CN101658598B - Method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma - Google Patents
Method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma Download PDFInfo
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Abstract
The invention provides a method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma, which comprises the following steps of: pretreating alisma medicinal materials into granules; extracting the granules by using ethanol solvent; adjusting the ethanol concentration of extracting solution to be 20%-50%; adsorbing the extracting solution by using non-polar or low-polar macroporous resin; eluting by using the ethanol solvent; concentrating eluent; and preparing finished product after drying. The method avoids using organic solvents such as chloroform, ethyl acetate and the like in the production process, enhances the safety of the product and production process, and has the outstanding advantages of simple process, low cost, easy operation, high contents of the finished product of the alisma total triterpenic ketone alcohol components and 23-acetyl alisol B and the like.
Description
Technical field
The present invention relates to from the Chinese crude drug Rhizoma Alismatis, extract the method for total triterpene ketols component, relate in particular to a kind of method of utilizing total triterpene ketols component in the macroporous resin separation and purification Rhizoma Alismatis medical material.
Background technology
With 23-alisol acetyl B is that the total triterpene ketols of the Rhizoma Alismatis component of index property composition is the main effective ingredient in the Rhizoma Alismatis; The total triterpene ketols of Rhizoma Alismatis component has the basic framework of protostane (protostane) type tetracyclic triterpene more, and mostly its structure is protostane (protostane) type tetracyclic triterpene.According to existing bibliographical information, from Rhizoma Alismatis, isolated about more than 30 triterpenoid compound, mainly comprise 23-alisol acetyl B, 24-alisol acetyl A etc.The structure of part Rhizoma Alismatis tetracyclic triterpene ketols composition is following in the Rhizoma Alismatis:
The total triterpene ketols of Rhizoma Alismatis component has blood fat reducing, atherosclerosis, anti-diabetic and multiple efficacies such as anticancer, is with a wide range of applications in fields such as pharmaceutical raw material, health foods.Along with the quickening of modernization of Chinese medicine process and the needs that go to the world, particularly American-European countries's pharmacy corporation, require active site and active component content in the Chinese medicine extract high more good more, so that qualitative and quantitative analysis, use.For this reason, carry out the Study on Preparation of effective extraction and the total triterpene ketols of enrichment Rhizoma Alismatis component,, have active operation significance to produce the total triterpene ketols of the Rhizoma Alismatis hypolipidemic activity position Rhizoma Alismatis constituent content high product that meets market demands.
At present, the Rhizoma Alismatis traditional extraction technique adopts decocting cooking method, heat reflow method, percolation etc. morely, and modern new technology adopts supercritical fluid processes.The decocting cooking method is relatively fat-soluble because of triterpenes components in these methods, thereby the stripping of triterpene ketols effective ingredient is few; Organic solvent heat reflow method, percolation, ultrasonic extraction etc. all exist the impurity component in the extract many, the low shortcoming of the total triterpene ketols of Rhizoma Alismatis constituent content; And modern supercritical extraction method requires to have supercritical fluid equipment, and investment is big, and cost is high, the extract obtained target that all is difficult to reach the total triterpene ketols of the Rhizoma Alismatis component in effective extraction and the enrichment Rhizoma Alismatis of above method.
In the article that the inventor delivers " experimentation of the total terpene alcohol of the Amberlyst process purification Rhizoma Alismatis constituents " literary composition following purification step has been proposed on " Colleges Of Traditional Chinese Medicine Of Fujian's journal " the 2nd phase in 2007: earlier with alcoholic solution leaching Rhizoma Alismatis medical material; Then extracting solution is concentrated into solution concentration of alcohol 15%; Re-using macroporous resin adsorbs; Macroporous resin after adopting alcoholic solution to absorption carries out eluting, the eluent of collection.But still there are following two subject matters in this method:
1. the total triterpene ketols of Rhizoma Alismatis constituent content is still not high enough in the finished product that makes;
2. the feed liquid of macroporous resin column absorption is because of the problem of dissolubility in the preparation after concentrating, and has the total triterpene ketols of part Rhizoma Alismatis compound precipitation to separate out and causes waste, and influence absorption because of precipitate stops up macroporous resin.
Summary of the invention
The technical problem that the present invention solves provides a kind of also method of the total triterpene ketols of enrichment Rhizoma Alismatis component of from Rhizoma Alismatis, extracting; This method for preparing can overcome the still not high enough technical problem of the total triterpene ketols of Rhizoma Alismatis constituent content in the finished product that makes in the said method, and the feed liquid of macroporous resin column absorption in the preparation after concentrating can reach higher concentration and do not have the total triterpene ketols of Rhizoma Alismatis compound precipitation and separate out.
