CN114224920A - Bee therapy product with sterilizing and inflammation diminishing functions and preparation method thereof - Google Patents
Bee therapy product with sterilizing and inflammation diminishing functions and preparation method thereof Download PDFInfo
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- CN114224920A CN114224920A CN202210076365.7A CN202210076365A CN114224920A CN 114224920 A CN114224920 A CN 114224920A CN 202210076365 A CN202210076365 A CN 202210076365A CN 114224920 A CN114224920 A CN 114224920A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
- A61K35/64—Insects, e.g. bees, wasps or fleas
- A61K35/644—Beeswax; Propolis; Royal jelly; Honey
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- 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]
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- 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
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Abstract
The invention discloses a bee therapy product with sterilization and inflammation diminishing functions and a preparation method thereof, wherein the preparation method comprises the following steps: s1: removing impurities from propolis raw materials, removing dust by blowing, freezing for 8-12 hours at-10 to-20 ℃, then crushing, and sieving by a sieve of 80-100 meshes to obtain fine powder for later use; s2: adding a first extracting agent, a second extracting agent and a third extracting agent into the fine powder according to the proportion of the fine powder to an extracting solvent of 1 (8-22) W/V for solid-liquid extraction, collecting the part of the third extracting agent, separating propolis extract, and leaving a first extract and a second extract. In the invention, by adopting a separation and extraction process, the optimal extraction medium and extraction conditions are respectively selected for each extraction stage, so that the active ingredients are promoted to be dissolved out and impurities are separated to the maximum extent, and finally substances enriched with effective ingredients are mixed to obtain the propolis extract, wherein each mars component has high Haniang and the product effect is obvious.
Description
Technical Field
The invention relates to the technical field of bee therapy products, in particular to a bee therapy product with sterilization and inflammation diminishing functions and a preparation method thereof.
Background
Propolis is a colloidal substance prepared by collecting plant gum by bee, mixing with frontal gland secretion and beeswax, and chewing by bee. The propolis mainly comprises flavonoid, terpene, aromatic acid and ester of aromatic acid, phenol, alcohol, aldehyde, ketone, and trace amount of protein and enzyme. Propolis is proved to have wide pharmacological activities of resisting bacteria, fungi, viruses, inflammation, oxidation and cancer at present. The discovery of the effects directly provides a basis for developing special nutritional health care and medicines.
An antibacterial experiment is carried out on 100 strains of 14 bacterins by using the alcohol immersion liquid of the propolis capsule in Hetian seal and the like (1980) which are experts in China, and the propolis capsule is proved to have high antibacterial effect on gram-positive bacteria and also has strong antibacterial effect on meningococcus which is negative to the gram-positive bacteria, vibrio coagulans and highly pathogenic halophilic bacteria. Experiments prove that the propolis has better anti-inflammatory effect. The propolis ethanol extract and propolis water extract can be used for resisting pleural effusion increase of pleurisy rats. The prior art discloses several methods for preparing propolis extracts, for example, a patent ZL200710111189.1 of the applicant discloses a water-soluble propolis extract and a light-colored propolis extract having biological activity. The method comprises pulverizing propolis raw materials, extracting with 10-60% ethanol solution for 2-4 times (not less than four times of solvent amount per time), mixing extractive solutions, filtering, recovering ethanol under reduced pressure to obtain concentrated solution, standing to obtain precipitate and supernatant, washing the precipitate with water, and drying to obtain solid pale propolis extract. However, the extract obtained by the technical scheme is mainly light-colored propolis, and the antibacterial and anti-inflammatory components cannot be effectively enriched, so that the extract has a good pertinence, and the antibacterial and anti-inflammatory effects are slow in effect and large in dosage.
In view of the above, the present problems are improved and researched, and a sterilization and inflammation-diminishing bee therapy product and a preparation method thereof are provided to solve the problem of low concentration of active ingredients of the existing sterilization and inflammation-diminishing bee therapy product, and the purpose of solving the problems and improving the practical value is achieved through the technology.
Disclosure of Invention
The present invention is directed to solving one of the technical problems of the prior art or the related art.
