CN112138131A - Preparation method and application of total gingerol external preparation - Google Patents
Preparation method and application of total gingerol external preparation Download PDFInfo
- Publication number
- CN112138131A CN112138131A CN202011080586.9A CN202011080586A CN112138131A CN 112138131 A CN112138131 A CN 112138131A CN 202011080586 A CN202011080586 A CN 202011080586A CN 112138131 A CN112138131 A CN 112138131A
- Authority
- CN
- China
- Prior art keywords
- gingerol
- macroporous resin
- total
- extraction
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9068—Zingiber, e.g. garden ginger
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Rheumatology (AREA)
- Engineering & Computer Science (AREA)
- Oncology (AREA)
- Physical Education & Sports Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Dermatology (AREA)
- Pain & Pain Management (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a preparation method of a total gingerol external preparation, which comprises the steps of supercritical extraction, macroporous resin column chromatography, HPLC purification and preparation of a gel preparation, wherein the gel preparation comprises 100 parts of refined gingerol, 5-15 g of carbomer, 20-30 g of glycerol and 0.1-0.2 g of polysorbate; and (4) distilled water. The invention prepares the total gingerol into an external preparation, on one hand, the invention can utilize the antipyretic and analgesic effects and other drug effects of the total gingers, and simultaneously avoids the serious side effect of long-term oral administration on the digestive tract.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a preparation method and application of a total gingerol external preparation.
Background
Rheumatoid arthritis and the like (RA) are chronic systemic diseases mainly caused by inflammatory synovitis with unknown etiology. Its onset may be related to genetics, infection, sex hormones, etc. The pathology of RA arthritis is mainly synovial lining cell hyperplasia, interstitial massive inflammatory cell infiltration, microvascular neogenesis, pannus formation, cartilage and bone tissue destruction, and the like. Rheumatoid arthritis (rheumatoid arthritis) is a common acute or chronic inflammation of connective tissue. Generally, the rheumatic arthritis is one of the main manifestations of rheumatic fever, and is clinically characterized by joint and muscle wandering soreness, red swelling and pain. It is associated with group A type B hemolytic streptococcal infection, and cold, dampness and other factors can induce the disease. The large joints of the lower limbs such as the knee joints and the ankle joints are most frequently affected. The manifestations of the two diseases are chronic diseases which are difficult to cure radically, and pain is also a common manifestation of the two diseases, because of long-term and intractable publication, the symptomatic treatment analgesic with obvious and small side effect is very important, and a proper administration route is also equally important.
Gingerol is a series of phenolic compounds with longer alkyl side chains found in ginger, has very good anti-inflammatory and analgesic effects, and modern pharmacology shows that gingerol is a very strong cyclooxygenase inhibitor, thereby showing the activity of non-steroidal anti-inflammatory drugs. The gingerol soft capsules are oral preparations prepared by taking ginger oleoresin with lower content as a raw material, and have the common stimulation effect of non-steroidal anti-inflammatory drugs on gastrointestinal tracts after long-term administration, even serious complications such as bleeding or perforation and the like.
Therefore, one of the problems to be solved by the present invention is: improving the stimulation effect of gingerol oral medicine on intestinal tract.
The gingerol extract has ginger odor mainly caused by volatile oil components such as limonene, citronellal, terpinene, zingiberene, p-cymene and the like, and the substances form the peculiar smell of ginger.
Reference 1: liu Dan and Zhang Cheng Hui; the research progress of extracting and applying the main bioactive components of ginger.
Document 1 describes ginger essential oil, gingerol, and diphenylheptane compounds, which are main bioactive components of ginger. From document 1, it is known that ginger essential oil is a water-insoluble yellowish or yellowish green oily liquid, and the main component thereof is monoterpene substances and the like. The essential oil component is the source of main odor in ginger or gingerol, and total gingerol or gingerol is the main active ingredient of ginger. Thus, the odor in ginger is mainly due to the presence of ginger essential oil. When the molecular distillation method is adopted to separate the monoterpene component of the ginger essential oil, the content of the zingiberene and the content of the caryophyllene respectively reach more than 20 percent of 55 percent. It follows that zingiberene is a major source of the odour of ginger.
Reference 2: lu Qu Jiang; (ii) an omming; summary of chemical analysis of ginger
Document 2 discloses: the ginger oil obtained by the supercritical extraction method is obviously different from the ginger oil obtained by the steam distillation method in components, and the ginger oil obtained by the supercritical extraction method contains both volatile oil components and higher gingerol components; the ginger oil obtained by distillation contains volatile oil component, but contains no gingerol component.
