CN108685951A - A kind of stingless bee Mel extract and its extracting method and application - Google Patents
A kind of stingless bee Mel extract and its extracting method and application Download PDFInfo
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- CN108685951A CN108685951A CN201810750208.3A CN201810750208A CN108685951A CN 108685951 A CN108685951 A CN 108685951A CN 201810750208 A CN201810750208 A CN 201810750208A CN 108685951 A CN108685951 A CN 108685951A
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- stingless bee
- honey
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- bee
- mel extract
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of extracting method of stingless bee Mel extract and its applications;The method includes:1) stingless bee honey is dissolved in the aqueous solution that pH value is 1-3, forms honey solution;2) macroreticular resin after activation is added in honey solution, the macroreticular resin of adsorption of effective component is obtained after stirring;3) macroreticular resin that step 2) obtains is loaded into elution column and is eluted, eluted and collect eluent;4) eluent low temperature is concentrated, redissolved in water, extraction, collected organic layer to obtain the final product.The present invention proposes a kind of more excellent stingless bee Mel extract, the active ingredient group of the extract has good synergistic effect, and have been surprisingly found that the stingless bee Mel extract for colitis especially ulcerative colitis, especially the disease pathogenic because of the acute ulcer colitis of dextran sulfate induction has especially pronounced application activity, equally has broad application prospects for the colitis treatment caused by other many reasons.
Description
Technical field
The present invention relates to biochemical fields, and in particular to stingless bee honey, which is used to prepare, treats or prevents colitis
Application in drug, food, health products.
Background technology
The close object for having connected and composed intestinal mucosa between the intestinal epithelial cell and intestinal epithelial cell of enteron aisle chamber side
Barrier function is managed, Gut barrie r can effectively prevent the harmful substances such as intestinal bacterium and endotoxin from entering blood across intestinal mucosa to follow
Ring.Wherein, close connection to maintain the integrity and stability of enteron aisle mechanical barrier be it is particularly important, by occludin,
The peripheries such as claudins and connection 3 kinds of memebrane proteins of adhesion molecule and closure small cyclase protein (ZO-1, ZO-2, ZO-3) packet slurry albumen
Composition.Close-connected destruction and dysfunction can lead to the increase of cell permeability, make the noxious materials such as bacterium and toxin
Easily into intestinal mucosa lamina propria and immune cell activated, cause enterogastric diseases and systemic disease.For example, inflammatory
Enteropathy, acute pancreatitis, alcohol induced liver injury etc..In addition, enteron aisle oxidative stress is also cause gut barrier injury important
One of mechanism.Not only only same-action is combined the excessive active oxygen generated during oxidative stress with the sulfydryl of enteric epithelium mucous membrane
Cause protein denaturation and enzyme to inactivate, and can with the polyunsaturated fatty acid in cell membrane in conjunction with and lead to lipid peroxidation,
It is further exacerbated by intestinal cell damage.Some current results of study have shown that MAPK signal paths can regulate and control close connection
Protein expression and close-connected permeability play weight in terms of the integrality of cell tight junction and gut barrier function
It acts on.Have studies have shown that the destruction of gut barrier plays an important roll in the pathogenesis of Bowel inflammatory disease, therefore
Improving gut barrier can be as a kind of new method of prevention and treatment intestines problem.
Stingless bee (Stingless bees) is a kind of insect sought group life and can made honey, and is under the jurisdiction of Hymenoptera
(Hymenoptera), Apoidea (Apoidea), Apidae (Apidae), honeybee subfamily (Apinae), Mai Feng races
(MeliPonini).Main stingless bee honey collection belongs to stingless bee in Melipona, Scaptotrigona belong to stingless bee and
Trigona belongs to stingless bee.Stingless bee body is in black, and small body is flexibly the main pollination honeybee kind of torrid areas plant, main point
Subtropical and tropical zones are distributed in, China is only distributed in the field forest of Yunnan, Hainan and Taiwan.Stingless bee honey
It is that the nectar that stingless bee is adopted from plant nectary or other positions is stored in the happy substance of lair, with west through fully making
Square honeybee honey is compared, and has the characteristics that moisture is high, acidity is high, enzyme values are low, viscosity is low.Honey is already by people's conduct
A kind of natural sweetener is also widely used for clinical practice due to its existing active constituent.Having been reported proves bee
Honey has multiple biological activities, and the extraction and Separation Research progress to the active constituent of stingless bee honey is also fewer.