For realizing the object of the invention, technical scheme provided by the invention may further comprise the steps:
A. pretreatment: with Rhizoma Alismatis pulverizing medicinal materials or section;
B. extract: it is that (concentration of alcohol described in the present invention is volume fraction (V/V) like no specified otherwise for the ethanol water of 60%-95% (volume) that pretreated medical material is placed concentration; The ethanol that adds described in the present invention or use is ethanol water like no specified otherwise, down with) in soak, the extraction soluble component;
C. adjust the extracting solution concentration of alcohol: concentration of ethanol in the extracting solution is adjusted to 20%-50%;
D. absorption: with carrying out adsorption treatment through the macroporous resin of pretreated or the nonpolar or low pole that regeneration is good extracting solution after to the adjustment concentration of alcohol;
E. eluting: the macroporous resin after will adsorbing is with the form of resin column; Elder generation's feeding concentration is that the ethanol of 0%-50% carries out eluting to said macroporous resin; Discard this part eluent, the ethanol that feeds 70% above concentration again carries out eluting to said macroporous resin, collects this part eluent;
F. concentrate: the eluent to collecting carries out concentration, obtains concentrated solution;
G. dry: as concentrated solution to be carried out dried, obtain finished product.
Compare the technical scheme that provides in the article that the inventor delivers " experimentation of the total terpene alcohol of Amberlyst process purification Rhizoma Alismatis constituents " on " Colleges Of Traditional Chinese Medicine Of Fujian's journal " the 2nd phase in 2007; This programme has been adjusted the concentration of alcohol in the extracting solution before the absorption in step c, be adjusted into 20%-50% by original 15%.
The inventor finds in experimentation; When utilizing the total triterpene ketols of the macroporous resin adsorption Rhizoma Alismatis chemical compound of nonpolar or low pole; Difference with concentration of alcohol in the solution; The macroporous resin of nonpolar or low pole is also different with the absorption situation that alcoholic solution extracts the extracting solution after handling to the Rhizoma Alismatis medical material, and indication absorption situation had both comprised the total triterpene ketols of Rhizoma Alismatis in extracting solution chemical compound is also comprised other impurity that contain in the extracting solution.Concentration of alcohol is under the situation of 20%-50% in solution; Impurity be not easy into the macroporous resin of nonpolar or low pole adsorbed, thereby improve target component adsorption rate and with the separating effect of impurity component, this point can be referring to embodiment 6 adsorption conditionses experiment 1; On the other hand; Reduce the water used in the eluting impurity step before the eluting effective ingredient and the consumption of low-concentration ethanol, saved link, time and cost, improved efficient.
In addition because the dissolving more easily in the non-polar solven environment of the total triterpene ketols of Rhizoma Alismatis chemical compound; So in the higher aqueous solution of ethanol content; Identical feed liquid can be dissolved the total triterpene ketols of more Rhizoma Alismatis chemical compound; Thereby avoid in the adsorption process parsing the waste that causes and, improve yield because of precipitate stops up macroporous resin owing to the total triterpene ketols of Rhizoma Alismatis chemical compound is insoluble.
As optimal technical scheme of the present invention, step c is adjusted to 20%-40% with concentration of ethanol in the extracting solution.Not only separating effect is better for impurity and target product under these process conditions, and the macroporous resin of nonpolar or low pole is better to the adsorption effect of the total triterpene ketols of Rhizoma Alismatis chemical compound, and promptly absorbability is stronger, leaks still less.This point can be referring to embodiment 7 adsorption conditionses experiment 2.
As more preferably scheme of the present invention; Concentration of alcohol is to the basis of 20%-40% in said step c adjustment extracting solution; Steps d is improved to: described resin absorption is specially, and the extracting solution of adjustment behind the concentration of alcohol resin column of flowing through is done dynamically absorption, and be adsorbed to saturated.The static adsorption of comparing, the resin absorption saturation of dynamic adsorption is higher, and large-scale production more easily on industry.
As more preferably scheme of the present invention, concentration of alcohol is to 20%-40% in said step c adjustment extracting solution; Steps d is improved to: described resin absorption is specially; The extracting solution of adjustment behind the concentration of alcohol resin column of flowing through is done dynamically absorption; And be adsorbed on the saturated basis; Described resin absorption step also comprises: the effluent of the resin column of will flowing through repeats through resin column 1-2 time, and the absorption number of times is total up to 2-3 time.Can further adsorb the total triterpene ketols of Rhizoma Alismatis residual in effluent chemical compound like this, improve yield.