Therefore, gingerol and gingerol components are lost by a steam distillation method, and gingerol can be obtained by a supercritical extraction method, but ginger volatile oil components cannot be removed.
The chemical components of ginger mainly comprise volatile oil, gingerol and diphenyl heptane with antioxidant activity. Wherein the volatile oil mainly comprises alpha-pinene and beta-phellandrene of monoterpene; alpha-zingiberene and beta-bisabolene of sesquiterpenes; these substances have a strong odor.
Gingerol is a pungent ingredient in ginger, mainly 6-gingerol, and is a main active ingredient of ginger.
Reference 3: qiu qin, zhangguo english; a comparison between supercritical CO2 fluid extraction method and steam distillation method for extracting volatile oil from rhizoma Zingiberis recens slice is provided.
The use of supercritical CO is disclosed in document 32Extracting volatile oil from rhizoma Zingiberis recens by fluid extraction method and steam distillation method, determining percentage content by normalization method, and identifying chemical components by gas chromatography-mass spectrometry. The supercritical CO2 extraction method identifies 47 components in total, and accounts for more than 72% of the total components of the volatile oil, and the steam distillation method identifies 53 components in total, and accounts for more than 89% of the total components of the volatile oil.
The water vapor distillation method is used for extracting the volatile oil of the dried ginger slices to separate 59 compounds, and 53 compounds are identified. The compound fractions were identified to account for more than 89% of the total distillate fraction and more than 97% of the chromatographic total distillate peak area. In the water vapor distillation method for extracting the volatile oil of the dried ginger slices, the component with the highest content is the zingiberene with the relative content of 9.12 percent, and the component with the highest content is the sesquiterpenoid with the relative content of 7.99 percent and the phellandrene with the relative content of 7.71 percent. The relative contents of camphene (7.32%), farnesene (6.35%), bisabolene (5.14%), curcumene (4.95%), pinene (4.10%), citral (3.13%) and the like are also high.
65 compounds are separated from the volatile oil of the dried ginger slices extracted by the supercritical CO2 extraction method, 47 compounds are identified, and the identified compound components account for more than 72 percent of the total distillate components and account for more than 84 percent of the area of the chromatographic total distillate peaks. Of the identified fractions, the highest content was zingiberene, at a relative content of 10.79%, followed by-sesquiphellandrene, at a relative content of 8.78%. The relative contents of curcumene (7.11%), shogaol (6.60%), bisabolene (6.40%), farnesene (5.05%) and the like are higher.
It is known from documents 1 and 2 that the main source of the odor of ginger is ginger volatile oil, which is extracted by supercritical and distillation methods, and zingiberene is the highest content of components in both volatile oils, and sesquiterpenene is the second.
Ginger has a certain pungent odor which is not acceptable to all patients, and the higher odor volatile oil component also reduces shelf life. Furthermore, children are often more sensitive to smell when taking the medicine, and most children reject the external medicine with special smell.
Secondly, the volatile oil component is easy to generate irritation to the skin, and the red, swollen and burning phenomenon is easy to occur after the medicine is taken, although most of the volatile oil component can relieve the red, swollen and burning phenomenon by itself, the volatile oil component still has certain discomfort, and the phenomenon is easy to occur after children take the medicine.
Therefore, one of the other technical problems to be solved by the present invention is: after the total gingerol is prepared into the external preparation, how to reduce the special smell of ginger, how to reduce the volatile oil component of the ginger and how to reduce the inconvenience of the external preparation.
Disclosure of Invention
The invention aims to provide a preparation method of a total gingerol external preparation.
The problems to be solved by the invention are as follows: improving the stimulation effect of gingerol oral medicine on intestinal tract.
The other technical problem to be solved by the invention is as follows: after the total gingerol is prepared into the external preparation, how to reduce the special smell of ginger, reduce the content of ginger volatile oil in the supercritical extraction process, reduce the irritation to the external preparation and improve the acceptance of children medicine.
