CN101491567A - Method for separating Huperzia serrata Thunb. total alkaloid in Huperzia serrata Thunb. extract liquid - Google Patents
Method for separating Huperzia serrata Thunb. total alkaloid in Huperzia serrata Thunb. extract liquid 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 Huperzia serrata Thunb, and belongs to the field of traditional Chinese medicines. The method comprises the following technical proposal: taking the extracting solution of the Huperzia serrata Thunb; 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; collecting eluent; adding alkali to the eluent for neutralization; performing desalting treatment on the mixed solution; and obtaining the Huperzia serrata Thunb 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 Huperzia serrata Thunb total alkaloid from the extracting solution of the Huperzia serrata Thunb.
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 Lycopodii serrati extracting solution the method for extraction separation huperzia serrata 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.Herba Lycopodii serrati Huperzia serrata, have another name called serrate clubmoss herb, feet added to a snake by an ignorant artist grass, Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) etc., be the dry herb of Huperziaceae plants of Huperzia Herba Lycopodii serrati (Huperz ia serrata (Thunb.) Trev.), property is flat, acrid in the mouth, has effects such as dehumidifying, repercussive, hemostasis.Herba Lycopodii serrati is a kind of Chinese herbal medicine among the people commonly used, and existing Huperzine Serrate P.E is sold on the market now.Studies show that, huperzia serrata total alkaloids is its main effective site, mainly comprises huperzine A (Huperzine A), huperzine B (Huperzine B), lycoclavanin alkali (Lycodavine), serrate clubmoss herb alkali (Huperzinine), serratinine (Serratimine), serratine (Serratin) etc.Wherein, huperzine A is a kind of potent, reversible cholinesterase inhibitor, can improve senile memory function decline, can significantly improve memory, cognition and the behavioral competence of patients of senile dementia, has good prospect on the treatment senile dementia.Herba Lycopodii serrati is mainly used in diseases such as treatment pneumonia, lung abscess, over-strained hemoptysis, hemorrhoidal hemorrhage, leucorrhea, traumatic injury, toxic swelling.Therefore, huperzia serrata 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).
Though the method for extraction separation Chinese medicine total alkaloids is arranged in textbook and various document, and these methods only stay in breadboard research level more, majority is to transform back organic solvent extraction alkaloid by soda acid; Or adopt the macroporous resin separation and purification to get alkaloid; Or after will containing alkaloidal solution and crossing post,, collect behind the eluent again with the organic solvent extraction alkaloid with ammoniacal liquor or NaOH eluant solution; The resin that perhaps will go up sample is with dipping by lye, and then with organic solvent reflux, extract, alkaloid.Shortcomings such as but these methods have complex steps, and consumption of organic solvent is big, production cost is too high, large equipment can't be realized and the alkaloid rate of transform is low, thereby be the laboratory research level, can not in big production, use. always" Chinese herbal medicine extract " (Chinese Medicine science and technology publishing house the Hou Tuanzhang chief editor, 2004) under the Huperzine Serrate P.E item in the book, various extracting method have been enumerated, wherein having related to a kind of up-to-date technology of extraction separation total alkaloids from the Chinese medicine Herba Lycopodii serrati is the spent ion exchange resin method, basic process is: the sour water percolate of Herba Lycopodii serrati medical material is passed through 732 type cation exchange resin columns, with ammonia ethanol (5% ammonia, 80% ethanol) eluting, collect eluent, reclaim ethanol, the dry huperzia serrata total alkaloids that gets.This method has been avoided the use of a large amount of organic solvents, has certain progress, but still has major defect: though can find in the test that this method can improve alkaloidal purity, the rate of transform is still not high; Free huperzia serrata total alkaloids need just have dissolubility preferably under the alkali condition in alcoholic solution, but the use of alcoholic solution can be brought the residual of resin monomer composition such as styrene, divinylbenzene etc. in eluent, influence preparation safety; Owing to use ethanol in the technology, must increase the Ethanol Treatment step when also requiring the resin pre-treatment simultaneously, the later stage also will be reclaimed ethanol, has increased processing step, has improved production cost.So in order to adapt to market and production demand, press for a kind of safe, low-cost, extraction separation method that yield is high.
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 huperzia serrata total alkaloids.
This goal of the invention realizes by following technology: get the Herba Lycopodii serrati 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata total alkaloids greatly, making cation exchange resin be used for refining separating huperzia serrata 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 Herba Lycopodii serrati 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 Lycopodii serrati decoction pieces 1500g, the sour water that adds pH=4 decocts 2 times, and each 10 times of amounts decocted 2 hours, merged decoction liquor, and centrifugal, filtrate is diluted to 3000ml and regulates pH=3, is divided into three parts, handles according to the following steps respectively:
1. according to prior art, get said extracted liquid 1000ml, last cation exchange resin (001 * 7, Hydrogen, blade diameter length ratio 1: 5), with 5% ammonia, 80% ethanol elution, elution speed be 5 times of column volumes/hour, collect eluent, check negative to alkaloid, reclaim ethanol, concentrate drying gets huperzia serrata total alkaloids.