As more preferably scheme of the present invention; Concentration of alcohol is to the basis of 20%-40% in said step c adjustment extracting solution; The said eluting of step e is improved to: the concentration that the feeding 1-5 of elder generation doubly measures the resin volume is the ethanol elution of 0%-50%; This part eluent discards, and feeding the concentration that 3-12 doubly measures the resin volume again is the ethanol aqueous wash deresination post of 70% above concentration, collects this part eluent.The resin of doubly measuring with 1-5 early stage like this after the washing with alcohol absorption of 0%-50% concentration of resin volume can wash away by the impurity that is adsorbed on the macroporous resin that some polarity are stronger; Simultaneously be unlikely to again the total triterpene ketols of the Rhizoma Alismatis chemical compound that is adsorbed on the macroporous resin is eluted in a large number, further reach the purpose of purification.And then use the ethanol-eluting resin column of 70% above concentration, and collect this part eluent, the total triterpene ketols of the Rhizoma Alismatis chemical compound overwhelming majority that this moment, we needed is included in this part eluent.
As preferred version of the present invention, the macroporous resin of the nonpolar or low pole described in the steps d is: among D4020, D101, the AB-8 any one.The inventor finds that through after a large amount of resin screening tests under process conditions of the present invention, these three kinds of resins have separating effect preferably; It is higher to be embodied in the finished product yield; The total triterpene ketols of the Rhizoma Alismatis of finished product compounds content is higher, with the D4020 best results, secondly is AB-8 in these three kinds of resins; And the D101 effect also better and be current unique macroporous resin through the food stage demonstration, has a wide range of applications.
As preferred version of the present invention; The method of the immersion described in the step b, extraction soluble component is: is the ethanol of 60%-95% with pretreated Rhizoma Alismatis medical material in the ratio adding concentration that the former weight of every kg medical material adds 6-12L; Soak 1-24 hour drenched to medical material; The lasting combination of carrying out one of ultrasound wave, diafiltration or hot reflux method or above method in 0.5-4 hour is extracted; Collecting extracting solution, is the ethanol of 60%-95% to remaining medicinal residues in the ratio adding concentration that the former weight of every kg medical material adds 6-12L, and the lasting combination of carrying out one of ultrasound wave, diafiltration or hot reflux method or above method in 0.5-4 hour is extracted once more; Collect extracting solution, with extracting solution merging, the filtration of twice collection.Such method for distilling can be guaranteed as much as possible the effective ingredient in the medical material to be leached out, has guaranteed that again leaching process is unlikely too very long simultaneously.
As preferred version of the present invention, step c described adjustment solution concentration of alcohol or f are described to be concentrated, and the method for taking is that distilling under reduced pressure concentrates.Because distilling under reduced pressure is one of process means commonly used in the Chemical Manufacture, and the total triterpene ketols of the Rhizoma Alismatis of our required extraction chemical compound is relatively stable to heat, can adopt this process means.In the process of the said adjustment solution of step c concentration of alcohol, adopt solution is carried out distilling under reduced pressure, both can reduce the upper prop liquor capacity, reduce the material liquid volume that needs processing; Simultaneously can improve solution concentration, help absorption; And the concentration of alcohol of solution is constantly to descend in the process of distilling under reduced pressure, so can reach the purpose of adjustment solution concentration of alcohol simultaneously.But the said adjustment solution concentration of step c also can be taked other modes; Such as directly in the feed liquid adding aqueous solution directly regulate concentration of alcohol and also allow; Possibly bring results such as the material liquid volume change is big although it is so; Perhaps adopt and be all the thin film concentration of evaporation and concentration and adjust the solution concentration of alcohol; Can select suitable ultrafilter membrane or NF membrane filtering and concentrating for use to feed liquid in addition, combine simultaneously to add the water top in the filter process and means such as wash and also can reach regulator solution concentration of alcohol and spissated purpose.In like manner the concentration process in step f also can adopt mode or ultrafiltration, the nanofiltration filtering and concentrating of thin film concentration.
As preferred version of the present invention, step a is specially: the Rhizoma Alismatis medical material is pulverized, and being crushed to particle size range is decoction pieces size to 60 order.The medical material of size is convenient to the follow-up soaking and extracting of medical material like this, and the also comparatively convenient processing of the medical material granule of yardstick is produced so simultaneously.