In order to achieve the above objects, one embodiment of the present invention provides a method for preparing a total gingerol external preparation, comprising the steps of:
step (1): supercritical extraction
Pulverizing Zingiberis rhizoma into fine powder, sieving the fine powder, placing into supercritical carbon dioxide extraction kettle, extracting with supercritical carbon dioxide as solvent, and adding entrainer during extraction; the entrainer comprises the following components in mass: 60-70% of ethanol, 5-8% of methyl hydroxybenzoate, 2-3% of triethanolamine and water;
the technological parameters of the supercritical extraction are as follows: the extraction pressure is 25 MPa-32 MPa, the extraction temperature is 52-60 ℃, the flow of carbon dioxide is controlled to be 5L/h-15L/h, and the extraction time is 2 h-4 h; the flow ratio of the entrainer to the carbon dioxide solvent is 2-5%; centrifuging the material in the extraction kettle of the supercritical extraction equipment, and removing impurities to obtain a total gingerol crude extract;
step (2): macroporous resin column chromatography
Subjecting the crude extract of total gingerol obtained after supercritical extraction to macroporous resin adsorption treatment; the macroporous resin adsorption treatment steps are as follows:
(A) soaking the macroporous resin in dilute alkali liquor with the pH value of 7.5-8.5 for 5-10 min, washing the macroporous resin with deionized water to be neutral, soaking the macroporous resin in ethyl acetate for 5-10 min, and washing the macroporous resin with deionized water to be neutral again to obtain pretreated macroporous resin;
(B) loading the pretreated macroporous resin into a column by a wet method, uniformly stirring the macroporous resin and deionized water, continuously adding the macroporous resin and the deionized water into a glass chromatographic column, and discharging excess water from the lower part to enable the macroporous resin to gradually settle to the middle part of the glass chromatographic column;
(C) dissolving the crude extract of total gingerol in methanol, performing centrifugal precipitation and filtration, adding the filtrate above a glass chromatographic column, and adding a mobile phase for elution after the mixed solution of the crude extract of total gingerol and methanol slowly falls to the same level as the stationary phase;
step (3) preparation of external preparation of total gingerol
Purifying the received eluent by HPLC, and evaporating to dryness under reduced pressure to obtain refined gingerol; adding refined gingerol into gel matrix, and mixing to obtain total gingerol gel preparation.
In one optimized scheme of the invention, the macroporous resin in the step (2) is AB-8 type macroporous resin or D4020 macroporous resin.
According to one optimized scheme of the invention, the volume ratio of the total gingerol crude extract to methanol in the step (C) is 1: 1-2; the mobile phase is an ethanol solution, and the mass fraction of the ethanol solution is 60-70%; the elution flow rate is 0.3mL/min to 1 mL/min.
In one optimization scheme of the invention, in the step (C), one tube of eluent is received every 5mL, and the content of the total gingerol in the eluent is detected until the eluent does not contain the total gingerol.
In the supercritical extraction process, an entrainer is added by adopting a continuous pumping method; separating and resolving after the supercritical extraction is finished, wherein the separating and resolving pressure is controlled to be 5 MPa-8 MPa; the temperature is controlled to be 35-45 ℃, and the flow rate during separation and analysis is controlled to be 10-15L/h.
In one of the optimization schemes of the invention, the total gingerol gel preparation comprises the following components by weight: 100 parts of refined gingerol, 5-15 g of carbomer, 20-30 g of glycerin and 0.1-0.2 g of polysorbate; and (4) distilled water.
The invention also discloses application of the total gingerol external preparation prepared by any one of the preparation methods in anti-mutagenesis, anti-inflammatory, antirheumatic and antibacterial medicines.
In summary, the invention has the following advantages:
1. by optimizing the extraction process, the extraction rate of the volatile oil component of the ginger is reduced in the extraction process, so that the main odor component in the refined gingerol is greatly reduced; is more beneficial to external preparations.
2. The invention prepares the total gingerol into an external preparation, on one hand, the invention can utilize the antipyretic and analgesic effects and other drug effects of the total gingers, and simultaneously avoids the serious side effect of long-term oral administration on the digestive tract.
