CN112442097A - Process for extracting glucopyranoside derivative from coptis chinensis - Google Patents
Process for extracting glucopyranoside derivative from coptis chinensis Download PDFInfo
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Abstract
The invention provides a preparation method of a compound shown as a formula IV, which comprises the following steps: 1) drying Coptidis rhizoma, and pulverizing to obtain medicinal powder; 2) extracting the medicinal material powder obtained in the step 1) by using an extraction solvent to obtain an extracting solution; the extraction solvent is selected from water, alcohol solvent or mixed solution of water and alcohol solvent; 3) concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate; 4) taking the filtrate obtained in the step 3), performing gradient elution by adopting a chromatographic column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a when the ethanol volume percentage is 20-60%; 5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting a chromatographic column and taking an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30-40%, concentrating and drying to obtain the eluent bAnd (5) obtaining the product. The method provided by the invention is simple to operate, the chromatographic column packing can be recycled, the overall cost is reduced, and the obtained product has high purity and high yield, and is suitable for commercial production.
Description
Technical Field
The invention belongs to the field of chemical purification, and particularly relates to a process for extracting glucopyranoside derivatives from coptis chinensis.
Background
Coptidis rhizoma is dried rhizome of Coptis chinensis Franch (Coptis chinensis Franch.) of Ranunculaceae. Weilian, also called Chuan Lian and Ji Zhang Huang, is cultivated in Hubei, Hunan, Shaanxi and Gansu provinces. Bitter in taste and cold in nature, and enters heart, spleen, stomach, liver, gallbladder and large intestine meridians. Has the effects of clearing heat, eliminating dampness, purging pathogenic fire and removing toxic substances. It is mainly used for treating damp-heat distention and fullness, emesis, acid regurgitation, dysentery, jaundice, hyperpyrexia, excessive heart-fire, vexation, insomnia, blood heat, conjunctival congestion, toothache, carbuncle, furuncle, eczema, and suppuration of auditory canal.
The literature "separation and identification of chemical components in coptis aqueous extract, plum blossom improvement and the like" discloses a method for separating compounds from coptis aqueous extract, and 22 compounds are extracted and identified, and the extraction and separation method is as follows: drying Coptidis rhizoma (1 kg), extracting with water under reflux for 2 times (8 times and 6 times) for 2 hr, mixing extractive solutions, recovering solvent under reduced pressure at 48 deg.C, and concentrating to obtain total extract (1l0 g). The water extract of rhizoma Coptidis is subjected to systematic separation by means of recrystallization, repeated open silica gel column chromatography, Sephadex LH-20 column chromatography, preparative thin layer chromatography, PHPLC, etc. to obtain 22 compounds, and their structures are identified. Among the 22 compounds obtained, a glucopyranoside derivative, i.e., compound 15: 3- (3 ', 4' -dihydroxyphenyl) - (2R) -lactic acid-4 '-O-beta-D-glucopyranoside (3- (3', 4 '-dihydroxyphe. ny1) - (2R) -lactic acid-4' -O-beta-D-glucopyranoside).
The application of Chinese patent 201610459981.5 of the applicant discloses the application of the compound 15 (namely the compound shown in formula IV) in the preparation of drugs for preventing and/or treating diabetes, wherein three dosage groups of the compound, namely 10mg/kg, 20mg/kg and 40mg/kg, have obvious reduction effect on blood sugar 60min after glucose administration in a normal mouse glucose tolerance test, 20mg/kg and 40mg/kg can also reduce AUC and increase the blood sugar inhibition rate, and no obvious toxic or side effect is seen, so that the drugs are safe and effective. Therefore, the compound shown in the formula IV has a very good application prospect in preparing medicines for preventing and/or treating diabetes.
However, the preparation method of the compound is very complex, high in cost and low in yield, and is not suitable for commercial production. Therefore, a method which is simple and short in time consumption, simple to operate and high in yield is found to prepare the compound, and the application prospect is good.
Disclosure of Invention
The invention aims to provide a process for extracting a compound shown as a formula IV from coptis chinensis.
The invention provides a preparation method of a compound shown as a formula IV, which comprises the following steps:
1) drying Coptidis rhizoma, and pulverizing to obtain medicinal powder;
2) extracting the medicinal material powder obtained in the step 1) by using an extraction solvent to obtain an extracting solution; the extraction solvent is selected from water, alcohol solvent or mixed solution of water and alcohol solvent;
3) concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
4) taking the filtrate obtained in the step 3), performing gradient elution by adopting a chromatographic column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a when the ethanol volume percentage is 20-60%;
5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting a chromatographic column and taking an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30-40%, concentrating and drying to obtain the eluent b;
wherein the structure of the compound shown in the formula IV is as follows:
further, in the step 4), the eluent a with 40% of ethanol volume percentage is collected; in step 5), the eluate b was collected with a volume percentage of 30% ethanol.
