CN102973679A - Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants - Google Patents
Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants Download PDFInfo
- Publication number
- CN102973679A CN102973679A CN2012105594365A CN201210559436A CN102973679A CN 102973679 A CN102973679 A CN 102973679A CN 2012105594365 A CN2012105594365 A CN 2012105594365A CN 201210559436 A CN201210559436 A CN 201210559436A CN 102973679 A CN102973679 A CN 102973679A
- Authority
- CN
- China
- Prior art keywords
- extract
- aconitum
- aqueous solution
- preparation
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a preparation method of total alkaloids extract in aconitum medical material or processed products of the aconitum medical material. The method comprises the following steps of: regulating pH value of a water solution containing the aconitum medical material or the processed product extract of the aconitum medical material to be 7-10, then, adsorbing with a macroporous absorption resin column, washing with water to remove impurities, eluting by at least the acid alcohol-water solution of which the pH is not more than 5 and the alcohol volume content is 50-90%, collecting the eluant, and removing the solvent to obtain the total alkaloids extract in aconitum medical material or processed products of the aconitum medical material. The method not only can completely remove the impurities, but also cannot lose and damage the needed alkaloid constituent, and completely elutes the corresponding alkaloid constituent; and the method is high in recovery ratio, effectively solves the problem of serious loss of the alkaloid in the conventional method, greatly saves the medical material resource, improves the utilization rate of the medical resources, and fully uses and gives play to the medical effect of the aconitum medical material or the processed products of the aconitum plants.
Description
Technical field
The present invention relates to the preparation method of total alkaloids extract in a kind of aconitum plants or its processed goods, the method that particularly aconitum plants that obtained by the whole bag of tricks or its processed goods total alkaloids extract crude product is prepared its purified extract product.
Background technology
Aconitum plant has abundant medicinal kind, according to record, but China has 36 kinds of aconitum plant hyoscines approximately, medicinal herbs most in use mainly contain Aconitum carmichjaelii Debx. (Radix Aconiti), Aconitum kusnezoffii Reichb (Radix Aconiti Kusnezoffii), Radix Aconiti Coreani, aconitum brachypodum, aconitum tanguticumStapf etc., and the Preparation process product, such as Radix Aconiti Kusnezoffii Preparata, Radix Aconiti Preparata, RADIX ACONITI LATERALIS PREPARATA, Radix Aconiti Lateralis Preparata, RADIX ACONITI LATERALIS PREPARATA, yellow Radix Aconiti Lateralis Preparata, big gun Radix Aconiti etc.Local and the among the people medicinal kind of commonly using has Radix aconiti szechenyiani, Dendrobium denneanum Kerr. crow, pawl leaf Aconitum carmichjaelii Debx. and poly-leaf scape monkshood root and Preparata thereof etc.Aconitum plant has the effect of expelling wind and removing dampness, antalgic more, can be used for anemofrigid-damp arthralgia, arthralgia, trusted subordinate's cold type of pain etc., be widely used in the clinical treatment, " national medical product complete works of " record contain Radix Aconiti, Radix Aconiti Lateralis Preparata, Radix Aconiti Kusnezoffii, aconitum brachypodum, Aconitum sinomontanum Nakai, Radix Aconiti Coreani Chinese patent medicine above 350 kinds.
The main pharmacodynamics composition is alkaloid in the Aconitum medicinal plants, comprises diester-type alkaloids (aconitine, mesaconitine, hypaconitine etc.), monoester alkaloid (benzoyl aconine, Benzoylmesaconine, benzoyl hypo-aconine etc.) and hydramine type alkaloid (aconine, new aconine, inferior aconine) etc.At present such alkaloid component is had been reported and/isolation and purification method that uses, be its decocting liquid or its aqueous solution after macroporous adsorptive resins absorption, wash first remove impurity with water, use again ethanol elution, collect ethanol elution, obtain Radix Aconiti Lateralis Preparata after separation and purification or the total alkaloids composition of Radix Aconiti.Such as reports such as Zhao Lei, Radix Aconiti Lateralis Preparata extract is passed through the D101 purification with macroreticular resin, water, volume fraction are 10%, 80% alcoholic solution gradient elution, the collected volume mark is 10% ethanol elution partial concentration drying, namely get the Radix Aconiti Lateralis Preparata total alkaloids (extraction and purification process of Radix Aconiti Lateralis Preparata total alkaloids, Shenyang Pharmaceutical University's journal, 2007,24 (7): 433).The reports such as Wan Lixiang, with D101 macroporous adsorptive resins on the Radix Aconiti Lateralis Preparata extract, with the distilled water remove impurity, use again 95% ethanol elution, concentrated evaporate to dryness eluent namely gets Radix Aconiti Lateralis Preparata total alkaloids (purifying process of aconitum carmichaeli debx. Total alkaloids and the research of the purification of mesaconitine, Agriculture of Anhui science, 2008,36 (12): 5044).Wu Ping has reported a kind of extraction separation and purification method of Radix Aconiti Lateralis Preparata alkaloid, with the Radix Aconiti Lateralis acid extraction, extracting solution adsorbs through macroporous resin column, wash first decontamination with water, use again 60% ethanol elution, collect eluent, Recycled ethanol, obtain Radix Aconiti Lateralis Preparata alkaloid extract (Chengdu University of Traditional Chinese Medicine's master thesis, 2007) through further processing.