Therefore, the technical scheme adopted by the invention is as follows: a preparation method of bee therapy product with antibacterial and antiinflammatory effects comprises the following steps:
s1: removing impurities from propolis raw materials, removing dust by blowing, freezing for 8-12 hours at-10 to-20 ℃, then crushing, and sieving by a sieve of 80-100 meshes to obtain fine powder for later use;
s2: adding a first extracting agent, a second extracting agent and a third extracting agent into fine powder according to the proportion of extraction solvent W/V of 1 (8-22) to perform solid-liquid extraction, adding the first extracting agent, the second extracting agent and the third extracting agent, extracting for 2 hours at 45 ℃ and at the rotating speed of 150rpmn, standing overnight at 4 ℃, then performing suction filtration and collection, mixing a silica gel adsorbent and the first extracting agent according to the volume ratio of 1:2-8, stirring and extracting for 1-3 times, filtering for 1-3 hours each time, naturally volatilizing filter residues to serve as a first extract, performing antibiotic removal treatment on the extract, and performing antibiotic removal by using macroporous adsorption resin; mixing the first extract and the second extractant according to a volume ratio of 1:2-8, stirring and extracting for 1-3 times, each time for 1-3 hours, filtering, and naturally volatilizing filter residues to obtain a second extract; mixing the second extract and the third extractant according to a volume ratio of 1:2-8, stirring and extracting for 1-3 times, each time for 1-3 hours, combining the third extracts after stirring and extraction, performing antibiotic removal treatment on the extracts, performing antibiotic removal by adopting macroporous adsorption resin, collecting the part of the third extractant, separating propolis extract, performing ultrasonic treatment on the rest of the first extract and the second extract at 15-35 ℃ for 30-60 min, filtering to obtain filter residue and filtrate, concentrating the filtrate under reduced pressure to obtain propolis refined extract, dispersing the propolis extract in an ethanol water solution with the ethanol weight percentage concentration of 60-95%, and performing ultrasonic treatment;
s3: adding an ethanol aqueous solution into the filter residue obtained in the previous step according to the W/V ratio of the filter residue to a 75% ethanol aqueous solution of 1 (4-8), extracting for 1-3 times at 65-75 ℃ for 1-3 h each time, and filtering the extracting solution to obtain a filtrate;
s4: and extracting the filtrate by using ultrahigh pressure extraction equipment, wherein the extraction pressure is 500-600 Mpa, the extraction time is 10-15 min, and the extract is concentrated and dried to obtain the compound.
The present invention in a preferred example may be further configured to: the first extractant is petroleum ether, the second extractant is a mixed solvent formed by mixing acetone and petroleum ether according to the volume ratio of less than 2:1, and the third extractant is a mixed solvent formed by mixing acetone and petroleum ether according to the volume ratio of (2-4) to 1.
The present invention in a preferred example may be further configured to: the silica gel adsorbent is of a silica gel adsorption powder structure, and the particle diameter of the silica gel adsorbent is 0.2-0.5 mm.
A bee therapy product with antibacterial and antiinflammatory effects comprises the extract of bee therapy product prepared by the above method, wherein the active ingredients include chrysin, pinocembrin, galangin, caffeic acid phenethyl ester, rhamnonin and benzoic acid.
The present invention in a preferred example may be further configured to: in the propolis extract, the weight percentage of the chrysin is 40-53%, the weight percentage of the pinocembrin is 11-15%, the weight percentage of the galangin is 7-10%, the weight percentage of the caffeic acid phenethyl ester is 8-112%, the weight percentage of the rhamnosin is 9-14%, and the weight percentage of the benzoic acid is 7-12%.
By adopting the technical scheme, the benzoic acid has the effects of inhibiting the growth of fungi, bacteria and mildew, the caffeic acid has wider antibacterial and antiviral effects and the effects of shortening the hemagglutination and bleeding time, and the types of the active ingredients are wide and high in content.
The beneficial effects obtained by the invention are as follows:
1. in the invention, by adopting a separation and extraction process, the optimal extraction medium and extraction conditions are respectively selected for each extraction stage, so that the active ingredients are promoted to be dissolved out and impurities are separated to the maximum extent, and finally substances enriched with effective ingredients are mixed to obtain the propolis extract, wherein each mars component has high Haniang and the product effect is obvious.
