RO129062B1 - Bee pollen tincture - Google Patents

Abstract

The invention relates to a bee pollen tincture as hydro-alcoholic extract. According to the invention, the tincture is extracted by maceration with 70% alimentary ethanol, in an association ratio of 20...30% bee pollen : 70...80% ethanol, the percentage being expressed by weight.

Description

The invention relates to a hydroalcoholic extract of the paste, intended for internal or external use for various therapeutic purposes, with antimicrobial, antifungal, anti-inflammatory, immunomodulatory, soothing and healing action.

There are known various tinctures obtained from natural products, used to prevent or treat various conditions: tincture of propolis, tincture of blueberries, tincture of echinacea, tincture of burns, etc. Tinctures are hydroalcoholic extracts obtained from various natural products, by extraction with ethyl alcohol of various concentrations. They have a complex composition. One of the most used in medical practice is propolis tincture. Propolis tincture has proven over time to be effective in treating infectious diseases, enteritis, pharyngitis and gingivitis.

It is known from the patent R0122619 B1, a preparation based on pollen and Lactobacillus sp., Which consists of 10 ... 40% pollen, 0.1 ... 10% honey, 0.1x105 ... 10x106 CFU / ml strains of Lactobacillus sp., 1 ... 10% lactic acid and 20 ... 80% distilled water, the percentages being expressed by weight. This preparation is assimilated to the shepherd, and has the therapeutic properties of a probiotic product through the addition of amino acids, enzymes and vitamins.

Patent document RO 83095 refers to a process for processing pollen for the production of sheep, by treating pollen with acetate buffer until a 66% pollen suspension is obtained, which, after incubation at 37 ° C, for 50 hours , is dried at a maximum temperature of 45 ° C for 30 hours.

The technical problem, which the invention solves, consists in using the active principles of the shepherd in the form of a product with therapeutic effects.

The invention removes the disadvantages mentioned by the fact that the hydroalcoholic extract is made up of paste and ethyl alcohol, in an association ratio of 20 ... 30% paste and 70 ... 80% food ethyl alcohol 70 °.

Propolis and tincture of propolis were studied as intermediates for obtaining products intended as adjuvants in the following treatments:

- in dental diseases and oral hygiene, for reducing the bacterial load in the mouth, in periodontitis, for accelerating the healing of inflammation and wounds, in foot-and-mouth ulcers, reducing the formation of tartar;

- as an adjuvant in the treatment of genital herpes, in the form of ointments;

- as an adjuvant in ionophoric procedures for the improvement of rheumatisms of different etiologies;

- as an antiseptic (antibacterial and antiparasitic) in digestive tract disorders, including gastrointestinal ulcers caused by Helicobacter pylorr,

- as an adjuvant in the treatment of damage caused by corneal herpes;

- as an adjuvant in the treatment of colds and nasal congestion.

The invention relates to a hydroalcoholic extract of the paste, respectively, of a paste tincture, obtained from the extraction by maceration with food ethyl alcohol 70 °, in an association report, parts by weight of: 20 ... 30% paste: 70 ... 80% ethyl alcohol 70 °.

The main advantages of the shepherd's tincture, according to the invention, are the following:

- is a natural product, with a complex composition, which enriches the range of bee preparations with multiple therapeutic effects;

- due to the diversity of active principles it contains, it presents a series of special therapeutic effects: it increases the defense capacity of the body (immunostimulator, immunomodulator); anti-inflammatory, antiallergic, healing, radioprotective, chemoprotective, detoxifying, heavy metal chelator, antiviral, antibacterial, remineralizing;

- the product has no toxicity;

- has nice organoleptic properties.

RO 129062 Β1

The therapeutic properties of the herring tincture are determined by the active principles extracted from the herd.

Bumblebee or bee bread is pollen stored in the honeycomb and covered with a layer of wax. The paste is obtained by fermenting the pollen under the bacterial microflora conditions of the hive. Pollen undergoes a series of biochemical transformations, under the influence of added bees, microorganisms, temperature and humidity in the hive.