Further, in the step 1), the drying mode comprises drying in the shade, drying in the sun and drying, and the weight of water in the dried product is below 14%; the crushing is to be crushed to 10-80 meshes;
preferably, the drying mode is drying under the following conditions: drying at 40-200 deg.C for 0.5-10 hr.
Further, in the step 2), the extraction mode is one or more than two of reflux extraction, ultrasonic extraction and soaking extraction; wherein the temperature for soaking and extracting is 60-90 ℃;
and/or the mass ratio of the medicinal material powder to the extraction solvent is 1: (2-10), preferably 1: (6-8);
and/or, the extraction times are 2-3 times, and the extraction time of each time is 1.5-2 h;
and/or the extraction solvent is selected from water or ethanol with the volume concentration of 30%.
Further, in the step 3), the acid is selected from one or more of hydrochloric acid, sulfuric acid and acetic acid, and the pH value of the acidified system is 1.5-6.5;
and/or the volume of the system after concentration is 15% of that before concentration.
Further, in step 4), the conditions of the gradient elution are as follows:
further, in step 5), the conditions of the gradient elution are as follows:
further, in the step 4), the chromatographic column is selected from HPD-400 resin column, D-101 resin column, HPD-100 resin column, MCL resin column, AB-8 resin column, ODS column; preferably an ODS column or an MCL resin column;
in the step 5), the filler of the chromatographic column is selected from an MCL resin column, an ODS column, an HPD-300 resin column, a D-301 resin column and an X-5 resin column; preferably an ODS column.
Further, in the step 5), the drying mode is one or more than two of spray drying, vacuum drying, freeze drying, near infrared drying and microwave drying, and the drying temperature is below 70 ℃;
and/or, the volume of the concentrated solution is 5% of the eluent a.
Further, in the step 1), the coptis is selected from one or more than two of coptis flowers, coptis leaves, coptis roots, coptis stems and coptis whiskers; preferably rhizoma Coptidis root or rhizoma Coptidis leaf.
And the ODS column is an octadecylsilane bonded silica gel column.
Experimental results show that the method provided by the invention can be used for extracting the compound shown in the formula IV from the coptis, and the method is simple to operate, short in process flow, less in time consumption, capable of recycling the chromatographic column filler, low in total production cost, high in product yield and purity, and suitable for commercial production.
In the invention, drying in the shade refers to natural drying in places without direct sunlight and with good ventilation; sun drying refers to drying under direct sunlight; drying refers to heating and drying.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
Example 1 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 6 times of water into the medicinal material powder obtained in the step (1), and performing ultrasonic extraction for 2 times at 60Hz for 1 hour each time to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by adopting an HPD-400 resin column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a when the ethanol volume percentage is 30%; the conditions for the gradient elution were as follows:
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting an MCL resin column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent b with the ethanol volume percentage of 40%, concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV:
the conditions for the gradient elution were as follows:
3. characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 91%; the compound was weighed out after drying to give a yield of 36%.
Example 2 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 8 times of 30% ethanol aqueous solution into the medicinal powder obtained in the step (1), and performing reflux extraction at 80 ℃ for 2 times, wherein each extraction time is 1.5 hours to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using a D-101 resin column and using an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, and collecting an eluent a with the volume percentage of ethanol of 20%, wherein the gradient elution condition is the same as that in the step (4) of the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an aqueous solution with the ethanol volume percentage of 0-80% as an eluent through an ODS column, collecting an eluent b with the ethanol volume percentage of 30% (the gradient elution conditions are the same as the step (5) in the example 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 92%; weighing the dried compound gave a yield of 35%.
Example 3 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 6 times of water into the medicinal material powder obtained in the step (1), soaking and extracting for 3 times at 90 ℃, wherein each time of extraction is 1 hour, so as to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an HPD-100 resin column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a with the ethanol volume percentage of 20%, wherein the gradient elution condition is the same as that in the step (4) of the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an HPD-300 resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30% (the gradient elution conditions are the same as the step (5) in the example 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 88%; the compound was weighed out after drying and calculated to give a yield of 34%.
Example 4 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 8 times of water into the medicinal material powder obtained in the step (1), soaking and extracting for 2 times at 70 ℃, and extracting for 2 hours each time to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an HPD-400 resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, and collecting an eluent a with the volume percentage of ethanol of 20%, wherein the gradient elution condition is the same as that in the step (4) of the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an MCL resin column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent b with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (5) of the embodiment 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 90%; the compound was weighed out after drying and calculated to give a yield of 32%.
