CN101278980A - A kind of quaternary ammonium total alkali and preparation method thereof - Google Patents

Abstract

The invention discloses a preparation method of total base of quaternary ammonium corydalis. The method comprises the steps of ultrasonic treatment when Corydalis is soaked, extraction of quaternary ammonium base components under reflux or microwave conditions, the resulting extract is recovered from ethanol, exchanged and adsorbed through a treated resin column, and washed with ammonia alcohol solution or aqueous ethanol. The eluate is concentrated under reduced pressure, dried, and the dried product is redissolved with ethanol containing water, filtered or centrifuged, the filtrate or supernatant is concentrated under reduced pressure, and dried to produce total base of quaternary ammonium corydalis. The quaternary ammonium total base of Corydalis prepared by the method has high extraction efficiency, less impurity content and low cost, and is especially suitable for mass production of medicinal material extracts, single prescriptions of traditional Chinese medicines or new compound preparations containing Corydalis Corydalis. The corydalis quaternary ammonium total alkaloid prepared by the invention has definite curative effect and can be used for the prevention and treatment of cardiovascular and cerebrovascular diseases.

Description

A kind of quaternary ammonium total alkali and preparation method thereof

technical field

The invention relates to a method for extracting and separating effective parts of traditional Chinese medicine, in particular to a method for extracting and separating effective parts of quaternary ammonium total alkaloids from Chinese medicine Corydalis Corydalis.

Background technique

Corydalis is commonly known as Yuanhu, and the Chinese Pharmacopoeia records that its authentic source is the dry tuber of Corydalis yanhusuo W.T.Wang, a plant of the family Papaveraeeae (Corydalis), which has the effects of promoting blood circulation, removing blood stasis, promoting qi and relieving pain. Corydalis Corydalis as a traditional analgesic and blood-activating and stasis-removing drug has been used in Chinese medicine for more than two thousand years. The chemical components of Corydalis Corydalis are mainly alkaloids, and there are many studies. Twenty-five species were found, which can be divided into protoberberine, prototropine, aporphenanthrene, benzophenanthridine, phthalide isoquinoline, benzyl isoquinoline, Several types of alkaloids such as isoquinoline benzimidazolines, most of which belong to protoberberines and aporberines. According to the nature of its amino nitrogen, it can be divided into two categories: tertiary amine base and quaternary ammonium base. At present, the research on Corydalis is still mostly limited to the research on tertiary amine bases such as tetrahydropalmatine or its derivatives. Modern research on Corydalis mainly focuses on the central nervous system. The tertiary amine bases in Corydalis mostly show analgesia and sedation and stable effect, tetrahydropalmatine is the representative compound; if the dehydrogenation of tetrahydropalmatine is converted into the corresponding quaternary ammonium base (palmitine), it will lose its analgesic and sedative effects. On the contrary, berberine will be hydrogenated into the corresponding tertiary amine Alkalines show strong analgesic and sedative effects, indicating that their pharmacological effects are closely related to the properties of their amino nitrogen. Ancient Chinese medical books often mentioned the use of Corydalis Corydalis to treat heartache and chest obstruction, and it is also widely used in the prescriptions for treating cardiovascular diseases in modern times. Modern pharmacological studies have proved that Corydalis Corydalis can significantly increase coronary blood flow, improve myocardial ischemia and hypoxia, thereby relieving angina. Corydalis Corydalis has a significant effect on the cardiovascular system, mainly the original berberine-type quaternary ammonium alkaloids represented by dehydrocorydine.