Detailed Description
The invention provides a preparation method of a total gingerol external preparation, which comprises the following steps:
step (1): supercritical extraction
Pulverizing Zingiberis rhizoma into fine powder, sieving the fine powder, placing into supercritical carbon dioxide extraction kettle, extracting with supercritical carbon dioxide as solvent, and adding entrainer during extraction; the entrainer comprises the following components in mass: 60-70% of ethanol, 5-8% of methyl hydroxybenzoate, 2-3% of triethanolamine and water;
the technological parameters of the supercritical extraction are as follows: the extraction pressure is 25 MPa-32 MPa, the extraction temperature is 52-60 ℃, the flow of carbon dioxide is controlled to be 5L/h-15L/h, and the extraction time is 2 h-4 h; the flow ratio of the entrainer to the carbon dioxide solvent is 2-5%; centrifuging the material in the extraction kettle of the supercritical extraction equipment, and removing impurities to obtain a total gingerol crude extract;
step (2): macroporous resin column chromatography
Subjecting the crude extract of total gingerol obtained after supercritical extraction to macroporous resin adsorption treatment; the macroporous resin adsorption treatment steps are as follows:
(A) soaking the macroporous resin in dilute alkali liquor with the pH value of 7.5-8.5 for 5-10 min, washing the macroporous resin with deionized water to be neutral, soaking the macroporous resin in ethyl acetate for 5-10 min, and washing the macroporous resin with deionized water to be neutral again to obtain pretreated macroporous resin;
(B) loading the pretreated macroporous resin into a column by a wet method, uniformly stirring the macroporous resin and deionized water, continuously adding the macroporous resin and the deionized water into a glass chromatographic column, and discharging excess water from the lower part to enable the macroporous resin to gradually settle to the middle part of the glass chromatographic column;
(C) dissolving the crude extract of total gingerol in methanol, performing centrifugal precipitation and filtration, adding the filtrate above a glass chromatographic column, and adding a mobile phase for elution after the mixed solution of the crude extract of total gingerol and methanol slowly falls to the same level as the stationary phase;
step (3) preparation of external preparation of total gingerol
Purifying the received eluent by HPLC, and evaporating to dryness under reduced pressure to obtain refined gingerol; adding refined gingerol into gel matrix, and mixing to obtain total gingerol gel preparation.
The HPLC purification process can adopt the following steps: centrifuging the eluate, filtering, adding ethanol with equal volume to obtain purified solution, and adding C18A 250X 30mm chromatography column; the sample introduction amount is 10 microliter, the column temperature is controlled at 28 ℃, and the flow rate is 5 mL/min; the mobile phase is a methanol-water system; isocratic elution.
Wherein the macroporous resin in the step (2) is AB-8 type macroporous resin or D4020 macroporous resin.
The volume ratio of the total gingerol crude extract to the methanol in the step (C) is 1: 1-2; the mobile phase is an ethanol solution, and the mass fraction of the ethanol solution is 60-70%; the elution flow rate is 0.3mL/min to 1 mL/min.
And (C) receiving one tube of eluent every 5mL in the step (C), and detecting the content of the total gingerol in the eluent until the eluent does not contain the total gingerol.
In the supercritical extraction process, an entrainer is added by adopting a continuous pumping method; separating and resolving after the supercritical extraction is finished, wherein the separating and resolving pressure is controlled to be 5 MPa-8 MPa; the temperature is controlled to be 35-45 ℃, and the flow rate during separation and analysis is controlled to be 10-15L/h.
The total gingerol gel preparation comprises the following components in parts by weight: 100 parts of refined gingerol, 5-15 g of carbomer, 20-30 g of glycerin and 0.1-0.2 g of polysorbate; and (4) distilled water.
Example 1: supercritical extraction
Crushing the dried ginger into fine powder, sieving the fine powder with a 50-mesh sieve, putting the fine powder into a supercritical carbon dioxide extraction kettle, extracting by using supercritical carbon dioxide as a solvent, and adding an entrainer in the extraction process; the entrainer comprises the following components in mass: 70% of ethanol, 7% of methyl hydroxybenzoate, 2% of triethanolamine and 20% of water.
The technological parameters of the supercritical extraction are as follows: the extraction pressure is 30Mpa, the extraction temperature is 55 ℃, the carbon dioxide flow is controlled to be 12L/h, and the extraction time is 2.3 h; the flow ratio of the entrainer to the carbon dioxide solvent is 2%; centrifuging the material in the extraction kettle of the supercritical extraction equipment, and removing impurities to obtain a total gingerol crude extract;
in the supercritical extraction process, an entrainer is added by adopting a continuous pumping method; separating and resolving after the supercritical extraction is finished, wherein the separating and resolving pressure is controlled to be 6 MPa; the temperature was controlled at 42 ℃ and the flow rate during separation and analysis was controlled at 13L/h.
Example 2: macroporous resin column chromatography
(A) Soaking the AB-8 type macroporous resin in dilute alkali solution with the pH value of 7.8 for 10min, washing the soaked resin to be neutral by using deionized water, soaking the soaked resin in ethyl acetate for 6min, and washing the resin to be neutral by using the deionized water again to obtain the pretreated macroporous resin.
(B) And (3) loading the pretreated macroporous resin into a column by a wet method, uniformly stirring the macroporous resin and deionized water, continuously adding the macroporous resin and the deionized water into a glass chromatographic column, and discharging excess water from the lower part to enable the macroporous resin to gradually settle to the middle part of the glass chromatographic column.
(C) Dissolving 1 part of the crude extract of the total gingerol in 2 parts of methanol by weight, performing centrifugal precipitation filtration, adding the mixture above a glass chromatographic column, slowly reducing the mixed solution of the crude extract of the total gingerol and the methanol to be level with the stationary phase, and adding 65% ethanol of a mobile phase for elution, wherein the elution flow rate is 0.6 mL/min.