2. according to technology of the present invention, get said extracted liquid 1000ml, last cation exchange resin (001 * 7, Hydrogen, blade diameter length ratio 1: 5), is washed till with deionized water that to outpour liquid colourless, reuse 3% hydrochloric acid solution eluting, elution speed be 5 times of column volumes/hour, collect eluent, negative to the alkaloid inspection, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
3. according to technology of the present invention, get said extracted liquid 1000ml, last cation exchange resin (001 * 7, Hydrogen, blade diameter length ratio 1: 8), sample on the adverse current, be washed till with deionized water that to outpour liquid colourless, 3% hydrochloric acid solution of NaCl that adds 0.1mol/L left standstill 2 hours to full post, the above-mentioned solution of reuse with 5 times of column volumes/hour the speed eluting, collect eluent, negative to the alkaloid inspection, be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Two, the result calculates: distinguish the weight of three kinds of technology gained of weighing huperzia serrata total alkaloids, and be divided by with the medical material amount, get the yield of total alkaloids; With acid-base titrations three 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 three kinds of technologies
| Technology 1 | Technology 2 | Technology 3 | |
| Yield | 0.3％ | 0.7％ | 0.6％ |
| Purity | 54％ | 78％ | 80％ |
| Cost | Step is many, and the ethanol consumption is big, the cost height | Without organic solvent, cost is low | Without organic solvent, and the minimizing of the consumption of sour eluent, cost is lower |
| Time | Cycle is long, needs 8 hours at least | Cycle is short, only needs 5 hours | Cycle is short, only needs 4.5 hours |
| Safety | Record with gas chromatogram and to contain styrene, divinylbenzene in the extract | No resin monomer residue | No resin monomer residue |
| Complexity | Need to use the alcohol pre-treatment resin column, also want the regenerating resin post behind the eluting; Eluted product need reclaim ethanol, and elution time is long, and elution efficiency is low | Process is simple, only needs normal pretreatment resin column, the resin column of also having regenerated when pickling is taken off | Process is simple, only needs normal pretreatment resin column, the resin column of also having regenerated when pickling is taken off |
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 Lycopodii serrati 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, collect eluent, check negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide to alkaloid, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 2:
Get Herba Lycopodii serrati 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 this part eluent, be neutralized to neutrality with calcium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 3:
Get Herba Lycopodii serrati 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, collect eluent, check negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide to alkaloid, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 4:
Get Herba Lycopodii serrati 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, collect eluent, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 5:
Get Herba Lycopodii serrati 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, collect eluent, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 6:
Get Herba Lycopodii serrati 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, collect eluent, check negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide to alkaloid, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 7:
Get Herba Lycopodii serrati 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, collect eluent, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 8:
Get Herba Lycopodii serrati 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, collect eluent, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Embodiment 9:
Get Herba Lycopodii serrati 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, collect eluent, to alkaloid inspection negative (effluent drips 10% silico-tungstic acid does not have precipitation), be neutralized to neutrality with sodium hydroxide, desalination, the filtrate concentrate drying gets huperzia serrata total alkaloids.
Claims (13)
1, a kind of from Chinese medicine Herba Lycopodii serrati extracting solution the method for separating huperzia serrata total alkaloids, it is characterized in that this method is: get the Herba Lycopodii serrati 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 huperzia serrata total alkaloids.
2, the method for separating huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata 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 huperzia serrata total alkaloids of any one described method preparation among the claim 1-12.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108191762A (en) * | 2018-01-24 | 2018-06-22 | 湖南华诚生物资源股份有限公司 | A kind of extracting method of pure natural heavy metal free Huperzine Serrate P.E |
| CN112402466A (en) * | 2020-11-30 | 2021-02-26 | 湖南华诚生物资源股份有限公司 | Method for separating total alkaloids from huperzia serrata extract |
-
2007
- 2007-04-18 CN CNA2007100984405A patent/CN101491567A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108191762A (en) * | 2018-01-24 | 2018-06-22 | 湖南华诚生物资源股份有限公司 | A kind of extracting method of pure natural heavy metal free Huperzine Serrate P.E |
| CN108191762B (en) * | 2018-01-24 | 2020-04-14 | 湖南华诚生物资源股份有限公司 | Extraction method of huperzia serrata extract without heavy metal |
| CN112402466A (en) * | 2020-11-30 | 2021-02-26 | 湖南华诚生物资源股份有限公司 | Method for separating total alkaloids from huperzia serrata extract |
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