The applicant finds after deliberation, decocting liquid to aconitum plants or its processed goods, or the aqueous solution of other solvent extractable matter such as its ethanol, when adopting the macroporous adsorptive resins separation and purification, after the water elution remove impurity, use again the aquiferous ethanol eluting (desorption) of 30 ~ 95 (v) %, all fully eluting reclaims corresponding alkaloid component, cause alkaloidal loss serious, loss rate can reach 30 ~ 50%, waste herb resource, also affected giving full play to of aconitum plants or its processed goods drug action.
Summary of the invention
Given this, the invention provides the preparation method of a kind of aconitum plants or its processed goods total alkaloids extract, the method that particularly aconitum plants that obtained by the whole bag of tricks or its processed goods total alkaloids extract crude product is prepared its purified extract product.
The preparation method of total alkaloids extract in aconitum plants of the present invention or its processed goods, that the aqueous solution that will contain aconitum plants or its processed goods extract is regulated pH value 7 ~ 10, after preferably regulating pH value 8 ~ 10, adsorb with macroporous adsorptive resins, then after washing the removal of impurity with water first, be 50 ~ 90% acidic ethanol-aqueous solution eluting with pH≤5 and alcohol volume content at least again, collect eluent, desolventizing namely obtains aconitum plants and processed goods total alkaloids extract thereof.Wherein, said macroporous adsorbent resin can operate according to the mode of present routine, as selects HPD100, the HPD300 that commonly uses, the macroporous resin of the various models such as D101, AB-8, wherein the macroporous adsorbent resin of the models such as preferred HPD100 or D101.
The above-mentioned said aqueous solution that contains aconitum plants or its processed goods extract, the water extraction solution that comprises aconitum plants or its processed goods, or the aqueous solution that contains aconitum plants or its processed goods extract component in other various sources, particularly by different organic solvents aconitum plants or its processed goods are extracted and desolventizing after the aqueous solution of extract preparation.
Wherein, when adopting the aqueous extract of aconitum plants or its processed goods, this aqueous extract except can the aqueous extract for routine, the acid water extracting liquid of pH≤5 more preferably, and preferably be adopted as aqueous solution with hydrochloric acid or sulfuric acid acidation.
To the said pH regulator that contains aconitum plants or its processed goods solution of extract as raw materials, can adopt the various alkaline components that allow use in the medicine, such as usually adopting alkali metal hydroxide or its carbonate, the bicarbonate etc. that comprise that sodium hydroxide, sodium carbonate or sodium bicarbonate etc. are the most commonly used.Experiment shows, after the aqueous solution of aconitum plants or its processed goods extract is adjusted to pH value 7 ~ 10, process with said absorption with macroporous adsorbent resin again, the adsorbing separation of hydramine type alkaloid component will more be conducive to, obviously improve the adsorption capacity of macroporous resin, improve separation efficiency, reduce cost.
When washing remove impurity with water after the said aqueous solution that contains aconitum plants or its processed goods extract adsorbs with macroporous adsorptive resins, can adopt respectively usual manner only with the mode of neutral water washing, preferred mode is to adopt alkaline water and the medium-sized water washing of using successively neutral water, pH 8 ~ 10; Perhaps adopt alkaline water washing and the neutral water washing of using successively pH 8 ~ 10.Experiment shows, uses first the soda lye wash of pH 8 ~ 10 behind the absorption with macroporous adsorbent resin, can better remove impurity, washes with water to nearly neutrality (pH 7 ~ 8) again.And then carry out said with acidic ethanol-aqueous solution eluting, thereby enriching and purifying total alkaloids composition better.
In the said method, be to improve treatment effeciency, said aqueous solution for macroporous adsorptive resins absorption, preferably wherein the extractive content of aconitum plants or its processed goods is no more than and is equivalent to 2 gram crude drug/milliliters.
As the said acid ethanol solution of eluent, except preferably carrying out the ethanol-water solution of acidify with hydrochloric acid the most commonly used and that be easy to get in the commercial production, also can use other acid ingredient that allows in the medicine to use to carry out acidify in the above-mentioned preparation method.
Another kind of optimal way during eluting is, can adopt and use first acidic ethanol-aqueous solution eluting, use again at last neutral alcohol-aqueous solution eluting, namely adopt first acid (particularly acid stronger) ethanol-water solution eluting, after making macroporous adsorptive resins be acidity, use again at last the neutral alcohol eluting instead.
To collected eluent, except can directly removing the ethanol equal solvent, obtain outside the said extract (or containing aconitum plants or the corresponding hydrochlorate of its processed goods total alkaloids), the alternate manner that can also adopt comprises, with the eluent of said collection first with the alkaline components that allows in the medicine to use (as equally can be preferably above-mentioned sodium hydroxide, sodium carbonate or sodium bicarbonate etc.) be adjusted to pH value 5 ~ 7 after, carry out again the operation of desolventizing.Another kind method is, can be with the eluent of said collection alkali type anion exchange resin, as select to have at present wide coverage and/use 717, after the alkali type anion exchange resin process of the models such as D201, D202,711, D301, D311, D318, carry out again the operation of desolventizing.