Receiving one tube of eluent every 5mL, and detecting the content of the total gingerol in the eluent until the eluent does not contain the total gingerol; all eluates were then pooled.
Example 3: preparation of external preparation of total gingerol
Purifying the received eluent by HPLC, and evaporating to dryness under reduced pressure to obtain refined gingerol; the total gingerol gel preparation comprises the following components in parts by weight: 100 parts of refined gingerol, 5-15 g of carbomer, 20-30 g of glycerin and 0.1-0.2 g of polysorbate; and (4) distilled water. Mixing refined gingerol, carbomer 940, glycerol, polysorbate 80 and appropriate amount of distilled water to obtain paste to obtain gel preparation.
The total gingerol external preparation prepared by the method disclosed by the invention can be used for resisting mutation, inflammation, rheumatism and bacteria.
Experimental example 1: influence of different entrainers on extraction degree of volatile oil components such as zingiberene and sesquioenanthe
The experimental method comprises the following steps: based on the supercritical extraction method disclosed in example 1, the entrainer component in example 1 was replaced with a different entrainer, and the remaining process steps and parameters remained the same as in example 1; the entrainer composition in each experimental example is shown in table 1 below:
| group of | Entrainer component |
| Experimental example 1 | 80% of ethanol and 20% of water |
| Experimental example 2 | N-octane |
| Experimental example 3 | Acetone (II) |
| Experimental example 4 | 80 percent of methyl hydroxybenzoate and 20 percent of water |
| Experimental example 5 | 80 percent of triethanolamine and 20 percent of water |
| Example 6 | 70 percent of methyl hydroxybenzoate, 20 percent of triethanolamine and 10 percent of water |
The experimental steps are as follows:
1. the entrainers of example 1 and experimental examples 1 to 6 were divided into 7 groups, and refined gingerols were prepared according to the methods disclosed in example 1, example 2 and example 3, as follows:
(1) supercritical extraction
Crushing the dried ginger into fine powder, sieving the fine powder with a 50-mesh sieve, putting the fine powder into a supercritical carbon dioxide extraction kettle, extracting by using supercritical carbon dioxide as a solvent, and adding an entrainer in the extraction process; entrainers were selected in turn from the entrainers disclosed in table 1.
The technological parameters of the supercritical extraction are as follows: the extraction pressure is 30Mpa, the extraction temperature is 55 ℃, the carbon dioxide flow is controlled to be 12L/h, and the extraction time is 2.3 h; the flow ratio of the entrainer to the carbon dioxide solvent is 2%; centrifuging the material in the extraction kettle of the supercritical extraction equipment, and removing impurities to obtain a total gingerol crude extract; in the supercritical extraction process, an entrainer is added by adopting a continuous pumping method; separating and resolving after the supercritical extraction is finished, wherein the separating and resolving pressure is controlled to be 6 MPa; the temperature was controlled at 42 ℃ and the flow rate during separation and analysis was controlled at 13L/h.
(2) Macroporous resin column chromatography
(A) Soaking the AB-8 type macroporous resin in dilute alkali solution with the pH value of 7.8 for 10min, washing the soaked resin to be neutral by using deionized water, soaking the soaked resin in ethyl acetate for 6min, and washing the resin to be neutral by using the deionized water again to obtain the pretreated macroporous resin.
(B) And (3) loading the pretreated macroporous resin into a column by a wet method, uniformly stirring the macroporous resin and deionized water, continuously adding the macroporous resin and the deionized water into a glass chromatographic column, and discharging excess water from the lower part to enable the macroporous resin to gradually settle to the middle part of the glass chromatographic column.
(C) Dissolving 1 part of the crude extract of the total gingerol in 2 parts of methanol by weight, performing centrifugal precipitation filtration, adding the mixture above a glass chromatographic column, slowly reducing the mixed solution of the crude extract of the total gingerol and the methanol to be level with the stationary phase, and adding 65% ethanol of a mobile phase for elution, wherein the elution flow rate is 0.6 mL/min.
Receiving one tube of eluent every 5mL, and detecting the content of the total gingerol in the eluent until the eluent does not contain the total gingerol; all eluates were then pooled.