Example 5 extraction Process of the invention
1. Extraction of
(1) Drying the extract part of the coptis leaves for 0.5-10 hours at 40-200 ℃ until the water content is below 14%, and then crushing the coptis leaves into 10-80 meshes to obtain medicinal powder;
(2) adding 6 times of water into the medicinal material powder obtained in the step (1), soaking and extracting for 2 times at 80 ℃, and extracting for 2 hours each time to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an MCL resin column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a with the ethanol volume percentage of 60%, wherein the gradient elution condition is the same as that in the step (4) of the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using a D-301 resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30% (the gradient elution conditions are the same as the step (5) of the embodiment 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 80%; the compound was weighed out after drying to give a yield of 15%.
Example 6 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 6 times of water into the medicinal material powder obtained in the step (1), soaking and extracting for 2 times at 70 ℃, and extracting for 2 hours each time to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by adopting an AB-8 resin column and using an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, and collecting an eluent a with the volume percentage of ethanol of 30%, wherein the gradient elution condition is the same as that of the step (4) in the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an X-5 resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30% (the gradient elution conditions are the same as the step (5) in the example 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 85%; the compound was weighed out after drying to give a yield of 28%.
Example 7 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 6 times of 30% ethanol aqueous solution into the medicinal material powder obtained in the step (1), and performing reflux extraction at 70 ℃ for 2 times, wherein each extraction is performed for 1.5 hours to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an ODS column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a with the ethanol volume percentage of 40%, wherein the gradient elution condition is the same as that in the step (4) of the example 1; (5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an aqueous solution with the ethanol volume percentage of 0-80% as an eluent through an ODS column, collecting an eluent b with the ethanol volume percentage of 30% (the gradient elution conditions are the same as the step (5) in the example 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 94%; the compound was weighed out after drying to give a yield of 38%.
Example 8 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 8 times of 30% ethanol aqueous solution into the medicinal material powder obtained in the step (1), and performing reflux extraction at 80 ℃ for 2 times, wherein each extraction is performed for 2 hours to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an MCL column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, and collecting an eluent a with the volume percentage of ethanol of 40%, wherein the gradient elution condition is the same as that of the step (4) in the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an aqueous solution with the ethanol volume percentage of 0-80% as an eluent through an ODS column, collecting an eluent b with the ethanol volume percentage of 30% (the gradient elution conditions are the same as the step (5) in the example 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 92%; the compound was weighed out after drying to give a yield of 40%.
Example 9 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 8 times of the medicinal powder obtained in the step (1), and performing reflux extraction for 3 times under 60HZ, wherein each extraction is performed for 1 hour to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an ODS column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a with the ethanol volume percentage of 30%, wherein the gradient elution condition is the same as that in the step (4) of the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, carrying out gradient elution on the concentrated solution by using an aqueous solution with the ethanol volume percentage of 0-80% as an eluent through an MCL resin column, collecting an eluent b with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (5) of the embodiment 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 90%; and calculating the content of the compound shown in IV in the raw medicinal materials, and weighing the dried compound to obtain the yield of 38%.
Example 10 extraction Process of the invention
1. Extraction of
(1) Drying the rhizoma Coptidis root at 40-200 deg.C for 0.5-10 hr until the water content is below 14%, and pulverizing into 10-80 mesh powder;
(2) adding 6 times of water into the medicinal material powder obtained in the step (1), soaking and extracting for 2 times at 90 ℃, and extracting for 2 hours each time to obtain an extracting solution;
2. purification of
(3) Concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
(4) taking the filtrate obtained in the step 3), performing gradient elution by using an MCL resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, and collecting an eluent a with the volume percentage of ethanol of 40%, wherein the gradient elution condition is the same as that of the step (4) in the example 1;
(5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an MCL resin column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent b with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (5) of the embodiment 1), concentrating to obtain an extract, and drying the extract to obtain the compound shown in the formula IV.
3. Characterization of
And comparing the nuclear magnetic detection with a nuclear magnetic spectrum of a reference substance to confirm that the compound is the compound shown as IV. Octadecylsilane chemically bonded silica is used as a filler through chromatographic conditions and system applicability tests; acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and the HPLC detection and analysis purity is 90%; the compound was weighed after drying and calculated to yield 33%.
TABLE 1 comparison of extraction Process, yield and purity of each sample
As can be seen from Table 1, the compounds represented by formula IV can be extracted from Coptidis rhizoma by the methods of examples 1-10 of the present invention. Particularly, the method of embodiment 7 or 8 is adopted, and the target product with high yield and high purity can be obtained, namely, after the reflux extraction is adopted and twice extraction is carried out, the water solution with 0-80% of ethanol volume percentage is taken as eluent, gradient elution is carried out sequentially through 2 times of ODS columns, or gradient elution is carried out sequentially through MCL resin columns and ODS columns, the eluents with 40% and 30% of ethanol volume percentage are respectively collected, and the yield of the obtained target product is up to 38% -40%, and the purity is up to 92% -94%.
