CN107674081B - Preparation method of bicuculline and protopine - Google Patents
Preparation method of bicuculline and protopine Download PDFInfo
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- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
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Abstract
The invention discloses a preparation method of bicuculline and protopine, which comprises the following steps of 1) extracting: pulverizing the medicinal materials, extracting with acidic methanol or ethanol solution to obtain extractive solution A, and concentrating under reduced pressure to obtain concentrated solution B. 2) Separation and enrichment: and (3) taking the concentrated solution B, adjusting the pH value of the concentrated solution B, adsorbing by using a reverse polymerization filler C, eluting by using water to obtain a water washing solution D, then, resolving by using an alcohol solution E, collecting a resolving solution F, and then, resolving by using an alcohol solution G to obtain a resolving solution H. 3) And (3) purification: taking the analytic solution H, concentrating under reduced pressure to adjust the pH value, standing, filtering and precipitating, mixing the precipitate with a sample, and performing silica gel column chromatography to obtain a solution J; concentrating the analysis solution F under reduced pressure, adjusting the pH value to 9-10, and separating out a precipitate K; adjusting pH of the water washing solution D to 9-10, and separating out precipitate L. 4) And (3) crystallizing and drying: concentrating, crystallizing and drying the solution J under reduced pressure to obtain pure bicuculline; and crystallizing and drying the precipitate K and the precipitate L to obtain the protopine pure product. The method has high yield and high purity, and can be used for industrial production.
Description
Technical Field
The invention relates to the field of natural pharmacy, in particular to a preparation method of biculin alkali and protopine.
Background
The molecular formula of the Bicuculline (+) -Bicuculline is C20H17NO6367.36 molecular weight, CAS number 485-49-4, is light yellow powder, is soluble in benzene, chloroform, ethyl acetate, and slightly soluble in ethanol and diethyl ether; protopine molecular formula is C20H19NO5The molecular weight is 353.37, CAS number is 130-86-9, it is white powder, it is soluble in benzene, chloroform, ethyl acetate, slightly soluble in ethanol and ether, both compounds are alkaloids, and its structural formula is shown below.
In the prior art, a preparation method capable of separating and purifying the bicuculline from plants does not exist, and the problems that the extraction method of the protopine is low in yield and difficult in industrial production exist.
Disclosure of Invention
The invention provides a preparation method of biculin alkali and protopine, and by using the method, the yield of the biculin alkali and the protopine is over 80 percent, and the purity of the biculin alkali and the protopine is over 99 percent.
In order to solve the technical problems, the invention provides a method for preparing biculin alkali and protopine, which is characterized by comprising the following steps:
1) extraction: pulverizing one or more of rhizoma corydalis Decumbentis, rhizoma corydalis Decumbentis or corydalis edulis, extracting with acidic methanol or ethanol solution to obtain extractive solution A, mixing extractive solutions, concentrating under reduced pressure to remove ethanol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5, adsorbing with reverse polymeric filler C, eluting with water to obtain water washing solution D, eluting with alcoholic solution E, collecting eluate F, and eluting with alcoholic solution G to obtain eluate H
3) And (3) purification: taking analysis liquid H, concentrating under reduced pressure, adjusting pH to 8-10, standing, filtering, precipitating, dissolving precipitate with methanol, mixing with silica gel to obtain column-passing raw material I, loading the sample I, eluting with a dichloromethane-methanol system with eluent volume percentage of 100:1-10:1, and collecting solution J with dichloromethane-methanol volume percentage of 80:1-40: 1; concentrating the analysis solution F under reduced pressure, adjusting pH to 9-10, and separating out a precipitate K; and (5) adjusting the pH of the water washing solution D to 9-10, and separating out a precipitate L.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain an extract M, and crystallizing the extract M twice to obtain a pure product of the bicolor sodium; crystallizing the precipitate K and the precipitate L once to obtain a crude sodium macleyite pure product, and drying the pure product by vacuum blast to obtain pure product powder of the series of products;
the reverse polymerization filler C in the step 2) is any one selected from AB-8 macroporous resin, D-101 macroporous resin and NM 100-reverse polymer chromatographic filler.
