CN100509713C - Method for preparing water-soluble bee glue product and use thereof - Google Patents
Method for preparing water-soluble bee glue product and use thereof Download PDFInfo
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- CN100509713C CN100509713C CNB2006100039952A CN200610003995A CN100509713C CN 100509713 C CN100509713 C CN 100509713C CN B2006100039952 A CNB2006100039952 A CN B2006100039952A CN 200610003995 A CN200610003995 A CN 200610003995A CN 100509713 C CN100509713 C CN 100509713C
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Abstract
The invention relates to a process for extracting active ingredients from bee glue and the use of the water-soluble bee glue product, wherein the extraction consists of disintegrating natural bee glue, chaging into water, sufficiently stirring, extracting through immersion, carrying out solid-liquid separation, subjecting the solid to backflow extraction with petroleum ether, distilling and seasoning, dissolving the obtained solid into aqueous solution of L-arginine or L-lycine, filtering liquid and subjecting the solution to freeze drying treatment.
Description
Technical field
The present invention relates to extraction of effective components in a kind of propolis, particularly the effective ingredient in the propolis is handled being converted into water miscible method, and the purposes of this water-soluble bee glue product.
Background technology
Propolis is that the resin secretory product of honeybee herborization and beeswax etc. are through chewing a kind of resinous substance that modulation forms.Propolis all has very long applicating history all over the world.Both at home and abroad to the chemical constitution of propolis, the physiological and pharmacological effect has many researchs in recent years, finds that propolis has effects such as antibiotic, antiviral, anti-inflammatory, reducing blood-fat, hypoglycemic and enhancing immunity.
At present, bee glue product both domestic and external all is to obtain effective constituent by the alcohol lixiviate basically, but the alcohol extract of propolis is not soluble in water, and degree of absorbing is low, and still is mixed with certain impurity beeswax composition.At present, utilize to change propolis extract acid-basicity (disclosed) though have, or add heat soaking propolis alcohol extract (disclosed) and prepare water-soluble bee glue with the method for kinds of surface promoting agent hydrotropy (disclosed) as patent of invention 03150648.8 as patent of invention 03141394.3,94118328.9 with glycerine, polyoxyethylene glycol as patent of invention 97111643.The water-soluble bee glue of gained also discloses its physiologically active in the prior art.Used non-natural hydrotropy composition to improve the water-soluble of propolis in the prior art in addition.By the water-soluble bee glue preparation method of inventions such as Bulgarian Nikolov is that the propolis that ethanol or methyl alcohol were slightly carried is dissolved in amino acid (L-Methionin) with 8% and heating, obtains the water-soluble bee glue powder.
Summary of the invention
The invention provides and a kind of natural propolis is converted into the method for water-soluble bee glue product, particularly more purified water-soluble bee glue product, and the purposes of this goods.
Method of the present invention is to add in the entry after natural propolis is pulverized, after fully stirring, soak extraction, carry out solid-liquid separation then, again solid is carried out refluxing extraction with sherwood oil, after removing beeswax, extracting solution is carried out refluxing extraction with ethanol and acetone mixing solutions again, ethanol wherein and acetone can be arbitrary proportions, will be through ethanol, the extracting solution that the acetone mixed-liquor return extracts distills drying treatment, the gained solid dissolves in the aqueous solution of L-arginine or L-Methionin in the ratio of 100 milliliters of every grams, the mass ratio of L-arginine or L-Methionin and water is 3～5% in its aqueous solution, treat to leach liquid behind the solid dissolution equilibrium, again the solution lyophilize is handled.
Ethanol is 1 to 1 with the optimum volume ratio of acetone in the method for the present invention, and ethanol, acetone mixed-liquor return solid extract preferably dissolve in the L-arginine aqueous solution that contains 3% (mass ratio).
System can be heated to 30～80 degree when being dissolved in solid in the amino acid among the preparation method of the present invention.
The resultant goods of the inventive method can be used for preparing the medicine of hypermnesis function.
The resultant goods of the inventive method can be used for preparing the medicine for the treatment of senile dementia.
Its purity of bee glue product that adopts method of the present invention to make is higher than prior art, its effective ingredient also is higher than prior art, with contained general flavone content in the propolis is example, and its general flavone content of the goods of prior art (in the Quercetin standard substance) is generally about 15%; And remove water-soluble impurity (accounting for total mass 4%), pesticide residue and heavy metal ion with the water extraction propolis among the present invention; Sherwood oil thermal backflow paraffin removal (about 35%); Refining propolis (accounting for total mass 40%) is extracted in ethanol and acetone thermal backflow, and the general flavone content of its end article is still in the Quercetin standard substance) can be up to 28.6%.On the other hand, the present invention is different from the prior art and adds strong acid and strong base and change the deliquescent way of propolis, and enrichment concentrates in the propolis and comprises total flavones in each interior effective ingredient effectively, and keeps its natural constituent.