(3) Preparation of refined gingerol
And (3) carrying out HPLC purification on the received eluent, and then carrying out reduced pressure evaporation to dryness after purification to obtain the refined gingerol. The purification process of HPLC can be adopted as follows: centrifuging the eluate, filtering, adding ethanol with equal volume to obtain purified solution, and adding C18A 250X 30mm chromatography column; the sample introduction amount is 10 microliter, the column temperature is controlled at 28 ℃, and the flow rate is 5 mL/min; the mobile phase is a methanol-water system; isocratic elution.
2. Detecting the content of ginger essential oil in 7 groups of refined gingerols.
The detection principle is as follows: because gingerol or gingerol can not be prepared by a steam distillation method, refined gingerol is treated by the steam distillation method, volatile oil components are collected, the weight difference between the collected volatile oil components and the refined gingerol is the weight of gingerol or gingerol, and thus the content of the volatile oil of the ginger essential oil and the content of gingerol or gingerol can be calculated.
The detection method comprises the following steps: weighing 100g of the 7 groups of refined gingerols, respectively adding into 5g of water, introducing water vapor under normal pressure to distill off the ginger volatile oil at a temperature lower than the boiling point of the ginger volatile oil, drying, weighing, and calculating the content of the ginger volatile oil; the results are shown in table 2 below:
| group of | Total g of refined gingerol | Distilled volatile oil amount g | The residual gingerol |
| Example 1 | 100 | 15 | 75 |
| Experimental example 1 | 100 | 55 | 45 |
| Experimental example 2 | 100 | 52 | 48 |
| Experimental example 3 | 100 | 58 | 42 |
| Experimental example 4 | 100 | 42 | 58 |
| Experimental example 5 | 100 | 50 | 50 |
| Experimental example 6 | 100 | 36 | 54 |
The refined gingerol is obtained by supercritical extraction process; after the treatment by the same process, the larger the residual gingerol content is, the lower the content of volatile oil components in the original refined gingerol is, and the lower the content of volatile oil components is, the special smell in the product can be reduced, the external irritation is reduced, and the medicine acceptance of children is improved.
From the above experimental data it can be seen that:
the content of the rest gingerol is higher than that of the embodiment 1; the remaining amount of gingerol in the remaining experimental examples differed slightly. Wherein, the embodiment 1 is the technical proposal of the invention, the residual gingerol content is far larger than that of the rest control groups, and the entrainer in the experimental example 6 comprises 70 percent of methylparaben, 20 percent of triethanolamine and 10 percent of water; the entrainer in example 1 is ethanol 70%, methylparaben 7%, triethanolamine 2% and 20% water; experimental example 4 and Experimental example 5 are 80% of methylparaben + 20% of water, 80% of triethanolamine + 20% of water, respectively.
The conventional entrainer is selected in the experimental examples 1 to 3, the entrainer selected in the experimental example 1 is the preferred scheme of the invention, and the residual gingerol is the highest in the experimental example 1, so that the extraction of the volatile oil is inhibited in the supercritical extraction process, and the dissolution degree of the volatile oil component in carbon dioxide is reduced; compared with the experimental example 1, the experimental example 1 comprises 80% of ethanol and 20% of water; in example 1, methyl hydroxybenzoate is added by 7 percent, and triethanolamine is added by 2 percent; due to the addition of the two components, the dissolution degree of the volatile oil component in the supercritical extraction process is reduced; further ensuring the highest residual gingerol content in the obtained refined gingerol.
Experimental examples 4 to 6 were conducted to verify the effect of methylparaben and triethanolamine when used alone, and from the experimental data, the effect of experimental examples 4 to 6 was not obvious, and the amount of gingerol remained was slightly increased, but the difference was very large compared with example 1, which is probably because the above two components need to be under the combined action of ethanol, a conventional entrainer, to exert the above effect of inhibiting the volatile oil from dissolving into the extractant.