The alcoholic solution E in the step 2) is methanol or ethanol solution, and the volume percentage of the methanol or the ethanol in the alcoholic solution E is 30-50%.
The alcoholic solution G in the step 2) is methanol or ethanol solution, and the volume percentage of the methanol or the ethanol in the alcoholic solution G is 80-95%.
Preferably, in step 1), the acidic methanol or ethanol solution contains 0.1-2% by volume of hydrochloric acid and 50-100% by volume of methanol or ethanol;
and 4) crystallizing the extract M in the step 4) by using a dichloromethane-methanol system, and crystallizing the precipitate K by using a dichloromethane-absolute ethanol system.
Preferably, the extraction method in step 1) comprises: reflux extraction, ultrasonic extraction and percolation extraction.
Preferably, the temperature for concentration under reduced pressure in step 1) is 50 to 70 ℃.
Preferably, the temperature at which the solution H and the solution F are concentrated under reduced pressure in step 3) is 50 to 60 ℃.
Preferably, ammonia water is used in step 3) to adjust the pH values of the analysis liquid H, the analysis liquid F and the washing liquid D.
In the invention, the medicinal materials are crushed and extracted in different modes, the extracting solution is adsorbed by a reverse packed column and then eluted by methanol or ethanol solutions with different volume percentages for resolution, the resolving solution is subjected to silica gel column chromatography and direct precipitation to obtain a crude product of a target product, and the crude product is recrystallized to obtain a pure product.
Detailed Description
In order that those skilled in the art will better understand the technical solutions of the present invention, the present invention will be further described in detail with reference to the following embodiments.
The invention discloses a preparation method of bicuculline and protopine, which comprises the following steps of 1) extracting: pulverizing the medicinal materials, extracting with acidic methanol or ethanol solution to obtain extractive solution A, and concentrating under reduced pressure to obtain concentrated solution B. 2) Separation and enrichment: and (3) taking the concentrated solution B, adjusting the pH value, adsorbing by using a reverse polymerization filler C, eluting by using water to obtain a water washing solution D, then, resolving by using an alcoholic solution E, collecting a resolving solution F, and then, resolving by using an alcoholic solution G to obtain a resolving solution H. 3) And (3) purification: taking the analytic solution H, concentrating under reduced pressure to adjust pH, standing, filtering, precipitating, mixing with sample, and performing silica gel column chromatography to obtain solution J; concentrating the analysis solution F under reduced pressure, adjusting pH to 9-10, and separating out a precipitate K; and (5) adjusting the pH of the water washing solution D to 9-10, and separating out a precipitate L. 4) And (3) crystallizing and drying: concentrating, crystallizing and drying the solution J under reduced pressure to obtain pure bicuculline; and crystallizing and drying the precipitate K and the precipitate L to obtain the protopine pure product. The method has high yield and high purity, and can be used for industrial production.
The foregoing is a detailed description of the invention and the following is an example of the invention.
Example one
1) Extraction: pulverizing one or more of rhizoma corydalis Decumbentis, rhizoma corydalis Decumbentis or corydalis edulis, extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid and 50-100 vol% methanol or ethanol under reflux to obtain extractive solution A, extracting for 1-2 hr each time for 3-6 times, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no ethanol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water solution D, eluting with 3 times of alcohol solution E (30-50% methanol or ethanol) to obtain an eluate F, and eluting with alcohol solution G (80-95% methanol or ethanol) to obtain an eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; and (5) adjusting the pH of the water washing solution D to 9-10 with ammonia water, separating out a precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: according to different medicinal materials used in the step 1), the experiment is divided into 3 groups, wherein a group is corydalis amabilis, b group is corydalis tuber, and c group is corydalis edulis. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 1
TABLE 1 Experimental results of a different grouping of examples
The experimental results show that the medicinal material corydalis amabilis is the preferred embodiment of the invention.