2. According to the invention, through extracting and enriching the flavonoid compounds in the propolis raw material, the propolis tea has different degrees of inhibition effects on various microorganisms, benzoic acid has the effects of inhibiting the growth of fungi, bacteria and molds, caffeic acid has wider antibacterial and antiviral effects and the effects of shortening the hemagglutination and bleeding time, and the variety of the active ingredients is wide and high in content.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to specific embodiments. It should be noted that the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.
It is to be understood that this description is made only by way of example and not as a limitation on the scope of the invention.
The following describes a bee therapy product with antibacterial and anti-inflammatory effects and a preparation method thereof according to some embodiments of the present invention.
The invention provides a preparation method of a bee therapy product with sterilization and inflammation diminishing functions, which comprises the following steps:
s1: removing impurities from propolis raw materials, removing dust by blowing, freezing for 10 hours at-10 to-20 ℃, then crushing, and sieving by a sieve of 80-100 meshes to obtain fine powder for later use;
s2: adding a first extracting agent, a second extracting agent and a third extracting agent into the fine powder according to the proportion of extraction solvent W/V of 1 (8-22) to perform solid-liquid extraction, adding the first extracting agent, the second extracting agent and the third extracting agent, extracting for 2 hours at 45 ℃ and 150rpmn at the rotating speed, standing for one night at 4 ℃, then performing suction filtration and collection, mixing a silica gel adsorbent and the first extracting agent according to the volume ratio of 1:5, stirring and extracting for 2 times, stirring and extracting for 2.5 hours each time, filtering, naturally volatilizing filter residues to serve as a first extract, performing antibiotic removal treatment on the extract, and performing antibiotic removal by adopting macroporous adsorption resin; mixing the first extract with a second extractant according to a volume ratio of 1:5, stirring and extracting for 2 times, each time for 2 hours, filtering, and naturally volatilizing filter residues to obtain a second extract; mixing the second extract with a third extracting agent according to a volume ratio of 1:5, stirring and extracting for 2 times, each time for 2 hours, combining the third extracts after stirring and extracting, performing antibiotic removal treatment on the extracts, performing antibiotic removal by adopting macroporous adsorption resin, collecting the part of the third extracting agent to obtain propolis extract for separation, remaining the first extract and the second extract, performing ultrasonic treatment for 40min at the temperature of 25 ℃, then filtering to obtain filter residue and filtrate, concentrating the filtrate under reduced pressure to obtain propolis refined extract, dispersing the propolis extract in an ethanol water solution with the weight percentage concentration of 80-90% of ethanol, and performing ultrasonic treatment;
s3: adding an ethanol water solution into the filter residue obtained in the above step according to the ratio of the filter residue to a 75% ethanol water solution of 1:5W/V, extracting for 2 times at 70 ℃ for 2h each time, and filtering the extracting solution to obtain a filtrate;
s4: extracting the filtrate by using ultrahigh pressure extraction equipment, wherein the extraction pressure is 500-600 Mpa, the extraction time is 15min, concentrating the extracting solution, and drying to obtain the compound.
In this embodiment, the first extractant is petroleum ether, the second extractant is a mixed solvent in which acetone and petroleum ether are mixed in a volume ratio of less than 2:1, and the third extractant is a mixed solvent in which acetone and petroleum ether are mixed in a volume ratio of 2, 7: 1.
In the embodiment, the silica gel adsorbent is in a silica gel adsorption powder structure, and the particle diameter of the silica gel adsorbent is 0.2-0.5mm, so that the propolis cream is easy to solidify into blocks and is convenient to separate.
A bee therapy product with antibacterial and antiinflammatory effects is the extract of bee therapy product prepared according to any one of claims 1-6, wherein the active ingredients include chrysin, pinocembrin, galangin, caffeic acid phenethyl ester, rhamnonin and benzoic acid.
In the embodiment, in the propolis extract, the weight percentage of the chrysin is 40-53%, the weight percentage of the pinocembrin is 11-15%, the weight percentage of the galangin is 7-10%, the weight percentage of the caffeic acid phenethyl ester is 8-112%, the weight percentage of the rhamnosin is 9-14%, and the weight percentage of the benzoic acid is 7-12%.
