alkali and neutralize unnecessary acid,, can obtain common camptotheca fruit total alkaloids that purity is higher again through conventional desalting processing.
This technical process compared with prior art, technological process is not simple, not with an organic solvent, residual, the alkaloid rate of transform height of no resin monomer composition; Its important contribution also is, has broken ion-exchange capacity order rule, that is: Ca that traditional theory is thought
2+＞K
+≈ NH
4 +＞Na
+＞H
+Just under the influence of this traditional theory, in the ionic exchange resin of common camptotheca fruit total alkaloids all is that alkali liquor or ammoniacal liquor carry out eluting, 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, though hydrionic exchange capacity is the most weak, but by improving hydrionic concentration in the sour eluent, thereby offset the weak shortcoming of its exchange capacity, simultaneously under acid condition, the alkaloid that exchanges is dissolved in the acid solution rapidly, and is rushed out pillar, guarantee alkaloidal elute effect thus.And, carry out eluting with acid solution, in eluting, cation exchange resin column is regenerated, thereby can recycle, reduced the process of regenerating resin post behind the alkali liquor eluting, reduced cost.Technology of the present invention has broken through in the prior art traditional view and the technical process with the alkali liquor eluting fully, obtained beyond thought effect, improved the yield of common camptotheca fruit total alkaloids greatly, making cation exchange resin be used for refining separating common camptotheca fruit total alkaloids in commercial production becomes possibility.
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 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, can according to circumstances be processed in Hydrogen, sodium type or the ammonium type any when reality is used.
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 used acid solution of eluting is preferably hydrochloric acid solution or sulfuric acid solution, and its concentration all is preferably 3%～6%.
7, the flow velocity of eluent is unsuitable too fast or slow excessively during eluting, if too fast, then the exchange of hydrion and alkaloid organic ion is insufficient, and acid solution is wasted more; If cross slowly, the alkaloid organic 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.
8, the inventor also finds, improves elution efficiency if think more the highland, can not carry out the acid solution eluting immediately after the water elution remove impurity, but is full of acid solution earlier in resin column and leaves standstill more than 2 hours, generally need not surpass 12 hours, carry out eluting again.Through after leaving standstill, a large amount of alkaloid organic ions are exchanged by hydrion, 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 further to improve elution efficiency, can also in the acid solution that eluting is used, add small amount of N H
4Cl, NaCl or CaCl
2, addition is unsuitable very few or too much, if very few, does not then have effect; If then easily saltouing because of salinity is too high influences the dissolubility of alkaloid organic ion too much.Experiment finds, in the eluent with NH
4 +, Na
+Or Ca
2+Be good when concentration is 0.1～1mol/L, further preferred, NH
4 +, Na
+Or Ca
2+Concentration best when being 0.1～0.3mol/L.
10, said desalination process can be the desalination method that 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 Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) 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 Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) medical material 5000g, add 8 times of amounts of sour water of pH=3, soak a night, percolation is collected percolate to do not have a precipitation with the silico-tungstic acid inspection till, merges percolate, and is centrifugal, supernatant; Supernatant is five parts, carries out alkaloidal separation by following five kinds of methods respectively and purify:
(1) ion exchange resin+acid solution eluting: supernatant is regulated pH=3, on handled strong cation-exchanging resin 001 * 7 (Hydrogen well, blade diameter length ratio is 1: 8), 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, it is colourless substantially that water is eluted to eluent, reuse 3% hydrochloric acid solution eluting, elution speed be 4 times of column volumes/hour the speed 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, be neutralized to neutrality with sodium hydroxide, desalination (standby), drying gets common camptotheca fruit total alkaloids.
(2) the saline solution eluting of ion exchange resin+acid: supernatant is regulated pH=3, on handled strong cation-exchanging resin 001 * 7 (Hydrogen well, blade diameter length ratio is 1: 8), 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, and 0.5% hydrochloric acid solution of reuse 2% calcium chloride soaks resin.0.5% hydrochloric acid solution eluting of reuse 2% calcium chloride, 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 calcium hydroxide, concentrate drying gets common camptotheca fruit total alkaloids.