The stingless bee honeybee kind that China has found has yellow line stingless bee (Trigona ventralis Smith), top stingless bee
(T.terminate Smith), dark wing stingless bee (T.vidua Lepeletier), nectarine stingless bee (T.lutea
Bingham), black chest stingless bee (T.pagdeni Sehwarz), the sufficient stingless bee (T.smithii Smith) of palm fibre, brown chest stingless bee
(T.thoracica Smith), black leg stingless bee (T.vidua Smith), black abdomen stingless bee (T.canifrons Smith),
Yellow instep stingless bee (T.iridipennis Smith).But the related protection intestinal health of no related China stingless bee honey is active
Research report.
Invention content
A kind of stingless bee Mel extract of present invention offer and its extracting method, while above-mentioned stingless bee honey being provided and is existed
Prepare the application in the drug, food and health products for inhibiting enteritis class.
The present invention adopts the following technical scheme that:
A kind of extracting method of stingless bee honey, includes the following steps:
1) stingless bee honey is dissolved in the solution that pH value is 1-3, forms honey solution;
2) macroreticular resin after activation is added in the honey solution, the big of adsorption of effective component is obtained after stirring
Hole resin;
3) by the macroreticular resin that step 2) obtains be loaded into elution column in, the hydrochloric acid for being first 1.0-3.0 with pH value it is water-soluble
Liquid elutes, then elutes to neutral (preferably with water wash), is finally eluted with methanol and collects eluent;
4) eluent is concentrated into after removal methanol to obtain solid content by low temperature, redissolved in water, with extracting n-butyl alcohol,
N-butanol layer is collected to get stingless bee Mel extract.
Extracting method of the present invention, in step 1), it is 3.0-4.0 that the stingless bee honey, which is pH, and amylase value is
20-40mL/ (gh), total sugar content 40-50g/100g;The acidity of stingless bee honey is significantly higher than common Apis honey.
Amylase value is to judge the important indicator of honey quality quality, and sugar content is less than apis mellifera (Apis mellifera) bee
Sweet (>60 g/100g), wherein secondly glucose content highest is fructose 20-25g/100g and sucrose for 20-30g/100g
0.1-0.5g/100g。
The stingless bee belongs to stingless bee from Melipona, Scaptotrigona belongs to stingless bee or Trigona belongs to nothing
Pierce bee.
Further, the stingless bee honey is Trigona, and the preferably described stingless bee honey is the bee of yellow line stingless bee
Honey.
And/or the nectariferous plant of the stingless bee honey includes one or more of lichee, longan, eucalyptus, it is above-mentioned
Nectariferous plant is the plant of tropical and subtropical region;It is preferred that Hainan Region.
And/or the stingless bee honey is the stingless bee that Northern Hemisphere tropical and subtropical region harvests between July in May-
Honey.
Preferably, the volume ratio of the aqueous solution and the stingless bee honey is (3-5):1;More preferable 5:1.To reach
The purpose of active material fully in dissolving honey, and this ratio Mel extract yield highest.
Preferably, to be carried out under ultrasound condition, ultrasound parameter is the dissolving operation in step 1):Ultrasonic power 40/
60KHz, time 30-60min, 40-50 DEG C of ultrasonic temperature.Extracting method of the present invention, in step 2),
Preferably, the macroreticular resin is Supelco SupeliteTMDAX-8;It is further preferred that the macroreticular resin and institute
The volume ratio for stating honey solution is 1:(1.5-3), preferably 1:1.5;This ratio Mel extract yield highest.
The concrete operations of the step 2) preferably weigh stingless bee honey 50g, and honey is put into 500mL beakers, is added
The solution for entering 250mL pH=2 obtains the aqueous solution of honey;It is preferred that mixing time 30 minutes.
Extracting method of the present invention, in step 3),
Preferably, the concrete operations of the step 3) are, by Supelco SupeliteTMDAX-8 macroreticular resins load
Into elution column, the aqueous solution of Ph 1.0-3.0 hydrochloric acid is first used to elute, then with water wash, finally eluted and collected with methanol and washed
De- liquid.
Extracting method of the present invention, in step 4),
Preferably, the low temperature concentration is that reduced vacuum concentrates, and temperature is 30-50 DEG C.
Further preferably, further include that liquid chromatography-tandem mass spectrometry is carried out to the stingless bee Mel extract that step 4) obtains
It is combined the operation of separation:Chromatographic condition:Using octadecylsilane chromatographic column, mobile phase is pure water-methanol;Flow velocity 0.2mL/
min;Sample size 2yL;30 DEG C of column temperature;Gradient is 0-2min, 5%;2-10min, 15%-30%;10-25min, 30%-
90%;25-30min, 90%;30-31min 5%;Mass Spectrometry Conditions:The sources ESI, carrier gas temperature:350 DEG C, dry gas stream speed:
11L/min, atomization air pressure:40psig.
Preferably, in step 4), the volume ratio of water phase and organic phase is 1:3-4.
The present invention also provides the preparation-obtained stingless bee Mel extracts of any of the above-described technical solution.
In the stingless bee Mel extract, chlorogenic acid equivalent is at least 90ug/g;And/or Quercetin equivalent is at least
15ug/g;Preferably, in the stingless bee Mel extract, the content of gallic acid is at least 200 μ g/100g;
And/or morin content is at least 200 μ g/100g.Under above-mentioned active component content, the stingless bee honey
Extract has more excellent application benefit.
Gained Mel extract is the solid that sepia has certain viscosity, is soluble in the organic solvents such as ethyl alcohol, insoluble
Yu Shui.
Meanwhile the present invention provides a kind of stingless bee Mel extract and is preparing drug, food and the health care for inhibiting enteritis class
Application in product.
Enteritis class of the present invention, refers mainly to colitis, wherein covers a variety of colitis, including ischemic colon
Inflammation, sigmoiditis, ulcerative colitis and pseudomembranous colitis etc..There is significantly more application for ulcerative colitis
Effect.
Colitis of the present invention particularly relates to lead to mucous membrane of colon barrier by local macrophage release lysosome
Damage,
Or, intestinal flora changes ulcerative colitis caused by the forfeiture with Intestinal mucosal barrier.
Drug of the present invention, food, health products are using the stingless bee Mel extract as single active ingredient
Or one of active constituent.
The present invention proposes a kind of more excellent stingless bee Mel extract, and the active ingredient group of the extract is with good
Good synergistic effect, and have been surprisingly found that the stingless bee Mel extract for colitis especially ulcerative colitis, especially
Being the disease pathogenic because of the acute ulcer colitis that dextran sulfate induces has especially pronounced application activity, for it
Colitis treatment caused by his many reasons equally has broad application prospects.Meanwhile finding stingless bee Mel extract
Tight junction protein ZO-1 and Occludin gene expression abundances can be dramatically increased.
Description of the drawings
Fig. 1:The acute ulcer colitis induced dextran sulfate from three kinds of stingless bee category stingless bee honey is big
The influence of mouse disease activity index.MST:Melipona belongs to stingless bee;SST:Scaptotrigona belongs to stingless bee;TST:
Trigona belongs to stingless bee.
Fig. 2:The acute ulcer colitis induced dextran sulfate from three kinds of stingless bee category stingless bee honey is big
The influence of mouse colon lengths.MST:Melipona belongs to stingless bee;SST:Scaptotrigona belongs to stingless bee;TST:Trigona
Belong to stingless bee.
Fig. 3:The acute ulcer colitis induced dextran sulfate from three kinds of stingless bee category stingless bee honey is big
The influence of mouse colon tight junction protein ZO-1 gene expression.MST:Melipona belongs to stingless bee;SST:Scaptotrigona
Belong to stingless bee;TST:Trigona belongs to stingless bee.
Fig. 4:The acute ulcer colitis induced dextran sulfate from three kinds of stingless bee category stingless bee honey is big
The influence of mouse colon Tight junction protein occludin gene expression.MST:Melipona belongs to stingless bee;SST:
Scaptotrigona belongs to stingless bee;TST:Trigona belongs to stingless bee.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Stingless bee honey described in following embodiment is the honey of yellow line stingless bee.
Embodiment 1
The present embodiment provides a kind of stingless bee Mel extract and its extracting methods, are as follows:
1) it weighs stingless bee honey 50g, is added 250ml hydrochloric acid solutions (pH 2.0), after stirring and dissolving, in 50kHz work(
Rate ultrasound 30min, cotton filtering, forms uniform honey solution;
2)Supelco SupeliteTMDAX-8 macroreticular resins weigh the macropore tree of 50g activation after ethyl alcohol impregnates for 24 hours
Fat, to pouring into stingless bee honey aqueous solution after tasteless, dynamic agitation adsorbs 1h for washing 2 times;
3) it stands, discards supernatant liquid, and macroreticular resin is inserted in glass column, use the hydrochloric acid water that the pH of 0.1L is 2 respectively
Solution, the elution of 0.15L pure water finally elute with 0.4L ethanol solutions and collect eluent to remove desaccharification isopolarity substance,
4) 40 DEG C of reduced vacuums are concentrated into solid content, redissolve in 10mL pure water, 10mL extracting n-butyl alcohols are used in combination.It collects just
Butanol layer, nitrogen drying, obtains stingless bee Mel extract.
Stingless bee Mel extract obtained by the present embodiment is measured (total in Forint phenol method measurement Mel extract
General flavone content in phenol content, aluminum nitrate colorimetric method for determining honey), it is as a result as follows:Total phenol acid content is 121.7ug/g (green originals
Acid equivalent);General flavone content is 20.6ug/g (Quercetin equivalent);Gallic acid and morin content, respectively at least 240 μ
G/100g and 225 μ g/100g.
Embodiment 2
The present embodiment provides a kind of stingless bee Mel extract and its extracting methods, are as follows:
1) it weighs stingless bee honey 50g, is added 150ml hydrochloric acid solutions ml (pH 3.0), after stirring and dissolving, in 40kHz
Power ultrasonic 60min, cotton filtering, forms uniform honey solution;
2)Supelco SupeliteTMDAX-8 macroreticular resins weigh the macropore tree of 50g activation after ethyl alcohol impregnates 48h
Fat, to pouring into stingless bee honey aqueous solution after tasteless, dynamic agitation adsorbs 2h for washing 3 times;
3) it stands, discards supernatant liquid, and macroreticular resin is inserted in glass column, use the hydrochloric acid water that the pH of 0.1L is 3 respectively
Solution, the elution of 0.15L pure water finally elute with 0.4L ethanol solutions and collect eluent to remove desaccharification isopolarity substance,
4) 60 DEG C of reduced vacuums are concentrated into solid content, redissolve in 10mL pure water, 10mL extracting n-butyl alcohols are used in combination.It collects just
Butanol layer, nitrogen drying, obtains stingless bee Mel extract.
Stingless bee Mel extract obtained by the present embodiment is measured (total in Forint phenol method measurement Mel extract
General flavone content in phenol content, aluminum nitrate colorimetric method for determining honey), it is as a result as follows:Total phenol acid content is 105ug/g (chlorogenic acids
Equivalent);General flavone content is 18ug/g (Quercetin equivalent);Gallic acid and morin content, respectively 220 μ g/100g and
218μ g/100g。
Embodiment 3
The present embodiment provides a kind of stingless bee Mel extract and its extracting methods, are as follows:
1) it weighs stingless bee honey 50g, is added 250ml hydrochloric acid solutions (pH 1.0), after stirring and dissolving, in 60kHz work(
Rate ultrasound 20min, cotton filtering, forms uniform honey solution;
2)Supelco SupeliteTMDAX-8 macroreticular resins weigh the macropore tree of 50g activation after ethyl alcohol impregnates 12h
Fat, to pouring into stingless bee honey aqueous solution after tasteless, dynamic agitation adsorbs 0.5h for washing 1 time;
3) it stands, discards supernatant liquid, and macroreticular resin is inserted in glass column, use the hydrochloric acid water that the pH of 0.1L is 1 respectively
Solution, the elution of 0.15L pure water finally elute with 0.4L ethanol solutions and collect eluent to remove desaccharification isopolarity substance,
4) 30 DEG C of reduced vacuums are concentrated into solid content, redissolve in 10mL pure water, 10mL extracting n-butyl alcohols are used in combination.It collects just
Butanol layer, nitrogen drying, obtains stingless bee Mel extract.
Stingless bee Mel extract obtained by the present embodiment is measured (total in Forint phenol method measurement Mel extract
General flavone content in phenol content, aluminum nitrate colorimetric method for determining honey), it is as a result as follows:Total phenol acid content is 95ug/g (chlorogenic acids
Equivalent);General flavone content is 15.5ug/g (Quercetin equivalent);Gallic acid and morin content, respectively at least 205 μ g/
100g and 201 μ g/100g.
Comparative example 1
A kind of acacia Mel extract of this comparative example offer and its extracting method, the extracting method are made with acacia honey
For raw material, remaining extraction step is same as embodiment 1.
Comparative example 2
This comparative example provides a kind of stingless bee Mel extract and its extracting method, the extracting method with
The mode of embodiment 1 extracts in CN107432877A.
Improve intestinal health being used to prepare below by way of test example the present invention is furture elucidated the stingless bee honey
Application effect in drug, food, health products.
Test example 1
This test example provides the determination of moisture of stingless bee honey.
Moisture is to evaluate a major criterion of honey quality, surveys three kinds of stingless bee category stingless bee honey samples
Moisture in (Melipona belongs to stingless bee honey, Scaptotrigona belongs to stingless bee honey and Trigona belongs to stingless bee honey)
Content is 25-27g/100g, due to regulation in honey national standard (GB 14963-2011):Ripe Apis (Apis categories) bee
The moisture of honey must not be higher than 24g/100g, therefore be compared to apis mellifera (Apis mellifera) honey, stingless bee
Honey has higher water content.PH value is an important factor for influencing honey shelf life, and stingless bee honey pH is 3.0-4.0, acid
Degree is significantly higher than common Apis honey.Amylase value is the important indicator for judging honey quality quality, Hainan stingless bee honey
Amylase value be 20-40 DN.The total sugar content of Hainan stingless bee honey is 40-50g/100g, is less than apis mellifera (Apis
Mellifera) honey (>60g/100g), wherein glucose content highest, is 20-30g/100g, is secondly fructose 20-25g/
100g and sucrose 0.1-0.5g/100g, lower sugar content keep stingless bee honey sweet taste thin.Protein present in honey
It is mainly derived from the enzyme of nectar, pollen and honeybee secretion.
Test example 2
This test example provides a kind of therapeutic effect of stingless bee Mel extract to ulcerative colitis.
(1) experimental animal:(45,8 week old, weight 190 ± 20 g) is purchased from Beijing dimension tonneau China to male SPF grades of SD rat
Experimental animal Technology Co., Ltd..Animal feeding is in Inst. of Traditional Chinese Medicine, Zhejiang Prov Experimental Animal Center, culture environment 12h
Illumination+12h is dark, 23 DEG C of temperature, relative humidity 50%~70%, is freely eaten and drinks water.
(2) experiment packet:Rat is randomly divided into 5 groups after the raising of one week adaptability:
Blank control group (n=5);
Model group (n=10);
Melipona belongs to stingless bee honey (MST) gavage group (n=10, daily gavage 5g/kg weight);
Scaptotrigona belongs to stingless bee honey (SST) gavage group (n=10, daily gavage 5g/kg weight);
Trigona belongs to stingless bee honey (TST) gavage group (n=10, daily gavage 5g/kg weight).
Diet Formula refers to U.S.'s AIN-93M standard recipes.
(3) experimental procedure:Experimental rat starts the induction of ulcerative colitis after 1 week experiment Diet.It removes
Blank control group is drunk outside distilled water, remaining 40 rat, which is drunk, drinks containing 3%DSS, continues 7d.It is replaced with after 7d normal
Distilled water.DSS drinking-water terminates after starting until testing, and carries out rat disease activity index (Disease Activity
Index, DAI) assessment.Daily observation 2 times, it is total according to weight loss, stool, situation of occulting blood and animal by 1 standard of table
The scoring of body situation is added, and obtains the disease activity index of every rat.
Table 1:Disease activity index
1 integral status includes:Vigor, behavior, posture etc.;
2 scorings are explained:0=is normal;1=is slight;2=moderates;3=is serious;
(4) experimental result:
As a result, it has been found that the acute ulcer knot induced dextran sulfate from three kinds of stingless bee category stingless bee honey
Attached drawing 1 is shown in the influence of enteritis rat disease activity index.As seen from the figure, with Normal group ratio, model group rats were from the 4th day
Start apparent ulcerative colitis symptom occur, including state of mind decline, diarrhea, the symptoms such as have blood in stool, DSS start after the 8th
Its DAI index peaks.Compared with DSS model groups, the processing of stingless bee honey has centainly rat disease activity index
It influences.From modeling the 7th day, stingless bee honey processing group can significantly reduce rat disease activity index.
From three kinds of stingless bee category stingless bee honey to the shadow of the DSS ulcerative colitis in rats colon lengths induced
It rings result and sees attached drawing 2.As seen from the figure, with Normal group ratio, model group rats colon lengths shorten.With DSS model group phases
Than three kinds of stingless bee category stingless bee honey processing groups can be obviously improved the colon lengths of rat.
Using fluorescence quantitative PCR detection ulcerative colitis in rats intestinal epithelial cell tight junction protein ZO-1 and
Occludin genes, as a result as shown in Figure 3,4.It can significantly inhibit ZO-1's and Occludin after DSS ulcerative colitis in rats
Gene expression.It compares and DSS damage groups, after the processing of stingless bee honey, the gene expression of ZO-1 and Occludin significantly carry
It is high.It follows that stingless bee honey can promote closely to connect the expression of ZO-1 and Occludin genes, and have to DSS damages
Certain antagonism can reduce close-connected degree of injury, have protective effect to gut barrier function.
Test example 3
The Mel extract that this test example provides embodiment 1, comparative example 1 is provided imitates the application of mouse colitis
Fruit detects.
Experimental subjects:Male ICR mouse (6-8 week old, 18-22g), the raising of 8/cage in illumination (12h day-night cycles) and
The room of (20-23 DEG C) stabilization of temperature.It is to filter without pathogen air, relative humidity 50% to supply air.Mouse is freely eaten
Drinking-water, diet are supplied as being commercialized pollution-free feed.
Experimental method:By 1 week laundering period, mouse was randomly divided into 5 groups (n=8):(1) blank control group, the group
Mouse is handled using solvent control;(2) model group, this group of mouse take the drinking-water of sulfur acid glucose, continue one week, induction
Acute ulcer colitis;(3) positive controls, this group of mouse take 2mg/kg aminosalicyclics before dextran sulfate stimulation
Acid;(4) stingless bee honey group, this group of Mouse oral 1000mg/kg stingless bees honey drink sulfur acid glucose after a week
Drinking-water;(5) acacia honey group, this group of Mouse oral 1000mg/kg acacias honey drink the drink of sulfur acid glucose after a week
Water.
The experimental results showed that acacia honey group can not alleviate mouse dextran sulfate induction acute ulcer colitis
Shape;Stingless bee honey works well, and it is similar with positive controls symptoms, the effect such as diarrhea can be effectively relieved, have blood in stool.
Test example 4
The Mel extract that this test example provides embodiment 1, comparative example 2 is provided imitates the application of mouse colitis
Fruit detects.
Experimental subjects:Male ICR mouse (6-8 week old, 18-22g), the raising of 8/cage in illumination (12h day-night cycles) and
The room of (20-23 DEG C) stabilization of temperature.It is to filter without pathogen air, relative humidity 50% to supply air.Mouse is freely eaten
Drinking-water, diet are supplied as being commercialized pollution-free feed.
Experimental method:By 1 week laundering period, mouse was randomly divided into 5 groups (n=8):(1) blank control group, the group
Mouse is handled using solvent control;(2) model group, this group of mouse take the drinking-water of sulfur acid glucose, continue one week, induction
Acute ulcer colitis;(3) positive controls, this group of mouse take 2mg/kg aminosalicyclics before dextran sulfate stimulation
Acid;(4) 1 stingless bee honey group of embodiment, the stingless bee honey extraction that this group of Mouse oral 1000mg/kg embodiment 1 obtains
Object drinks the drinking-water of sulfur acid glucose after a week;(5) control group, this group of Mouse oral 1000mg/kg comparative example 2 are provided
Mel extract drinks the drinking-water of sulfur acid glucose after a week.
The experimental results showed that:Stingless bee honey works well, and the symptoms such as diarrhea can be effectively relieved, have blood in stool, effect is the same as sun
Property control group is similar;Control group has certain alleviation diarrhea effect, but disease activity index is significantly higher than stingless bee honey group,
And it cannot effectively promote intestinal tight Connexin gene gene expression abundance.
Although above having used general explanation, specific implementation mode and experiment, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is aobvious and easy to those skilled in the art
See.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to that the present invention claims guarantors
The range of shield.
Claims (10)
1. a kind of extracting method of stingless bee Mel extract, includes the following steps:
1) stingless bee honey is dissolved in the aqueous solution that pH value is 1-3, forms honey solution;
2) macroreticular resin after activation is added in the honey solution, the macropore tree of adsorption of effective component is obtained after stirring
Fat;
3) macroreticular resin that step 2) obtains being loaded into elution column, pH value is the aqueous solution elution of the hydrochloric acid of 1.0-3.0, then
Elution is finally eluted with methanol to neutrality and collects eluent;
4) solid content is obtained after the eluent being concentrated removal methanol by low temperature, redissolves in water, with extracting n-butyl alcohol, collects just
Butanol layer is to get stingless bee Mel extract.
2. according to the method described in claim 1, it is characterized in that, the stingless bee honey be Trigona, belong to the bee of stingless bee
Honey, the honey of preferably yellow line stingless bee.
3. method according to claim 1 or 2, which is characterized in that the nectariferous plant of the stingless bee honey include lichee,
One or more of longan, eucalyptus;
And/or the stingless bee honey is the stingless bee honey that tropical and subtropical region harvests between July in May-.
4. according to claim 1-3 any one of them methods, which is characterized in that in step 1), the aqueous solution and the nothing
The volume ratio for piercing bee honey is (3-5):1;It is preferred that 5:1.
5. according to claim 1-4 any one of them methods, which is characterized in that the dissolving operation in step 1) is in ultrasonic item
It is carried out under part, ultrasound parameter is:Ultrasonic power 40/60KHz, time 30-60min, 40-50 DEG C of ultrasonic temperature.
6. according to claim 1-5 any one of them methods, which is characterized in that in step 2), the macroreticular resin is
Supelco SupeliteTMDAX-8;
It is preferred that the volume ratio of the macroreticular resin and the honey solution is 1:(1.5-3).
7. according to claim 1-6 any one of them methods, which is characterized in that in step 4),
The low temperature concentration is that reduced vacuum concentrates, and temperature is 30-50 DEG C;
It is preferred that further including carrying out Liquid Chromatography-tandem Mass separation to the stingless bee Mel extract that step 4) obtains
Operation:
Chromatographic condition:Using octadecylsilane chromatographic column, mobile phase is pure water-methanol;Flow velocity 0.2mL/min;Sample size
2yL;30 DEG C of column temperature;Gradient is 0-2min, 5%;2-10min, 15%-30%;10-25min, 30%-90%;25-
30min, 90%;30-31min 5%;Mass Spectrometry Conditions:The sources ESI, carrier gas temperature:350 DEG C, dry gas stream speed:11L/min, atomization
Air pressure:40psig.
8. a kind of stingless bee Mel extract, which is characterized in that extract to obtain by claim 1-7 any one of them methods;
Preferably, in the stingless bee Mel extract, chlorogenic acid equivalent is at least 90ug/g;And/or Quercetin equivalent is at least
For 15ug/g;
It is highly preferred that in the stingless bee Mel extract, the content of gallic acid is at least 200 μ g/100g;
And/or morin content is at least 200 μ g/100g.
9. stingless bee Mel extract according to claim 8 inhibits drug, food or the health products of enteritis class preparing
In application;
Preferably, the enteritis class, refers to colitis, wherein including ischemic colitis, sigmoiditis, ulcerative colitis
And pseudomembranous colitis.
10. application according to claim 9, which is characterized in that drug, food or the health products are with described stingless
Bee Mel extract is as one of single active ingredient or active constituent.
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|---|---|---|---|---|
| CN111838022A (en) * | 2020-07-15 | 2020-10-30 | 中国科学院西双版纳热带植物园 | Honey producing tray for stingless bees and honey taking method for producing stingless bees |
| CN114916494A (en) * | 2022-03-23 | 2022-08-19 | 中国农业大学 | Method for establishing bee enteritis model and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111838022A (en) * | 2020-07-15 | 2020-10-30 | 中国科学院西双版纳热带植物园 | Honey producing tray for stingless bees and honey taking method for producing stingless bees |
| CN114916494A (en) * | 2022-03-23 | 2022-08-19 | 中国农业大学 | Method for establishing bee enteritis model and application thereof |
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| CN108685951B (en) | 2019-11-26 |
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