阅读说明：本技术 一种复合酶解法提取蜂胶总黄酮装置及提取方法 (Device and method for extracting propolis total flavonoids by complex enzymolysis ) 是由 仲延龙 邵兴军 朱慧芬 沈井旺 周娟 马明明 毛日文 于 2021-08-24 设计创作，主要内容包括：本发明公开了一种复合酶解法提取蜂胶总黄酮装置及提取方法,包括机壳,机壳内部设置有下料管,下料管上方设置有真空干燥室,下料管下方设置有粉碎室,真空干燥室包括格网、插板阀、真空气泵和加热器,插板阀水平安装在机壳侧部,插板阀设置在下料管上方,格网设置在插板阀插板的上方,格网固定安装在机壳内部,真空气泵安装在机壳侧部,加热器安装在机壳侧部,粉碎室包括粉碎辊和盛料盒,粉碎辊成对设置,粉碎辊设置在下料管出料口的正下方,粉碎辊转动安装在机壳内壁上,盛料盒活动安装在机壳底部。本发明中通过设置格网,在混合液干燥时先一步处理成若干个小块,相较于整个大块物体直接破碎,小方块更方便上料和粉碎,提高了粉碎的效率。(The invention discloses a device and a method for extracting total flavonoids from propolis by a complex enzymatic hydrolysis method, and the device comprises a machine shell, wherein a blanking pipe is arranged in the machine shell, a vacuum drying chamber is arranged above the blanking pipe, a crushing chamber is arranged below the blanking pipe, the vacuum drying chamber comprises a grid, a gate valve, a vacuum air pump and a heater, the gate valve is horizontally arranged on the side part of the machine shell, the gate valve is arranged above the blanking pipe, the grid is arranged above a gate of the gate valve, the grid is fixedly arranged in the machine shell, the vacuum air pump is arranged on the side part of the machine shell, the heater is arranged on the side part of the machine shell, the crushing chamber comprises a crushing roller and a material containing box, the crushing rollers are arranged in pairs, the crushing rollers are arranged under a discharge port of the blanking pipe, the crushing rollers are rotatably arranged on the inner wall of the machine shell, and the material containing box is movably arranged at the bottom of the machine shell. According to the invention, the grid is arranged, the mixed liquid is firstly processed into a plurality of small blocks in one step when being dried, compared with the situation that the whole large object is directly crushed, the small blocks are more convenient to feed and crush, and the crushing efficiency is improved.)

1. The device for extracting propolis total flavonoids by the complex enzymolysis comprises a shell (100), and is characterized in that: a discharging pipe (130) is arranged in the machine shell (100), a vacuum drying chamber (200) is arranged above the discharging pipe (130), and a crushing chamber (300) is arranged below the discharging pipe (130);

the vacuum drying chamber (200) comprises a grid (210), a gate valve (220), a vacuum air pump (230) and a heater (240), wherein the gate valve (220) is horizontally arranged on the side part of the shell (100), the gate valve (220) is arranged above the blanking pipe (130), the grid (210) is arranged above a gate of the gate valve (220), the grid is fixedly arranged in the shell (100), the vacuum air pump (230) is arranged on the side part of the shell (100), and the heater (240) is arranged on the side part of the shell (100);

the crushing chamber (300) comprises crushing rollers (310) and a material containing box (320), the crushing rollers (310) are arranged in pairs, the crushing rollers (310) are arranged under a discharge hole of the discharging pipe (130), the crushing rollers (310) are rotatably installed on the inner wall of the machine shell (100), and the material containing box (320) is movably installed at the bottom of the machine shell (100).

2. The device for extracting total flavonoids in propolis by complex enzymolysis as claimed in claim 1, which is characterized in that: cover (110) are installed at casing (100) top, cover (110) downside is provided with the sealing washer, the sealing washer with casing (100) lateral wall laminating.

3. The device for extracting total flavonoids in propolis by complex enzymolysis as claimed in claim 1, which is characterized in that: the connection part of the blanking pipe (130) and the inner wall of the shell (100) is provided with a slope (140).