It is known that the toning, immunostimulatory and detoxifying properties of pollen are taken over by the shepherd [Dr. Biochem. C. Mateescu, Research Institute for Beekeeping Development, Rev. The medical week, no. 82; 94; 102/2010]. It even seems that the therapeutic action of so-called bee bread is faster and more intense than the action of pollen. The therapeutic properties of the pastry are due to the high content of essential amino acids, enzymes, vitamin K and simple sugars. Due to the high amount of lactic acid and the antibiotic properties, the oil may be kept for a long time, without any major, quantitative or qualitative changes being observed.

This product is currently enjoying increased attention from researchers, for the effects obtained by applying it in various conditions; At the present stage of experimentation it seems that there are sufficient premises that justify the assertion of the specialists that shepherd is a natural beekeeping product with great prospects of use for the purpose of human health.

It covers the entire range of pollen recommendations: liver disease, anemia, stress, rheumatism, regulation of intestinal transit, colon conditions, especially those accompanied by constipation (enterocolitis, fermentation or putrefaction colitis, chronic constipation of various etiologies), but the actions they are much faster and stronger. Paste is a natural product, with more valuable properties than pollen due to its higher content in simple sugars, vitamin K, enzymes and amino acids, as well as the increased acidity that makes it easily assimilable. Compared to pollen, the nutritional and antibiotic value is 3 times higher.

In order to comply with the physico-chemical and microbiological quality conditions for dyeing of sheep, required by the regulations in force, used sheep as raw material will be analyzed in accredited laboratories, before being purchased.

If the raw material corresponds qualitatively, it will be purchased, after which it will be stored properly, in a dry and cold place.

The anti-inflammatory action of the gel with 3% and 5% itch tincture was investigated, by two experimental methods that aimed at how they influence the inflammatory exudate from the experimental model of rat edema induced with different edemogenic substances, with the possibility of focusing on the mechanism. of their action (1,2).

Sheep tincture was incorporated in carbopol gel 1% in concentration of 3% and 5%. The base of incorporation of the propolis tincture, the carbopol hydrogel, was prepared from: carbopol 940 1 g, glycerine 5 g, triethanolamine 2.5 g, preservative solution up to 100 g.

The carbopol is triturated with glycerin, then 80 g of preservative solution is added and homogenized. The mixture was neutralized with 2.5 g of triethanolamine added gradually, and then supplemented with 100 g preservative. Leave to stand for 24 hours for complete gelation.

The tests were carried out by two experimental methods of acute inflammation: edema induced in rat paw with 10% kaolin suspension, and 6% dextran solution (1,2,3,4).

By injecting kaolin into the rat's paw, prostaglandin formation is stimulated, causing inflammation and local edema. The dextran-induced edema is mainly due to the release of histamine and serotonin, and is called anaphylactoid edema.

Edema was induced by intra-implant injection of 0.1 ml 10% kaolin suspension and 0.2 ml dextran solution.

RO 129062 Β1

Each batch of 10 male rats, the Wistar breed, weighing 170 ± 15 g, was used for each edemogen agent. One batch was the control batch, one batch was treated with 3% sheep dye gel, one batch was treated with 5% sheep dye gel, and one batch was treated with 4% phenylbutazone gel.

The animals brought from the farm were kept under the conditions of the laboratory for 2 days, in order to get used to the new habitat and the diet.

Edemogenic agent was administered to all animals. The test preparation was applied to the paw where the edema was induced (uniform application, in a thin layer -0.25 g gel).

The evaluation of the anti-inflammatory effect of the gels taken in the study was made against the 4% phenylbutazone preparation (on a similar basis) applied on the paw with edema, under the same conditions as above.

The determinations were made against control groups (individuals not treated).

Rat paw volume was measured plethysometrically, after intra-implant injection of the edemogenic agent, further pletismometric measurements were performed, at intervals: 2 h, 4 h, 6 h, 24 h (for the 10% kaolin suspension edemogenic agent) and at intervals: 30 min, 60 min, 90 min, 120 min after induction of edema (for the edemogenic agent 6% dextran solution).

The average value of the anti-inflammatory edema (expressed in ml), the standard error and the percentage of edema inhibition were calculated for each batch, according to the formula:

Edema inhibition,% = (1-X substance / X control) x 100 where:

X substance - the average value of the edema produced by the tested substance;

X control - the average value of the edema produced in the control within the same time frame after administration of the edemogenic agent.

The statistical evaluation of the results was done by the t test (Student) (5.6).