At present, the literature reports that the total alkaloids extracted from Corydalis Corydalis are mainly composed of tertiary amine bases; the method used involves supercritical carbon dioxide extraction technology (Liu Longzhong. Optimizing the Extraction Process of Corydalis Corydalis. Journal of Zunyi Medical College[J] , 2004, 27(6): 595-596.), ultrasonic extraction technology (Xie Caijuan, Zhang Zhiqi. Ultrasonic extraction of total alkaloids from Corydalis [J]. Northwest Pharmaceutical Journal, 2005, 20(4): 153-154.) and macroporous adsorption resin technology (Liu Junhong, Wei Junfeng, Wang Hongzhi, etc. Application of macroporous adsorption resin in the extraction and separation of Corydalis alkaloids [J]. Chinese Herbal Medicine, 2002, 33(1): 37-38; Zhang Lizhu, Zhu Zhu, Hao Xiaoyan. Comparison of two extraction methods of total alkaloids from Corydalis [J]. Journal of Guiyang Medical College, 2006, 31(3): 280-282.) and so on. Chinese Patent Application No. 200610049743.3 discloses a method for preparing total alkali extract from Corydalis Corydalis and various preparations prepared from the extract, which are used to treat acute and chronic pain, sedation, hypnosis, and antidepression. The technical method involved in this invention is: use 5-10% Na ₂ CO ₃ or NaHCO ₃ or NaOH to soak and alkalize, then extract with 10-85% ethanol, concentrate the extract under reduced pressure, and use 1 ～10% HCl or 1～10% H ₂ SO ₄ acidification, the acidified solution is subjected to resin column chromatography, and eluted with 0～90% ethanol, the eluate is concentrated under reduced pressure, and freeze-dried to obtain dry powder of total alkaloids. None of the existing methods mentions the extraction of the quaternary ammonium base components of Corydalis.

Contents of the invention

The purpose of the present invention is to provide a kind of preparation method of quaternary ammonium total alkaloids of Corydalis. The present invention aims at the problem that the content of quaternary ammonium alkaloids is low when extracting Corydalis, by adopting ultrasonic treatment in the soaking process, and utilizing macroporous adsorption resin and The adsorption properties of the weakly acidic cation exchange resin to the quaternary ammonium component of Corydalis, adding impurity removal procedures before and after desorption, not only greatly increases the extraction efficiency of the quaternary ammonium component, but also greatly increases its content.

The second object of the present invention is to provide a kind of total alkali of quaternary ammonium fumarium, and total alkali content is more than 55%, wherein the peak area of quaternary ammonium base composition accounts for 2% of the total peak area in the total peak area of quaternary ammonium fumarium in the high performance liquid chromatography More than 80% can be used for the prevention and treatment of cardiovascular and cerebrovascular diseases.

The purpose of the present invention is to realize through following technical scheme:

A kind of preparation method of quaternary ammonium fumarate total alkali is characterized in that it comprises the steps:

(1) mixing and soaking Corydalis Corydalis powder in hydrous ethanol and performing ultrasonic treatment at the same time, heating the mixed solution after soaking to reflux for extraction, or placing the mixed solution in microwave equipment for microwave radiation extraction;

(2) After the extract is filtered or centrifuged, the filtrate or supernatant is recovered from ethanol until it has no alcohol smell, and after being diluted with water, it is exchanged and adsorbed through a resin column that has been treated by a conventional method;

(3) After resin exchange and adsorption are completed, first elute with deionized water or dilute alkali solution, then elute with ammonia alcohol solution or aqueous ethanol, collect the eluate, concentrate under reduced pressure, and dry;

(4) The dried extract is redissolved with aqueous ethanol, suction filtered or centrifuged, and the filtrate or supernatant is recovered by conventional methods for ethanol and dried to obtain total base of quaternary ammonium corydalis.

In the step (1), the powder particle size of Corydalis Corydalis is 5-50 meshes; the concentration of hydrous ethanol used is 30%-95%;

The soaking time in the step (1) is 30-240 minutes; the ultrasonic frequency used is 20-40KHz.

The extraction time during the reflux extraction in the step (1) is 1 to 4 hours, and the extraction times are 2 to 4 times.

During the microwave extraction in the step (1), the microwave output power is 500-800W, the irradiation time is 10-30 minutes, the extraction temperature is 50-70° C., and the extraction times are 1-3 times.

The resin used in the step (2) is a weakly acidic cation exchange resin, the type of which is 724 or D151.

The resin used in the step (2) is a macroporous adsorption resin, and the model is AB-8 or X-5.

The dilute alkali solution used in the step (3) is 1-10% ammonia solution; the concentration of ammonia-alcohol solution is 5%-25%; and the concentration of aqueous ethanol is 30-95%.