4. The method for extracting the total flavonoids in the propolis by the complex enzymolysis is characterized by comprising the following specific steps of:

s1, selecting high-quality wool glue as a raw material, removing impurities by an impurity removing machine, freezing, crushing, and then homogenizing and mixing a certain amount of an enzymolysis agent;

s2, filling the mixed raw materials on a biological enzymolysis bed, and carrying out biological enzymolysis by adjusting parameters such as enzymolysis temperature, humidity, pH, time and the like;

s3, extracting flavonoid compounds in the propolis in a grading manner in an extraction tank by controlling factors such as alcoholic strength, material-liquid ratio, temperature, pressure, time and the like;

s4, controlling the concentration temperature and the vacuum degree to perform reduced pressure distillation to remove impurities and water;

s5, purifying the propolis total flavonoids from the extract by adopting an adsorption chromatography-gradient elution process.

5. The method for extracting total flavonoids from propolis by complex enzymolysis, as claimed in claim 4, wherein the removing impurities and purifying in S2 comprises: after the propolis virgin rubber is qualified, crushing the propolis virgin rubber by using an impurity removing machine to 8-10 meshes, dewaxing the propolis virgin rubber by using petroleum ether, leaching the propolis virgin rubber twice by using 75-85% ethanol at room temperature, wherein each time lasts for 47-49 hours, and the ratio of the ethanol added to the propolis, namely the liquid to the material, is 6: 1.

6. the method for extracting propolis total flavonoids by complex enzymatic hydrolysis as claimed in claim 4, wherein silica gel G for thin layer chromatography is selected as a chromatographic adsorbent in S5, and petroleum ether-ethyl acetate is selected as a gradient eluent.

7. The method for extracting propolis total flavonoids by complex enzymatic hydrolysis as claimed in claim 4, wherein the polarity change sequence of gradient elution in S5 is from small to large, and the volume ratio of petroleum ether to ethyl acetate is from 10: 0. 10: 1. 10: 2 …, gradually increasing the polarity of the eluent until the next 10: 10, stopping, and collecting the eluent for later use.

8. The method for extracting propolis total flavonoids by complex enzymatic hydrolysis as claimed in claim 4, wherein the method for purifying total flavonoids in S5 comprises separating out crystals from the evaporation concentration eluent at room temperature by using a vacuum centrifuge, collecting the crystals, repeating the crystallization for multiple times, combining the crystals, collecting and storing the crystals respectively, analyzing a small amount of the crystals by high performance liquid chromatography, and combining the crystals with similar components and high flavonoid content to obtain the prepared propolis total flavonoids.

Technical Field

The invention relates to the technical field of extracting total flavonoids in propolis, in particular to a device and a method for extracting total flavonoids in propolis by a complex enzymolysis method.

Background

Modern pharmacological research shows that propolis has wide pharmacological effects of resisting bacterial infection, resisting virus, resisting tumor, resisting oxidation lipid, reducing blood sugar, lowering blood pressure, enhancing immunity, promoting tissue regeneration and the like. Researches prove that flavonoid compounds and mushroom alkene substances in the propolis are the main substance basis of the propolis playing a wide range of pharmacological effects. The main flavones in propolis include flavanone, flavonols and dihydroflavonoids. The flavonoid compound is one of essential nutrients for human body, can not be synthesized by human body, and can only be ingested from food, and the current intake level of the flavonoid compound in the human body is far from reaching the beneficial intake. The flavonoids have effects in increasing blood flow of coronary artery and cerebral vessels, reducing myocardial oxygen consumption, resisting arrhythmia, softening blood vessel, lowering blood sugar, reducing blood lipid, reducing blood viscosity, resisting oxidation, scavenging free radicals, and resisting aging.

Because the artificially synthesized flavonoid compounds have the defects of large side effect, high price and the like, and the flavonoid compounds extracted from natural substances have better pharmacological action and small toxic and side effect, people gradually turn attention to obtaining the flavonoid compounds from plants, the variety and the quantity of the flavonoid compounds contained in the propolis are beyond the reach of other natural plants, various structures such as quercetin, chrysin, rutin and the like are separated and identified, and the extraction and the separation of the flavonoid compounds from the propolis and the research on the new pharmacological action of the flavonoid compounds are always valued by people.

