CN113121486A - Single-configuration extraction method of dihydromyricetin - Google Patents
Single-configuration extraction method of dihydromyricetin Download PDFInfo
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- CN113121486A CN113121486A CN202110289675.2A CN202110289675A CN113121486A CN 113121486 A CN113121486 A CN 113121486A CN 202110289675 A CN202110289675 A CN 202110289675A CN 113121486 A CN113121486 A CN 113121486A
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- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/32—2,3-Dihydro derivatives, e.g. flavanones
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- C—CHEMISTRY; METALLURGY
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- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
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Abstract
The invention discloses a single-configuration extraction method of dihydromyricetin, which comprises the steps of grinding vine tea samples into powder, extracting and purifying the single-configuration dihydromyricetin in the vine tea powder by using an ethanol-water mixed solution system at the temperature of not higher than 40 ℃ and the like, and the invention firstly provides a method for extracting and purifying the single-configuration dihydromyricetin sample by maintaining a low-temperature condition in the whole course of a purification process, so that the risk of racemization of the single-configuration dihydromyricetin due to temperature is avoided, and the invention does not need any high-temperature condition or reagent at all, and does not cause racemization of the dihydromyricetin when being applied to industrial production; the low-temperature extraction method provided by the invention has a guiding effect on the industrial production of the single-configuration dihydromyricetin, and is beneficial to realizing the large-scale preparation of the single-configuration dihydromyricetin.
Description
Technical Field
The invention belongs to the field of extraction of natural products, and particularly relates to a single-configuration extraction method of dihydromyricetin.
Background
Dihydromyricetin (DMY) is a polyphenol hydroxydihydroflavonol compound with the chemical structure of 3,5, 7-trihydroxy-2- (3,4, 5-trihydroxy phenyl) chroman-4-one, and can be extracted from plants of Ampelopsis in Vitaceae. Dihydromyricetin has pharmacological activity in the aspects of diminishing inflammation, relieving alcoholism, resisting oxidation, resisting thrombus, resisting tumor, inhibiting bacteria and the like, and can play multiple effects of protecting liver, reducing blood fat and the like.
At present, dihydromyricetin is generally extracted from vine tea. Vine tea, also known as Ampelopsis grossedentata, is called Gucha ramulus Uncariae cum uncis in the book Shijing President and is widely planted in Hunan, Guangdong, Guangxi, etc. Ampelopsis grossedentata has a long history of being used as a medicine, and has been widely used for treating infectious diseases such as skin diseases, furuncle, osteomyelitis, acute lymphadenitis and the like. The content of dihydromyricetin in Ampelopsis Grossdentata is extremely high, and the content of dihydromyricetin in stem and leaf parts exceeds 20%, and the content of dihydromyricetin in tender shoot parts can even exceed 30%.
Dihydromyricetin is generally white needle-shaped crystal, is easily soluble in organic solvents such as ethanol, methanol, tetrahydrofuran and the like and hot water, is slightly soluble in ethyl acetate and acetonitrile, and is hardly soluble in cold water, dichloromethane and n-hexane. Currently, most of the dihydromyricetin on the market is racemic samples of (2R,3R) and (2S,3S) configurations 1:1, while the dihydromyricetin contained in the ampelopsis grossedentata is a single configuration sample of (2R, 3R). This is because the racemization of dihydromyricetin is very easy, and the conditions of over-high temperature, alkaline pH value and transition metal ions contained in the solution are all important inducers of racemization of dihydromyricetin. In the current industrial production process of dihydromyricetin, a factory often adopts a mode of soaking and boiling ampelopsis grossedentata in hot water for a long time to extract the dihydromyricetin. This was done in the factory because the cost of the hot water immersion process is the least expensive, but it is also a significant reason for the large majority of racemic samples of commercially available dihydromyricetin. The currently reported single-configuration dihydromyricetin extraction method is to extract a dihydromyricetin crude product from ampelopsis grossedentata by using ethanol, and then purify the dihydromyricetin by dissolving the dihydromyricetin crude product in hot water and then cooling and recrystallizing. The method can be used for dissolving dihydromyricetin in boiling water and rapidly cooling to precipitate, and can shorten heating time to prevent racemization of dihydromyricetin. However, in large-scale industrial production, it is difficult for the existing industrial equipment to rapidly reduce the temperature of the dihydromyricetin after the dihydromyricetin is soaked and boiled in boiling water to the extent that racemization does not occur any more, so that the commercial dihydromyricetin is racemized.