As preferred version of the present invention, the said dried of step g is lyophilization or spray drying.These two kinds of drying modes all are convenient to implementation and operation on the industry, and the simultaneous spray drying heated time is extremely short, and lyophilization need not to be heated especially, and the total triterpene ketols of Rhizoma Alismatis chemical compound keeps stable chemical state in the finished product so these two kinds of drying modes all help.
Technological process after the present invention optimizes is as shown in Figure 9.
As the pretreatment or the regeneration scheme of the resin of selecting for use among the present invention, get final product according to usual manner pretreatment or regeneration.The pretreating scheme of resin is soak with ethanol, distilled water cleaning etc. in the embodiment of the invention.
The detection method that is adopted among the present invention is following:
One. the mensuration to the total triterpene ketols of Rhizoma Alismatis constituent content adopts metric measurement, and is specific as follows:
Instrument: 752 UV, visible light grating spectrophotometers, water-bath;
Reagent: perchloric acid (analytical pure; Shanghai gold deer chemical industry company limited); Glacial acetic acid (Shanghai chemical reagents corporation of Chinese Medicine group), 5% vanillin-glacial acetic acid developer (Shanghai chemical reagents corporation of Chinese Medicine group), 23-alisol acetyl B standard substance (self-control).
Method: adopt triterpenoid compound mass fraction in glacial acetic acid-vanillin-perchloric acid spectrophotometry Rhizoma Alismatis, adopt C (vanillin-glacial acetic acid)=5% solution 0.3ml, perchloric acid 0.8ml; 60 ℃ of water-bath 20min, ice bath cooling 5min adds glacial acetic acid to 5.0ml; Shake up; At the absorbance of 544nm place working sample, compare with the absorbance of 23-alisol acetyl B standard substance, calculate in the sample the total triterpene ketols of Rhizoma Alismatis constituent content in 23-alisol acetyl B.
The method of mentioning in the article that this detection method can be delivered on " Colleges Of Traditional Chinese Medicine Of Fujian's journal " the 2nd phase in 2007 referring to the inventor " experimentation of the total terpene alcohol of the Amberlyst process purification Rhizoma Alismatis constituents " literary composition.
Two, among the present invention 23-alisol acetyl B is adopted the high-efficient liquid phase color spectrometry, represent the absworption peak of 23-alisol acetyl B to appear between the 8.5min-9.5min, measuring method is specific as follows:
Instrument: Tianjin, island LC-20A high performance liquid chromatograph, SPD-20A type UV-detector, CBM-10A chromatographic work station.
Reagent: alisol B-23 acetas standard substance are self-control (pressing literature method), and acetonitrile is a chromatographically pure, and water is the WAHAHA pure water.
Method: chromatographic condition is a chromatographic column: Kromasil-C18 post (4.6mm * 250mm, 5 μ m);
Mobile phase: acetonitrile-water (9: 1); Detect wavelength: 210nm; Flow velocity: 0.8ml/min.
Description of drawings
Fig. 1 is that medicinal liquid high performance liquid chromatogram collection of illustrative plates is extracted in embodiment 6 said experiment Central Plains;
Fig. 2 is D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 15% the extracting solution for concentration of alcohol in the embodiment 6 said experiments;
Fig. 3 is D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 20% the extracting solution for concentration of alcohol in the embodiment 6 said experiments;
Fig. 4 is D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 30% the extracting solution for concentration of alcohol in the embodiment 6 said experiments;
Fig. 5 is D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 40% the extracting solution for concentration of alcohol in the embodiment 6 said experiments;
Fig. 6 is D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 50% the extracting solution for concentration of alcohol in the embodiment 6 said experiments;
Fig. 7 is with the high performance liquid chromatogram collection of illustrative plates of the eluent of 70% ethanol elution in the embodiment 8 said experiments;
Fig. 8 is with the high performance liquid chromatogram collection of illustrative plates of the eluent of 90% ethanol elution in the embodiment 8 said experiments;
Fig. 9 is the process flow diagram after optimizing.
The specific embodiment
Embodiment 1:
The present invention with the Rhizoma Alismatis medical material be raw material production have a blood fat reducing effect be rich in the total triterpene ketols of Rhizoma Alismatis component materials, mainly comprise the steps:
1. pretreatment
The Rhizoma Alismatis medical material is taken from the Rhizoma Alismatis bulb of the Rhizoma Alismatis GAP base production in Ji Yang town, Jian'ou, Fujian, after the Rhizoma Alismatis bulb is broken into pieces, selects 20 purpose sieve plates that Rhizoma Alismatis bulb fragment is pulverized.
