CN114276391A - Separation and purification method of verbascoside - Google Patents
Separation and purification method of verbascoside Download PDFInfo
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- CN114276391A CN114276391A CN202111608349.XA CN202111608349A CN114276391A CN 114276391 A CN114276391 A CN 114276391A CN 202111608349 A CN202111608349 A CN 202111608349A CN 114276391 A CN114276391 A CN 114276391A
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Abstract
The invention discloses a method for separating and purifying verbascoside, which relates to the technical field of biology and is separated and extracted from traditional Chinese medicine rehmannia root, and the method for separating the verbascoside comprises the following steps: preparing materials and reagents; carrying out sample treatment; separating and purifying acteoside. The method has the advantages that beta-glucosidase which is expensive and difficult in source is not used as an enzymolysis reagent, the process route is simpler, industrial production is easier to realize, the content of verbascoside prepared by the method is higher than that of verbascoside prepared by the prior art, the requirement of new reported medicines on the content can be met, the verbascoside product is derived from Scrophulariaceae plants, the plant source and the medicine source of the verbascoside are expanded, the extraction and preparation method of the verbascoside is optimized and improved, the operation is simple and convenient, the cost is low, only ethanol is used as an organic reagent in the operation, toxic reagents such as methanol and acetone are not used, and the method is environment-friendly.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a method for separating and purifying verbascoside.
Background
Rehmannia glutinosa (Rehmannia glutinosa) is fresh or dried root tuber of Rehmannia glutinosa belonging to Scrophulariaceae family, and is Scrophulariaceae family. Widely distributed in Henan, Hubei, Shanxi, Sichuan, Zhejiang, inner Mongolia, Hunan, Shandong, Liaoning and other provinces and cities in China. The fresh rehmannia root is cold in nature, sweet and bitter in taste, and has the effects of clearing heat, promoting fluid, cooling blood and stopping bleeding; radix rehmanniae is cold in nature and sweet in taste, and has the effects of clearing heat and cooling blood, nourishing yin and promoting the production of body fluid; prepared rehmannia root is slightly warm in nature and sweet in taste, and has the effects of nourishing yin and supplementing blood, and replenishing vital essence and marrow. The radix rehmanniae Preparata mainly contains iridoid components, phenethanol glycosides, monoterpenes, amino acids, saccharides, 5-hydroxyaeginetic acid, succinic acid, dihydroxy-beta-ionone, uracil, etc. Modern researches have proved that the product has the effects of promoting hematopoiesis, reducing blood sugar, improving memory and immunity, and can be widely used in medicines and health foods. Through research summary in recent 30 years, it has been roughly determined that the main components of rehmannia include glycosides, saccharides and amino acids, the glycosides are the most abundant and abundant, wherein Verbascoside belongs to phenylethanoid glycosides, which are one of the quality control indexes of rehmannia root drugs, and the alias of Verbascoside is Acteoside, Verbascoside and Acteoside, which are also named as Acteoside, Verbascoside and Kusaginin, the molecular formula of which is C29H36O15, the molecular weight of which is 624.59, and the structural formula of which is shown in formula 1: formula 1, consulting the literature, finds that there are many methods for separating and purifying verbascoside compounds, such as extracting and separating verbascoside from callicarpa nudiflora, cistanche salsa, osmanthus fragrans and paulownia tomentosa, and the separation and purification processes mainly adopt a method of repeated column chromatography to obtain a high-purity verbascoside sample, and the repeated chromatography process has the defects of long time consumption, large solvent consumption and the like; in addition, there are a number of reports in the literature of high performance liquid chromatography for the analysis of verbascoside content in these plants.
Acteoside (Verbascoside), also known as acteoside, etc., is a phenylpropanoid glycoside compound belonging to natural polyphenol component, with molecular formula of C29H36O15 and molecular weight of 624.59. The acteoside mainly exists in cistanche, purple plants, campsis, rehmannia and the like, and modern researches find that the acteoside has strong biological activities of resisting bacteria, resisting virus, resisting inflammation, resisting oxidation, resisting tumors, protecting nerves and the like. For example, Funari et al found that verbascoside has an effect against cryptococcus neoformans, and zhao et al found that verbascoside can inhibit pneumolysin-mediated cytotoxicity and eliminate pneumococcal pneumonia symptoms.