Example two
1) Extraction: pulverizing rhizoma corydalis Decumbentis, extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid under reflux to obtain extractive solution A, extracting for 3-6 times for 1-2 hr each time, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no ethanol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water solution D, eluting with 3 times of alcohol solution E (30-50% methanol or ethanol) to obtain an eluate F, and eluting with alcohol solution G (80-95% methanol or ethanol) to obtain an eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; and (5) adjusting the pH of the water washing solution D to 9-10 with ammonia water, separating out a precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: according to the difference of the percentage of methanol or ethanol in the acidic methanol or ethanol in the step 1), dividing the experiment into 4 groups, wherein the group d is the acidic methanol with the volume percentage of 1-50% of methanol, the group e is the acidic methanol with the volume percentage of 50-100% of methanol, the group f is the acidic ethanol with the volume percentage of 1-50% of ethanol, and the group g is the acidic ethanol with the volume percentage of 50-100% of ethanol. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 2
TABLE 2 Experimental results for two different groups of examples
The above experimental results show that the percentage of methanol or ethanol in acidic methanol or ethanol is 50-100% is the preferred mode.
EXAMPLE III
1) Extraction: pulverizing rhizoma corydalis Decumbentis, and extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid and 50-100 vol% methanol or ethanol. Extracting with one of reflux extraction, ultrasonic extraction or percolation extraction to obtain extractive solution A, extracting for 3-6 times for 1-2 hr each time, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no alcohol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water solution D, eluting with 3 times of alcohol solution E (30-50% methanol or ethanol) to obtain an eluate F, and eluting with alcohol solution G (80-95% methanol or ethanol) to obtain an eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; and (5) adjusting the pH of the water washing solution D to 9-10 with ammonia water, separating out a precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: dividing the experiment into three groups of h, i and j according to different extraction modes in the step 1), wherein the h adopts a reflux extraction mode, the i adopts an ultrasonic extraction mode, and the j adopts a percolation extraction mode. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 3
TABLE 3 grouping test results of example III
The experimental results show that the yield of reflux extraction, ultrasonic extraction and percolation extraction is over 80 percent, the purity is over 99 percent, and the results have no significant difference, so the three extraction modes are the preferred modes of the invention.
Example four
1) Extraction: pulverizing rhizoma corydalis Decumbentis, and extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid and 50-100 vol% methanol or ethanol. Reflux extracting to obtain extractive solution A, extracting for 3-6 times each for 1-2 hr, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no alcohol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water washing solution D, eluting with 3 column volumes of alcohol solution E (30-50% methanol or ethanol) containing ethanol 30-50%, collecting the eluate F, and eluting with alcohol solution G (80-95% methanol or ethanol) to obtain eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; adjusting pH of the water washing solution D to 9-10 with ammonia water, separating out precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: dividing the experiment into three groups of k, l and m according to the difference of the reverse polymerization fillers C in the step 2), wherein the k adopts AB-8 macroporous resin, the l adopts D-101 macroporous resin, and the m adopts NM 100-reverse polymer chromatographic filler. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 4
TABLE 4 grouping test results of example four
The experimental results show that the reverse polymerization filler C adopts AB-8 macroporous resin, l adopts D-101 macroporous resin, m adopts NM 100-reverse polymer chromatographic filler, the yield is more than 80%, the purity is more than 99%, and the results have no significant difference, so the three extraction modes are the preferred modes of the invention.
EXAMPLE five
1) Extraction: pulverizing rhizoma corydalis Decumbentis, and extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid and 50-100 vol% methanol or ethanol. Reflux extracting to obtain extractive solution A, extracting for 3-6 times each for 1-2 hr, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no alcohol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water washing solution D, eluting with 3 times column volume of alcoholic solution E, collecting the eluate F, eluting with alcoholic solution G, and eluting with 80-95% methanol or ethanol to obtain eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; and (5) adjusting the pH of the water washing solution D to 9-10 with ammonia water, separating out a precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: dividing the experiment into n, o, p, q, r and s groups according to the different volume percentages of methanol or ethanol in the alcohol solution E in the step 2). n group is 10-30% methanol, o group is 30-50% methanol, p group is 50-70% methanol, q group is 10-30% ethanol, r group is 30-50% ethanol, and s group is 50-70% ethanol. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 5
TABLE 5 grouping test results of EXAMPLE five
The above experimental results show that the alcohol E solution with 30-50% by volume of methanol or ethanol is the preferred embodiment of the present invention.