Gel with tincture of 3% had the most intense anti-inflammatory effect after 2 h after the induction of edema, the inflammation produced with kaolin suspension 10% being decreased by 33.19%.

The gel with 5% itch tincture showed the most intense anti-inflammatory effect after 2 h after the induction of edema, the inflammation produced with kaolin suspension 10% being reduced by 42.85%.

The phenylbutazone gel showed the most intense anti-inflammatory effect after 2 h after the induction of edema, the inflammation produced with kaolin suspension 10% being reduced by 68.57% (table 1).

Table 1

Anti-inflammatory effect of gum tincture studied on inflammatory edema produced with kaolin suspension 10%

Product tested Edem 2 h (ml) (x + SD.) Edema 4 h (ml) (x + SD.) Edema 6 h (ml) (R&D) Edem 24 h (ml) (x + SD.) witnesses 0.238 + 0.01 0.276 ± 0.03 0.340 ± 0.02 0.293 ± 0.01 Sheep tincture gel 5% 0.136 ± 0.04 ** 0.173 ± 0.01 ** 0.229 ± 0.03 ** 0.201 ± 0.02 ** Effect% -42.85 -37.31 -32.64 -31.39

RO 129062 Β1

Table 1 (continued)

Product tested Edem 2 h (ml) (x + SD.) Edema 4 h (ml) (Â + SD.) Edema 6 h (ml) (x + SD.) Edem 24 h (ml) (x + SD.) Sheep tincture gel 3% 0.159 ± 0.01 ** 0.189 ± 0.01 ** 0.231 ± 0.02 ** 0.211 ± 0.03 ** Effect% -33.19 -31.52 -32.05 -27.98 Phenylbutazone gel 0.076 ± 0.01 ** 0.103 ± 0.03 ** 0.135 ± 0.02 ** 0.104 ± 0.04 ** Effect% - 68.57 -62.65 - 60.05 -64.50

X ± SD = mean ± standard deviation; ** p <0.05.

In the experimental model of edema produced with 6% dextran solution, the gel with tincture of 5% showed the most intense anti-inflammatory effect after 90 minutes after the induction of edema (46.82%).

Gel with tincture of 3% had the most intense anti-inflammatory effect after 60 minutes after the induction of edema, the inflammation produced with 6% dextran solution being reduced by 37.00%.

The phenylbutazone gel showed the most intense anti-inflammatory effect after 30 minutes after the induction of edema, the inflammation produced with 6% dextran solution being reduced by 72.41%) (Table 2).

Table 2

Anti-inflammatory effect of gum tincture studied in inflammatory edema produced with 6% dextran solution

Product tested Edem 30 min (ml) (x + SD.) Edema 60 min (ml) (X ± 5D.) Edema 90 min (ml) (X ± 5D.) Edem 120 min (ml) (x + SD.) witnesses 0.203 ± 0.02 0.227 ± 0.02 0.252 ± 0.03 0.265 ± 0.03 Sheep tincture gel 5% 0.114 ± 0.01 ** 0.128 ± 0.01 ** 0.134 ± 0.01 ** 0.151 ± 0.01 ** Effect% -43.84 -43.61 -46.82 -43.01 Sheep tincture gel 3% 0.129 ± 0.01 ** 0.143 ± 0.02 ** 0.163 ± 0.01 ** 0.177 ± 0.03 ** Effect% -36.45 -37.00 -35.31 -33.73 Gel with phenylbutazone 0.056 ± 0.04 ** 0.065 ± 0.01 ** 0.074 ± 0.02 ** 0.074 ± 0.01 ** Effect% / initial -72.41 -71.36 -70.63 -72.07

X ± SD = mean ± standard deviation; ** p <0.05.

By testing the anti-inflammatory action of the gels in the study, it was found that the gel with 5% itch tincture has significant anti-inflammatory effects on both the edema produced by the 10% kaolin suspension and the edema produced by the 6% dextran solution. This is due to the complex composition of the shepherd's tincture (polyphenols, flavones).

In case of edema produced with 10% kaolin suspension, the most intense anti-inflammatory effect occurred within 2 h after the induction of edema, for all gels studied, as well as for phenylbutazone.

in the case of edema produced with 6% dextran solution, the most intense anti-inflammatory effect was observed after 90 minutes after induction of edema for 5% stain gel, after 60 minutes after induction of edema for 3% propolis tincture gel, and after 30 min for the phenylbutazone gel.