Specifically, benzoic acid has the effect of inhibiting the growth of fungi, bacteria and moulds, while caffeic acid has the broad antibacterial and antiviral effects and the effect of shortening the blood coagulation and bleeding time, and the variety of the effective components is wide and the content is high.
The bee therapy product has the following effects:
for use in the treatment of bacterial infections caused by bacteria in the following group: (1) gram-positive bacteria: staphylococcus species: staphylococcus aureus, methicillin-resistant staphylococcus aureus (MRSA), methicillin-sensitive staphylococcus aureus (MSSA), staphylococcus intermedius, staphylococcus pseudointermedius, coagulase-negative staphylococcus: staphylococcus epidermidis, staphylococcus saprophyticus, staphylococcus suis; streptococcus genus: streptococcus uberis, Streptococcus bovis, Streptococcus dysgalactiae, Streptococcus agalactiae, Streptococcus canis, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus oralis, Streptococcus thermophilus; the genus Streptococcus; corynebacterium: corynebacterium bovis; steinernema pyogenes; nocardia; b, bacillus subtilis; bacillus cereus;
enterococcus: enterococcus faecium, enterococcus faecalis; vancomycin-resistant enterococci (enterococcus casseliflavus); and, (ii) gram-negative bacteria: e.coli; acinetobacter baumannii; pseudomonas aeruginosa; haemophilus influenzae; salmonella choleraesuis; yersinia enterocolitica; enterobacter: enterobacter cloacae; klebsiella bacteria: klebsiella pneumoniae, klebsiella oxytoca; shigella flexneri; burkholderia cepacia; proteus mirabilis; proteus vulgaris; actinobacillus actinomycetemcomitans; actinomycetes in Israel; bacteroides fragilis; helicobacter pylori; e.coli; campylobacter jejuni; porphyromonas gulila;
porphyromonas salivarius; porphyromonas dentata; prevotella intermedia; treponema; a Paenibacillus sp;
treating a fungal infection caused by a fungus selected from the group consisting of: candida species: candida albicans, Candida ducreyi, Candida glabrata, Candida kruensis, Candida tropicalis, Candida paratyphi; the genus Aspergillus: aspergillus niger, aspergillus versicolor; penicillium pinophilum; a variant paecilomyces; trichoderma viride; chaetomium globosum; marylaria pachydermatis.
Treating mastitis in an animal.
Treating functional gastrointestinal inflammation, said functional gastrointestinal disorder being selected from: oesophagus, stomach, duodenum, small intestine and colon inflammation, anorectal inflammation, childhood and adolescent specific gastrointestinal inflammation.
In the description herein, the description of the terms "one embodiment," "some embodiments," "specific embodiments," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the claims and their equivalents.
Claims (8)
1. The preparation method of the bee therapy product with the functions of sterilization and inflammation diminishing is characterized by comprising the following steps:
s1: removing impurities from propolis raw materials, removing dust by blowing, freezing for 8-12 hours at-10 to-20 ℃, then crushing, and sieving by a sieve of 80-100 meshes to obtain fine powder for later use;
s2: adding a first extracting agent, a second extracting agent and a third extracting agent into the fine powder according to the proportion of an extracting solvent (W/V, 1-22) for solid-liquid extraction, collecting the part of the third extracting agent to obtain a propolis extract, separating the propolis extract, remaining the first extract and the second extract, carrying out ultrasonic treatment for 30-60 min at the temperature of 15-35 ℃, then filtering to obtain filter residue and filtrate, concentrating the filtrate under reduced pressure to obtain a propolis refined extract, dispersing the propolis extract into an ethanol water solution with the weight percentage concentration of 60-95% of ethanol, and carrying out ultrasonic treatment;
s3: adding an ethanol aqueous solution into the filter residue obtained in the previous step according to the W/V ratio of the filter residue to a 75% ethanol aqueous solution of 1 (4-8), extracting for 1-3 times at 65-75 ℃ for 1-3 h each time, and filtering the extracting solution to obtain a filtrate;
s4: and extracting the filtrate by using ultrahigh pressure extraction equipment, wherein the extraction pressure is 500-600 Mpa, the extraction time is 10-15 min, and the extract is concentrated and dried to obtain the compound.