The verbascoside is a phenylethanoid glycoside component, modern researches prove that the phenylethanoid glycoside component is closely related to the pharmacological action of prepared rehmannia root, and the current verbascoside extract is mainly treated by methods of heating extraction, membrane separation, resin purification, biotransformation, chemical catalysis and the like, and the methods have the defects of complex process, inconvenient operation, long period, low efficiency and the like.
Disclosure of Invention
The invention provides a method for separating and purifying verbascoside, which aims to solve the problems in the background technology.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a method for separating and purifying verbascoside is separated and extracted from a traditional Chinese medicine rehmannia root, and is characterized in that: the method for separating acteoside comprises the following steps:
step one, preparing materials and reagents;
step two, sample treatment is carried out;
step three, separating and purifying verbascoside.
The technical scheme of the invention is further improved as follows: the preparation of materials and reagents in the first step further comprises: 80% ethanol, adsorption resin and 5-25% ethanol water solution with volume concentration.
The technical scheme of the invention is further improved as follows: the sample processing in the second step further comprises:
A. pretreating radix rehmanniae, cutting into small pieces, and placing in an extraction tank;
B. introducing steam from the bottom of the pot for 40 minutes, and scalding the small pieces of rehmannia with saturated steam;
C. extracting with ethanol, concentrating the extractive solution to remove ethanol, and diluting the concentrated solution with water;
D. the diluted extracts were combined and filtered to obtain a filtrate.
The technical scheme of the invention is further improved as follows: the separation and purification of verbascoside in the third step also comprises the following steps,
A. preparing crude verbascoside;
B. purifying the crude product of acteoside;
C. the purified verbascoside is stored.
The technical scheme of the invention is further improved as follows: the process for preparing the crude verbascoside comprises the following steps:
a1, combining and diluting the filtrates, and adsorbing the combined and diluted filtrates by polyamide resin;
a2, eluting with ethanol after polyamide resin adsorption, collecting eluate rich in acteoside, concentrating, and gradient eluting, wherein the procedure comprises: pure water, ethanol water solution with volume concentration of 5%, 10% and 20%;
a3, concentrating and drying the eluent rich in verbascoside to obtain crude verbascoside.
The technical scheme of the invention is further improved as follows: the process for purifying the crude verbascoside is as follows:
b1, dissolving the crude product with water, and adsorbing with macroporous adsorption resin;
b2, eluting with ethanol, collecting eluate rich in acteoside, and concentrating;
b3, drying to obtain refined acteoside.
The technical scheme of the invention is further improved as follows: the process of storing the purified verbascoside is as follows:
c1: sterilizing the medicinal plastic bag, loading purified acteoside into the medicinal plastic bag, weighing,
c2: filling a certain amount of nitrogen into the medicinal plastic bag, sealing with a plastic packaging machine, labeling, and placing in a normal temperature drying area.
The technical scheme of the invention is further improved as follows: the ethanol extraction in the step c is reflux extraction or ultrasonic extraction; the extraction times are 2-5 times, each time for 2-5 hours, the addition amount of ethanol for each extraction is 1.5-12 times of the weight of the crude drug, the concentration of the ethanol is 0% -95%,
the technical scheme of the invention is further improved as follows: the macroporous resin model is selected from: d101 resin, AB-8 macroporous adsorption resin, HPD100 resin, HPD-200A resin and HPD-400 resin.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:
1. the invention provides a method for separating and purifying verbascoside, which does not use beta-glucosidase which is expensive and difficult in source as an enzymolysis reagent, has a simpler process route, is easier to realize industrial production, avoids the defects of complex process, inconvenient operation, long period, low efficiency and the like of the traditional verbascoside extract mainly by heating extraction, membrane separation, resin purification, biotransformation, chemical catalysis and other methods, simultaneously determines key process parameters of operation procedures of each intermediate and quality standards of each intermediate, ensures the synthesis quality of the intermediate, is beneficial to improving the quality of a final product after the purification of dezocine, and can conveniently perform selective inspection on the quality of the verbascoside finished product by classified packaging, determines product bodies to ensure the quality of the product, thereby being more beneficial to large-scale production.
2. The invention provides a method for separating and purifying verbascoside, which is characterized in that crude verbascoside is prepared, purified and stored, synthesis steps and reaction conditions are optimized, the synthesis period is shortened, key process parameters and the quality standard of a product are determined, so that a key control point can be quickly mastered in the process of purifying dezocine, the purified verbascoside is efficiently and stably prepared, the operation is convenient and fast, the content of the verbascoside prepared by the method is higher than that of the verbascoside prepared by the prior art, and the requirement of a new reported medicine on the content can be met.
