CN112442099A - Process for extracting glucopyranoside derivative from crassula argentea - Google Patents
Process for extracting glucopyranoside derivative from crassula argentea Download PDFInfo
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- 241001660917 Crassula ovata Species 0.000 title description 6
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- 238000001035 drying Methods 0.000 claims abstract description 72
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- 239000011347 resin Substances 0.000 claims description 18
- 229920005989 resin Polymers 0.000 claims description 18
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- 235000002991 Coptis groenlandica Nutrition 0.000 claims description 11
- 230000005484 gravity Effects 0.000 claims description 11
- 238000002791 soaking Methods 0.000 claims description 8
- 238000002137 ultrasound extraction Methods 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000007603 infrared drying Methods 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 238000001291 vacuum drying Methods 0.000 claims description 2
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- 238000001514 detection method Methods 0.000 description 20
- 239000000284 extract Substances 0.000 description 13
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 11
- 238000000746 purification Methods 0.000 description 11
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- RLDKDWYKNIHFKF-UHFFFAOYSA-N 3-(3',4'-dihydroxyphenyl)-(2R)-lactic acid 4'-O-beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(CC(O)C(O)=O)C=C1O RLDKDWYKNIHFKF-UHFFFAOYSA-N 0.000 description 1
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- 241001517299 Bulbophyllum Species 0.000 description 1
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- 206010020741 Hyperpyrexia Diseases 0.000 description 1
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- 239000008103 glucose Substances 0.000 description 1
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- AGDRDFJCYMPNFZ-UHFFFAOYSA-N methane;methanol Chemical compound C.OC AGDRDFJCYMPNFZ-UHFFFAOYSA-N 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Diabetes (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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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 and pulverizing radix Coptidis 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 an alcohol solvent, precipitating, filtering, removing solids, and keeping filtrate; 4) concentrating the filtrate obtained in the step 3) 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 the eluent with the volume percentage of ethanol of 20-60%, concentrating and drying to obtain the product. The method provided by the invention is simple to operate, low in cost, high in product purity and high in yield, and the used reagents are ethanol and water, so that the influence on the environment and operators is reduced, and the reagent residue is reduced; the filler of the chromatographic column can be recycled, so that the overall production cost is reduced, and the pollution of filler waste to the environment is also reduced; in addition, the invention utilizes the residual part (the coptis chinensis whiskers) of the coptis chinensis after being harvested, thereby enhancing the comprehensive utilization of the coptis chinensis.
Description
Technical Field
The invention belongs to the field of chemical purification, and particularly relates to a process for extracting a glucopyranoside derivative from crassula argentea.
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 in a formula IV from crassula argentea.
The invention provides a preparation method of a compound shown as a formula IV, which comprises the following steps:
1) drying and pulverizing radix Coptidis 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 an alcohol solvent, precipitating, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting a chromatographic column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent with the ethanol volume percentage of 20-60%, concentrating and drying to obtain the compound preparation;
wherein the structure of the compound shown in the formula IV is as follows:
further, in the step 3), the concentration is carried out until the specific gravity of the system is 1.00-1.15;
the alcohol solvent is ethanol;
the mass ratio of the system obtained by concentration to the alcohol solvent is 1: (1-10), preferably 1 (4-8);
the precipitation time is 1-72 hours, and the temperature is 0-25 ℃.
Further, in the step 3), the ethanol is 50-80% by volume of ethanol aqueous solution, and preferably 50-70% by volume of ethanol aqueous solution.
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 extraction temperature of the soaking extraction 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 step 4), the eluent with 40 to 45 volume percent of ethanol is collected, and the eluent with 40 volume percent of ethanol is preferred.
Further, in step 4), the conditions of the gradient elution are as follows:
further, in step 4), the chromatography column is selected from a D-101 macroporous resin column, an MCI resin column, an ODS column, an AB-8 resin column, an HPD-400 resin column, a D3520 resin column, and an MCL resin column, preferably an ODS column.
Further, in the step 4), 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 filtrate.
Further, in the step 1), the coptis root whiskers are selected from one or two of coptis root short whiskers and coptis root long whiskers.
Further, the coptis root is selected from one or a combination of more than two of the group consisting of the rhizoma coptidis, the elegans and the cissus oblonga.
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.
Huanglian palpus refers to the root palpus of Huanglian.
The short fibrous root of coptis refers to the part close to the fibrous root of the main root.
The long fibrous root of coptis refers to the part of the long fibrous root far away from the main root.
And the ODS column is an octadecylsilane bonded silica gel column.
The reagents used in the method are ethanol and water, so that the reagent residue is reduced, and the influence on the environment and operators is reduced.
The filler of the chromatographic column can be recycled, so that the overall production cost is reduced, and the pollution of filler waste to the environment is reduced.
The invention utilizes the residual part (the coptis chinensis whiskers) of the harvested coptis chinensis to carry out secondary utilization, thereby enhancing the comprehensive utilization rate of the coptis chinensis, reducing the cost and reducing the waste of data.