In the field of medicine, different enantiomers of the same compound are highly likely to show distinct pharmacological activities, and thus how to obtain a single configuration of a drug molecule has been a very important research topic. At present, no work of maintaining the low temperature of the dihydromyricetin with a single configuration in the whole extraction process is reported. The patent discloses a method for extracting single-configuration dihydromyricetin at low temperature in the whole process for the first time, and if the method can be applied to the industrial field, the large-scale production of the single-configuration dihydromyricetin can be realized.
Disclosure of Invention
The invention aims to provide a method for extracting and purifying a dihydromyricetin single-configuration sample under a low-temperature condition.
The invention is realized by the following technical scheme for solving the technical problems:
the invention discloses a single-configuration extraction method of dihydromyricetin, which comprises the following steps:
1) grinding the vine tea sample into powder;
2) and extracting and purifying the single-configuration dihydromyricetin in the ampelopsis grossedentata powder by using an ethanol-water mixed solution system under the condition of not higher than 40 ℃.
As a further improvement, the method specifically comprises the following steps:
1) grinding the vine tea sample into powder;
2) adding the vine tea powder into ethanol at the temperature of not higher than 40 ℃, soaking under the condition of not higher than 40 ℃, performing ultrasonic treatment, and then filtering; in the traditional extraction method and industrial production, hot water is often adopted to extract dihydromyricetin from ampelopsis grossedentata in a long-time leaching manner, and although the method can save cost, the dihydromyricetin is racemized, and a product with a single configuration cannot be obtained.
3) Volatilizing the filtrate to dryness at the temperature of not higher than 40 ℃; in the traditional extraction method and industrial production, a high-temperature heating and drying method or a cooling and separating method after dissolving in hot water is often adopted, so that the dihydromyricetin is in a high-temperature environment for a long time, and racemization is finally caused.
4) Adding the obtained dihydromyricetin solid into ethanol for complete dissolution, then adding deionized water, and cooling for crystallization; the method provided by the patent for extracting the dihydromyricetin with a single configuration reported at present is to add boiling water to dissolve the dihydromyricetin and then rapidly cool the solution to separate out the dihydromyricetin. Although the method can shorten the time of the dihydromyricetin in a high-temperature environment under the laboratory condition, the dihydromyricetin is difficult to be rapidly cooled to the extent that racemization does not occur in the practical industrial production after being soaked and boiled in boiling water. The method provided by the patent does not need any high temperature condition or reagent at all, and does not cause racemization of dihydromyricetin when being applied to industrial production.
5) And filtering the precipitated solid, and repeating the crystallization step to obtain a white solid product. The purity of the dihydromyricetin is improved by repeated crystallization, and high temperature conditions are not involved in the crystallization process, so that racemization of the dihydromyricetin is avoided.
The temperature is strictly controlled, and the temperature is maintained below 40 ℃ in the whole process of extracting dihydromyricetin, including the reagent added in the extraction process, and the temperature is also maintained at not higher than 40 ℃.
The invention has the beneficial effects that:
1) in the traditional extraction method and the industrial production, hot water is often adopted to extract dihydromyricetin from the ampelopsis grossedentata in a long-time leaching manner, and although the method can save the cost, the dihydromyricetin is racemized, and a product with a single configuration cannot be obtained. The invention provides a method for extracting and purifying a dihydromyricetin single-configuration sample by maintaining a low-temperature condition in the whole purification process for the first time, and avoids the risk of racemization of the dihydromyricetin with a single configuration due to temperature.
2) The method provided by the patent for extracting the dihydromyricetin with the single configuration reported at present is to add boiling water to dissolve the dihydromyricetin and then rapidly cool the solution to separate out the dihydromyricetin. Although the method can shorten the time of the dihydromyricetin in a high-temperature environment under the laboratory condition, the dihydromyricetin is difficult to be rapidly cooled to the extent that racemization does not occur in the practical industrial production after being soaked and boiled in boiling water. The method provided by the patent does not need any high temperature condition or reagent at all, and does not cause racemization of dihydromyricetin when being applied to industrial production.