Experimental result shows, the above-mentioned preparation method of the present invention compares with present traditional method, when particularly adopting the remove impurity of alkaline aqueous solution eluting, can fully remove impurity, again can significantly sacrificing or destroy needed alkaloid component, and with acidic ethanol-aqueous solution adsorbent resin is carried out eluting, can make corresponding alkaloid component by abundant eluting, the response rate is high, the alkaloid loss serious problems of traditional method have effectively been avoided, greatly saved herb resource, improved the utilization rate of medical material, the drug action of aconitum plants or its processed goods is utilized more fully and brought into play.
The specific embodiment by the following examples is described in further detail foregoing of the present invention again.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.Do not breaking away from the above-mentioned technological thought situation of the present invention, various replacements or change according to ordinary skill knowledge and customary means are made all should comprise within the scope of the invention.
The specific embodiment
Embodiment 1
Get Radix Aconiti Lateralis 10kg, add 10 times in water, soaked 4 hours, decocted 2 hours, filter, decocting liquid is for subsequent use; Medicinal residues add 8 times in water again, decoct 1 hour, filter, merge decocting liquid, regulate pH value 9 ~ 10 with sodium hydroxide solution, go up according to a conventional method HPD100 type macroporous adsorptive resins, wash remove impurity with water first after, use again 70 (v) %(of pH 2 ~ 3 as follows) acidic alcohol-aqueous solution eluting, collect ethanol elution, regulate pH value to 6 ~ 7, decompression recycling ethanol with sodium hydroxide solution, concentrated, dry, namely get the Radix Aconiti Lateralis total alkaloids extract.
Embodiment 2
Get and fry Radix Aconiti Lateralis Preparata 10kg, add 12 times in water, soaked 2 hours, decocted 3 hours, filter, decocting liquid is for subsequent use; Medicinal residues add 10 times in water again, decoct 2 hours, filter, merge decocting liquid, regulate pH value 9 ~ 10, upper HPD100 type macroporous adsorptive resins with sodium carbonate liquor, respectively sodium hydroxide solution and the water elution remove impurity of water, pH value 9 ~ 10, use again 70% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, collect ethanol elution, upper 717 alkali type anion-exchange resin columns, collect the ethanol stream fluid, decompression recycling ethanol, concentrated, dry, namely get and fry the Radix Aconiti Lateralis Preparata total alkaloids extract.
Embodiment 3
Get Radix aconiti szechenyiani (system) 10kg, add 10 times in water, decoct 2 times, each 2 hours, filter, merge decocting liquid, regulate pH value 8 ~ 9 with sodium hydroxide solution, upper D101 type macroporous adsorptive resins is used respectively sodium carbonate liquor and the water elution remove impurity of pH value 9 ~ 10, uses 90% acidic alcohol of pH 3 ~ 4-aqueous solution eluting again, collect ethanol elution, decompression recycling ethanol, concentrated, dry, namely get Radix aconiti szechenyiani total alkaloids extract processed.
Embodiment 4
Get RADIX ACONITI LATERALIS PREPARATA 10kg, 12 times of hydrochloric acid solutions that add pH value 2 ~ 3, decoct 2 times, each 3 hours, filter, merge decocting liquid, regulate pH value 7 ~ 8 with sodium carbonate liquor, upper D101 type macroporous adsorptive resins, after using respectively the sodium hydroxide solution and water elution remove impurity of pH value 8 ~ 9, use again 60% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, when being acidity to effluent, use neutral 60% alcohol-water eluting instead, collect sour ethanol and ethanol elution, the hydro-oxidation sodium solution is regulated pH value to 5 ~ 7, decompression recycling ethanol, concentrated, dry, namely get the RADIX ACONITI LATERALIS PREPARATA total alkaloids extract.
Embodiment 5
Get Radix Aconiti Lateralis Preparata 10kg, add 10 times of 70 (v) % ethanol, reflux, extract, 2 times each 2 hours, filters, merge ethanol extract, reclaim under reduced pressure eliminates ethanol, obtains the extract of Radix Aconiti Lateralis Preparata, is dissolved in water, be mixed with the aqueous solution that content is equivalent to 1 grams per milliliter crude drug, regulate pH value 7 ~ 8 with sodium bicarbonate solution, filter upper HPD300 type macroporous adsorptive resins, the difference water, the sodium carbonate liquor of pH value 8 ~ 9 and water elution remove impurity, use again 80% acidic alcohol of pH 4 ~ 5-aqueous solution eluting, collect ethanol elution, decompression recycling ethanol, concentrated, drying namely gets the Radix Aconiti Lateralis Preparata total alkaloids extract.
Embodiment 6
Get RADIX ACONITI LATERALIS PREPARATA 10kg, 12 times of hydrochloric acid solutions that add pH value 4 ~ 5, decoct 2 times, each 2 hours, filter, merge decocting liquid, regulate pH value 8 ~ 9 with sodium carbonate liquor, upper HPD300 type macroporous adsorptive resins, after using respectively the sodium hydroxide solution and water elution remove impurity of pH value 9 ~ 10, use again 60% acidic alcohol of pH 3 ~ 4-aqueous solution eluting, when being acidity to effluent, use neutral 60% alcohol-water eluting instead, collect sour ethanol and ethanol elution, the hydro-oxidation sodium solution is regulated the pH value near neutral, decompression recycling ethanol, concentrated, drying namely gets the RADIX ACONITI LATERALIS PREPARATA total alkaloids extract.