(3) ion exchange resin+reflux, extract: supernatant is regulated pH=3, on handled strong cation-exchanging resin 001 * 7 (Hydrogen, blade diameter length ratio are 1: 8) 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 common camptotheca fruit total alkaloids that gets to the dried molassed pulpous state.
(4) macroporous adsorbent resin method: supernatant 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 common camptotheca fruit 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 common camptotheca fruit 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; Adopt the quantitative approach of acid base titration respectively, five kinds of method gained total alkaloidss are carried out assay, calculate the purity of total alkaloids.
Three, result's contrast, see the following form:
The effect assessment table of five 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 Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 1000g, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merged decoction liquor, centrifugal, regulate pH=3, centrifugal, supernatant is with 4 times of column volumes/hour by cation exchange resin (001 * 7, Hydrogen, blade diameter length ratio 1: 8), being washed to effluent does not have color, reuse 15% hydrochloric acid 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 common camptotheca fruit total alkaloids.
Embodiment 2:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 1000g, the sour water percolation that adds pH=5, merge the percolate thin up to 10000ml, centrifugal, regulate pH=1, centrifugal, supernatant is so that 5 times of column volumes/hour (Doulex 50 by cation exchange resin, the sodium type, blade diameter length ratio 1: 5), being washed to effluent does not have color, reuse 5% hydrochloric acid solution eluting, elution speed be 4 times of column volumes/hour, soaked behind 2 times of column volumes of eluting 5 hours, 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 calcium hydroxide, desalination, the filtrate concentrate drying gets common camptotheca fruit total alkaloids.
Embodiment 3:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) 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 is with 2 times of column volumes/hour by last cation exchange resin (001 * 10, the sodium type, blade diameter length ratio 1: 7), being washed to effluent does not have color, reuse 1% hydrochloric acid solution (containing 1% sodium chloride) 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 common camptotheca fruit total alkaloids.
Embodiment 4:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 10kg, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 70%, leave standstill, supernatant reclaims ethanol, adds water to 3000ml, regulates pH=4, centrifugal, supernatant is with 3 times of column volumes/hour by last cation exchange resin (AmberliteIR--120, ammonia type, blade diameter length ratio 1: 9), be washed to effluent and do not have color, reuse 10% hydrochloric acid solution eluting, elution speed be 6 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 common camptotheca fruit total alkaloids.
Embodiment 5:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 10kg, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 70%, leave standstill, supernatant reclaims ethanol, adds water to 5000ml, regulates pH=5, centrifugal, supernatant is with 3 times of column volumes/hour by last cation exchange resin (D001 * 4, Hydrogen, blade diameter length ratio 1: 10), be washed to effluent and do not have color, reuse 3% hydrochloric acid solution eluting, elution speed be 3 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 common camptotheca fruit total alkaloids.
Embodiment 6:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 2000g, 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, filters, the filtrate thin up is to 1600ml, regulate pH=6, centrifugal, supernatant is so that 2 times of column volumes/hour (Doulex 50 by last cation exchange resin, Hydrogen, blade diameter length ratio 1: 6), being washed to effluent does not have color, reuse 2% hydrochloric acid solution (ammonia chloride that contains 0.2mol/L) 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 common camptotheca fruit total alkaloids.
Embodiment 7:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 10kg, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 70%, leave standstill, supernatant reclaims ethanol, adds water to 4000ml, regulates pH=4, centrifugal, supernatant is with 0.5 times of column volume/hour by last cation exchange resin (AmberliteIR--120, Hydrogen, blade diameter length ratio 1: 6), be washed to effluent and do not have color, reuse 0.5% sulfuric acid solution (sodium chloride that contains 1mol/L) 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 common camptotheca fruit total alkaloids.
Embodiment 8:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 10kg, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 70%, leave standstill, supernatant reclaims ethanol, adds water to 10000ml, regulates pH=3, centrifugal, supernatant is with 3 times of column volumes/hour by last cation exchange resin (001 * 5, Hydrogen, blade diameter length ratio 1: 8), be washed to effluent and do not have color, reuse 2% hydrochloric acid solution (calcium chloride that contains 0.3mol/L) eluting, elution speed be 6 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 common camptotheca fruit total alkaloids.