3) The temperature is strictly controlled, the condition of below 40 ℃ is maintained in the whole process of extracting the dihydromyricetin, including the reagent added in the extracting process, and the temperature of not higher than 40 ℃ is also maintained. The purity of the dihydromyricetin is improved by repeated crystallization, and high temperature conditions are not involved in the crystallization process, so that racemization of the dihydromyricetin is avoided.
4) The low-temperature extraction method provided by the invention has a guiding effect on the industrial production of the single-configuration dihydromyricetin, and is beneficial to realizing the large-scale preparation of the single-configuration dihydromyricetin. A
Drawings
FIG. 1 is a liquid chromatography spectrum of a commercially available racemic dihydromyricetin;
FIG. 2 is a liquid chromatography spectrum of non-racemic dihydromyricetin extracted from Ampelopsis grossedentata.
Detailed Description
The invention discloses a single-configuration extraction method of dihydromyricetin, which comprises the following specific steps:
1) grinding the vine tea sample into powder;
2) extracting and purifying dihydromyricetin in the vine tea powder by using an ethanol-water system under a low temperature condition;
the specific process of the step 2) is as follows: adding Ampelopsis grossedentata powder into 5 deg.C ethanol (the ethanol is cooled to 5 deg.C in a refrigerator in advance, and the volume mass ratio of the liquid to the material is 5mL:1g), soaking overnight at 5 deg.C, and filtering; rotatably steaming the filtrate at 30 ℃ until the filtrate is dried, adding 10mL of ethanol at 5 ℃ into the solid obtained by the rotary steaming for complete dissolution, then adding 250mL of deionized water at 5 ℃ (the deionized water is cooled to 5 ℃ in a refrigerator in advance, and when the solid cannot be separated out), and placing the mixture into the refrigerator at 5 ℃ for crystallization; the precipitated solid was filtered and the crystallization step was repeated to obtain the product as a white solid.
The vine tea is produced from Zhang Jiajie.
The technical solution of the present invention is further illustrated by the following specific examples:
examples
Extracting single-configuration dihydromyricetin from Ampelopsis grossedentata at low temperature
1) Grinding a 10g vine tea sample produced from Zhang Jiajie into powder;
2) adding Ampelopsis grossedentata powder into 50mL of 5 deg.C ethanol (ethanol is cooled to 5 deg.C in refrigerator in advance), and soaking at 5 deg.C overnight;
3) filtering the overnight soaked ampelopsis grossedentata system; rotatably steaming the filtrate at 30 ℃ until the filtrate is dried, adding 10mL of ethanol at 5 ℃ into the solid obtained by the rotary steaming for complete dissolution, then adding 250mL of deionized water at 5 ℃ (the deionized water is cooled to 5 ℃ in a refrigerator in advance, and when the solid cannot be separated out), and placing the mixture into the refrigerator at 5 ℃ for crystallization;
4) the precipitated solid was filtered and the crystallization step was repeated to obtain 2.4g of a white solid product.
And then, identifying the extracted dihydromyricetin single-configuration sample by using a High Performance Liquid Chromatography (HPLC) technology.
The liquid chromatography uses the following instruments and conditions: agilent 1260 high performance liquid chromatograph with octadecylsilane chemically bonded silica as filler; the method comprises the following steps of mixing methanol: beta-cyclodextrin (taking 18g of beta-cyclodextrin, 10ml of phosphoric acid and adding 1000ml of water for dissolving) 5: 95 is mobile phase, the flow rate is 1ml per minute; the column temperature is 30 ℃; the ultraviolet detection wavelength is 290 nm.
Weighing the extracted non-racemic dihydromyricetin, and precisely preparing a solution of 100 mug/mL by using a chromatographic pure methanol solvent to serve as a test sample; commercially available racemic dihydromyricetin was weighed and prepared as a control sample in a 100. mu.g/mL solution using a chromatographically pure methanol solvent.
The test results are shown in FIG. 1, where 10. mu.L of the control sample was injected into the liquid chromatograph. Through detection, the peak of the (2S,3S) -DMY enantiomer is found to be in the front, and the retention time is 13.416 min; after the peak of the (2R,3R) -DMY enantiomer, the retention time was 14.949 min. The peak area ratio of the (2S,3S) -DMY enantiomer to the (2R,3R) -DMY enantiomer was 1:1.097, and the optical purity of the dihydromyricetin sample was calculated using the following formula;
ee％＝[(A1-A2)/(A1+A2)]*100％(a)
in the formula (a), A1Is the peak area of the (2R,3R) -DMY enantiomer, A2The ee value was calculated to be 4.657% for the peak area of the (2S,3S) -DMY enantiomer.