2. extract
Above-mentioned Rhizoma Alismatis coarse powder 20kg packed into extract to be concentrated in the extraction vessel of unit, adds 85% ethanol of 160L, soaks 2h, extracts 70 ℃ of temperature, and 2h is extracted in hot reflux, filters extracting solution; 85% ethanol that adds 120L again extracts 70 ℃ of temperature, and hot reflux is extracted 1h and carried out secondary extraction and filtration.
3. adjust the solution concentration of alcohol.
Merge extracted twice liquid, reclaim ethanol, be about at 30% o'clock to concentration of alcohol and stop to concentrate, it is subsequent use to get its supernatant.
4. absorption
Select nonpolar D4020 macroporous resin; After conventional treatment before the uses such as soak with ethanol, distilled water cleaning, the dress post is with the Rhizoma Alismatis ethanol extraction supernatant upper prop for preparing early stage; Carry out dynamic adsorption, effluent is saturated with the abundant absorption that reaches resin after post 2 times.
5. desorption
(" BV " represents the resin volume in the description with 5 times of amount BV at normal temperatures and pressures with the macroporous resin that is adsorbed with the total triterpene ketols of Rhizoma Alismatis component; 30% ethanol elution down together); To remove non-targeted composition, discard the above-mentioned steps eluent, 12 times of amounts of reuse BV's; 70% ethanol elution is collected this part eluent and is placed basin.
6. concentrate
With the eluent of above-mentioned collection, decompression recycling ethanol, and be concentrated into to a certain degree and get final product (density is 1.1g/m3).
7. dry
The concentrated solution of above-mentioned gained had both been got finished product 436g through DC-1500 spray drying instrument drying, and yield is 2.2%, and 23-alisol acetyl B is about 32.56% in the finished product; Total triterpene ketols material is about 68.20%.The spray drying actual conditions is: inlet temperature is 180-200 ℃, and outlet temperature is 85-90 ℃, and inlet amount is 300ml/h.Above-mentioned finished product is put into the airtight package container, place shady and cool dry place to deposit.
Embodiment 2:
The present invention with the Rhizoma Alismatis medical material be raw material production have a blood fat reducing effect be rich in the total triterpene ketols of Rhizoma Alismatis component materials, mainly comprise the steps:
1. pretreatment
The Rhizoma Alismatis medical material is available from the Rhizoma Alismatis decoction pieces of pharmaceuticals.
2. extract
Above-mentioned Rhizoma Alismatis decoction pieces 20kg packed into extract to be concentrated in the extraction vessel of unit, adds 95% the ethanol of 160L, soaks 12h, extracts 70 ℃ of temperature, and 2h is extracted in hot reflux, filters extracting solution; 95% ethanol that adds 120L again extracts 70 ℃ of temperature, and hot reflux is extracted 30min and carried out secondary extraction and filtration.
3. adjust the solution concentration of alcohol
Merge extracted twice liquid, reclaim ethanol, be about at 30% o'clock to concentration of alcohol and stop to concentrate, it is subsequent use to get its supernatant.
4. absorption
Select nonpolar D101 macroporous resin; After conventional treatment before the uses such as soak with ethanol, distilled water cleaning, the dress post is with the Rhizoma Alismatis ethanol extraction supernatant upper prop for preparing early stage; Carry out dynamic adsorption, effluent is saturated with the abundant absorption that reaches resin after post 1 time.
5. eluting
With the macroporous resin that is adsorbed with the total triterpene ketols of Rhizoma Alismatis component at normal temperatures and pressures with 5 times the amount BV 30% ethanol elutions; To remove non-targeted composition; Discard this part eluent, 95% ethanol elution of 8 times of amounts of reuse BV is collected this part eluent and is placed basin.
6. concentrate
With the eluent of above-mentioned collection, decompression recycling ethanol, and be concentrated into to a certain degree and get final product (density is 1.1g/m3).
7. dry
The concentrated solution of above-mentioned gained had both been got finished product 412g through DC-1500 spray drying instrument drying, and yield is 2.1%.23-alisol acetyl B is about 30.73% in the finished product; Total triterpene ketols material is about 67.75%, and the spray drying actual conditions is: inlet temperature is 180-200 ℃, and outlet temperature is 85-90 ℃, and inlet amount is 400ml/h.Above-mentioned finished product is put into the airtight package container, place shady and cool dry place to deposit.