Embodiment 7
Get and steam Radix Aconiti Lateralis Preparata 10kg, 8 times of sulfuric acid solutions that add pH value 1 ~ 2, decoct 3 times, each 1.5 hours, filter, merging filtrate is regulated pH value 9 ~ 10, upper AB-8 type macroporous adsorptive resins with sodium hydroxide solution, respectively behind the sodium hydroxide solution and water elution remove impurity of water, pH value 9 ~ 10, use again 70% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, collect ethanol elution, upper 717 alkali type anion-exchange resin columns, collect the ethanol stream fluid, decompression recycling ethanol, concentrated, dry, namely get and steam the Radix Aconiti Lateralis Preparata total alkaloids extract.
Embodiment 8
Get Radix Aconiti Preparata 10kg, add 12 times of the hydrochloric acid solutions of pH value 3 ~ 4, decoct 2 times, each 2 hours, filter, merge decocting liquid, regulate pH value 9 ~ 10 with sodium hydroxide solution, filter upper HPD100 type macroporous adsorptive resins, wash first remove impurity with water, use again 50% acidic alcohol of pH 3 ~ 4-aqueous solution eluting, collect ethanol elution, the hydro-oxidation sodium solution is regulated pH value to 5 ~ 6, decompression recycling ethanol, concentrated, dry, namely get the Radix Aconiti Preparata total alkaloids extract.
Embodiment 9
Get Radix Aconiti 10kg, add 10 times in water, decoct 2 times, each 5 hours, filter, merge decocting liquid, regulate pH value 8 ~ 9 with sodium carbonate liquor, upper HPD300 type macroporous adsorptive resins, use respectively the sodium hydroxide solution and water elution remove impurity of pH value 9 ~ 10 after, use again 70% acidic alcohol of pH 1 ~ 2-aqueous solution eluting, collect ethanol elution, upper D201 alkali type anion-exchange resin column, collect the ethanol stream fluid, decompression recycling ethanol, concentrated, dry, namely get the Radix Aconiti total alkaloids extract.
Embodiment 10
Get Radix Aconiti Kusnezoffii Preparata 10kg, add 8 times of 80 (v) % ethanol, reflux, extract, 3 times, each 2 hours, filter, merge ethanol extract, reclaim under reduced pressure eliminates ethanol, obtains the extract of Radix Aconiti Kusnezoffii Preparata, be dissolved in water, be mixed with pH 8 ~ 9 with sodium hydroxide solution, be mixed with content and be no more than the aqueous solution that is equivalent to 2 grams per milliliter crude drugs, filter upper AB-8 type macroporous adsorptive resins, after using respectively the sodium hydroxide solution and water elution remove impurity of pH value 9 ~ 10, use again 60% acidic alcohol of pH 0.5-aqueous solution eluting, collect ethanol elution, upper D301 alkali type anion-exchange resin column, collect the ethanol stream fluid, decompression recycling ethanol, concentrated, drying namely gets the Radix Aconiti Kusnezoffii Preparata total alkaloids extract.
Below content and drug action by effective ingredient be evaluation index, further specify the beneficial effect that the preparation method of Radix Aconiti Lateralis Preparata of the present invention or total alkaloid of common monkshood root extract has.
1, the assay of effective ingredient
Get Radix Aconiti Lateralis 4kg, add 10 times in water, soaked 4 hours, decocted 2 hours, filter, decocting liquid is for subsequent use; Medicinal residues add 8 times in water again, decoct 1 hour, filter, merge decocting liquid, regulate pH value 9 ~ 10 with sodium hydroxide solution, filter, be divided into 4 equal portions, the HPD100 type macroporous adsorptive resins (A, B, C, D) that to go up respectively 4 identical bed volumes according to a conventional method be 1.5L:
A post: adopt usual manner, use first the water elution remove impurity of 3 times of bed volumes, use again 70% ethanol-water solution eluting, collect the ethanol elution of 3 times of bed volumes, (A liquid) for subsequent use;
B post: the mode that adopts above-described embodiment 1, use first the water elution remove impurity of 3 times of bed volumes, use again 70% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (B liquid) for subsequent use with sodium hydroxide solution;
C post: the mode that adopts above-described embodiment 2, use respectively the water of 3 times of bed volumes, sodium hydroxide solution and the water elution remove impurity of pH value 9 ~ 10, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, upper 717 alkali type anion-exchange resin columns, collect the ethanol stream fluid, (C liquid) for subsequent use.
D post: the mode that adopts above-described embodiment 3, use respectively sodium hydroxide solution and the water elution remove impurity of the pH value 9 ~ 10 of 3 times of bed volumes, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (D liquid) for subsequent use with sodium hydroxide solution.
Other gets RADIX ACONITI LATERALIS PREPARATA 4kg, 12 times of hydrochloric acid solutions that add pH value 2 ~ 3, decoct 2 times, each 3 hours, filter merging filtrate, regulate pH value 7 ~ 8 with sodium carbonate liquor, filter, be divided into 4 equal portions, the D101 type macroporous adsorptive resins (E, F, G, H) that to go up respectively 4 identical bed volumes according to a conventional method be 1.5L:
E post: with A post mode, use first the water elution remove impurity of 3 times of bed volumes, use again 70% ethanol-water solution eluting, collect the ethanol elution of 3 times of bed volumes, (E liquid) for subsequent use;
F post: with B post mode, use first the water elution remove impurity of 3 times of bed volumes, use again 70% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (F liquid) for subsequent use with sodium hydroxide solution;
G post: with C post mode, use respectively the water of 3 times of bed volumes, sodium hydroxide solution and the water elution remove impurity of pH value 9 ~ 10, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, upper 717 alkali type anion-exchange resin columns, collect the ethanol stream fluid, (G liquid) for subsequent use.
