Method of separating biologically active compounds from propolis by vacuum ultrasonic extraction

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing dry propolis. Said method includes grinding raw material, extraction with 96% ethyl alcohol at temperature of +20-25°C using a vacuum-ultrasonic device, filtering, purification from heavy metals and other impurities using a carbon sorbent, followed by evaporation.

EFFECT: disclosed method increases output of biologically active substances and enables purification of propolis from wax, mechanical impurities, high-molecular weight substances, heavy metals and pesticides.

2 dwg, 3 tbl, 1 ex

 

The invention relates to medicine and perfumery industry and relates to a method of cleaning propolis from heavy metals, toxins and radioactive substances to produce organic active groups, used as medicines.

The closest to the proposed method to the technical nature, can be considered as a method of producing the alcoholic extract of propolis vacuum ultrasonic extraction (patent No. 23729280, however, he does not receive dry biologically active compounds, to prepare a standardized medicines for various applications.

The purpose of the invention is the obtaining of purified dry substance of propolis with simultaneous cleaning from heavy metals such as si, Pb, Zn, Cd) and from residues of medical drugs with toxicity used in treatment of bees and pesticides from entering the hive in the processing of agricultural land, while reducing the retrieval time of extractives and concentrating them to dry condition with moisture content of 10-12%, using for this purpose the vacuum-ultrasonic extraction continuous extraction of propolis.

The goal of obtaining a dry purified substance of propolis was achieved by grinding propolis raw �predvaritelno frozen at a temperature of 16-20°C and extraction with ethyl alcohol 96° at a temperature of 20-25°C.

In a glass vacuum ultrasonic device of continuous action, ethanolic extract of propolis treated multifunctional carbon sorbent (know-how) to remove heavy metals, radioactive substances, pesticides with high toxicity.

Purified ethanolic extract of propolis was subjected to vacuum evaporation to a moisture content of 9-12%. Finished dry substance propolis is readily soluble in saline solution. From the dry substance can be prepared standardized alcohol-dosage form, as well as to prepare ointments for medical use.

Example 1. Method for producing of dry propolis.

We have developed a simple method for pre-cleaning of propolis-raw, allowing to remove wax and contaminants (see figure No. 1). Chopped cooked propolis is loaded into glass columns No. 1 and No. 3 of 20 kg of propolis in each. The column is equipped with an ultrasonic device under No. 11 and No. 12.

Column No. 1, No. 2 and No. 3 have a diameter of 0, 2 m, height 1.2 m with filter unit above and below (#13 and # 14).

All columns are sealed. Centered within a column mounted ultrasonic device with the consuming power of 5 volts, and having a mass of 200 g, which may be included in electric network of alternating current with the help of �stantsionnogo management. Column No. 1 and No. 3 is provided above and below the multi-way opening and closing valves (No. 15). To the upper crane columns No. 1 and No. 3 is attached the vacuum hose connected to a vacuum pump (No. 9) and the bottom valve of the columns No. 1 and No. 3 is connected via a hose to a centrifugal pump (No. 10) and the lower valves of the columns No. 1 and No. 3.

From the tank (No. 6), where the prepared alcohol 96° concentration pump (No. 18) fill in the column (No. 2) 35-40 liters of ethyl alcohol, include the vacuum in the columns No. 1 and No.). In the upper part of the column (No. 1) installed a vacuum gauge (No. 19), which controls the negative pressure in the columns No. 1 and No.).

Ultrasonic device include when the temperature of ethanol will reach 20-25°C and the vacuum becomes equal to 0,9-0,95 Athy, then fill with the prepared column alcohol from the column (No. 2), open the bottom valve of the column (No. 1 and No. 2) and through the centrifugal pump through the bottom of the crane column (No. 1) regulate the flow of ethanol manually, so that the pressure in the column (No. 1) did not fall below 0.65 in Athy.

When filling out column No. 1 of the alcohol to the top of the drain fitting and the drain of the extract in column # 2, speed drain, reduce and within 1.5-2 hours to conduct the extraction at a temperature of 20-25°C in a mobile alcohol layer enabled device. This leads to increased diffusion of molecules of biologically activediary from the depths of pieces of propolis.

We achieve this reduction of resistance free path of the active compounds to the surface of the extract at low temperatures, without losing a large amount of energy for heating, thereby getting rid of the remainder of the wax and other substances in the extract. Ultrasonic device affects propolis and leads him to powder state, thereby increasing the completeness of extraction of biologically active substances. After 1,5-2 hours of extraction shut off the vacuum and ultrasonic device and under a pressure of 0.5 ATM through the bottom valve of column No. 1 of the ethanolic extract of propolis move under pressure into the container No. 4, washed the meal after extraction of propolis 5 liters of 96% ethanol through the top fitting of column No. 1, and then removed from the column propolisnoe cake to download new propolis.