Embodiment 3:
The present invention with the Rhizoma Alismatis medical material be raw material production have a blood fat reducing effect be rich in the total triterpene ketols of Rhizoma Alismatis component materials, mainly comprise the steps:
1. pretreatment
The Rhizoma Alismatis medical material selects 20 purpose sieve plates that the Rhizoma Alismatis decoction pieces is pulverized available from the Rhizoma Alismatis decoction pieces of pharmaceuticals.
2. extract
Above-mentioned Rhizoma Alismatis coarse powder 20kg packed into extract to be concentrated in the extraction vessel of unit, adds 70% the ethanol of 180L, soaks 1h, extracts 70 ℃ of temperature, and 2h is extracted in hot reflux, filters extracting solution; 70% ethanol that adds 140L again extracts 70 ℃ of temperature, and hot reflux is extracted 3h and carried out secondary extraction and filtration.
3. adjust the solution concentration of alcohol
Merge extracted twice liquid, reclaim ethanol, be about at 30% o'clock to concentration of alcohol and stop to concentrate, it is subsequent use to get its supernatant.
4. absorption
Select non polarity A B-8 macroporous resin; After conventional treatment before the uses such as soak with ethanol, distilled water cleaning, the dress post is with the Rhizoma Alismatis ethanol extraction supernatant upper prop for preparing early stage; Carry out dynamic adsorption, effluent is saturated with the abundant absorption that reaches resin after post 2 times.
5. eluting
The macroporous resin that is adsorbed with the total triterpene ketols of Rhizoma Alismatis component is cleaned with the distilled water of 10 times of amount BV at normal temperatures and pressures; Again with 3 times of 50% ethanol elutions of measuring BV; To remove non-targeted composition; Discard the above-mentioned steps eluent, 95% ethanol elution of 6 times of amounts of reuse BV is collected this part eluent and is placed basin.
6. concentrate
With the eluent of above-mentioned collection, decompression recycling ethanol, and be concentrated into to a certain degree and get final product (density is 1.1g/m3).
7. dry
The concentrated solution of above-mentioned gained had both been got finished product 401g through the lyophilization drying, and yield is 2.0%.23-alisol acetyl B is about 30.51% in the finished product; Total triterpene ketols material is about 67.28%, and above-mentioned finished product is put into the airtight package container, places shady and cool dry place to deposit.
Embodiment 4:
The present invention with the Rhizoma Alismatis medical material be raw material production have a blood fat reducing effect be rich in the total triterpene ketols of Rhizoma Alismatis component materials, mainly comprise the steps:
1. pretreatment
The Rhizoma Alismatis medical material selects 60 purpose sieve plates that the Rhizoma Alismatis decoction pieces is pulverized available from the Rhizoma Alismatis decoction pieces of pharmaceuticals.
2. extract
Above-mentioned Rhizoma Alismatis coarse powder 20kg packed into extract to be concentrated in the extraction vessel of unit, adds 60% the ethanol of 240L, soaks 24h, extracts 70 ℃ of temperature, and 4h is extracted in hot reflux, filters extracting solution; 70% ethanol that adds 120L again extracts 70 ℃ of temperature, and hot reflux is extracted 30min and carried out secondary extraction and filtration.
3. adjust the solution concentration of alcohol
Merge extracted twice liquid, reclaim ethanol, be about at 20% o'clock to concentration of alcohol and stop to concentrate, it is subsequent use to get its supernatant.
4. absorption
Select nonpolar D4020 macroporous resin; After conventional treatment before the uses such as soak with ethanol, distilled water cleaning, the dress post is with the Rhizoma Alismatis ethanol extraction supernatant upper prop for preparing early stage; Carry out dynamic adsorption, effluent is saturated with the abundant absorption that reaches resin after post 2 times.
5. eluting
The macroporous resin that is adsorbed with the total triterpene ketols of Rhizoma Alismatis component is cleaned with the distilled water of 1BV at normal temperatures and pressures; Again with 2 times of 40% ethanol elutions of measuring BV; To remove non-targeted composition; Discard the above-mentioned steps eluent, 80% ethanol elution of 10 times of amounts of reuse BV is collected this part eluent and is placed basin.
6. concentrate
With the eluent of above-mentioned collection, decompression recycling ethanol, and be concentrated into to a certain degree and get final product (density is 1.1g/m3).
7. dry
The concentrated solution of above-mentioned gained had both been got finished product 420g through the lyophilization drying, and yield is 2.1%.23-alisol acetyl B is about 23% in the finished product; Total triterpene ketols material is about 68%, and above-mentioned finished product is put into the airtight package container, places shady and cool dry place to deposit.