H post: with D post mode, use respectively sodium hydroxide solution and the water elution remove impurity of the pH value 9 ~ 10 of 3 times of bed volumes, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (H liquid) for subsequent use with sodium hydroxide solution.
Get again Radix Aconiti Kusnezoffii Preparata 4kg, add 8 times of amounts of hydrochloric acid solution that pH 2 ~ 3 contains 80 (v) % ethanol, reflux, extract, 3 times, each 2 hours, filter, merge ethanol extract, reclaim under reduced pressure eliminates ethanol, obtain the extract of Radix Aconiti Kusnezoffii Preparata, add 4 L water dissolutioies, regulate pH 8 ~ 9 with sodium hydroxide solution, filter, be divided into 4 equal portions, the AB-8 type macroporous adsorptive resins (L, M, N, O) that to go up respectively 4 identical bed volumes according to a conventional method be 1.5L:
L post: with A post mode, use first the water elution remove impurity of 3 times of bed volumes, use again 70% ethanol-water solution eluting, collect the ethanol elution of 3 times of bed volumes, (L liquid) for subsequent use;
M post: with B post mode, use first the water elution remove impurity of 3 times of bed volumes, use again 70% acidic alcohol of pH 2 ~ 3-aqueous solution eluting, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (M liquid) for subsequent use with sodium hydroxide solution;
N post: with C post mode, use respectively the water of 3 times of bed volumes, sodium hydroxide solution and the water elution remove impurity of pH value 9 ~ 10, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, upper 717 alkali type anion-exchange resin columns, collect the ethanol stream fluid, (N liquid) for subsequent use.
O post: with D post mode, use respectively sodium hydroxide solution and the water elution remove impurity of the pH value 9 ~ 10 of 3 times of bed volumes, use again the 70% ethanol solution hydrochloride eluting of pH 2 ~ 3, collect the ethanol elution of 3 times of bed volumes, regulate pH value to 6 ~ 7, (O liquid) for subsequent use with sodium hydroxide solution.
Press the HPLC chromatographic process under 177 pages of Radix Aconiti Lateralis Preparata items of Chinese Pharmacopoeia version in 2010, measure respectively the content (in crude drug) of alkaloid component in above-mentioned A liquid～O liquid, the results are shown in Table 1.
Assay result (the mgg of alkaloid component in table 1 A～O eluent
-1, in crude drug)
| Alkaloid | Benzoyl aconine | Benzoyl hypo-aconine | Benzoylmesaconine | Aconitine | Hypaconitine | Mesaconitine |
| A liquid | 0.113 | 0.341 | 0.837 | — | — | — |
| B liquid | 0.158 | 0.425 | 1.562 | — | — | — |
| C liquid | 0.152 | 0.414 | 1.530 | — | — | — |
| D liquid | 0.157 | 0.431 | 1.578 | — | — | — |
| E liquid | 0.075 | 0.146 | 0.217 | 0.019 | 0.104 | 0.038 |
| F liquid | 0.102 | 0.168 | 0.412 | 0.027 | 0.125 | 0.070 |
| G liquid | 0.097 | 0.160 | 0.393 | 0.025 | 0.120 | 0.067 |
| H liquid | 0.104 | 0.165 | 0.419 | 0.028 | 0.126 | 0.071 |
| L liquid | 0.063 | 0.130 | 0.194 | 0.031 | 0.152 | 0.049 |
| M liquid | 0.096 | 0.173 | 0.351 | 0.036 | 0.183 | 0.104 |
| N liquid | 0.092 | 0.167 | 0.340 | 0.034 | 0.175 | 0.098 |
| O liquid | 0.098 | 0.170 | 0.355 | 0.037 | 0.181 | 0.0103 |
Annotate: "-" be not for detecting
By as seen from Table 1, the effect of sour ethanol elution (desorbing) significantly is better than the neutral alcohol eluting, and the remove impurity of aqueous alkali eluting has no significant effect alkaloid component.
2, cold resistance, anoxia enduring experiment
With above-mentioned E liquid, F liquid, G liquid, H liquid decompression recycling ethanol, be concentrated into clear paste, for subsequent use.(2005,21 (6): 31-33) method is carried out effect experiment to reference literature for Li Liji etc., Pharmacology and Clinics of Chinese Materia Medica.
2.1 cold resistance experiment
100 male and female half and half of mice, be divided at random 5 groups, successive administration 5 days, put into freezer compartment of refrigerator after time administration in 1 hour,-10 ℃ of half an hour, observe its dead number of elements, the results are shown in Table 2, show that the RADIX ACONITI LATERALIS PREPARATA total alkaloids extract (F, G, H extract) that preparation method of the present invention obtains has better cold resistance effect.