The traditional method for extracting flavonoid compounds from propolis mainly comprises solution or solvent extraction according to the principle of similarity and compatibility, and the methods comprise hot water extraction, alkali liquor extraction, acetone extraction, alcohol extraction, enzymolysis and the like. The hot water extraction method is mainly suitable for extracting flavone shake, but the polarity of water is large, so that components such as protein, saccharides and the like are easy to extract, the extracting solution is easy to decay and deteriorate, and the subsequent separation is difficult;

the alkaline solution extraction method comprises the steps of utilizing a multi-light-base compound taking phenyl chromone as a parent nucleus to open a chalcone type structure between carbons of the multi-light-base compound under an alkaline condition, dissolving the opening into water for extraction, but heating under a strong alkaline condition can destroy the parent nucleus of the flavonoid compound;

the acetone extraction method has certain advantages when being used for extracting flavonoid substances with dominant fat-soluble groups. The ethanol extraction method utilizes the different polarity of flavonoid compounds and the mixed impurities, selects ethanol for extraction, can extract the flavonoid compounds, and can achieve the purpose of separating the ligand and the aglucon or polar ligand and nonpolar ligand, and the ethanol extraction method is divided into an alcohol immersion method, a Soxhlet extraction method, a reflux hot extraction method and an alcohol percolation method, and has the defects of overlong extraction time, large solvent consumption, troublesome operation and the like of the percolation method;

supercritical fluid extraction is a novel physical separation and purification technology of substances, and is widely applied to the extraction and separation of various effective components of traditional Chinese medicine volatile oil, alkaloid, flavonoid and the like by utilizing the characteristics of high diffusion coefficient, good flowing and transferring performances and strong dissolving capacity of the supercritical fluid through methods of regulating pressure and temperature, adding a proper entrainer and the like. The separation of biomacromolecule, heat-sensitive and chemical unstable substances can be solved, but the application of the supercritical fluid extraction technology to the extraction of propolis flavone is questionable.

Therefore, a new device and a new method for extracting the propolis total flavonoids by the complex enzymolysis method are needed.

Disclosure of Invention

The invention aims to provide a device and a method for extracting total flavonoids in propolis by a complex enzymatic hydrolysis method, which aim to solve the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme:

on one hand, the invention provides a device for extracting total flavonoids from propolis by a complex enzymatic hydrolysis method, which comprises a shell, wherein a blanking pipe is arranged in the shell, a vacuum drying chamber is arranged above the blanking pipe, and a crushing chamber is arranged below the blanking pipe;

the vacuum drying chamber comprises a grid, a gate valve, a vacuum air pump and a heater, wherein the gate valve is horizontally arranged on the side part of the shell, the gate valve is arranged above the blanking pipe, the grid is arranged above a gate of the gate valve, the grid is fixedly arranged in the shell, the vacuum air pump is arranged on the side part of the shell, and the heater is arranged on the side part of the shell;

the crushing chamber comprises crushing rollers and material containing boxes, the crushing rollers are arranged in pairs, the crushing rollers are arranged under the discharge port of the discharge pipe, the crushing rollers are rotatably arranged on the inner wall of the machine shell, and the material containing boxes are movably arranged at the bottom of the machine shell.

In one embodiment of the invention, a cover is installed on the top of the casing, and a sealing ring is arranged on the lower side of the cover and is attached to the side wall of the casing.

In one embodiment of the invention, the connection part of the blanking pipe and the inner wall of the shell is provided with a slope.

In a second aspect, the invention provides a method for extracting propolis total flavonoids by a complex enzymolysis method, which specifically comprises the following steps:

s1, selecting high-quality wool glue as a raw material, removing impurities by an impurity removing machine, freezing, crushing, and then homogenizing and mixing a certain amount of an enzymolysis agent;

s2, filling the mixed raw materials on a biological enzymolysis bed, and carrying out biological enzymolysis by adjusting parameters such as enzymolysis temperature, humidity, pH, time and the like;

s3, extracting flavonoid compounds in the propolis in a grading manner in an extraction tank by controlling factors such as alcoholic strength, material-liquid ratio, temperature, pressure, time and the like;

s4, controlling the concentration temperature and the vacuum degree to perform reduced pressure distillation to remove impurities and water;

s5, purifying the propolis total flavonoids from the extract by adopting an adsorption chromatography-gradient elution process.