In columns # 2 and # 3 are the same process operations as in column No. 1, the combined alcoholic extracts of propolis obtained by extraction in columns # 1 and # 3 act in the capacity of No. 4, equipped with a stirrer. Ethanolic extract of propolis treated multifunctional coal-sorbent (know-how) to remove from propolis heavy metals, pesticides and residues of therapeutic drugs used for the treatment of bees, which have toxicity.

2 g multifunction Corben�and add to 1 liter of alcoholic extract of propolis, stirring occasionally, for 3 hours, then filtered or separated in the separator. The obtained purified extract from tank No. 4, freed from impurities, is collected in the tank No. 5. From tank No. 5 the extract is fed to a vacuum evaporator No. 7 and under reduced pressure with the degree of dilution (0.9 to 10-1)is evaporated to the dry dilution lumps. Discharged from the evaporation vessel and dried in a vacuum Cabinet at a baking sheet at 45-50°C, adjusted to a moisture content of 9-12%. Powdered propolis in the amount of 2170,0 g packaged in one-litre pitcher glass or two-liter jars of dark glass with an airtight seal screw caps or plastic bags and tightly wrapped, removing the air.

Warranty period of storage of dry propolis is 4 years from the manufacturing date.

For comparison, I took propolis from six regions of the Russian Federation and perform the vacuum-ultrasonic extraction and purification of multifunctional sorbent.

Next, in table.1 presents the results of the microbiological purity of the dry substance of propolis, which was obtained after vacuum ultrasonic processing device described above.

Propolis taken from the Orel region (No. 1), Krasnodar territory (No. 2), Pskov oblast (No. 3), Mordovia (No. 4), Tula region (No. 5) and the Kursk region (No. 6), which are listed in the table under the solicitation�existing room: 1, 2, 3, 4, 5, 6.

Table 1.
No. samples from different regionsBacteria and fungi in total, 1 ml not more than 102Enterobakteraceae and some other gram-negative bacteria in 1 g not more than 10-1Pseudomonas, Aeruginaosa. Staphylococcus, Freus should be no
1.366Missing
2.408-"-
3.454-"-
4.376-"-
5.405-"-
6.353-"-

All investigated samples of dry purified propolis meets the requirements of the global Fund XI, issue.2 p. 193, amendment No. 3, cat.2.

Table 2.
Antimicrobial activity of dry substances of propolis obtained from six different regions of the Russian Federation.
Nos. of samples of regions of the Russian FederationAntimicrobial effect Breeding 1:300 1:600 1:1200The content of heavy metals not more than 0.001%Note
1:3001:6001:1200
1.-++<0,001ACC. FS
2.++<0,001-"-
3.-++<0,001-"-
4.-++<0,001-"-
-++<0,001-"-
6.-++<0,001-"-

From the data obtained in table.2 that all of the sample inhibit the growth of test microorganisms in a dilution of 1: 300

Table 3.
Results content the amount of phenolic compounds of dry purified propolis vacuum ultrasonic extraction Cm. Figure No. 2
No. samples of regions of the Russian FederationNumber of propolis dry cleaned, taken in gramsNumber of added alcohol 80% mlContents. the amount of phenolic compounds, %Note: the content of phenolic compounds 2-6%
1.10up to 10005,24ACC. FS
2.10up to 1000 4,26- "-
3.10up to 1000Of 5.06- "-
4.10up to 10004,70- "-
5.10up to 1000The 4.25- "-
6.10up to 10004,93- "-

Upon receiving the dry substance of propolis with cleaning multifunction sorbitol, studies have shown that the biological activity of propolis is interconnected with the purity of identified compounds in it and, therefore, results in a higher quality dry substance and propolis tincture, relevant regulatory documents.

Thus, the developed vacuum-ultrasonic technology in a mobile alcoholic extract of receipt of propolis tincture, which increases the yield of biologically active substances, separates propolis waxes, solids, resinous lipidrich and other niskama�molecular and high-molecular substances, heavy metals and pesticides entering the propolis.

Test the microbiological purity of the dry substance of propolis was performed in accordance with the requirements of the global Fund XI, issue.2 p. 193, amendment No. 3, category 2. Microbiological purity was carried out in 6 series obtained from different regions of the Russian Federation.

Determination of phenolic compounds of propolis are presented in samples 1, 2, 3, 4, 5, 6, that speaks to the high quality of dry propolis.

Method for producing of dry propolis for the preparation of medicinal preparations containing propolis, which consists of grinding of raw material, extraction with ethyl alcohol 96% at a temperature of 20-25°C with the use of vacuum-ultrasonic device for filtration as well as purification from heavy metals and other impurities with the use of carbon sorbent and subsequent evaporation.



 

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