Embodiment 5:
The present invention with the Rhizoma Alismatis medical material be raw material production have a blood fat reducing effect be rich in the total triterpene ketols of Rhizoma Alismatis component materials, mainly comprise the steps:
1. pretreatment
The Rhizoma Alismatis medical material selects 30 purpose sieve plates that the Rhizoma Alismatis decoction pieces is pulverized available from the Rhizoma Alismatis decoction pieces of pharmaceuticals.
2. extract
Above-mentioned Rhizoma Alismatis coarse powder 20kg packed into extract to be concentrated in the extraction vessel of unit, adds 95% the ethanol of 120L, soaks 3h, extracts 70 ℃ of temperature, and 30min is extracted in hot reflux, filters extracting solution; 70 ℃ of 60% ethanol extraction temperature that add 240L again, 4h is extracted in hot reflux, carries out secondary extraction and filtration.
3. adjust the solution concentration of alcohol
Merge extracted twice liquid, reclaim ethanol, be about at 40% o'clock to concentration of alcohol and stop to concentrate, it is subsequent use to get its supernatant.
4. absorption
Select nonpolar D4020 macroporous resin; After conventional treatment before the uses such as soak with ethanol, distilled water cleaning, the dress post is with the Rhizoma Alismatis ethanol extraction supernatant upper prop for preparing early stage; Carry out dynamic adsorption, effluent is saturated with the abundant absorption that reaches resin after post 1 time.
5. eluting
With the macroporous resin that is adsorbed with the total triterpene ketols of Rhizoma Alismatis component at normal temperatures and pressures with 1 times the amount BV 50% ethanol elution; Discard the above-mentioned steps eluent; To remove non-targeted composition, 95% ethanol elution of 3 times of amounts of reuse BV is collected eluent and is placed basin.
6. concentrate
With the eluent of above-mentioned collection, decompression recycling ethanol, and be concentrated into to a certain degree and get final product (density is 1.1g/m3).
7. dry
The concentrated solution of above-mentioned gained had both been got finished product 402g through the lyophilization drying, and yield is 2.0%.23-alisol acetyl B is about 24% in the finished product; Total triterpene ketols material is about 67%, and above-mentioned finished product is put into the airtight package container, places shady and cool dry place to deposit.
Embodiment 6
Adsorption conditions experiment 1:
Can observe intuitively from scheming to go up us; The concentration of alcohol that Fig. 3, Fig. 4, Fig. 5, Fig. 6 characterized is a D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on the extracting solution of 20%-50%; It with respect to the concentration of alcohol that Fig. 2 characterized D4020 resin column absorption debris high performance liquid chromatogram collection of illustrative plates on 15% the extracting solution; Impurity absorption between 1min-2.5min obviously increases, and it is that resin is adsorbed that these these impurity of explanation are difficult in the ethanol environment of higher concentration.
Embodiment 7
Adsorption conditions experiment 2:
Experiment 2 processes that the inventor carries out are following: 6 parts of extracting solution getting equal volume V1; With wherein 5 parts be adjusted to different concentration of alcohol and the final identical V2 of each part extracting liquid volume respectively; Said 5 parts of extracting solution concentration of alcohol are respectively: 15%, 20%, 30%, 40%, 50%; Use same D4020 resin column to adsorb respectively these feed liquids, and amount of resin is collected to the effusive absorption residual liquid of each resin column enough to guarantee absorption back resin column unsaturation; With remaining 1 part of extracting solution, equally also adjusting volume is V2 simultaneously, and this part extracting solution is following with liquid-phase chromatographic analysis respectively with 5 parts of absorption residual liquids collecting before:
Can know by above table; Concentration of alcohol is 20%, 30%, 40% extracting solution in the extracting solution; The slip of its effective ingredient 23-alisol acetyl B is that 15% extracting solution is low than concentration of alcohol in the extracting solution; Especially the slip of effective ingredient 23-alisol acetyl B is minimum when the extracting solution concentration of alcohol is 30%, and adsorption effect is best.
Embodiment 8
The eluting experiment
The experiment that the inventor carries out is specific as follows: get the saturated D4020 resin of a absorption; 30% ethanol with 5 times of amounts is washed, and then resin is divided into two parts of equivalent, loads the same resin column; Use 70% ethanol of 8 times of amounts of resin volume and 90% ethanol elution respectively; Collect eluent, the eluent of collecting is used the liquid chromatograph qualitative analysis, like Fig. 7, Fig. 8; We can find in the eluent under the alcoholic solution eluting of high concentration impurity content still less, and 23-alisol acetyl B content is higher in the eluent.