3. The invention provides a method for separating and purifying verbascoside, the verbascoside product is derived from Scrophulariaceae plants, the plant source of the verbascoside is expanded, the medicine source is rich, the method for extracting and preparing the verbascoside is optimized and improved, the operation is simple and convenient, the used cost is low, only ethanol is used as an organic reagent in the operation, toxic reagents such as methanol, acetone and the like are not adopted, the method is nontoxic, harmless, green and environment-friendly, the yield and the purity of the verbascoside are high, the popularization and the use are easy, and the recommendation is worthy.
Detailed Description
The present invention will be described in further detail with reference to the following examples:
example 1
The invention provides a method for separating and purifying verbascoside, which is separated and extracted from traditional Chinese medicine rehmannia root, and comprises the following steps:
step one, preparing materials and reagents, comprising: 80% ethanol, adsorption resin and 5-25% ethanol water solution;
step two, sample treatment:
A. pretreating radix rehmanniae, cutting into small pieces, and placing in an extraction tank;
B. introducing steam from the bottom of the pot for 40 minutes, and scalding the small pieces of rehmannia with saturated steam;
C. extracting with ethanol, concentrating the extractive solution to remove ethanol, and diluting the concentrated solution with water;
D. the diluted extracts were combined and filtered to obtain a filtrate.
The separation and purification of verbascoside in the three steps also comprises the following steps,
A. preparation of crude verbascoside:
A. combining the diluted filtrates, and adsorbing by polyamide resin;
B. eluting with ethanol after polyamide resin adsorption, collecting eluate rich in acteoside, concentrating,
C. concentrating and drying the eluent rich in verbascoside to obtain crude verbascoside;
B. and (3) purifying a crude verbascoside product:
A. dissolving the crude product with water, and adsorbing with macroporous adsorbent resin;
B. eluting with ethanol, collecting eluate rich in acteoside, and concentrating;
C. drying to obtain refined verbascoside;
C. storage of purified verbascoside:
a: sterilizing the medicinal plastic bag, loading purified acteoside into the medicinal plastic bag, weighing,
b: filling a certain amount of nitrogen into the medicinal plastic bag, sealing with a plastic packaging machine, labeling, and placing in a normal temperature drying area.
Example 2
On the basis of embodiment 1, the invention provides a technical scheme that: preferably, the macroporous resin type is selected from: d101 resin, AB-8 macroporous adsorption resin, HPD100 resin, HPD-200A resin and HPD-400 resin, and the gradient elution procedure comprises the following steps: eluting with 5%, 10%, 20%, 25% ethanol water solution, wherein the ethanol extraction is reflux extraction or ultrasonic extraction, the extraction time is 2-5 times, each time is 2-5 hr, the amount of ethanol added in each extraction is 1.5-12 times of the weight of the crude drug, and the ethanol concentration is 0% -95%.
Has the advantages that: the product of acteoside is derived from Scrophulariaceae plants, the plant source and the medicine source of the acteoside are expanded, the extraction and preparation method of the acteoside is optimized and improved, the operation is simple and convenient, the cost is low, only ethanol is used as an organic reagent in the operation, toxic reagents such as methanol, acetone and the like are not adopted, the method is nontoxic, harmless, green and environment-friendly, the yield and the purity of the acteoside are high, the method is easy to popularize and use and is worthy of recommendation, beta-glucosidase which is expensive and difficult in source is not used as an enzymolysis reagent, the process route is simpler, the industrial production is easier to realize, the defects of complex process, inconvenient operation, long period, low efficiency and the like existing in the conventional methods such as heating extraction, membrane separation, resin purification, biotransformation, chemical catalysis and the like of the acteoside extract are avoided, and simultaneously the key process parameters of the operation procedures of each intermediate and the quality standard of each intermediate are clear, the synthesis quality of the intermediate is ensured, the final quality of the finished product after dezocine purification is favorably improved, and the classified package can conveniently carry out spot inspection on the quality of the finished product of the verbascoside, the product body is determined to ensure the quality of the product, thereby being more favorable for large-scale production.
The present invention has been described in general terms in the foregoing, but it will be apparent to those skilled in the art that modifications and improvements can be made thereto based on the present invention. Therefore, modifications or improvements are within the scope of the invention without departing from the spirit of the inventive concept.