The concentration of the aqueous ethanol used in the step (4) is 10-95%.

The invention relates to a total quaternary ammonium corydalis, which is prepared by a preparation method of the total quaternary corydalis ammonium.

The preparation method of the present invention is suitable for the extraction of quaternary ammonium base components of Corydalis. The process of the present invention first crushes the medicinal material of Corydalis Corydalis, and uses ultrasonic treatment in the soaking process, which directly affects the infiltration speed and degree of the medicinal material of Corydalis Corydalis, and then affects the quaternary ammonium base The extraction effect of ingredients; the extraction process adopts microwave extraction, which can greatly shorten the extraction time, reduce costs, and improve production efficiency. In addition, aqueous ethanol extraction can also prevent excessive water-soluble impurities in Corydalis Corydalis from being brought into the extract. After the ethanol is recovered from the extract, it is directly treated with a resin column, and the quaternary ammonium base components of Corydalis can be well separated and enriched, and a large amount of non-alkaloid impurities can be removed by elution with a dilute alkali solution. Redissolution and further removal of impurities reduce the amount of solids and increase the content of the quaternary ammonium fumarium alkaloids substantially, thereby ensuring the purity and quality of the total alkaloid extract of the quaternary ammonium fumatillon.

Identification of quaternary ammonium total bases of Corydalis:

Quantitative analysis by the acid dye method: with dehydrodrycardine as the reference substance, potassium dihydrogen phosphate-sodium hydroxide buffer solution with pH 8.0 and 0.1% bromothymol blue solution as the chromogen, and 50% methanol solution as the extraction agent. The solvent of the sample and the reference substance, the concentration of the extract sample is set at 0.2mg/ml, the detection wavelength is set at 415nm, the regression equation is Y=5.7549X+0.0117, r=0.9991; the linear range is 0～0.102mg. The total alkaloid content (calculated as dehydrodrygosine) in all the extracts extracted by this process is above 55%.

High-performance liquid chromatography qualitative analysis: select acetonitrile and water (containing 0.8% glacial acetic acid and 0.2% triethylamine) as organic and inorganic mobile phases, the ratio of the two phases is 23: 77, and the mobile phase flow rate is 1.0ml/min, The column temperature was set at 30 °C, the detection wavelength was set at 335 nm; the injection volume was 10 μl. The reference substance used in the experiment includes dehydrodrytidine, berberine, coptisine, palmatine and African tetrandrine, and 50% methanol solution is selected as the solvent of the extract sample and the reference substance, and the concentration of the extract sample is set at 2mg/ ml, taking the HPLC chromatogram of the reference substance as a comparison, there are at least 7 chromatographic peaks in all the extracts extracted by this process, and there are dehydrodrycardine, berberine, coptisine, palmatine and African tetrandrine , the peak areas of the five quaternary ammonium base components accounted for more than 80% of the total peak area.

The preparation method of total quaternary ammonium corydalis of the present invention can be made into oral dosage form or injection dosage form with one or more pharmaceutically acceptable carriers, and can be used for the preparation of prevention and treatment of heart and brain Drugs for vascular disease.

The pharmaceutically acceptable carrier mentioned above refers to the conventional drug carrier in the pharmaceutical field, for example: diluents such as lactose, starch, dextrin, powdered sugar, microcrystalline cellulose, etc.; binders such as cellulose derivatives, gelatin, Polyvinylpyrrolidone, etc.; wetting agents such as water, ethanol, etc.; disintegrants such as dry starch, sodium carboxymethyl starch, sodium alginate, etc.; flavoring agents such as stevioside, xylitol, etc.; lubricants such as talcum powder, hard Magnesium fatty acid, micropowder silica gel, etc.; substrates such as vegetable oil, polyethylene glycol, beeswax, etc.; preservatives such as benzoic acid, paraben, etc.

The preparation method of the total base of quaternary ammonium corydalis of the present invention can be administered to patients who need such treatment through oral administration or injection. The total base of quaternary ammonium corydalis prepared in the present invention and pharmaceutically acceptable carriers can be made into conventional solid or liquid preparations such as tablets, hard capsules, soft capsules, dropping pills, granules, oral liquids, injections and the like.