Experimental results show that the method provided by the invention can be used for extracting the compound shown in the formula IV from the crassula argentea, and the method is simple to operate, low in cost, high in product yield and purity, more environment-friendly and suitable for commercial production.
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 short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) extracting the medicinal material powder obtained in the step 1) with 8 times of water at 80 ℃ for 2 times, and each time for 2 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to a specific gravity of 1.1 to obtain an extract, adding 90% ethanol in an amount which is 4 times that of the extracting solution, precipitating for 24 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using a D-101 macroporous resin column and using an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent with the volume percentage of ethanol of 20%, concentrating, and drying to obtain a compound shown in a 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; the purity of HPLC detection analysis is 78% by taking acetonitrile as a mobile phase A and water as a mobile phase B; the compound was weighed out after drying to give a yield of 28%.
Example 2 extraction Process of the invention
1. Extraction of
(1) Drying the extracted part of the radix coptidis rhizoma, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) performing ultrasonic extraction on the medicinal material powder obtained in the step 1) with 6 times of water at 60HZ for 2 times, wherein each time lasts for 1 hour to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 4 times of 80% ethanol, precipitating for 36 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) 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 MCI resin column, collecting an eluent with the ethanol volume percentage of 45% (the gradient elution conditions are the same as the step (4) in the example 1), concentrating, and drying 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 30%.
Example 3 extraction Process of the invention
1. Extraction of
(1) Drying the short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) reflux-extracting the medicinal powder obtained in the step 1) with 30% ethanol aqueous solution of which the amount is 8 times that of the medicinal powder at 80 ℃ for 2 times, wherein each time lasts for 1.5 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to a specific gravity of 1.1 to obtain an extract, adding 70% ethanol with 4 times of volume, precipitating for 48 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) 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 with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (4) in the example 1), concentrating, and drying 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 91%; the compound was weighed out after drying to give a yield of 28%.
Example 4 extraction Process of the invention
1. Extraction of
(1) Drying the short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) soaking and extracting the medicinal material powder obtained in the step 1) with 6 times of water at 90 ℃ for 2 times, wherein each time lasts for 2 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 4 times of 65% ethanol by volume, precipitating for 72 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting an AB-8 resin column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent with the ethanol volume percentage of 30% (the gradient elution condition is the same as the step (4) of the embodiment 1), concentrating, and drying 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; the purity of HPLC detection analysis is 79 percent by taking acetonitrile as a mobile phase A and water as a mobile phase B; the compound was weighed after drying and calculated to yield 33%.
Example 5 extraction Process of the invention
1. Extraction of
(1) Drying the extracted part of the radix coptidis rhizoma, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) carrying out ultrasonic extraction on the medicinal material powder obtained in the step 1) by using 8 times of water at 60HZ for 3 times, wherein each time is 1 hour, so as to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 5 times of 70% ethanol, precipitating for 72 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an HPD-400 resin column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent with the ethanol volume percentage of 30% (the gradient elution conditions are the same as the step (4) of the embodiment 1), concentrating, and drying 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 76%; the compound was weighed out after drying to give a yield of 26%.
Example 6 extraction Process of the invention
1. Extraction of
(1) Drying the extracted part of the radix coptidis rhizoma, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) soaking and extracting the medicinal material powder obtained in the step 1) with 8 times of water at 70 ℃ for 2 times, and each time for 2 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.1 to obtain an extract, adding 65% ethanol with 4 times of volume amount, precipitating for 48 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using a D3520 resin column and an aqueous solution with the volume percentage of ethanol of 0-80% as an eluent, collecting an eluent with the volume percentage of ethanol of 20% (the gradient elution conditions are the same as the step (4) of the embodiment 1), concentrating, and drying 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; the purity of HPLC detection analysis is 77 percent by taking acetonitrile as a mobile phase A and water as a mobile phase B; 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 short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) reflux-extracting the medicinal powder obtained in the step 1) with 30% ethanol aqueous solution of which the amount is 8 times that of the medicinal powder at 70 ℃ for 2 times, wherein each time lasts for 2 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 60% ethanol with the volume of 10 times, precipitating for 72 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) 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 MCL resin column, collecting an eluent with the ethanol volume percentage of 45% (the gradient elution conditions are the same as the step (4) in the example 1), concentrating, and drying 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 86%; the compound was weighed after drying and calculated to yield 35%.
Example 8 extraction Process of the invention
1. Extraction of
(1) Drying the short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) soaking and extracting the medicinal material powder obtained in the step 1) with 8 times of water at 90 ℃ for 3 times, wherein each time lasts for 1.5 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to 1.1 to obtain an extract, adding 50% ethanol with 4 times of volume amount, precipitating for 36 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) 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 with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (4) in the example 1), concentrating, and drying 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 to give a yield of 36%.