Claims (7)
1. A preparation method of a total gingerol external preparation is characterized by comprising the following steps:
step (1): supercritical extraction
Pulverizing Zingiberis rhizoma into fine powder, sieving the fine powder, placing into supercritical carbon dioxide extraction kettle, extracting with supercritical carbon dioxide as solvent, and adding entrainer during extraction; the entrainer comprises the following components in mass: 60-70% of ethanol, 5-8% of methyl hydroxybenzoate, 2-3% of triethanolamine and water;
the technological parameters of the supercritical extraction are as follows: the extraction pressure is 25 MPa-32 MPa, the extraction temperature is 52-60 ℃, the flow of carbon dioxide is controlled to be 5L/h-15L/h, and the extraction time is 2 h-4 h; the flow ratio of the entrainer to the carbon dioxide solvent is 2-5%; centrifuging the material in the extraction kettle of the supercritical extraction equipment, and removing impurities to obtain a total gingerol crude extract;
step (2): macroporous resin column chromatography
Subjecting the crude extract of total gingerol obtained after supercritical extraction to macroporous resin adsorption treatment; the macroporous resin adsorption treatment steps are as follows:
(A) soaking the macroporous resin in dilute alkali liquor with the pH value of 7.5-8.5 for 5-10 min, washing the macroporous resin with deionized water to be neutral, soaking the macroporous resin in ethyl acetate for 5-10 min, and washing the macroporous resin with deionized water to be neutral again to obtain pretreated macroporous resin;
(B) loading the pretreated macroporous resin into a column by a wet method, uniformly stirring the macroporous resin and deionized water, continuously adding the macroporous resin and the deionized water into a glass chromatographic column, and discharging excess water from the lower part to enable the macroporous resin to gradually settle to the middle part of the glass chromatographic column;
(C) dissolving the crude extract of total gingerol in methanol, performing centrifugal precipitation and filtration, adding the filtrate above a glass chromatographic column, and adding a mobile phase for elution after the mixed solution of the crude extract of total gingerol and methanol slowly falls to the same level as the stationary phase;
step (3) preparation of external preparation of total gingerol
Purifying the received eluent by HPLC, and evaporating to dryness under reduced pressure to obtain refined gingerol; adding refined gingerol into gel matrix, and mixing to obtain total gingerol gel preparation.
2. The method for preparing a total gingerol external preparation according to claim 1, wherein: the macroporous resin in the step (2) is AB-8 type macroporous resin or D4020 macroporous resin.
3. The method for preparing a total gingerol external preparation according to claim 1, wherein: the volume ratio of the total gingerol crude extract to the methanol in the step (C) is 1: 1-2; the mobile phase is an ethanol solution, and the mass fraction of the ethanol solution is 60-70%; the elution flow rate is 0.3mL/min to 1 mL/min.
4. The method for preparing a total gingerol external preparation according to claim 1, wherein: and (C) receiving one tube of eluent every 5mL in the step (C), and detecting the content of the total gingerol in the eluent until the eluent does not contain the total gingerol.
5. The method for preparing a total gingerol external preparation according to claim 1, wherein: in the supercritical extraction process, an entrainer is added by adopting a continuous pumping method; separating and resolving after the supercritical extraction is finished, wherein the separating and resolving pressure is controlled to be 5 MPa-8 MPa; the temperature is controlled to be 35-45 ℃, and the flow rate during separation and analysis is controlled to be 10-15L/h.
6. The method for preparing a total gingerol external preparation according to claim 1, wherein: the total gingerol gel preparation comprises the following components in parts by weight: 100 parts of refined gingerol, 5-15 g of carbomer, 20-30 g of glycerin and 0.1-0.2 g of polysorbate; and (4) distilled water.
7. Use of the total gingerol external preparation prepared by the preparation method according to any one of claims 1 to 6 in antimutagenic, antiinflammatory, antirheumatic and bacteriostatic medicaments.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011080586.9A CN112138131A (en) | 2020-10-11 | 2020-10-11 | Preparation method and application of total gingerol external preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011080586.9A CN112138131A (en) | 2020-10-11 | 2020-10-11 | Preparation method and application of total gingerol external preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112138131A true CN112138131A (en) | 2020-12-29 |
Family
ID=73952971
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011080586.9A Pending CN112138131A (en) | 2020-10-11 | 2020-10-11 | Preparation method and application of total gingerol external preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112138131A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023060914A1 (en) * | 2021-10-14 | 2023-04-20 | 重庆文理学院 | Method for extracting gingerol from ginger |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1762956A (en) * | 2005-09-26 | 2006-04-26 | 闫东海 | Supercritical column chromatography method for preparing general gingerol |