The usage amount of the total alkaloids of quaternary ammonium corydalis prepared by the preparation method of total quaternary ammonium corydalis of the present invention can be according to the patient's body weight, the type and severity of the disease to be treated, and its 24-hour dose can be 0.5～5mg/ kg body weight.

Description of drawings

Fig. 1 is the high performance liquid phase chromatogram of a kind of Corydalis quaternary ammonium total alkali extract of the present invention.

Wherein No. 1 peak is African tetrandrine (molecular weight 338), No. 2 peak is blue cordyline (molecular weight 370), No. 3 peak is coptisine (molecular weight 320), No. 4 peak (molecular weight 352), No. 5 peak (molecular weight 352 ), the No. 6 peak is palmatine (molecular weight 352), the No. 7 peak is berberine (molecular weight 336), and the No. 8 peak is dehydrodrycodine (molecular weight 366).

Detailed ways

The present invention will be further described below in conjunction with specific examples, but the present invention is not limited in any way.

Embodiment 1,

Corydalis medicinal material is crushed, passed through a 5-mesh sieve, soaked in 6 times the amount of 80% ethanol and 20KHz ultrasonic treatment, soaking time is 60 minutes, the soaked Corydalis medicinal material reflux extraction method is extracted 3 times (6 times, 4 times, 4 times) , 4 hours each time, combine the extracts, filter to remove the medicinal residues, recover the ethanol from the filtrate until it has no alcohol smell, dilute the medicinal solution with deionized water, pass through the X-5 macroporous adsorption resin, and the medicinal solution is absorbed by the resin column, The flow rate is 2 ml/min. After the adsorption is completed, first wash off the non-adsorbed impurities with 2% ammonia solution, then elute with 50% ethanol solution, collect the alcohol eluate, and concentrate the eluate under reduced pressure. Vacuum drying, redissolving the dried product with 95% ethanol, filtering, concentrating the filtrate under reduced pressure, and vacuum drying below 60°C to obtain the total base of quaternary ammonium corydalis.

Embodiment 2,

Corydalis medicinal material is crushed, passed through a 5-mesh sieve, soaked in 6 times the amount of 80% ethanol and 20KHz ultrasonic treatment, soaking time is 60 minutes, the soaked Corydalis medicinal material reflux extraction method is extracted 3 times (6 times, 4 times, 4 times) , 4 hours each time, combine the extracts, filter to remove the medicinal residues, reclaim the ethanol from the filtrate until there is no alcohol smell, the medicinal liquid passes through the 724 type weakly acidic cation exchange resin, and the filtrate is adsorbed and exchanged by the resin column at a flow rate of 0.5 ml/min; After the exchange is completed, first wash away the non-adsorbed impurities with water, then elute with 20% ammonia alcohol solution, collect the ammonia alcohol eluate, use alkaloid reagent to detect the elution process, stop adding eluent until the end of elution, Concentrate the eluate under reduced pressure, dry it in vacuum below 60°C, redissolve the dried product in 50% ethanol, filter, recover the ethanol from the filtrate under reduced pressure, and dry it under vacuum below 60°C to obtain the total base of quaternary ammonium corydalis.

Embodiment 3,

Corydalis medicinal material is crushed, passed through a 50-mesh sieve, soaked in 10 times the amount of 70% ethanol, and then subjected to 20KHz ultrasonic treatment for 120 minutes. The soaked medicinal material Corydalis Corydalis is placed in a microwave extraction tank for microwave extraction. The output power of the microwave is 650W. 65°C, irradiated for 10 minutes, extracted 3 times, combined the extracts, filtered to remove the medicinal residues; the filtrate recovered ethanol until it had no alcohol smell, the medicinal liquid passed through the AB-8 macroporous adsorption resin, and the filtrate was adsorbed by the resin column; the flow rate was 1.5 mL/min, after the adsorption is completed, first wash away the non-adsorbed impurities with 5% ammonia solution, then elute with 50% ethanol solution, collect the eluate, concentrate the eluate under reduced pressure, and dry it below 60°C. The dry matter is redissolved with 95% ethanol, filtered, the filtrate is concentrated under reduced pressure, and dried at a temperature of 60°C to obtain the total base of quaternary ammonium corydalis.