In an embodiment of the present invention, the removing and purifying in S2 specifically includes: the propolis virgin rubber is inspected to be qualified, crushed into 8-10 meshes, dewaxed by petroleum ether, extracted twice by using 75-85% ethanol at room temperature, each time lasts for 47-49 hours, and the ratio of the ethanol added to the propolis in each time is 6: 1.

in one embodiment of the present invention, silica gel G for thin layer chromatography is selected as the chromatographic adsorbent in S5, and petroleum ether-ethyl acetate is selected as the gradient eluent.

In one embodiment of the present invention, the polarity of the gradient elution in S5 is changed from small to large in order, and the volume ratio of petroleum ether to ethyl acetate is from 10: 0. 10: 1. 10: 2 …, gradually increasing the polarity of the eluent until the next 10: 10, stopping, and collecting the eluent for later use.

In an embodiment of the present invention, the method for purifying total flavonoids in S5 includes separating out crystals from the evaporation-concentration eluent at room temperature by using a vacuum centrifuge, collecting the crystals, repeating the crystallization for a plurality of times, combining the crystals, collecting and storing the crystals respectively, analyzing a small amount of the crystals by using high performance liquid chromatography, and combining the crystals with similar components and high flavonoid content to obtain the prepared propolis total flavonoids.

In summary, due to the adoption of the technology, the invention has the beneficial effects that:

in the invention, the grid is arranged in the vacuum drying chamber, so that the mixed liquid is solidified in blocks during drying, and compared with the solidification of a large block, the small block is more convenient for subsequent crushing;

the crushing roller is arranged, so that the small mixture is conveniently crushed into powder;

in the method for extracting the total flavonoids, the total flavonoids of the propolis are extracted by adopting an alcohol extraction and ester extraction method, and the propolis flavonoid glycoside is subjected to enzymolysis by a complex enzyme system to improve the content of flavonoid aglycone such as myricetin, quercetin, apigenin, pinocembrin and the like in the propolis extract, improve the total flavonoids of the propolis, enhance the antioxidant activity of the propolis extract and better apply the propolis to the fields of medicines, health-care foods, cosmetics and the like.

Drawings

FIG. 1 is a schematic sectional front view of the present invention;

FIG. 2 is a schematic top view of a grid according to the present invention;

FIG. 3 is a flow chart of an apparatus used in the method of the present invention.

In the figure: 100. a housing; 110. a machine cover; 130. a discharging pipe; 140. a slope; 200. a vacuum drying chamber; 210. grid forming; 220. a gate valve; 230. a vacuum air pump; 240. a heater; 300. a crushing chamber; 310. a crushing roller; 320. a material containing box.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

In the description of the present invention, it is to be understood that the terms indicating an orientation or positional relationship are based on the orientation or positional relationship shown in the drawings only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art from the specification.

Example 1

Referring to fig. 1-3, the present invention provides a device for extracting total flavonoids from propolis by a complex enzymatic hydrolysis method, including a casing 100, a feeding pipe 130 is disposed inside the casing 100, a vacuum drying chamber 200 is disposed above the feeding pipe 130, a pulverizing chamber 300 is disposed below the feeding pipe 130, a cover 110 is mounted on the top of the casing 100, a sealing ring is disposed on the lower side of the cover 110, and the sealing ring is attached to the side wall of the casing 100, such that when the cover 110 is closed, the casing 100 forms a closed space.

Meanwhile, in order to facilitate the mixture to slide down, a slope 140 is provided at the connection of the blanking pipe 130 and the inner wall of the casing 100.

The vacuum drying chamber 200 comprises a grid 210, a gate valve 220, a vacuum air pump 230 and a heater 240, wherein the gate valve 220 is horizontally arranged at the side part of the casing 100, the gate valve 220 is arranged above the blanking pipe 130, as shown in fig. 2, the grid 210 is arranged above a gate of the gate valve 220, the grid is fixedly arranged inside the casing 100, the vacuum air pump 230 is arranged at the side part of the casing 100, and the heater 240 is arranged at the side part of the casing 100.