Table 2 RADIX ACONITI LATERALIS PREPARATA extract is on the impact of mice cold resistant
| Group | Dosage (the g crude drug/Kg) | Test Mus number (only) | Dead Mus number (only) | Survival rate (%) |
| Contrast | — | 20 | 13 | 35 |
| The E extract | 20 | 20 | 9 | 55 |
| The F extract | 20 | 20 | 5 | 75 |
| The G extract | 20 | 20 | 6 | 70 |
| The H extract | 20 | 20 | 5 | 75 |
2.2 anoxia enduring experiment
60 of mices, male and female half and half are divided into 5 groups at random, successive administration 5 days, after time administration 1 hour, the time of its mouth breathing of broken end observation, the results are shown in Table 3, show that the RADIX ACONITI LATERALIS PREPARATA total alkaloids extract that preparation method of the present invention obtains has better resisting oxygen lack.
Table 3 Radix Aconiti Lateralis Preparata extract is on the impact of mice broken end mouth breathing time (x ± s)
| Group | Dosage (the g crude drug/Kg) | Test Mus number (only) | Breathing time (second) |
| Contrast | — | 12 | 18.9±4.05 |
| The E extract | 20 | 12 | 20.7±3.16 |
| The F extract | 20 | 12 | 25.4±3.65 ** |
| The G extract | 20 | 12 | 24.1±3.43 * |
| The H extract | 20 | 12 | 25.8±3.72 ** |
Annotate: compare with matched group
*P＜0.05,
*P＜0.01.
3, analgesia, antiinflammatory experiment
With above-mentioned A liquid, B liquid, C liquid, D liquid decompression recycling ethanol, be concentrated into clear paste, for subsequent use, carry out following effect experiment.
Get 50 of mices, male and female half and half are divided into 5 groups at random, i.e. Normal group, A extract group, B extract group, C extract group and D extract group.Each group was pressed table 4 dosed administration 3 days, 1h lumbar injection 1% sodium tartrate 0.2ml/ only after the not inferior administration, observe its writhing number of times, the results are shown in Table 4, show that the Radix Aconiti Lateralis Preparata total alkaloids extract (B, C, D extract) that preparation method of the present invention obtains has better analgesic activity.
Table 4 Radix Aconiti Lateralis Preparata extract analgesic experiment result
| Group | Dosage (the g crude drug/Kg) | Test Mus number (only) | Writhing number of times (inferior) |
| Normal control | — | 10 | 28±8.1 |
| The A extract | 12 | 10 | 18±7.8 * |
| The B extract | 12 | 10 | 10±4.5 *** |
| The C extract | 12 | 10 | 12±6.9 *** |
| The D extract | 12 | 10 | 10±5.3 *** |
Annotate: compare with matched group
*P＜0.05,
*P＜0.01,
* *P＜0.001.
Other gets male little 50, is divided at random 5 groups, i.e. Normal group, A extract group, B extract group, C extract group and D extract group.Each group was pressed table 5 dosed administration 3 days, only evenly be coated with dimethylbenzene 0.05ml/ for the left ear of mice in 40 minutes after the last administration, causing scorching rear 30 minutes execution animals, be that the rustless steel punching pin of 8mm takes off left and right sides same area auricle with diameter, weigh, be calculated as follows swelling and inhibitory rate of intumesce, the results are shown in 3, show that the Radix Aconiti Lateralis Preparata total alkaloids extract that preparation method of the present invention obtains has better antiinflammatory action.
Swelling=(left auricle Chong – auris dextra sheet is heavy),
Table 5 Radix Aconiti Lateralis Preparata extract antiinflammatory action experimental result
| Group | Dosage (the g crude drug/Kg) | Test Mus number (only) | Swelling (mg) | Suppression ratio (%) |
| Normal control | — | 10 | 10.85±2.54 | — |
| The A extract | 12 | 10 | 7.98±3.73 * | 26.5 |
| The B extract | 12 | 10 | 6.23±2.51 *** | 42.6 |
| The C extract | 12 | 10 | 6.58±2.85 *** | 39.4 |
| The D extract | 12 | 10 | 6.37±2.68 *** | 41.3 |
Annotate: compare with matched group
*P＜0.05,
*P＜0.01,
* *P＜0.001.
4, be adsorbed the different pH value of solution to the impact of adsorbing separation effect
Trial test shows, the pH value of the aqueous solution of aconitum plants or its processed goods extract, macroporous resin static adsorption effect is had a significant impact, transfer to respectively pH 4, pH 7, pH 10 such as Radix Aconiti Lateralis decocting liquid, adding waits the HPD100 type macroporous resin adsorption of crude drug amount, jolting 4 hours, the HPLC method is measured the content of benzoyl aconine, Benzoylmesaconine, benzoyl hypo-aconine, aconine, new aconine and inferior aconine in the water liquid of absorption front and back, calculate adsorption rate, the result shows that adsorption rate obviously increases from pH 4 to pH 10.Following research experiment has shown that further the pH value of solution is on the impact of resin dynamic adsorption.