Embodiment 4,

Corydalis medicinal material is crushed, passed through a 50-mesh sieve, soaked in 10 times the amount of 70% ethanol, and then subjected to 20KHz ultrasonic treatment for 120 minutes. The soaked medicinal material Corydalis Corydalis is placed in a microwave extraction tank for microwave extraction. The output power of the microwave is 650W. 65°C, irradiated for 10 minutes, extracted 3 times, combined the extracts, filtered to remove the medicinal residues; the filtrate recovered ethanol until it had no alcohol smell, and the liquid passed through the D151 weakly acidic cation exchange resin, and the flow rate of the liquid was absorbed and exchanged through the resin column 1.0 ml/min; after the exchange is completed, wash away the non-adsorbed impurities with water, elute with 20% ammonia alcohol solution, collect the ammonia alcohol eluate, use alkaloid reagent to detect during the elution process, and stop at the end of the elution Add the eluent, concentrate the eluent under reduced pressure, dry it in vacuum below 60°C, redissolve the dried product in 50% ethanol, filter, recover the ethanol from the filtrate under reduced pressure, and dry it under vacuum below 60°C to obtain the total quaternary ammonium corydalis. alkali.

Claims (10)

1, a kind of preparation method of Rhizoma Corydalis quaternary ammonium total alkali is characterized in that carrying out according to the following steps:

(1) the corydalis tuber medicinal material powder is placed in the aquiferous ethanol to mix and soaks and carry out ultrasonic Treatment simultaneously, the mixed solution heating and refluxing extraction after the immersion is maybe put this mixed solution and is carried out microwave radiation extraction in the microwave equipment;

(2) with after extracting solution filtration or the centrifugal sedimentation, filtrate or supernatant reclaim ethanol and do not distinguish the flavor of to there being alcohol, exchange, adsorb through the resin column of having handled well according to a conventional method behind the thin up;

(3) finish through resins exchange, absorption, earlier with deionized water or dilute alkaline soln eluting, reuse ammonia alcoholic solution or aquiferous ethanol eluting are collected eluent, eluent concentrating under reduced pressure, drying;

(4) extract after drying redissolves with aquiferous ethanol, sucking filtration or centrifugal, and filtrate or supernatant reclaim ethanol according to a conventional method, and drying promptly gets Rhizoma Corydalis quaternary ammonium total alkali.

2, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: the corydalis tuber medicinal material powder diameter is 5～50 orders in the described step (1); The aquiferous ethanol concentration of using is 30%～95%; Amount and the Rhizoma Corydalis that adds aquiferous ethanol count 1: 2 by weight～and 20.

3, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: soak time is 30～240 minutes in the described step (1); The ultrasonic frequency of using is 20～40KHz.

4, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: extraction time be 1～4 hour during reflux, extract, in the described step (1), extraction time is 2～4 times.

5, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: in the described step (1) during microwave extraction microwave output power be 500～800W, exposure time is 10～30 minutes, extracting temperature is 50～70 ℃; Extraction time is 1～3 time.

6, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: the resin that uses in the described step (2) is weak-acid cation-exchange resin, and model is 724 types or D151 type.

7, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: the resin that uses in the described step (2) is macroporous adsorbent resin, and model is AB-8 type or X-5 type.

8, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: employed dilute alkaline soln is 1～10% ammonia spirit in the described step (3); Ammonia alcoholic solution concentration is 5%～25%; Aquiferous ethanol concentration is 30～95%.

9, by the preparation method of the described a kind of Rhizoma Corydalis quaternary ammonium total alkali of claim 1, it is characterized in that: employed aquiferous ethanol concentration is 10～95% in the described step (4).

10, a kind of Rhizoma Corydalis quaternary ammonium total alkali is characterized in that making with the method for one of claim 1～9.

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