In a specific configuration, the heater 240 may be a heating wire or a hot air blower, which heats the air in the casing 110 through heat conduction, and further dries the mixed liquid.

When the mixed liquid is poured into the grid 210, the grid 210 and the gate of the gate valve 220 form a plurality of independent small spaces, the mixed liquid is solidified into a plurality of small blocks by heating of the heater 240, at this time, the gate of the gate valve 220 is moved to be opened, and the small blocks fall into the crushing chamber 300. Compared with the method that the small blocks are solidified into a large block, the small blocks are more convenient to crush, and the crushing efficiency is improved.

Crushing room 300 includes crushing roller 310 and flourishing magazine 320, and crushing roller 310 sets up in pairs, and crushing roller 310 sets up under the discharge gate of unloading pipe 130, and crushing roller 310 rotates and installs on casing 100 inner wall, and the rotation motor is still installed to casing 100 lateral wall, and the output that rotates the motor passes through gear connection with crushing roller 310 for two crushing rollers 310 are inwards rotated simultaneously, and then carry out the extrusion breakage to the small square.

In order to facilitate receiving materials, the material containing box 320 is movably installed at the bottom of the casing 100, the material containing box 320 is slidably connected with the casing 100 through a sliding rail, when the mixed block is crushed into powder, the material containing box 320 is drawn out, and subsequent operations of extracting the propolis total flavonoids are facilitated.

The working principle is as follows: during the use, pour into graticule mesh 210 with the propolis extract who has mixed chromatography adsorbent and gradient eluant, graticule mesh 210 and the little space that push-pull valve 220 picture peg formed separate into a plurality of portions with mixed liquid, through the vacuum drying of vacuum air pump 230 and heater 240, mixed liquid becomes the mixture, and the picture peg of push-pull valve 220 removes to open this moment, and the mixture that forms the dice drops whereabouts, rotates the motor and drives crushing roller 310 and rotate, extrudes the breakage to the mixture for the mixture is smashed into the powder, and the powder drops to flourishing magazine 320 in. This device is through setting up graticule mesh 210, and one step earlier is handled into a plurality of fritter when mixing the liquid drying, compares in whole bold object, and the fritter more makes things convenient for the material loading and smashes, has improved kibbling efficiency.

It should be noted that: the model specifications of the vacuum air pump 230 and the heater 240 need to be determined by model selection according to the actual specification of the device, and the specific model selection calculation method adopts the prior art, so detailed description is omitted.

The power supply of the vacuum pump 230 and the heater 240 and the principle thereof will be apparent to those skilled in the art and will not be described in detail herein.

A method for extracting propolis total flavonoids by a complex enzymolysis method comprises the following specific steps:

s1, selecting high-quality wool glue as a raw material, removing impurities by an impurity removing machine, freezing, crushing, and then homogenizing and mixing a certain amount of an enzymolysis agent;

s2, filling the mixed raw materials on a biological enzymolysis bed, and carrying out biological enzymolysis by adjusting parameters such as enzymolysis temperature, humidity, pH, time and the like;

s3, extracting flavonoid compounds in the propolis in a grading manner in an extraction tank by controlling factors such as alcoholic strength, material-liquid ratio, temperature, pressure, time and the like;

s4, controlling the concentration temperature and the vacuum degree to perform reduced pressure distillation to remove impurities and water;

s5, purifying the propolis total flavonoids from the extract by adopting an adsorption chromatography-gradient elution process.

The bioavailability of flavonoid glycoside in animal bodies is far lower than that of aglycone flavone, the utilization rate of flavonoid glycoside substances in human bodies is very low, and the antioxidant activity is not ensured. Therefore, improving the configuration of the flavone and increasing the absorption rate of the flavone in blood is an important way for improving the bioavailability of flavonoid products. At present, most of flavone glycosides are hydrolyzed by a chemical method, and the characteristics of the flavone are changed, such as fat solubility improvement, through chemical derivatization reactions of etherification, esterification, acylation and the like of the flavone, but the reaction processes usually shield the main functional group, namely phenolic hydroxyl group, of the flavonoid compound, so that adverse effects are generated on the antioxidation, and environmental pollution is easily generated in the processing. The propolis flavone molecules are modified by a biological enzymolysis method to be converted into aglycone type flavone, so that the biological value of the propolis flavone can be greatly improved.