Claims (9)
1. one kind is extracted the also method of the total triterpene ketols of enrichment Rhizoma Alismatis component from Rhizoma Alismatis, it is characterized in that this method may further comprise the steps:
A. pretreatment: with Rhizoma Alismatis pulverizing medicinal materials or section;
B. extract: it is ethanol immersion, the extraction soluble component of 60%-95% (volume) that pretreated medical material is placed concentration, obtains extracting solution;
C. adjust the extracting solution concentration of alcohol: concentration of ethanol in the extracting solution is adjusted to 20%-40% (volume);
D. absorption: with carrying out adsorption treatment through the macroporous resin of pretreated or the nonpolar or low pole that regeneration is good extracting solution after to the adjustment concentration of alcohol;
E. eluting: the macroporous resin after will adsorbing is with the form of resin column; Elder generation's feeding concentration is that the ethanol of 0%-50% (volume) carries out eluting to said macroporous resin; Discard this part eluent; The ethanol that feeds the above concentration of 70% (volume) again carries out eluting to said macroporous resin, collects this part eluent;
F. concentrate: the eluent to collecting carries out concentration, obtains concentrated solution;
G. dry: as concentrated solution to be carried out dried, obtain finished product.
2. according to the method for claim 1, it is characterized in that wherein the described resin absorption of steps d is: will adjust extracting solution behind the concentration of alcohol resin column of flowing through and do dynamically absorption, and be adsorbed to saturated.
3. according to the method for claim 2, it is characterized in that wherein said resin absorption step also comprises: the effluent of the resin column of will flowing through repeats through resin column 1-2 time, and the absorption number of times is total up to 2-3 time.
4. according to the method for claim 1; It is characterized in that; Wherein the said eluting of step e is: the concentration that the feeding 1-5 of elder generation doubly measures the resin volume is the ethanol elution of 0%-50% (volume); This part eluent discards, and feeding the concentration that 3-12 doubly measures the resin volume again is the ethanol aqueous wash deresination post of the above concentration of 70% (volume), collects this part eluent.
5. according to the method for claim 1, it is characterized in that wherein the described macroporous resin nonpolar or low pole of steps d is: any one among D4020, D101, the AB-8.
6. according to the method for claim 1; It is characterized in that; The method of the wherein described immersion of step b, extraction soluble component is: is the ethanol of 60%-95% (volume) with pretreated Rhizoma Alismatis medical material in the ratio adding concentration that the former weight of every kg medical material adds the 6-12L alcoholic solution; Soak 1-24 hour drenched to medical material, the combination that continues to carry out in 0.5-4 hour one of ultrasound wave, diafiltration or hot reflux method or above method is extracted, and collects extracting solution; Is the ethanol of 60%-95% (volume) to remaining medicinal residues in the ratio adding concentration that the former weight of every kg medical material adds the 6-12L alcoholic solution; The lasting combination of carrying out one of ultrasound wave, diafiltration or hot reflux method or above method in 0.5-4 hour is extracted once more, collects extracting solution, with extracting solution merging, the filtration of twice collection.
7. according to the method for claim 1, it is characterized in that wherein step c described adjustment solution concentration of alcohol or step f are described concentrates, the method for being taked is that distilling under reduced pressure concentrates.
8. according to any described method of claim 1 to 7, it is characterized in that wherein step a is specially: the Rhizoma Alismatis medical material is pulverized, and being crushed to particle size range is decoction pieces size to 60 order.
9. according to any described method of claim 1 to 7, it is characterized in that wherein the said dried of step g is lyophilization or spray drying.
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| CN102372759A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Purification method for alisol A |
| CN104497092B (en) * | 2014-12-24 | 2016-08-24 | 陕西嘉禾生物科技股份有限公司 | A kind of method extracting 23-alisol acetyl C from Rhizoma Alismatis |
| CN110368438A (en) * | 2019-07-01 | 2019-10-25 | 厦门医学院 | A kind of rhizoma alismatis alcohol extracting optimization technique based on property material changes of contents |
| CN113105522B (en) * | 2021-04-06 | 2022-04-01 | 神威药业集团有限公司 | Oriental water plantain triterpenes compound, preparation method, structure characterization method and application thereof |
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| 吴水生等.大孔树脂法纯化泽泻总萜醇类成分的实验研究.《福建中医学院学报》.2007,第17卷(第2期),43-45. * |
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