Get Radix Aconiti Lateralis 750g, add 12 times in water, soaked 2 hours, decocted 3 hours, filter, decocting liquid is for subsequent use; Medicinal residues add 10 times in water again, decoct 2 hours, filter, and merge decocting liquid, are divided into three equal parts, are adjusted to respectively about pH 4, pH 7, pH 10, add water move to total amount and are 4 L., D101 type macroporous resin column that 80ml be housed identical by 3 respectively collected the water liquid that flows out again, and every part of 160ml is equivalent to the 10g crude drug, and every resin column is collected respectively 18 parts.Get about 2ml for every part, centrifugal, supernatant injects hplc determination as need testing solution.Chromatographic condition: chromatographic column is Eclipse XDB C
18Post (4.6 mm * 250 mm, 5 microns); Take 0.1% triethylamine as mobile phase A, take methanol as Mobile phase B, gradient elution program: 0～20 min: mobile phase A is by 80%～40%, and 20～35 min: mobile phase A is 40%.Evaporative light scattering detector detects; Column temperature: 35 ℃; Flow velocity: 1 mLmin
-1, sample size 100 microlitres.The peak area of benzoyl aconine, Benzoylmesaconine, benzoyl hypo-aconine, aconine, new aconine and inferior aconine is as shown in table 6 in the water liquid of upper prop front and back.
Experimental result shows: (1) macroporous adsorbent resin obviously is better than hydramine type alkaloid to the ester alkaloid adsorption effect, and namely after the loading overload, what at first outflow or leakage were analysed is hydramine type alkaloid.
(2) under the condition of neutral meta-alkalescence (pH 7-10), the adsorption efficiency of macroporous resin obviously is better than acid condition, and heterogeneity or different pH value have significant impact to the macroporous resin adsorption performance.。
For above-mentioned experimental phenomena, replace experiment medical material such as Radix Aconiti, Radix Aconiti Lateralis Preparata, Radix aconiti szechenyiani etc., or change macroporous resin such as HPD300, D101, AB-8 etc., all can obtain similar experimental result.Sufficient proof, the aqueous solution of aconitum plants or its processed goods extract is behind adjust pH to 7 ~ 10, go up again macroporous adsorbent resin, more be conducive to the absorption of hydramine type alkaloid component, obviously improve the macroporous resin adsorption capacity, therefore, improve production efficiency, reduced production cost.
Above-mentioned experimental result fully shows, adopt the total alkaloids extract of the aconitum plants such as Radix Aconiti Lateralis Preparata that the inventive method prepares, Radix Aconiti or its processed goods, can keep well alkaloid component in its extract, avoid the heavy losses of art methods to alkaloid component.Further pharmacodynamic experiment clearly illustrates that, the total alkaloids extract that adopts the inventive method to prepare, at aspects such as antiinflammatory, analgesia, cold resistance, anoxia enduring, can have better drug action, can be applicable to clinically arthralgia due to cold-dampnes, quasi-wind gateway and heart failure, recuperating depleted YANG and rescuing the patient from collapse etc., have broad application prospects.
Claims (10)
1. the preparation method of total alkaloids extract in aconitum plants or its processed goods, it is characterized in that to contain the aqueous solution adjusting pH value 7 ~ 10 of aconitum plants or its processed goods extract, after being preferably pH value 8 ~ 10, adsorb with macroporous adsorptive resins, then after washing the removal of impurity with water first, be 50 ~ 90% acidic ethanol-aqueous solution eluting with pH≤5 and alcohol volume content at least again, collect eluent, desolventizing namely obtains aconitum plants and processed goods total alkaloids extract thereof.
2. preparation method as claimed in claim 1 is characterized in that the said aqueous solution that contains aconitum plants or its processed goods extract, is the aqueous extract of aconitum plants or its processed goods, or the aqueous solution of aconitum plants or its processed goods extract.
3. preparation method as claimed in claim 2 is characterized in that the aqueous extract of said aconitum plants or its processed goods is the aqueous extract of pH≤5, is preferably the aqueous solution with hydrochloric acid or sulfuric acid acidation.
4. preparation method as claimed in claim 1, after it is characterized in that the said aqueous solution that contains aconitum plants or its processed goods extract adsorbs with macroporous adsorptive resins, when washing remove impurity with water, adopt alkaline water and the neutral water washing of using successively neutral water, pH 8 ~ 10; Perhaps adopt alkaline water washing and the neutral water washing of using successively pH 8 ~ 10.
5. preparation method as claimed in claim 1 is characterized in that said extractive content for aconitum plants or its processed goods in the aqueous solution of macroporous adsorptive resins absorption is no more than and is equivalent to 2 gram crude drug/milliliters.
6. preparation method as claimed in claim 1 is characterized in that said acidic ethanol-aqueous solution is the ethanol-water solution with hcl acidifying.
7. preparation method as claimed in claim 1 is characterized in that saidly during with acidic ethanol-aqueous solution eluting, adopts and uses first acidic ethanol-aqueous solution eluting, uses at last neutral alcohol-aqueous solution eluting.
8. preparation method as claimed in claim 1, after the eluent that it is characterized in that said collection is adjusted to first pH value 5 ~ 7, again desolventizing.
9. preparation method as claimed in claim 1 after it is characterized in that the eluent of collecting used first alkali type anion exchange resin process, is carried out the operation of desolventizing again.