2. The method for preparing bee-therapy product with antibacterial and antiinflammatory effects as claimed in claim 1, wherein silica gel adsorbent and the first extractant are mixed at a volume ratio of 1:2-8, and are stirred and extracted for 1-3 times, each time for 1-3 hours, and then filtered, and the filter residue is naturally volatilized to obtain the first extract; mixing the first extract and the second extractant according to a volume ratio of 1:2-8, stirring and extracting for 1-3 times, each time for 1-3 hours, filtering, and naturally volatilizing filter residues to obtain a second extract; mixing the second extract and the third extractant according to a volume ratio of 1:2-8, stirring and extracting for 1-3 times, each time for 1-3 hours, and combining the third extracts after stirring and extracting.
3. The method for preparing bee-therapy product with antibacterial and antiinflammatory effects as claimed in claim 1, wherein the first extractant is petroleum ether, the second extractant is mixed solvent of acetone and petroleum ether at volume ratio of less than 2:1, and the third extractant is mixed solvent of acetone and petroleum ether at volume ratio of (2-4: 1).
4. The method for preparing bee-therapy product with antibacterial and anti-inflammatory effects as claimed in claim 1, wherein in step S2, the extract is subjected to antibiotic removal treatment by using macroporous adsorbent resin.
5. The method for preparing bee-therapy product with antibacterial and antiinflammatory effects as claimed in claim 1, wherein the first, second and third extractants are added, extracted at 45 deg.C and 150rpmn for 2h, left at 4 deg.C overnight, and collected by suction filtration.
6. The method for preparing bee-therapy product with antibacterial and antiinflammatory effects as claimed in claim 2, wherein the silica gel adsorbent is of silica gel adsorbent powder structure, and has particle diameter of 0.2-0.5 mm.
7. A bee therapy product with antibacterial and anti-inflammatory effects, which is the extract of bee therapy product prepared according to any one of claims 1-6, wherein the active ingredients comprise chrysin, pinocembrin, galangin, caffeic acid phenethyl ester, rhamnonin and benzoic acid.
8. The bee therapy product with antibacterial and anti-inflammatory effects as claimed in claim 7, wherein the propolis extract comprises 40-53 wt% of chrysin, 11-15 wt% of pinocembrin, 7-10 wt% of galangin, 8-112 wt% of caffeic acid phenethyl ester, 9-14 wt% of rhamnosin and 7-12 wt% of benzoic acid.
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Citations (7)
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| CN1465569A (en) * | 2002-06-25 | 2004-01-07 | 杨汝伟 | Method for raising total flavone content in propolis |
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| CN104557834A (en) * | 2015-01-21 | 2015-04-29 | 聊城大学 | Method for separating and purifying pinocembrin, chrysin and galangin from Chinese propolis aqueous extract |
| CN106892889A (en) * | 2017-02-07 | 2017-06-27 | 浙江大学 | A kind of Chinese propolis flavone extract and preparation method and application |
| CN110638837A (en) * | 2019-10-31 | 2020-01-03 | 广东国源国药制药有限公司 | Propolis extract and preparation method and application thereof |
| CN113750120A (en) * | 2021-10-11 | 2021-12-07 | 北京德蜂堂健康科技有限公司 | Propolis extract and production process and application thereof |
-
2022
- 2022-01-24 CN CN202210076365.7A patent/CN114224920A/en active Pending
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| CN1465569A (en) * | 2002-06-25 | 2004-01-07 | 杨汝伟 | Method for raising total flavone content in propolis |
| US20130129808A1 (en) * | 2011-11-23 | 2013-05-23 | Korea Institute Of Ceramic Engineering And Technology | Medical product containing active component of propolis or grapefruit seed extract and manufacturing method thereof |
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| CN110638837A (en) * | 2019-10-31 | 2020-01-03 | 广东国源国药制药有限公司 | Propolis extract and preparation method and application thereof |
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