EXAMPLE six
1) Extraction: pulverizing rhizoma corydalis Decumbentis, and extracting with acidic methanol or ethanol solution containing 0.1-2 vol% hydrochloric acid and 50-100 vol% methanol or ethanol. Reflux extracting to obtain extractive solution A, extracting for 3-6 times each for 1-2 hr, mixing extractive solutions, concentrating under reduced pressure at 50-70 deg.C until there is no alcohol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5 with 0.1-2% hydrochloric acid solution, adsorbing with reverse polymeric filler C, eluting with 3 column volumes of water to obtain water solution D, eluting with 3 column volumes of alcohol solution E, which is 30-50% methanol or ethanol, collecting eluate F, and eluting with alcohol solution G to obtain eluate H.
3) And (3) purification: taking an analytic solution H, concentrating the analytic solution H at 50-60 ℃ under reduced pressure until no alcohol exists, adjusting the pH value of ammonia water to 8-10, standing for 2 hours, filtering and precipitating, dissolving the precipitate with methanol, mixing the precipitate with 60-80-mesh silica gel to obtain a column raw material I, wherein the I is yellow dispersed particles, loading the I into a column, eluting with a dichloromethane-methanol system, collecting an eluent, namely dichloromethane-methanol, solution J with the volume percentage of 80:1-40:1, wherein in the elution process, the product has three color bands of brown, gray black and black, the black color band is mainly bichalysine, collecting eluent of the color band, tracking and detecting by TLC, and the TLC detection condition is that dichloromethane: methanol (60: 1); concentrating the resolution solution F at 50-60 deg.C under reduced pressure, adjusting pH to 9-10 with ammonia water, separating out precipitate K, and filtering for further purification; and (5) adjusting the pH of the water washing solution D to 9-10 with ammonia water, separating out a precipitate L, and filtering for further purification.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain extract M, crystallizing the extract M with dichloromethane-methanol system twice to obtain pure product of Bifunoline, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products; crystallizing the precipitate K and the precipitate L with dichloromethane-anhydrous ethanol system to obtain protopine pure product, and vacuum blast drying at 40-50 deg.C to obtain pure powder of the series of products.
Grouping: dividing the experiment into t, u, v and w groups according to the different volume percentages of the methanol or the ethanol in the alcohol solution G in the step 2). t group is 65-80% methanol, u group is 80-95% methanol, v group is 65-80% ethanol, and w group is 80-95% ethanol. The yield and product purity of each group of bicuculline and protopine were determined by HPLC, and the results are shown in Table 6
TABLE 6 grouping test results of EXAMPLE six
The above experimental results show that the alcohol G solution with 80-95% by volume of methanol or ethanol is the preferred embodiment of the present invention.
The detection conditions of bicuculline and protopine in the above examples are as follows:
mobile phase: acetonitrile: triethylamine acetic acid solution (water 500ml + acetic acid 15ml + triethylamine 4ml) ═ 25: 75;
wavelength: 289 nm.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and improvements can be made without departing from the spirit and scope of the invention, and such modifications and improvements should be considered within the scope of the invention.
Claims (6)
1. A preparation method of bicuculline and protopine is characterized by comprising the following steps:
1) extraction: pulverizing one or more of rhizoma corydalis Decumbentis, rhizoma corydalis Decumbentis or corydalis edulis, extracting with acidic methanol or ethanol solution to obtain extractive solution A, mixing extractive solutions, concentrating under reduced pressure to remove ethanol, and collecting concentrated solution B.