Experiment shows adopts the L-arginine aqueous solution can significantly improve the water-soluble bee glue product productive rate among the present invention.
Show that according to correlated test goods of the present invention have special activity, can be used for preparing the medicine of hypermnesis and the medicine of treatment senile dementia.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.
Get propolis raw material, pulverize the back and add in the distilled water, stir under the room temperature and spend the night; Leave standstill after-filtration, filter residue changes in the Soxhlet refluxing extraction device, is earlier that beeswax was removed in the solvent thermal backflow in 6 hours with the sherwood oil; Extracted 12 hours with ethanol (or ethanol: acetone is with the arbitrary proportion mixed solvent) thermal backflow afterwards, extracting solution obtains refining propolis through underpressure distillation.
The contrast experiment shows that the L-lysine solution of employing 3% dissolves the refining propolis of aforementioned gained, dissolve the refining propolis of aforementioned gained during perhaps to adopt total amount be 3% L-Methionin with the L-arginine mixes the aqueous solution, can improve the yield of water-soluble bee glue product, referring to table 1.
Therefore most preferred embodiment of the present invention should be to get aforementioned refining propolis 10 grams, adds in 1000 milliliter 3% the L-arginine aqueous solution, is heated to 30～80 ℃ and continue stirrings; After treating dissolving fully, filter; Can get 36.2 gram brown dry powder water-soluble bee glue products after the filtrate lyophilize.
The present invention is used to improve the example of learning and memory function
Scopolamine hydrobromide is a kind of nervus centralis anticholinergic, the scopolamine hydrobromide that gives the mouse doses continuously can cause the mouse nervus centralis to suppress, the space learning memory capability descends, show as the administration mouse and find the ability of hiding platform in the water to compare obvious decline with normal mouse by identical training in the water maze test, required time prolongs.Experiment by scopolamine hydrobromide induced mice ability of learning and memory obstacle also is one of research neurologic agent study of behaviour model commonly used.
Water maze laboratory
Get 60 of healthy adult mouse (male, 25～30 grams), be equally divided into 6 groups at random.Be normal group, damage group, positive controls, arginine control group, propolis small dose group, the heavy dose of group of propolis.Except that normal group, all the other each groups are all from first day abdominal injection scopolamine hydrobromide (0.1 mg/kg body weight sky), and normal group is then injected equal-volume physiological saline.Every day, the injection beginning was preceding 30 minutes, give medicine, normal group, damage group are irritated stomach physiological saline, positive controls is irritated stomach piracetam (the clinical piracetam of using, 100 mg/kg body weight * days), the arginine control group is irritated stomach L-arginine (100 mg/kg body weight sky), and the propolis small dose group is irritated stomach gained water-soluble bee glue product of the present invention (50 mg/kg body weight * days), and the heavy dose of group of propolis is irritated stomach gained water-soluble bee glue product of the present invention (100 mg/kg body weight * days).Injection finishes the test of back 30 minutes beginning water mazes.Test was carried out 4 days altogether, before the test every mouse is numbered mark, observe record it finds the time (is unit with the second) of hiding platform in the water in 2 tests of every day, averages as the observed data of this sample, and standard error and variance analysis are then from 10 samples of every group.The result is as shown in Table 2:
*Variance analysis is to compare with the damage group by each group of administration to obtain;
*Represent the p value less than 0.05, significant difference;
*Expression p value is less than 0.01, and difference is extremely remarkable.
Above result shows, after gained water-soluble bee glue product of the present invention (100 mg/kg body weight) gives mouse, can significantly improve the memory dysfunction that causes by scopolamine hydrobromide, may certain treatment and improvement effect be arranged symptoms such as senile dementias.Its effect is better than piracetam.The effect of goods of the present invention may be produced by activeconstituents in the propolis, also may be produced by activeconstituents in the propolis and L-arginine synergy.But use the L-arginine in relevant experiment, not observe and the identical effect of gained water-soluble bee glue product of the present invention merely.
Modern brain science studies show that vagusstoff participates in mammiferous learning and memory activity as a kind of major neurotransmitter involved.Therefore, the important lytic enzyme-acetylcholine esterase active of regulation and control levels of acetylcholine has just reflected the ability that animal learning is remembered to a certain extent in the animal brain.Generally speaking, learning and memory in rats and mice and acetylcholinesterase vigor are negative correlation to a certain extent.Acetyl choline content is closely related with memory in the existing known brain, and the vagusstoff amount reduces in old age or the dementia human brain.Research thinks that anticholinesterase can postpone the metabolism of vagusstoff and decompose, thus the excitement that prolongs postsynaptic receptor.Therefore anticholinesterase is considered to a kind of active drug of alleviating old age or dull-witted patient.