Example 9 extraction Process of the invention
1. Extraction of
(1) Drying the short beard of the extracted part of the coptis chinensis, drying at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing by 10-80 meshes to obtain medicinal powder;
(2) soaking and extracting the medicinal material powder obtained in the step 1) with 6 times of water at 70 ℃ for 2 times, wherein each time lasts for 1.5 hours to obtain an extracting solution;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 60% ethanol with the volume of 5 times, precipitating for 24 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by using an MCL (methanol-methane) column and an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent with the ethanol volume percentage of 45% (the gradient elution conditions are the same as the step (4) of the example 1), concentrating, and drying 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; the purity of HPLC detection analysis is 87% by taking acetonitrile as a mobile phase A and water as a mobile phase B; the compound was weighed out after drying to give a yield of 30%.
Example 10 extraction Process of the invention
1. Extraction of
(1) Drying the extractive part of the coptis chinensis short hair at 40-200 ℃ for 0.5-10 hours until the water content is below 14%, and then crushing into 10-80 meshes to obtain medicinal powder;
(2) reflux-extracting the medicinal powder obtained in the step 1) with 30% ethanol aqueous solution 6 times the amount of the medicinal powder at 80 ℃ for 2 times, and obtaining an extracting solution each time for 2 hours;
2. purification of
3) Concentrating the extracting solution obtained in the step 2) to the specific gravity of 1.15 to obtain an extract, adding 8 times of 50% ethanol by volume, precipitating for 48 hours, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) 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 with the ethanol volume percentage of 40% (the gradient elution conditions are the same as the step (4) in the example 1), concentrating, and drying 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 34%.
TABLE 1 comparison of extraction Process, yield and purity of each sample
As can be seen from Table 1, the compounds of formula IV can be extracted from the root of common Bulbophyllum root by the methods of examples 1-10 of the present invention. Particularly, the method of example 3, 8 or 10 is adopted, firstly the extract is precipitated in 50-70% ethanol, then the water solution with the ethanol volume percentage of 0-80% is used as eluent, gradient elution is carried out through an ODS column, the eluent with the ethanol volume percentage of 40% is collected, and the purity of the obtained target product is as high as more than 90%.
In conclusion, the method provided by the invention can be used for extracting the compound shown in the formula IV from the crassula argentea, is simple to operate, low in cost, high in product purity and high in yield, and the used reagents are ethanol and water, so that the reagent residue is reduced, and the influence on the environment and operators is reduced; the filler of the chromatographic column can be recycled, so that the overall production cost is reduced, and the pollution of filler waste to the environment is reduced; the invention utilizes the residual part (the coptis chinensis whiskers) of the harvested coptis chinensis to carry out secondary utilization, thereby enhancing the comprehensive utilization rate of the coptis chinensis, reducing the cost and reducing the waste of data.
Claims (11)
1. A process for preparing a compound of formula IV, comprising: the method comprises the following steps:
1) drying and pulverizing radix Coptidis 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 an alcohol solvent, precipitating, filtering, removing solids, and keeping filtrate;
4) concentrating the filtrate obtained in the step 3) to obtain a concentrated solution, performing gradient elution on the concentrated solution by adopting a chromatographic column and taking an aqueous solution with the ethanol volume percentage of 0-80% as an eluent, collecting an eluent with the ethanol volume percentage of 20-60%, concentrating and drying to obtain the compound preparation;
wherein the structure of the compound shown in the formula IV is as follows:
2. the method of claim 1, wherein: in the step 3), the concentration is carried out until the specific gravity of the system is 1.00-1.15;
the alcohol solvent is ethanol;
the mass ratio of the system obtained by concentration to the alcohol solvent is 1: (1-10), preferably 1 (4-8);
the precipitation time is 1-72 hours, and the temperature is 0-25 ℃.
3. The method of claim 2, wherein: in the step 3), the ethanol is 50-80% by volume of ethanol aqueous solution, and preferably 50-70% by volume of ethanol aqueous solution.
4. A method according to any one of claims 1-3, 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.
5. A method according to any one of claims 1-3, 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 extraction temperature of the soaking extraction 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%.
6. A method according to any one of claims 1-3, characterized in that: in step 4), the eluent with 40-45% of ethanol volume percent is collected, and the eluent with 40% of ethanol volume percent is preferred.
7. A method according to any one of claims 1-3, characterized in that: in the step 4), the conditions of gradient elution are as follows:
8. a method according to any one of claims 1-3, characterized in that: in step 4), the chromatographic column is selected from a D-101 macroporous resin column, a MCI resin column, an ODS column, an AB-8 resin column, a HPD-400 resin column, a D3520 resin column and a MCL resin column, and is preferably an ODS column.
9. A method according to any one of claims 1-3, characterized in that: in the step 4), 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 filtrate.
10. The method according to any one of claims 1-9, wherein: in the step 1), the coptis root whiskers are selected from one or two of coptis root short whiskers and coptis root long whiskers.
11. The method of claim 10, wherein: the said radix Scutellariae is selected from one or more of radix Melandrii Szechuanensis, radix Aristolochiae Paraquilegiae or radix seu folium Cayratiae Japonicae.
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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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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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| SHOJI YAHARA,等: "Isolation and Characterization of Phenolic Compounds from Coptidis Rhizoma" * |
| 李雪改,等: "黄连水提液化学成分的分离与鉴定" * |
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