The liquid chromatograph was injected with 10. mu.L of the test sample, and the test results are shown in FIG. 2. Through detection, the peak of the (2S,3S) -DMY enantiomer is found to be in the front, and the retention time is 12.917 min; after the peak of the (2R,3R) -DMY enantiomer, the retention time was 14.368 min. The peak area ratio of the (2S,3S) -DMY enantiomer to the (2R,3R) -DMY enantiomer is 1:1267.2, and the optical purity of the dihydromyricetin sample is calculated by adopting the following formula;
ee％＝[(A1-A2)/(A1+A2)]*100％(a)
in the formula (a), A1Is the peak area of the (2R,3R) -DMY enantiomer, A2The ee value was calculated to be 99.842% for the peak area of the (2S,3S) -DMY enantiomer.
The test result of High Performance Liquid Chromatography (HPLC) proves that the dihydromyricetin extracted from the vine tea is a sample with a single configuration.
The above description is only exemplary of the present invention and should not be taken as limiting, and any modifications, equivalents, and improvements made within the spirit and scope of the present invention should be included in the present invention.
Claims (2)
1. A single-configuration extraction method of dihydromyricetin is characterized by comprising the following steps:
1) grinding the vine tea sample into powder;
2) and extracting and purifying the single-configuration dihydromyricetin in the ampelopsis grossedentata powder by using an ethanol-water mixed solution system under the condition of not higher than 40 ℃.
2. The method for extracting a single configuration of dihydromyricetin according to claim 1, comprising the following steps:
1) grinding the vine tea sample into powder;
2) adding the vine tea powder into ethanol at the temperature of not higher than 40 ℃, soaking under the condition of not higher than 40 ℃, performing ultrasonic treatment, and then filtering;
3) volatilizing the filtrate to dryness at the temperature of not higher than 40 ℃;
4) adding the obtained dihydromyricetin solid into ethanol for complete dissolution, then adding deionized water, and cooling for crystallization;
5) and filtering the precipitated solid, and repeating the crystallization step to obtain a white solid product.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114163411A (en) * | 2021-11-16 | 2022-03-11 | 宁波大学 | Preparation method of enantiomer pure dihydromyricetin |
| CN114223762A (en) * | 2021-11-04 | 2022-03-25 | 宁波大学 | Racemization protection method of dihydromyricetin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140141082A1 (en) * | 2012-11-16 | 2014-05-22 | Song Gao | Compositions Containing Enriched Natural Crocin and/or Crocetin, and Their Therapeutic or Nutraceutical Uses |
| CN104496955A (en) * | 2014-12-01 | 2015-04-08 | 华中科技大学同济医学院附属同济医院 | Five different crystal substances of dihydromyricetin |
| CN104628695A (en) * | 2015-02-05 | 2015-05-20 | 华中科技大学同济医学院附属同济医院 | Method for controlling racemization rate of dihydromyricetin |
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2021
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140141082A1 (en) * | 2012-11-16 | 2014-05-22 | Song Gao | Compositions Containing Enriched Natural Crocin and/or Crocetin, and Their Therapeutic or Nutraceutical Uses |
| CN104496955A (en) * | 2014-12-01 | 2015-04-08 | 华中科技大学同济医学院附属同济医院 | Five different crystal substances of dihydromyricetin |
| CN106083792A (en) * | 2014-12-01 | 2016-11-09 | 华中科技大学同济医学院附属同济医院 | dextrorotatory dihydromyricetin monohydrate crystal |
| CN104628695A (en) * | 2015-02-05 | 2015-05-20 | 华中科技大学同济医学院附属同济医院 | Method for controlling racemization rate of dihydromyricetin |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114223762A (en) * | 2021-11-04 | 2022-03-25 | 宁波大学 | Racemization protection method of dihydromyricetin |
| CN114163411A (en) * | 2021-11-16 | 2022-03-11 | 宁波大学 | Preparation method of enantiomer pure dihydromyricetin |
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