10. preparation method as claimed in claim 1 is characterized in that said macroporous adsorbent resin is HPD100 or D101 type macroporous adsorbent resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210559436.5A CN102973679B (en) | 2012-12-21 | 2012-12-21 | Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210559436.5A CN102973679B (en) | 2012-12-21 | 2012-12-21 | Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102973679A true CN102973679A (en) | 2013-03-20 |
| CN102973679B CN102973679B (en) | 2014-06-11 |
Family
ID=47848214
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210559436.5A Active CN102973679B (en) | 2012-12-21 | 2012-12-21 | Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102973679B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2019212668B2 (en) * | 2018-01-24 | 2021-04-15 | Gooddoctor Pharmaceutical Group Co., Ltd. | Crystalline forms of mesaconine and preparation method therefor |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5852273A (en) * | 1981-09-21 | 1983-03-28 | Taiho Yakuhin Kogyo Kk | Aconitine-alkaloid, its preparation, and antiphlogistic anodyne containing it |
| RU2039568C1 (en) * | 1992-08-25 | 1995-07-20 | Институт химии растительных веществ АН Республики Узбекистан | Process for manufacturing antiarrythmic composition |
| CN1969971A (en) * | 2006-12-01 | 2007-05-30 | 哈尔滨商业大学 | Transdermal absorption formulation of total alkaloid of common monkshood root and preparation process thereof |
| CN101468076A (en) * | 2007-12-26 | 2009-07-01 | 上海玉森新药开发有限公司 | Chinese medicine gel for treating arthritis and preparation method thereof |
| CN101829201A (en) * | 2010-05-19 | 2010-09-15 | 重庆市中药研究院 | Method for extracting alkaloid from monkshood medicinal material |
-
2012
- 2012-12-21 CN CN201210559436.5A patent/CN102973679B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5852273A (en) * | 1981-09-21 | 1983-03-28 | Taiho Yakuhin Kogyo Kk | Aconitine-alkaloid, its preparation, and antiphlogistic anodyne containing it |
| RU2039568C1 (en) * | 1992-08-25 | 1995-07-20 | Институт химии растительных веществ АН Республики Узбекистан | Process for manufacturing antiarrythmic composition |
| CN1969971A (en) * | 2006-12-01 | 2007-05-30 | 哈尔滨商业大学 | Transdermal absorption formulation of total alkaloid of common monkshood root and preparation process thereof |
| CN101468076A (en) * | 2007-12-26 | 2009-07-01 | 上海玉森新药开发有限公司 | Chinese medicine gel for treating arthritis and preparation method thereof |
| CN101829201A (en) * | 2010-05-19 | 2010-09-15 | 重庆市中药研究院 | Method for extracting alkaloid from monkshood medicinal material |
Non-Patent Citations (1)
| Title |
|---|
| 李文兰等: "X-5大孔树脂纯化新乌头碱和乌头总碱", 《中国中药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2019212668B2 (en) * | 2018-01-24 | 2021-04-15 | Gooddoctor Pharmaceutical Group Co., Ltd. | Crystalline forms of mesaconine and preparation method therefor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102973679B (en) | 2014-06-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102451235B (en) | Preparation method of olive leaf extract | |
| CN103550273B (en) | A kind of method extracting the residual ginsenoside of low agriculture from Radix Ginseng | |
| CN103304613B (en) | A kind of method of separation and purification 4 kinds of ucleosides chemical compositions from Snakegourd Peel | |
| CN102423329B (en) | A kind of discoloration method of panax notoginsenoside extract | |
| CN102631414B (en) | SepHaniadelavayi Diels total alkaloid extraction and purification technology | |
| CN101348474A (en) | Method for preparing salvianolic acid B and tanshinol from Salvia miltiorrhiza stem | |
| CN102875450A (en) | Technological method for extracting 1-deoxynojirimycin from mulberry leaf | |
| CN109879919B (en) | Method for separating and preparing three flavonoid glycosides from spina date seeds | |
| CN104610401B (en) | A kind of method for extracting baicalin, baicalin and wogonin from Radix Scutellariae simultaneously | |
| CN102228515B (en) | Separation and enrichment method of total flavones and total alkaloids of Lotus Plumule | |
| WO2012019373A1 (en) | Method for preparing paeoniflorin and albiflorin | |
| CN102617669A (en) | Method for separating and purifying mangiferin from mango pericarp | |
| CN103304490A (en) | Method for separating and purifying five purine and pyrimidine bases from trichosanthes bark | |
| CN102895338B (en) | Preparation method of monkshood or radix aconite total alkaloid extractive | |
| CN105175426B (en) | A kind of method of the extraction purification Bergenin from treebine stem | |
| CN106674312A (en) | Separation and purification method for high-purity monomer sweroside series components | |
| CN102973679B (en) | Preparation method of total alkaloids extract in aconitum medical material or processed products of aconitum plants | |
| CN107722080A (en) | A kind of method that ursin is extracted in the leaf from purple bergenia herb | |
| CN102172361A (en) | Method for separating traditional Chinese medicine total alkaloids by ion exchange resin | |
| CN104189073B (en) | The preparation method of salvianolic acid | |
| CN103381200A (en) | White mulberry root-bark total alkaloid extract and preparation and application thereof | |
| CN102920785A (en) | Method for extracting total flavone from guava leaves | |
| CN102920727B (en) | Method for preparing extracts rich in vitexin rhamnoside and vitexin glucoside | |
| CN104725449A (en) | Paeoniflorin extracting method | |
| CN103083392A (en) | Method for separating and enriching part with prenylated flavonoid in subprostrate sophora |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |