CN1682770A - Process for preparing bee glue water solution and use - Google Patents
Process for preparing bee glue water solution and use Download PDFInfo
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Abstract
The water solution of bee glue includes the following steps: mixing bee glue and 95 % concentration alcohol solution via stirring, adding to water at 50-100 deg.c to obtain mixed water solution of bee glue and alcohol with alcohol content of 20-60 %, cooling to room temperature, letting stand and filtering; recovering alcohol and filtering to obtain filtrate; repeated alcohol dissolving and filtering of the filter residue and deposited matter to obtain filtrate; merging filtrate, concentration at 80 deg.c or adding water to density of 1.01-1.04, cooling, filtering to obtain the water solution of bee glue. The alcohol dissolving and water extracting process is simple, and the water solution of bee glue has high flavone content, no organic solvent and additive, low preparation cost and good biological effect. The water solution of bee glue may be used in preparing products for lowering blood sugar, lowering blood fat, resisting inflammation and regulating immunity.
Description
Technical field
The present invention relates to medicament preparation technology, especially relate to a kind of preparation technology and application in blood sugar lowering, blood fat reducing, antiinflammatory, immunoregulation medicament preparation that does not contain the propolis solution of organic solvent.
Background technology
The propolis resin that to be Apis gather from positions such as the tree bud of plant, barks mixes glandular secretion things such as lingual gland with Apis, wax gland again, a kind of colloid substance that is transformed through Apis processing.According to reported in literature, propolis has pharmacological action widely, comprising antitumor, resisting pathogenic microbes, hepatoprotective, immunomodulating, antioxidation, improve effects such as cardiovascular, antiinflammatory antiallergic action, radioprotective, antiulcer, blood fat reducing, blood sugar lowering, existing both at home and abroad many is the health food of raw material with the propolis.
The fundamental component of propolis is resinae, Cera Flava, pollen etc.From chemical compound, mainly contain chemical compound, ester, aldehyde, alcohol compound and alkene, hydrocarbon, terpenoid of flavonoids, organic acid etc.The water solublity of propolis is very poor, usually adopt organic solvent and emulsifying agent to prepare propolis solution at present, as dissolving such as ethanol, Polyethylene Glycol, chloroform, ethyl acetate, dichloromethane, glycerol or by emulsifyings such as tween 80, glyceride, soybean phospholipid, lecithins, in these chemical substances, the all non-people of other material material commonly used except that ethanol, normal conditions adopt down, as the propolis ethanol water that solvent prepares a kind of stimulation taste that is difficult to accept is arranged with ethanol, thereby limit its application.Add glycerol, the oily dilution when preparing propolis solution again after adopting emulsifying agent emulsifying, the adding of these organic solvents has increased toxic and side effects and product cost, all there are strict regulation and restriction in country to the application of these materials, and therefore preparing the green propolis solution that does not conform to organic solvent is the current problem that presses for solution.
Publication number is that the application for a patent for invention of CN1460512A discloses a kind of water-soluble bee glue liquid and preparation method thereof, this water-soluble bee glue system will just carry and the raw material propolis of deleading remove impurity joins in the aqueous solution of Polyethylene Glycol through ethanol, be heated to 80 ℃, and remain on that this temperature obtains after 2-3 hour.This invention adopts Polyethylene Glycol to prepare water-soluble bee glue liquid as solvent, eats for a long time as health food and exists toxic and side effects and production cost increase equally.
Summary of the invention
The invention provides a kind of preparation technology of propolis solution, this process using dissolve in alcohol and extract with water method is carried out, and technology is simple, flavones content is high, does not conform to organic solvent or additive, and preparation cost is low, and biological effect is good.
The present invention also provides the application of above-mentioned propolis solution in preparation blood sugar lowering, blood fat reducing, antiinflammatory, immunomodulating goods, and it has a extensive future.
A kind of propolis solution preparation technology comprises:
(1) propolis is added in 95% ethanol, abundant stirring slowly joins temperature bath in the 50-100 ℃ of hot water after making its dissolving while stirring, gets propolis-alcohol-water mixed liquor, and ethanol content is 20%-60% in the mixed liquor, is cooled to room temperature, leaves standstill, filtration; Filtrate recycling ethanol does not extremely have the alcohol flavor, filters, and gets filtrate;
(2) filtering residue in the said process and sediment are added in 95% ethanol again, abundant stirring slowly joins temperature bath in the 50-100 ℃ of hot water after making its dissolving while stirring, gets propolis-alcohol-water mixed liquor, and ethanol content reaches 20%-60% in the mixed liquor, be cooled to room temperature, leave standstill, filter; Filtrate recycling ethanol does not extremely have the alcohol flavor, filters, and gets filtrate;
(3) repeat above-mentioned dissolve in alcohol and extract with water process, the filtrate that obtains merged, 80 ℃ concentrate or water to transfer to the solution relative density be 1.01-1.04, cooling, filter, promptly get propolis solution.
Described propolis-alcohol-water mixed liquor ethanol content is 20%-60%.
Described temperature bath temperature is 80 ℃.
Described dissolve in alcohol and extract with water process is 2-4 time.
The application of described propolis solution in preparation blood sugar lowering, blood fat reducing, antiinflammatory, immunomodulating goods.
The present invention adopts the method for dissolve in alcohol and extract with water to prepare propolis solution, can not get not conforming to the difficult problem of the propolis solution of organic solvent before having solved always, its flavones content of propolis solution that extracts by this technology reaches more than the 8mg/ml, higher more than 7 times than simple flooding (1mg/ml), in the propolis solution finished product, do not contain any organic solvent, production technology is extremely simple, with low cost, without any side effects, the mice maximum tolerated dose reaches 120ml/kg, is 364 times of human body recommended dose (0.33ml/kg body weight).
The propolis solution of prepared of the present invention is carried out efficacy studies such as animal blood sugar lowering, blood fat reducing, antiinflammatory, immunomodulating, and the result shows that this propolis solution has effects such as obvious functions of blood sugar, blood fat reducing, antiinflammatory, immunomodulating.
Description of drawings
Fig. 1 is preparation technology's flow chart of the present invention.
The specific embodiment
The propolis solution preparation:
1. get pure propolis 100g, add 5 times of amount 95% edible ethanols, fully stir and make its dissolving.
2. the molten propolis liquid of above-mentioned alcohol is slowly joined in the 688ml80 ℃ of hot water while stirring, be cooled to room temperature, leave standstill 2h, filter.Filtrate recycling ethanol filters to there not being the alcohol flavor, filtrate (I) is standby.
3. filtering residue and sediment in inciting somebody to action 2. add 95% ethanol 500ml again, fully stir and make its dissolving.
4. the pure molten propolis liquid in inciting somebody to action 3. slowly joins in the 688ml80 ℃ of hot water while stirring, is cooled to room temperature, leaves standstill 2h, filters.Filtrate recycling ethanol does not extremely have the alcohol flavor, filters, and gets filtrate (II).
5. filtrate (I) and filtrate (II) are merged, add to 1000ml or 80 ℃ with distilled water and be concentrated into 1000ml, promptly relative density 1.02, cooling, filter, and promptly get propolis solution.
Concrete test:
1. different concentration ethanol is extracted the influence to flavone amount in the propolis solution
1. material: pure propolis, 1kg/ bag, total flavones 20%, 95% edible ethanol.
2. total flavones assay method: with the rutin is standard substance, adopts spectrophotography.
Pure propolis is dissolved in quintuple 95% ethanol, fully after the dissolving, the propolis alcoholic solution is poured in 80 ℃ of hot water, making the ethanol final concentration respectively and be 0% (directly adds propolis in 80 ℃ of hot water, warm macerating 16h), 20%, 40%, 60% solution is after fully stirring, be cooled to room temperature, leave standstill 2h, filter.Filtrate recycling ethanol is not to there being the alcohol flavor, and 80 ℃ are concentrated into 0.1g crude drug/ml concentration, sampling and measuring total flavones (the results are shown in Table 1).
Table 1 different concentration ethanol is extracted the influence to the flavone amount
| Concentration of alcohol (%) | Total flavones amount (mg/ml) |
| ????0 ????20 ????40 ????60 | ????0.8 ????2.1 ????8.8 ????9.0 |
By table as seen, flavones content is the highest in the propolis solution of 40～60% ethanol extractions.
2.40% ethanol extraction number of times is to the influence of flavone amount in the propolis solution
After getting pure propolis and adding quintuple 95% ethanol and fully dissolve, pour in 80 ℃ of hot water, make into 40% ethanol final concentration, stir after 10 minutes, be cooled to room temperature, filter, filtrate recycling ethanol is not to there being the alcohol flavor, 80 ℃ concentrate or water transfers to 0.1g crude drug/ml concentration.After sediment and filtering residue add five times of 95% ethanol again and fully dissolve, repeat above-mentioned steps and extract propolis solution, and sediment and filtering residue are repeated to extract once again.Then three extracting solution are merged, 80 ℃ are concentrated into 0.1g crude drug/ml, sampling and measuring total flavones (the results are shown in Table 2)
Table 2 40% ethanol extraction number of times is to the influence of flavone amount
| Number of times | Total flavones amount (mg/ml) |
| Merge concentrated solution three times for the third time for the second time for the first time | ????6.2 ????6.0 ????4.1 ????9.2 |
By table as seen, the total flavones amount is apparently higher than the 3rd time in the 1st time and the 2nd extracting solution, and three extracting solution merge and concentrate back flavone amounts and increase to some extent, but excessive concentration is prone to more precipitation, so the solution relative density is that 1.01-1.04 is the optimum working condition when concentrated.
3.40% ethanol extraction time is to the influence of flavone amount in the propolis solution
Propolis alcoholic solution preparation method is the same, the alcohol amount of will containing is that 40% propolis solution is put 80 ℃ of 5 minutes, 10 minutes, 30 minutes, 60 minutes and 1h, 2h, 16h, after each sample is put the room temperature cooling, filter, 80 ℃ concentrate or water adds to equal volume (concentration 0.1g crude drug/ml), shake up, total flavones (the results are shown in Table 3) is surveyed in sampling.
Table 3 40% ethanol different extraction times are to the influence of flavone amount
| Time | Total flavones amount (mg/ml) |
| 5 minutes 10 minutes 30 minutes 60 minutes 1h 2h 16h | ????6.2 ????8.3 ????8.2 ????8.5 ????8.3 ????8.2 ????8.0 |
By table as seen, 80 ℃ of temperature are bathed 10 fens to 16h, and the flavone amount is close in the propolis solution, therefore the molten propolis liquid of alcohol is slowly joined in 80 ℃ of water while stirring, stir the back time of repose and are getting final product more than 10 minutes.
4. different temperatures is to the influence of 40% ethanol extraction flavone amount
After getting pure propolis and adding quintuple 95% ethanol and fully dissolve, pour in 50 ℃, 80 ℃, the 100 ℃ hot water, make into 40% ethanol final concentration, stir after 10 minutes, be cooled to room temperature, filter, filtrate recycling ethanol is not to there being the alcohol flavor, 80 ℃ concentrate or water transfers to 0.1g crude drug/ml concentration.After sediment and filtering residue add five times of 95% ethanol again and fully dissolve, repeat above-mentioned steps and extract propolis solution, after each sample is put the room temperature cooling, filter, 80 ℃ concentrate or water adds to equal volume (concentration 0.1g crude drug/ml), shake up, total flavones (the results are shown in Table 4) is surveyed in sampling.
Table 4 different temperatures is to the influence of 40% ethanol extraction flavone amount
| Temperature (℃) | Total flavones amount (mg/ml) |
| ????50 ????80 ????100 | ????3.2 ????8.6 ????9.0 |
By table as seen, 50 ℃, 80 ℃, 100 ℃ temperature were bathed 10 fens, the flavone amount increases along with the rising of temperature in the propolis solution, wherein temperature is close 80-100 ℃ of gained flavone amount, consider that the too high meeting of temperature has influence on the activity of some composition in the propolis, therefore select for use 80 ℃ of temperature as optimum process condition.
The propolis solution effect test of the present invention's preparation:
(1), the experiment test of propolis solution hypoglycemic activity
1. material and condition test sample: propolis solution milk yellow liquid, flavones content is 8.5mg/ml, by this prepared in laboratory; Animal: the ICR mice, body weight 20～22g, male and female half and half, Zhejiang Province's Experimental Animal Center provides; Alloxan: Sigma company produces, and faces the time spent to be made into 6mg/ml; Test instrunment adopts 721 type spectrophotometers.
2. method
After the mice fasting 24 hours, tail vein injection alloxan solution 60mg/kg.Fasting 15h after 4 days, eye socket is got blood, and separation of serum is used the determination of glucose oxidase blood glucose value, moulding animal blood glucose value is higher than 10mmol/L hyperglycemia model mice is divided into four groups at random, 10 every group, establishes propolis solution high dose group (30ml/kg); Middle dosage group (20ml/kg); Low dose group (10ml/kg); Distilled water matched group (20ml/kg).Each treated animal equal every day of gastric infusion once, continuous 30 days.Intact animal's group of not doing any processing is established in experiment simultaneously.Each treated animal fasting 24h after the not inferior administration, eye socket is got blood, and separation of serum is used the determination of glucose oxidase blood glucose value.Then the equal per os of each treated animal is irritated stomach glucose 2.0g/kg, surveys and gives behind the glucose 0,0.5,2 hour blood glucose value, to observe the carbohydrate tolerance of animal.Blood glucose value and distilled water group blood glucose value by each group of propolis solution compare, and observe the blood sugar reducing function of propolis solution.
3. result
(1) propolis solution to blood sugar reducing function (seeing Table 5) table 5 propolis solution of alloxan induced hyperglycemia mice to the influence of alloxan induced mice blood glucose (X ± SD)
| Group and dosage | Example number (only) | Body weight (g) | Blood sugar content mmol/L | ||
| Before | After | Before | After | ||
| Propolis solution 30ml/kg 20ml/kg 10ml/kg water matched group 20ml/kg intact animal group | ????10 ????10 ????10 ????10 ????10 | ????20.3±1.47 ????20.2±1.24 ????21.1±1.04 ????21.3±0.99 ????20.8±1.23 | ??27.3±2.67 ??25.8±2.58 ??26.8±1.81 ??26.0±2.19 ??29.7±0.97 | ??17.93±4.68 ??17.54±4.85 ??17.87±5.05 ??17.70±3.88 ??5.89±0.76 *** | ??12.59±2.32 *??15.32±3.38 ??16.93±3.89 ??16.27±4.54 ??5.65±0.86 *** |
Annotate:
*,
* *Expression is compared P＜0.05,0.001 with the water matched group.
By table 5 as seen, the blood glucose value of moulding mice show the moulding success, and propolis solution high dose group blood glucose value and water matched group relatively has or not obvious decline (P＜0.05) apparently higher than intact animal (P＜0.001).Show that propolis solution has obvious decline effect to the mice hyperglycemia.
(2) propolis solution is to the influence (seeing Table 2) of the anti-sugar amount of hyperglycemia model mice
The influence that table 6 propolis solution is measured the anti-sugar of hyperglycemia model mice (X ± SD)
| Group and dosage | Example number (only) | Glucose (g) | Blood sugar content mmol/L | ||
| 0h 0.5h 2h behind the oral glucose | |||||
| Propolis solution 30ml/kg 20ml/kg 10ml/kg water matched group 20ml/kg intact animal group | ????10 ????10 ????10 ????10 ????10 | ????2g/kg ????2g/kg ????2g/kg ????2g/kg ????2g/kg | ????12.59±2.32 *????15.32±3.38 ????16.93±3.89 ????16.27±4.54 ????5.65±0.86 *** | ????29.72±5.24 ????30.23±3.74 ????31.54±5.23 ????32.82±4.70 ????11.57±1.68 *** | ????15.56±4.21 *????16.63±5.29 ????17.69±4.24 ????19.98±4.31 ????8.32±0.98 *** |
Annotate:
*,
* *Expression is compared P＜0.05,0.001 with the water matched group.
By table 6 as seen, intact animal's group all has notable difference (P＜0.001) with the blood glucose value of water matched group day part, and propolis solution high dose group and water matched group comparison 2h period blood glucose value have obvious decline (P＜0.05).The prompting propolis solution has obvious decline effect to the anti-sugar amount of hyperglycemia mice.
4. conclusion
Propolis solution has hypoglycemic activity to the hyperglycemia mice.
(2), the experiment test of the antiinflammatory action of propolis solution
1. material and condition propolis solution: milk yellow liquid, flavones content is 8.5mg/ml, by this prepared in laboratory.Animal: the ICR mice, the male and female dual-purpose, body weight 20 ± 2g is provided by the Zhejiang Experimental Animal Center; Medicine and reagent: acetic acid (analytical pure), reagent one factory in Shanghai is mixed with 0.7% solution with sterile saline.Aspirin is provided by Ningbo Medicine Mfg. Factory.Azo-Blue, Shanghai reagent purchasing and supply station.
2, method and result
2.1 capillary permeability test: get 50 of mices, be divided into 5 groups at random, 10 every group, establish propolis solution high dose group (30ml/kg); Middle dosage group (20ml/kg); Low dose group (10ml/kg); Aspirin (0.2g/kg) positive controls and water matched group (20ml/kg).Each treated animal is irritated stomach 1 time every day, continuous 5 days, 30 minutes tail vein injection 10% Azo-Blue normal saline solution 0.2ml/ only after last administration, lumbar injection 0.7% acetic acid 1ml/ only again, get peritoneal fluid after 30 minutes in 620nm place photometry density (721 spectrophotometer), with statistical procedures between abdominal cavity supernatant optical density value work group, the comparable group differences the results are shown in Table 7.
The antiinflammatory action of table 7 propolis solution (capillary permeability test)
| Group and dosage | Number of animals (only) | Optical density (X ± SD) | The P value |
| Propolis solution 30ml/kg | ????10 | ????0.250±0.046 | ????＜0.05 |
| ??????????20ml/kg | ????10 | ????0.272±0.047 | ????＞0.05 |
| ??????????10ml/kg | ????10 | ????0.281±0.055 | ????＞0.05 |
| Aspirin 0.2g/kg | ????10 | ????0.222±0.053 | ????＜0.01 |
| Water contrast 20ml/kg | ????1?0 | ????0.297±0.042 | ????/ |
Annotate: P value representation and water matched group are relatively
By table 1 as seen, the optical density of three dosage groups of propolis solution all is lower than the water matched group, wherein high dose group and water matched group relatively have notable difference (P＜0.05), show that propolis solution can obviously suppress the increase of capillary permeability, and inflammation is oozed out good inhibitory effect.
2.2 the mice ear test: get 50 of mices, be divided into 5 groups at random, 10 every group, group and dosage are the same.Each treated animal is irritated stomach 1 time every day, continuous 5 days, after last administration 30 minutes, get dimethylbenzene 20ul and drip and be applied to left ear, after 15 minutes animal is taken off neck and put to death, the mice ears are downcut with the position homalographic with card punch, electronic balance (BS210S type) is weighed, difference with left ear weight and auris dextra weight is the swelling degree, respectively organizes difference, and obtains inhibitory rate of intumesce (%).The results are shown in Table 8.
The antiinflammatory action of table 8 propolis solution (mice ear test) (X ± SD)
| Group and dosage | Number of animals (only) | Auris dextra weight (mg) | Left side ear weight (mg) | Swelling weightening finish (mg) | Inhibitory rate of intumesce (%) |
| Propolis solution 30ml/kg | ????10 | ??14.3±2.1 | ??19.1±2.2 | ??4.8±3.0 * | ????45.5 |
| ??????????20ml/kg | ????10 | ??13.2±1.0 | ??21.0±5.8 | ??7.8±5.6 | ????11.4 |
| ??????????10ml/kg | ????10 | ??12.8±1.0 | ??20.0±4.9 | ??7.2±5.3 | ????18.2 |
| Aspirin 0.2g/kg | ????10 | ??14.4±2.3 | ??20.4±4.6 | ??4.0±2.2 ** | ????54.5 |
| Water contrast 20ml/kg | ????10 | ??13.8±1.5 | ??22.6±4.3 | ??8.8±4.1 | ????/ |
Annotate:
*,
*Expression is compared P＜0.05,0.01 with the water matched group
By table 8 as seen, propolis solution is respectively organized xylol and is caused the ear swelling that inflammation causes inhibitory action is all arranged, inhibitory rate of intumesce is 11.4～45.5%, wherein high dose group (30ml/kg) and water matched group relatively have notable difference (P＜0.05), show the obviously swelling that causes of inflammation-inhibiting of propolis solution.
3, conclusion
Show that by above test the propolis solution high dose group all has obvious suppression effect (P＜0.05) to mice capillary permeability and mice ear, show that propolis solution has antiinflammatory and detumescence effect preferably.
(3) propolis solution is to the experiment test of Immune Function
1. material and condition test sample: propolis solution: milk yellow liquid, flavones content is 8.5mg/ml, by this prepared in laboratory.Animal: the ICR mice, body weight 17-20g, the male and female dual-purpose, Zhejiang Province's Experimental Animal Center provides.Sheep red blood cell (SRBC): adopt certainly, be kept in the Alsever solution, face, be mixed with desired concn with preceding normal saline washing 3 times.Reagent: canavaline element (ConA) is produced by Sigma company, is distributed into 1mg/ml, deposits for-20 ℃; RPMI-1640 complete culture solution Japan produces, and every 1000ml contains Hepes 25mM, glutamine 2mM, 2-ME 50 μ M, calf serum 10%, penicillin 100 μ/ml, streptomycin 100 μ/ml; Tritiated thymidine (
3H-TdR) Shanghai Atomic Nucleus Inst., Chinese Academy of Sciences produces, and concentration 1mCi/ml is than degree 42Ci/mM; Scintillation solution PPO 4 grams, POPOP 0.4 gram (Shanghai reagent one factory product) add to 1000ml with toluene, and 40 ℃ of water-baths are spent the night, and the darkroom is preserved standby.
2. method
Get 40 of mices, be divided into 4 groups at random, 10 every group, establish propolis solution high dose group (30ml/kg); Middle dosage group (20ml/kg); Low dose group (10ml/kg) and water matched group (20ml/kg).And numbering is weighed respectively, each treated animal gastric infusion every day once, continuous 12 days, in experiment the 8th day lumbar injection 3: 5 (v/v) sheep red blood cell (SRBC)s 0.2ml sensitization.Drug withdrawal is respectively organized mice next day and is plucked eyeball and get blood, collects serum, carries out hemolysin with colorimetry and measures, and mice is put to death in the cervical vertebra dislocation then, and the aseptic spleen of getting makes 1 * 10 with RPMI-1640
7/ ml cell suspension adopts the inductive MTT of the ConA method of mixing to detect the T lymphocyte transformation function].
3. result's (seeing Table 9)
Table 9 propolis solution is to normal effect of immunologic function (X ± SD)
| Group and dosage | Example number (only) | Body weight (gram) | T drenches changes (SI) | Hemolysin (HC 50) | |
| Before | After | ||||
| Propolis solution 30ml/kg 20ml/kg 10ml/kg water contrast 20ml/kg | ?10 ?10 ?10 ?10 | ?18.7±0.85 ?19.2±0.72 ?18.8±0.77 ?18.8±0.78 | ?27.7±1.6 ?26.4±2.1 ?26.7±1.7 ?27.0±2.2 | ????8.25±1.68 ????8.62±1.53 ????7.70±1.47 ????8.08±1.65 | ??101.6±13.7 *??97.3±12.4 ??94.1±15.0 ??87.0±13.2 |
Annotate:
*Expression is compared P＜0.05 with the water matched group.
By table 1 as seen, three dosage groups of propolis solution and water matched group comparison serum hemolysin content all have obvious rising, wherein high dose group propolis solution and water matched group relatively there were significant differences (P＜0.05=.And three dosage groups of propolis solution do not have obvious influence (P＞0.05) to normal mouse T lymphocyte transformation function.
4. conclusion
The be significantly improved effect of humoral immune function of propolis solution, and the pair cell immunologic function does not have tangible influence.
(4) the effect for reducing blood fat experiment test of propolis solution
1. material and condition test sample: propolis solution: milk yellow liquid, flavones content is 8.5mg/ml, by this prepared in laboratory.Animal: the SD rat, body weight 200-250g, the male and female dual-purpose is provided by Zhejiang Province's Experimental Animal Center.Test instrunment: 721 type spectrophotometers, cholesterol (TC); Triglyceride (TG) is measured test kit, produces by Ningbo Ci Cheng biochemical reagents factory.
Rat high lipid food preparation: high lipid food prescription cholesterol 1.5%, Adeps Sus domestica 10%, methylthiouracil 0.2%, normal feedstuff 88.3%., join in the Adeps Sus domestica of fusing earlier behind cholesterol and the methylthiouracil mixing, again with the normal feedstuff mixing after standby.
2. method
Get 50 of SD rats, be divided into 5 groups at random, 10 every group, establish first group, propolis solution high dose group (30ml/kg); Second group, dosage group (20ml/kg) in the propolis solution; The 3rd group, propolis solution low dose group (10ml/kg); The 4th group, model group (20ml/kg); The 5th group, normal group.All the other each groups are fed high lipid food respectively except that the 5th group of hello normal feedstuff, irritate the stomach relative medicine simultaneously.Once a day, continuous 30 days.Each treated animal fasting 16h after the inferior administration, femoral artery is got blood, and separation of serum is pressed TC, TG and is measured test kit explanation survey TC, TG content, the results are shown in Table 10.
3. assay (seeing Table 10)
Table 10, propolis solution are to the effect for reducing blood fat of experimental hyperlipidemia rat (X ± SD)
| Group and dosage | Example number (only) | Body weight (gram) | ????TC ????mmol/L | ??TG ??mmol/L | |
| Before | After | ||||
| Propolis solution 30ml/kg propolis solution 20ml/kg propolis solution 10ml/kg model group 20ml/kg normal group | ????10 ????10 ????10 ????10 ????10 | ????201±8.5 ????198±7.6 ????199±8.5 ????203±9.2 ????201±7.8 | ????290±5.8 ????293±41.7 ????289±38.7 ????286±39.5 ????298±42.9 | ????4.68±1.32 *????4.96±1.45 ????5.26±1.33 ????6.08±1.22 ????1.39±0.21 *** | ??0.95±0.25 *??1.05±0.30 ??1.11±0.36 ??1.25±0.27 ??0.79±0.19 *** |
Annotate:
*,
* *Expression is compared P＜0.05,0.001 with the water matched group.
By table 10 visible river bend high lipid food rat TC, TG apparently higher than normal rat (P＜0.001), show the moulding success, give propolis solution simultaneously and can reduce TC in the serum, TG content and feed the high lipid food rat, wherein propolis solution high dose group and model group relatively have significant difference (P＜0.05).
4. conclusion
Show blood fat the significantly decrease effect of propolis solution to the experimental hyperlipidemia rat.
(5) propolis solution acute toxicity test
Propolis solution is carried out acute toxicity test, i.e. the mensuration of the mensuration of median lethal dose(LD 50) (LD50), maximum tolerated dose (MTD).Be subjected to the reagent thing: propolis solution, the propolis solution milky white liquid, flavones content is 8.5mg/ml, by this prepared in laboratory.Laboratory animal: ICR kind mice, body weight 18-22g, male and female half and half, Inst. of Traditional Chinese Medicine, Zhejiang Prov laboratory animal room provides.Experimental technique: propolis solution was observed 5 days with the disposable gastric infusion 40ml/kg of the dosage of maximum volume, and 30 mices all do not have death.So can't measure LD50, change maximum tolerance determination into.
Maximum tolerance determination: get 30 of mices, male and female half and half, fasting was irritated stomach and is given propolis solution after 12 hours, and three times on the one, accumulated dose is 120ml/kg, observes activity and the death condition of 7 days mices continuously.
Experimental result: the no abnormal reaction of mice behind the gastric infusion, mice does not all have death in 7 days, and as calculated: the mice maximum tolerated dose is 120ml/kg, recommend amount (0.33ml/kg) for human body 364 times.
Claims (8)
1. propolis solution preparation technology comprises:
(1) propolis is added in 95% ethanol, fully stirring slowly joins in the 50-100 ℃ of hot water after making its dissolving while stirring, gets propolis-alcohol-water mixed liquor, and ethanol content is 20%-60% in the mixed liquor, is cooled to room temperature, leaves standstill, and filters; Filtrate recycling ethanol does not extremely have the alcohol flavor, filters, and gets filtrate;
(2) filtering residue in the said process and sediment are added in 95% ethanol again, fully stirring slowly joins in the 50-100 ℃ of hot water after making its dissolving while stirring, gets propolis-alcohol-water mixed liquor, and ethanol content reaches 20%-60% in the mixed liquor, be cooled to room temperature, leave standstill, filter; Filtrate recycling ethanol does not extremely have the alcohol flavor, filters, and gets filtrate;
(3) repeat above-mentioned dissolve in alcohol and extract with water process several times, the filtrate that obtains merged, 80 ℃ concentrate or water to transfer to the solution relative density be 1.01-1.04, cooling, filter, promptly get propolis solution.
2. propolis solution preparation technology according to claim 1 is characterized in that: ethanol content is 40%-60% in described propolis-alcohol-water mixed liquor.
3. propolis solution preparation technology according to claim 1 is characterized in that: described dissolve in alcohol and extract with water process is 2-4 time.
4. propolis solution preparation technology according to claim 1 is characterized in that: described temperature bath temperature is 80 ℃.
5. the application of propolis solution according to claim 1 in preparation blood sugar lowering goods.
6. the application of propolis solution according to claim 1 in preparation blood fat reducing goods.
7. the application of propolis solution according to claim 1 in preparation antiinflammatory goods.
8. the application of propolis solution according to claim 1 in preparation immunomodulating goods.
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| JP2008050301A (en) * | 2006-08-24 | 2008-03-06 | Prima Meat Packers Ltd | Pancreatic lipase inhibitor |
| CN100497599C (en) * | 2006-12-30 | 2009-06-10 | 中国科学院长春应用化学研究所 | Plant medium composition containing bee glue bacteriostatic agent and preparation process thereof |
| CN101099547B (en) * | 2007-06-18 | 2011-01-05 | 中国农业科学院蜜蜂研究所 | Water soluble propolis extract with bioactivity and light-colored propolis extract |
| CN104996540A (en) * | 2014-05-12 | 2015-10-28 | 浙江海洋学院 | Fresh keeping method for shrimp meat |
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| CN100497599C (en) * | 2006-12-30 | 2009-06-10 | 中国科学院长春应用化学研究所 | Plant medium composition containing bee glue bacteriostatic agent and preparation process thereof |
| CN101099547B (en) * | 2007-06-18 | 2011-01-05 | 中国农业科学院蜜蜂研究所 | Water soluble propolis extract with bioactivity and light-colored propolis extract |
| CN104996540A (en) * | 2014-05-12 | 2015-10-28 | 浙江海洋学院 | Fresh keeping method for shrimp meat |
| CN105169376A (en) * | 2015-10-19 | 2015-12-23 | 淄博维希尔生物技术有限公司 | Nystatin aqueous solution and preparation method thereof |
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| CN106943435A (en) * | 2017-03-07 | 2017-07-14 | 浙江大学 | A kind of water-soluble bee glue and preparation method thereof |
| CN113679742A (en) * | 2021-08-30 | 2021-11-23 | 神农架林区蜜蜂天堂食品有限公司 | Preparation method and application of honeycomb alcohol extract |
| CN117017889A (en) * | 2023-09-04 | 2023-11-10 | 吉林大学 | Composition for relieving xerostomia |
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