The following is an example of an embodiment of the invention:

Example

The paste is subjected to extraction by maceration with food ethyl alcohol 70 °, in the quantity ratio 20 ... 30% paste and 70 ... 80% extraction agent. The extraction of active principles from the shepherd is cold, 15 ... 18 ° C, for 15 days. The mixture subjected to extraction is stirred daily 2 to 3 times throughout the extraction.

The product resulting from the extraction is stored cold and, before packing, subjected to a filtering operation, in order to remove any mechanical impurities. The filtrate is packed, in compliance with the sanitary-veterinary norms, in individual packages, made of brown glass, sealed.

The storage is carried out under specific conditions: in dry and light-free premises.

The organoleptic characteristics of the shepherd's tincture are:

Appearance, consistency and color Clear, brown liquid The smell Characteristically, masked by the smell of alcohol used as an extraction agent The taste Characteristically, masked by the taste of alcohol used as an extraction agent

The physical-chemical characteristics are:

solubility Completely soluble in water Density (g / cm ³ ) 0.8 pH 4.5 ... 5.5 Water content (% vol) 20 ... 25% Ethanol content (% vol) 50 ... 55% Mechanical impurities Lack

Use:

Claims (2)

1. In the normal life regime, autumn and spring, for strengthening immunity, 3-4 weeks internal cures: adults: 2 x 1 ml / day; children under 14 years: 2 x 0.5-1 ml / day; children under 3 years: 1 drop / kg body / day, administered in 2 shots. It is administered mixed with honey bee or dripped on wheat bran, brown sugar, bread, biscuits, etc., in the morning and evening, on an empty stomach. 2. For people with health problems: the doses are increased according to the indications of an apifitotherapist doctor. External use: disinfectant, healing agent; mouth water: rinse with 20 to 30 drops solution tincture to 20 ... 30 ml water (also useful in periodontics). RO 129062 Β1 The antimicrobial and antifungal action was tested by the diffusimetric method: As culture media, the following were used: - for bacteria - nutritional jealousy, distributed in Petri dishes; - for fungi - solid Sabouraud medium, distributed in Petri dishes. inoculum: - for bacteria - 24 h bacterial cultures in nutritional broth, diluted 1/100; - for fungi - cultures of 24 hours in liquid Sabouraud medium, diluted 1/100 for yeasts of the genus Candida; culture for 48 hours in liquid Sabouraud medium, diluted 1/100 for Cryptococcus neoformans', suspension in distilled water of spores of Aspergillus niger. Product analyzed: - a sample of shepherd's tincture was evaporated to dryness at room temperature, after which the residue was dissolved in distilled water at 1 to 2 p distilled water; - the solvent, the distilled water, was used as a control. Working technique: The test microorganisms were seeded by microincorporation in the respective fluidized environment and cooled to 56 ... 60 ° C. The procedure was as follows: 1 ml of the inoculum was aseptically distributed in a sterile Petri dish, over which 15 ml of fluidized medium was cooled and cooled to 56 ... 60 ° C. The inoculum was homogenized by circular and perpendicular movements of the plate horizontally. The solidification of the seeded medium was then expected, then with a sterile glass tube, 8 mm in diameter, the wells were cut in the middle layer. With sterile spatula, the samples to be analyzed were introduced into wells. Plates were incubated at 37 ° C for bacteria and yeast for 48 h, and at 28 ° C for 5 days for Aspergillus niger filamentous fungus. In order to interpret the results, the diameter (mm) of the microbial growth inhibition zones around the washers impregnated with the tested products was measured. The diameter of the growth inhibition zones of the test microorganisms (mm) is shown in the following table: Micro-organism 10% tincture 20% tincture 30% tincture Staphylococcus aureus 15 28 32 Bacilus subtilus 19 32 36 Mycobacterium smegmatis 20 24 30 Candida albicans 3 22 24 Candida tropicalis 8 22 25 Candida pseudotropicalis 16 26 30 Candida krusei 16 28 31 Cryptococcus neoformans 18 34 38 Aspergillus niger 16 32 35 There is a strong antimicrobial and antifungal action for sheep dye with a concentration of 30%.