In mammiferous brain, there are several important positions, with learning and memory confidential relation is arranged, the structure-hippocampus and the amygdaloid body that wherein are arranged in both sides hemicerebrum face are the key structures of memory process.And in mammiferous dominance memory process, hippocampus plays an important role, and hippocampus has the plasticity-that the dominance memory needs.Therefore, detect the variation of acetylcholinesterase level in hippocampus and some cerebral tissue, can verify effectively that goods of the present invention improve the Mechanism of Physiological and Biochemical of Scopolamine damage ability of learning and memory in mice, and with water maze laboratory the result corresponding.
The enzyme assay experiment
Get 12 of healthy adult mouse (male, 25 ± 2 grams), be equally divided into 2 groups at random.The damage group is irritated stomach physiological saline, and the propolis group is irritated stomach gained water-soluble bee glue product of the present invention (100 mg/kg body weight * days), whole experiment mice abdominal injection scopolamine hydrobromides (0.1 mg/kg body weight * days) after the administration.Behind the successive administration four days, put to death mouse, strip the mouse brain tissue, separate cerebral hippocampal district and pallium part, measure each several part acetylcholinesterase level and corresponding protein content behind the tissue homogenate respectively.Can hydrolysis specific substrate iodo vagusstoff (Acetylcholine iodide) with every milligram of cerebral tissue albumen thus cause that photoabsorption (OD value) represents the height of enzymic activity in the variation at 410nm place.
The result is as shown in table 2
*(
* *Expression p value is less than 0.001, and difference is extremely remarkable).
Above result shows, give mouse doses gained water-soluble bee glue product of the present invention (100 mg/kg body weight) after, can significantly suppress the rising of the acetylcholinesterase level of the crucial memory of mouse brain position-hippocampus of causing by Scopolamine.This shows that also gained water-soluble bee glue product of the present invention can improve the amnesia learning and memory abilities in aging mice, also can be used as the medicine of a kind of treatment or alleviation senile dementia.
Claims (5)
1, a kind of preparation method of water-soluble bee glue product, it is characterized in that and to add in the entry after the natural propolis pulverizing, after fully stirring, soak extraction, carry out solid-liquid separation then, again solid is carried out refluxing extraction with sherwood oil, after removing beeswax, extracting solution is carried out refluxing extraction with ethanol again, to distill drying treatment through the extracting solution of alcohol reflux, the gained solid dissolves in the arginic aqueous solution of L-in the ratio of 100 milliliters of every grams, wherein the mass ratio of L-arginine and water is 3～5%, treats to leach liquid behind the solid dissolution equilibrium, the solution lyophilize is handled again.
2, according to the preparation method of the described a kind of water-soluble bee glue product of claim 1, the extracting solution that it is characterized in that alcohol reflux distills dry gained solid and dissolves in the L-arginine aqueous solution, and the mass ratio of L-arginine and water is 3%.
3,, it is characterized in that solid is dissolved in the process of the arginic aqueous solution of L-system to be heated to 30～80 degree according to the preparation method of claim 1 or 2 described a kind of water-soluble bee glue products.
4, get the medicine of the hypermnesis function of water-soluble bee glue product preparation according to the described either party's legal system of claim 1 to 3.
5, according to the described either party's legal system of claim 1 to 3 get water-soluble bee glue product preparation have in the treatment senile dementia medicine in application.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2006100039952A CN100509713C (en) | 2006-01-27 | 2006-01-27 | Method for preparing water-soluble bee glue product and use thereof |
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| CNB2006100039952A CN100509713C (en) | 2006-01-27 | 2006-01-27 | Method for preparing water-soluble bee glue product and use thereof |
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| CN1839890A CN1839890A (en) | 2006-10-04 |
| CN100509713C true CN100509713C (en) | 2009-07-08 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104687066B (en) * | 2015-03-23 | 2017-01-18 | 宁波大学 | Betel nut brine containing arginine |
| CN106213387A (en) * | 2016-08-26 | 2016-12-14 | 山东华康蜂业有限公司 | A kind of method of propolis wax removing |
| CN108991453A (en) * | 2018-08-06 | 2018-12-14 | 重庆市畜牧科学院 | A kind of preparation method and products thereof of compound nano-propolis |
| CN110538128B (en) * | 2019-09-24 | 2020-12-08 | 江苏蜂奥生物科技有限公司 | Method for recovering and extracting propolis effective components from beeswax |
| CN110638837B (en) * | 2019-10-31 | 2021-05-07 | 广东国源国药制药有限公司 | Propolis extract and preparation method and application thereof |
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2006
- 2006-01-27 CN CNB2006100039952A patent/CN100509713C/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 蜂产品的保健功能. 郭芳彬.养蜂科技，第6期. 2000 |
| 蜂产品的保健功能. 郭芳彬.养蜂科技，第6期. 2000 * |
| 蜂胶中有效成分提取工艺研究. 顾青等.食品工艺，第2期. 2001 |
| 蜂胶中有效成分提取工艺研究. 顾青等.食品工艺，第2期. 2001 * |
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