Claims (9)
1. A method for separating and purifying verbascoside is separated and extracted from a traditional Chinese medicine rehmannia root, and is characterized in that: the method for separating acteoside comprises the following steps:
step one, preparing materials and reagents;
step two, sample treatment is carried out;
step three, separating and purifying verbascoside.
2. The method for separating and purifying verbascoside as claimed in claim 1, wherein: the preparation of materials and reagents in the first step further comprises: 80% ethanol, adsorption resin and 5-25% ethanol water solution with volume concentration.
3. The method for separating and purifying verbascoside as claimed in claim 1, wherein: the sample processing in the second step further comprises:
A. pretreating radix rehmanniae, cutting into small pieces, and placing in an extraction tank;
B. introducing steam from the bottom of the pot for 40 minutes, and scalding the small pieces of rehmannia with saturated steam;
C. extracting with ethanol, concentrating the extractive solution to remove ethanol, and diluting the concentrated solution with water;
D. the diluted extracts were combined and filtered to obtain a filtrate.
4. The method for separating and purifying verbascoside as claimed in claim 1, wherein: the separation and purification of verbascoside in the third step also comprises the following steps,
A. preparing crude verbascoside;
B. purifying the crude product of acteoside;
C. the purified verbascoside is stored.
5. The method for separating and purifying verbascoside as claimed in claim 4, wherein: the process for preparing the crude verbascoside comprises the following steps:
a1, combining and diluting the filtrates, and adsorbing the combined and diluted filtrates by polyamide resin;
a2, adsorbing with polyamide resin, eluting with ethanol, collecting eluate rich in acteoside, and concentrating;
a3, concentrating and drying the eluent rich in verbascoside to obtain crude verbascoside.
6. The method for separating and purifying verbascoside as claimed in claim 4, wherein: the process for purifying the crude verbascoside is as follows:
b1, dissolving the crude product with water, and adsorbing with macroporous adsorption resin;
b2, eluting with ethanol, collecting eluate rich in acteoside, and concentrating;
b3, drying to obtain refined acteoside.
7. The method for separating and purifying verbascoside as claimed in claim 4, wherein: the process of storing the purified verbascoside is as follows:
c1: sterilizing the medicinal plastic bag, loading purified acteoside into the medicinal plastic bag, weighing,
c2: filling a certain amount of nitrogen into the medicinal plastic bag, sealing with a plastic packaging machine, labeling, and placing in a normal temperature drying area.
8. The method for separating and purifying verbascoside as claimed in claim 3, wherein: the ethanol extraction in the step c is reflux extraction or ultrasonic extraction; the extraction times are 2-5 times, each time for 2-5 hr, the amount of ethanol added for each extraction is 1.5-12 times of the weight of the crude drug, and the ethanol concentration is 0% -95%.
9. The method for separating and purifying verbascoside as claimed in claim 2, wherein: the macroporous resin model is selected from: d101 resin, AB-8 macroporous adsorption resin, HPD100 resin, HPD-200A resin and HPD-400 resin.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115708845A (en) * | 2022-12-15 | 2023-02-24 | 河南省纳普生物技术有限公司 | Method for extracting rehmanniae radix extract and its application in preparing antiinflammatory medicine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103896997A (en) * | 2014-04-14 | 2014-07-02 | 青海伊纳维康生物科技有限公司 | Method for separating and purifying verbascoside from desertliving cistanche |
| CN106065024A (en) * | 2015-10-30 | 2016-11-02 | 浙江工业大学 | A kind of method extracting separation verbascoside from Chinese medicine Radix Rehmanniae |
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- 2021-12-27 CN CN202111608349.XA patent/CN114276391A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103896997A (en) * | 2014-04-14 | 2014-07-02 | 青海伊纳维康生物科技有限公司 | Method for separating and purifying verbascoside from desertliving cistanche |
| CN106065024A (en) * | 2015-10-30 | 2016-11-02 | 浙江工业大学 | A kind of method extracting separation verbascoside from Chinese medicine Radix Rehmanniae |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115708845A (en) * | 2022-12-15 | 2023-02-24 | 河南省纳普生物技术有限公司 | Method for extracting rehmanniae radix extract and its application in preparing antiinflammatory medicine |
| CN115708845B (en) * | 2022-12-15 | 2023-09-08 | 河南省纳普生物技术有限公司 | Extraction method of rehmannia root extract and application of rehmannia root extract in preparation of anti-inflammatory drugs |
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