2) Separation and enrichment: adjusting pH of the concentrated solution B to 3-5, adsorbing with reverse polymeric filler C, eluting with water to obtain water washing solution D, eluting with alcoholic solution E, collecting eluate F, and eluting with alcoholic solution G to obtain eluate H
3) And (3) purification: taking analysis liquid H, concentrating under reduced pressure, adjusting pH to 8-10, standing, filtering, precipitating, dissolving precipitate with methanol, mixing with silica gel to obtain column-passing raw material I, loading the sample I, eluting with a dichloromethane-methanol system with eluent volume percentage of 100:1-10:1, and collecting solution J with dichloromethane-methanol volume percentage of 80:1-40: 1; concentrating the analysis solution F under reduced pressure, adjusting pH to 9-10, and separating out a precipitate K; and (5) adjusting the pH of the water washing solution D to 9-10, and separating out a precipitate L.
4) And (3) crystallizing and drying: concentrating the solution J under reduced pressure to obtain an extract M, and crystallizing the extract M twice to obtain a pure product of the bicolor sodium; crystallizing the precipitate K and the precipitate L once to obtain a crude sodium macleyite pure product, and drying the pure product by vacuum blast to obtain pure product powder of the series of products;
in the step 2), the reverse polymerization filler C is any one selected from AB-8 macroporous resin, D-101 macroporous resin and NM 100-reverse polymer chromatographic filler;
the alcoholic solution E in the step 2) is methanol or ethanol solution, and the volume percentage of the methanol or the ethanol in the alcoholic solution E is 30-50%;
the alcoholic solution G in the step 2) is methanol or ethanol solution, and the volume percentage of the methanol or the ethanol in the alcoholic solution G is 80-95%;
and 4) crystallizing the extract M in the step 4) by using a dichloromethane-methanol system, and crystallizing the precipitate K by using a dichloromethane-absolute ethanol system.
2. The method according to claim 1, wherein the acidic methanol or ethanol solution in step 1) contains 0.1-2% by volume of hydrochloric acid and 50-100% by volume of methanol or ethanol.
3. The method of claim 2, wherein the step 1) extraction method comprises: reflux extraction, ultrasonic extraction and percolation extraction.
4. The method according to claim 3, wherein the temperature at which the concentration under reduced pressure in step 1) is carried out is 50 to 70 ℃.
5. The method according to claim 1, wherein the temperature at which the solution H and the solution F are concentrated under reduced pressure in step 3) is 50 to 60 ℃.
6. The method according to claim 5, wherein the pH values of the eluent H, the eluent F and the rinsing solution D are adjusted by using ammonia water in the step 3).
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| CN1535969A (en) * | 2003-04-11 | 2004-10-13 | 湖南医药工业研究所 | Total alkaloid of xiatianwu (a Chinese medicinal material), its preparation method and application |
| CN1785261A (en) * | 2005-11-15 | 2006-06-14 | 河北智源医药科技有限公司 | Hotseason grow corydaline, its preparation method and its medicinal composition |
| CN101054377A (en) * | 2006-04-13 | 2007-10-17 | 上海医药工业研究院 | Total alkaloids extraction of corydalis, its preparation method, medicine composition containing the total alkaloids extraction and application thereof |
| CN101805345A (en) * | 2009-12-04 | 2010-08-18 | 南京泽朗医药科技有限公司 | Method for purifying protopine from corydalis amabilis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1535969A (en) * | 2003-04-11 | 2004-10-13 | 湖南医药工业研究所 | Total alkaloid of xiatianwu (a Chinese medicinal material), its preparation method and application |
| CN1785261A (en) * | 2005-11-15 | 2006-06-14 | 河北智源医药科技有限公司 | Hotseason grow corydaline, its preparation method and its medicinal composition |
| CN101054377A (en) * | 2006-04-13 | 2007-10-17 | 上海医药工业研究院 | Total alkaloids extraction of corydalis, its preparation method, medicine composition containing the total alkaloids extraction and application thereof |
| CN101805345A (en) * | 2009-12-04 | 2010-08-18 | 南京泽朗医药科技有限公司 | Method for purifying protopine from corydalis amabilis |
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