The propolis raw gum is inspected to be qualified, crushed into 8-10 meshes, dewaxed by petroleum ether, and extracted twice by 80% ethanol at room temperature, wherein each time is 48 hours, and the ratio of ethanol added to propolis, namely liquid material, is 6: 1, combining the two leaching solutions, carrying out suction filtration, concentrating the filtrate in vacuum to obtain a propolis ethanol extract, extracting the propolis ethanol extract with ethyl acetate for 3 times, and transferring most of flavone substances into the ethyl acetate. Vacuum concentrating under negative pressure, and recovering ethyl acetate to obtain black brown substance as propolis extract, i.e. propolis ethanol extract.

Silica gel G for thin layer chromatography is selected as a chromatographic adsorbent, and petroleum ether-ethyl acetate is selected as a gradient eluent. Accurately weighing 100G of propolis extract each time, placing the propolis extract in a 500ml beaker, adding 200ml of ethyl acetate to fully dissolve the propolis extract, mixing 100G of silica gel G for thin layer chromatography, uniformly stirring, fully soaking and dissolving the silica gel, using the vacuum drying chamber 200 of the invention to adjust the temperature to 45 ℃ for vacuum drying, using the crushing chamber 300 to crush the mixture into powder, and using 500ml of petroleum ether to dissolve the crushed mixture for later use.

Adding 1500ml petroleum ether into 400G of silica gel G for thin layer chromatography, stirring into homogenate, soaking for 4-6 hours, and fully dissolving.

The bottom of the glass chromatographic column is plugged with medical absorbent cotton, 400ml of petroleum ether is added, a piston below the column is opened, the silica gel G dissolved by the petroleum ether is shaken and homogenized, and the silica gel G homogenized slurry is directly and slowly poured into the glass chromatographic column by a funnel. After the natural sedimentation is finished, the silica gel column bed is compacted by increasing the amount of petroleum ether until the flow rate is constant.

And (3) adopting a wet column loading method, and slowly adding the dissolved and standby propolis extract silica gel G mixture into the glass chromatographic column to avoid damaging the flatness of the upper silica gel surface of the silica gel bed in the chromatographic column. After the sample is loaded, 500ml of petroleum ether is added, and a mass of degreased cotton is plugged to be close to the surface of the sample silica gel after the sample naturally settles in the chromatographic column so as to avoid adding eluent to destroy the flatness of the surface of the sample silica gel during column filling. Gradient elution is carried out by adopting petroleum ether-ethyl acetate solution, the polarity change sequence of the gradient elution is from small to large, and the volume ratio of the petroleum ether to the ethyl acetate is from 10: 0. 10: 1. 10: 2 …, gradually increasing the polarity of the eluent until the next 10: 10, stopping, and collecting the eluent for later use.

The color of the former eluent is close to that of the solvent, the eluent is concentrated by vacuum negative pressure rotary evaporation, and a small amount of milky white paraffin-like substances obtained by recovering the solvent are discarded. Collecting 500ml eluate each time, rotary evaporating under vacuum negative pressure (negative pressure of 0.03-0.05 MPa, water temperature of 45 deg.C) to concentrate eluate, recovering solvent, ultrasonically dissolving the rest with small amount of heated ethyl acetate to form supersaturated solution, precipitating crystals at room temperature, collecting crystals, repeatedly crystallizing, mixing crystals, collecting and storing respectively. And (3) analyzing a small amount of crystallized substances by adopting high performance liquid chromatography, and combining the substances with similar components and high flavone content to obtain the prepared propolis total flavone.

The content of flavonoid aglycone such as myricetin, quercetin, apigenin, pinocembrin and the like in the propolis extract prepared by the method is improved, the antioxidant activity of the propolis extract is enhanced, and the propolis can be better applied to the fields of medicines, health-care foods, cosmetics and the like.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.