Embodiment 9:
Get Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) decoction pieces 1000g, decoct with water 2 times, each 10 times of amounts decocted 2 hours, merge decoction liquor, filter, it is 1.1 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol to concentration is 70%, leave standstill, supernatant reclaims ethanol, adds water to 1000ml, regulates pH=3, centrifugal, supernatant is with 3 times of column volumes/hour by last cation exchange resin (Doulex 50, Hydrogen, blade diameter length ratio 1: 8), be washed to effluent and do not have color, reuse 4% sulfuric acid solution eluting, elution speed be 4 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 common camptotheca fruit total alkaloids.
Claims (13)
1, a kind of from Chinese medicine Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) extracting solution the method for separating common camptotheca fruit total alkaloids, it is characterized in that this method is: get the Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) extracting solution, adjust pH value 1 to 7, filter, get filtrate and be added on the cation exchange resin column, earlier with the water elution remove impurity, reuse 0.5%～15% acid solution eluting is collected eluent, adds the alkali neutralization, through desalting processing, promptly get common camptotheca fruit total alkaloids.
2, the method for separating common camptotheca fruit total alkaloids as claimed in claim 1 is characterized in that the preceding pH value scope of adjusting medicinal liquid of upper prop is 2 to 5.
3, the method for separating common camptotheca fruit total alkaloids as claimed in claim 1 is characterized in that used cation exchange resin can be macroporous type or gel-type strong acid ion exchange resin.
4, the method for separating common camptotheca fruit total alkaloids as claimed in claim 3 is characterized in that the post footpath of used cation exchange resin column: post height=1: 5～1: 10.
5, the method for separating common camptotheca fruit total alkaloids as claimed in claim 1 is characterized in that in this method that the concentration of sample medicinal liquid is that every ml is equivalent to crude drug 0.1～3g, last sample speed be 0.5～5 times of column volume/hour.
6, the method for separating common camptotheca fruit total alkaloids as claimed in claim 5 is characterized in that sample loading mode is preferably sample on the adverse current.
7, the method for separating common camptotheca fruit total alkaloids as claimed in claim 1 is characterized in that eluting is used in this method acid solution is 3%～6% hydrochloric acid solution or 3%～6% sulfuric acid solution.
8, the method for separating common camptotheca fruit total alkaloids as claimed in claim 7, it is characterized in that elution speed be 2～6 times of column volumes/hour.
9, the method for separating common camptotheca fruit total alkaloids as claimed in claim 8, it is characterized in that elution speed be 4～6 times of column volumes/hour.
10, the method for separating common camptotheca fruit total alkaloids as claimed in claim 1 is characterized in that after the water elution remove impurity, can be full of eluting earlier with acid solution and left standstill 2～12 hours in resin column, carries out eluting again.
11, the method for separating common camptotheca fruit total alkaloids as claimed in claim 7 is characterized in that can also adding in the acid solution that eluting uses the NH of 0.1～1mol/L
4Cl, NaCl or CaCl
2
12, the method for separating common camptotheca fruit total alkaloids as claimed in claim 11 is characterized in that the concentration of salt is preferably 0.1～0.3mol/L in the acid solution.
13, the common camptotheca fruit total alkaloids of any one described method preparation among the claim 1-12.
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| CNA2007100984301A CN101491555A (en) | 2007-04-18 | 2007-04-18 | Camptotheca acuminata total alkaloids separation method in Camptotheca acuminata extract |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2017155834A1 (en) | 2016-03-08 | 2017-09-14 | Cheng Lab & Company, Llc | Herbal formulation to raise the human body's ph level from one that is acidic to one that is more alkaline |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017155834A1 (en) | 2016-03-08 | 2017-09-14 | Cheng Lab & Company, Llc | Herbal formulation to raise the human body's ph level from one that is acidic to one that is more alkaline |
| EP3426043A4 (en) * | 2016-03-08 | 2019-10-23 | Cheng Lab&Company, LLC | Herbal formulation to raise the human body's ph level from one that is acidic to one that is more alkaline |
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