In conclusion, the experimental results show that the method provided by the invention can extract the compound shown in the formula IV from the coptis chinensis, the method is simple to operate, the chromatographic column packing can be recycled, the overall cost is reduced, the product yield is high, the purity is high, and the method is suitable for commercial production.
Claims (10)
1. A process for preparing a compound of formula IV, comprising: the method comprises the following steps:
1) drying Coptidis rhizoma, and pulverizing to obtain medicinal powder;
2) extracting the medicinal material powder obtained in the step 1) by using an extraction solvent to obtain an extracting solution; the extraction solvent is selected from water, alcohol solvent or mixed solution of water and alcohol solvent;
3) concentrating the extracting solution obtained in the step 2), adding acid, acidifying, separating out solid, then filtering, removing the solid, and keeping the filtrate;
4) taking the filtrate obtained in the step 3), performing gradient elution by adopting a chromatographic column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, and collecting an eluent a when the ethanol volume percentage is 20-60%;
5) concentrating the eluent a obtained in the step 4) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting a chromatographic column and taking an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent b with the volume percentage of ethanol of 30-40%, concentrating and drying to obtain the eluent b;
wherein the structure of the compound shown in the formula IV is as follows:
2. the method of claim 1, wherein: in the step 4), collecting the eluent a with the ethanol volume percentage of 40 percent; in step 5), the eluate b was collected with a volume percentage of 30% ethanol.
3. The method according to claim 1 or 2, characterized in that: in the step 1), the drying mode comprises drying in the shade, drying in the sun and drying, and the weight of water in the dried product is below 14%; the crushing is to be crushed to 10-80 meshes;
preferably, the drying mode is drying under the following conditions: drying at 40-200 deg.C for 0.5-10 hr.
4. The method according to claim 1 or 2, characterized in that: in the step 2), the extraction mode is one or more than two of reflux extraction, ultrasonic extraction and soaking extraction; wherein the temperature for soaking and extracting is 60-90 ℃;
and/or the mass ratio of the medicinal material powder to the extraction solvent is 1: (2-10), preferably 1: (6-8);
and/or, the extraction times are 2-3 times, and the extraction time of each time is 1.5-2 h;
and/or the extraction solvent is selected from water or ethanol with the volume concentration of 30%.
5. The method according to claim 1 or 2, characterized in that: in the step 3), the acid is selected from one or more of hydrochloric acid, sulfuric acid and acetic acid, and the pH value of the acidified system is 1.5-6.5;
and/or the volume of the system after concentration is 15% of that before concentration.
6. The method according to any one of claims 1 to 5, wherein: in the step 4), the conditions of gradient elution are as follows:
7. the method according to any one of claims 1 to 5, wherein: in step 5), the gradient elution conditions are as follows:
8. the method according to any one of claims 1 to 5, wherein: in the step 4), the chromatographic column is selected from HPD-400 resin column, D-101 resin column, HPD-100 resin column, MCL resin column, AB-8 resin column and ODS column; preferably an ODS column or an MCL resin column;
in the step 5), the filler of the chromatographic column is selected from an MCL resin column, an ODS column, an HPD-300 resin column, a D-301 resin column and an X-5 resin column; preferably an ODS column.
9. The method according to any one of claims 1 to 5, wherein: in the step 5), the drying mode is one or more than two of spray drying, vacuum drying, freeze drying, near infrared drying and microwave drying, and the drying temperature is below 70 ℃;
and/or, the volume of the concentrated solution is 5% of the eluent a.
10. The method according to any one of claims 1-9, wherein: in the step 1), the coptis is selected from one or more than two of coptis flowers, coptis leaves, coptis roots, coptis stems and coptis whiskers; preferably rhizoma Coptidis root or rhizoma Coptidis leaf.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107519191A (en) * | 2016-06-22 | 2017-12-29 | 成都中创蜀洋生物科技有限公司 | Glucoside compound is preparing the purposes in treating diabetes medicament |
| CN108440617A (en) * | 2018-06-22 | 2018-08-24 | 成都中创蜀洋生物科技有限公司 | A method of extracting glucoside compound from the coptis |
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2019
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107519191A (en) * | 2016-06-22 | 2017-12-29 | 成都中创蜀洋生物科技有限公司 | Glucoside compound is preparing the purposes in treating diabetes medicament |
| CN108440617A (en) * | 2018-06-22 | 2018-08-24 | 成都中创蜀洋生物科技有限公司 | A method of extracting glucoside compound from the coptis |
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| YAHARA, SHOJI等: "Isolation and characterization of phenolic compounds from coptidis rhizome", 《CHEMICAL & PHARMACEUTICAL BULLETIN》 * |
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