| CN102875347A (en) * | 2012-10-05 | 2013-01-16 | 南通四海植物精华有限公司 | Manufacturing process for extracting gingerol from gingers |
| CN102976909A (en) * | 2012-12-19 | 2013-03-20 | 中国科学院西北高原生物研究所 | Method for extracting and purifying 6-gingerol from ginger |
| CN103432562A (en) * | 2013-09-12 | 2013-12-11 | 南京泽朗农业发展有限公司 | Method for extracting fresh ginger polyphenol from fresh ginger |
| CN105695101A (en) * | 2016-03-17 | 2016-06-22 | 西双版纳版纳药业有限责任公司 | Zingiber purpureum Rosc. volatile oil extracted through supercritical carbon dioxide and preparation method and application of Zingiber purpureum Rosc. volatile oil |
| CN107778158A (en) * | 2016-08-30 | 2018-03-09 | 天津中新药业研究中心 | A kind of high-purity general gingerol, preparation method and the usage |
-
2020
- 2020-10-11 CN CN202011080586.9A patent/CN112138131A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1762956A (en) * | 2005-09-26 | 2006-04-26 | 闫东海 | Supercritical column chromatography method for preparing general gingerol |
| CN102875347A (en) * | 2012-10-05 | 2013-01-16 | 南通四海植物精华有限公司 | Manufacturing process for extracting gingerol from gingers |
| CN102976909A (en) * | 2012-12-19 | 2013-03-20 | 中国科学院西北高原生物研究所 | Method for extracting and purifying 6-gingerol from ginger |
| CN103432562A (en) * | 2013-09-12 | 2013-12-11 | 南京泽朗农业发展有限公司 | Method for extracting fresh ginger polyphenol from fresh ginger |
| CN105695101A (en) * | 2016-03-17 | 2016-06-22 | 西双版纳版纳药业有限责任公司 | Zingiber purpureum Rosc. volatile oil extracted through supercritical carbon dioxide and preparation method and application of Zingiber purpureum Rosc. volatile oil |
| CN107778158A (en) * | 2016-08-30 | 2018-03-09 | 天津中新药业研究中心 | A kind of high-purity general gingerol, preparation method and the usage |
Non-Patent Citations (2)
| Title |
|---|
| 林慧敏: "《现代分离纯化技术在水产品中的应用》", 31 December 2015, 海洋出版社 * |
| 范素琴等: "生姜风味物质及其应用", 《粮食与油脂》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023060914A1 (en) * | 2021-10-14 | 2023-04-20 | 重庆文理学院 | Method for extracting gingerol from ginger |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Huang et al. | Anti-inflammatory effects of Brucea javanica oil emulsion by suppressing NF-κB activation on dextran sulfate sodium-induced ulcerative colitis in mice | |
| US20170333503A1 (en) | Process for extraction, separation and purification of cannabinoids, flavonoids and terpenes from cannabis | |
| US11458087B2 (en) | Method of isolating cannabidiol from Cannabis plant and use thereof | |
| US20210100864A1 (en) | Method of continuously producing cannabidiol from cannabis plant and use thereof | |
| KR20130089305A (en) | Composition for preventing or treating of arthrits comprising herbal extract | |
| KR102287355B1 (en) | Method of isolating tetrahydrocannabinol from cannabis sp and uses thereof | |
| CN112138131A (en) | Preparation method and application of total gingerol external preparation | |
| Chen et al. | Identification of multiple constituents in the traditional Chinese medicine formula GuiZhiFuLing-Wan by HPLC–DAD–MS/MS | |
| Zhong et al. | Pharmacological effects of different ginger juices on the concurrent symptoms in animal models of functional dyspepsia: a comparative study | |
| CN110538223B (en) | Method for enriching polygonum hydropiper volatile oil by adopting macroporous resin | |
| JPH07330624A (en) | Antiinflammatory and antipruritic agent | |
| Hoshino et al. | Extraction of Functional components from freeze-dried Angelica furcijuga leaves using supercritical carbon dioxide | |
| JPH04266828A (en) | Plant extract and its use | |
| JP2017061501A (en) | Novel pharmacological use of cimicifuga extract | |
| CN107669728B (en) | Rape pollen total sterol for treating prostatic hyperplasia and preparation method thereof | |
| JP2000236843A (en) | Production of curcumins | |
| JP2007320958A (en) | Prophylactic and/or curative agent for fatty liver | |
| CN106631940B (en) | A kind of preparation method of high-purity garlic Ailitridin | |
| Abu-Shandi et al. | A Quick GC/MS Method correlated with LC/MS/MS for the Identification of Medicinal Natural Products in Convolvulus arvensis: An Injury Healing Plant. | |
| AU2019319172A1 (en) | Formulations containing lipophilic extracts of spicy edible plants useful in controlling pain and inflammation. | |
| WO2019211772A1 (en) | Obtaining extracts in a solid form | |
| CN110664962B (en) | Pure traditional Chinese medicine oil for relaxing tendons and activating collaterals and preparation method thereof | |
| EP3481408A1 (en) | A natural composition to inhibit beta-secretase and enhance bioavailability comprising of turmeric and sesame seed extract | |
| JP7270305B2 (en) | A pharmaceutical composition for prevention or treatment of inflammatory bowel disease, containing a mixed extract of Huanglian and radish as an active ingredient | |
| RU2394587C1 (en) | Medication possessing hypocholesterolemic, hypolipidemic and cholagogic action |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201229 |