RU2552291C2 - Method of separating biologically active compounds from propolis by vacuum ultrasonic extraction - Google Patents

Abstract

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

Description

The invention relates to medicine and the perfume industry and relates to a method for purifying propolis from heavy metals, toxins and radioactive substances to obtain biologically pure active groups used as medicines.

Closest to the proposed method by technical essence, we can recognize the method of producing alcoholic propolis extract by vacuum-ultrasonic extraction (patent No. 23729280, however, it does not produce dry biologically active compounds from which standardized drugs for various applications can be prepared.

The purpose of the invention is to obtain a purified dry substance of propolis while cleaning them from heavy metals, such as (Cu, Pb, Zn, Cd) and from the remnants of therapeutic drugs with toxicity used in the treatment of bees, as well as from pesticides that enter the hive when cultivation of agricultural land, while reducing the time of extraction of extractive substances and concentrating them to a dry state with a moisture content of 10-12%, using for this purpose vacuum-ultrasonic extraction of continuous extraction and propolis.

The goal of obtaining a dry purified substance of propolis was achieved by grinding raw propolis, previously frozen at a temperature of -16-20 ° C and extraction with ethanol 96 ° at a temperature of + 20-25 ° C.

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

The purified propolis alcohol extract is vacuum evaporated to a moisture content of 9-12%. Ready dry substance of propolis is easily dissolved in isotonic solution. From a dry substance, you can prepare a standardized alcohol dosage form, as well as prepare ointments for medical therapeutic use.

Example 1. A method of obtaining dry propolis.

We have developed a simple method for preliminary purification of raw propolis, which allows to significantly eliminate wax and mechanical impurities (see Fig. No. 1). The ground cooked propolis is loaded into glass columns No. 1 and No. 3, 20 kg of propolis each. The speakers are equipped with an ultrasonic device nos. 11 and 12.

Columns No. 1, No. 2 and No. 3 have a diameter of 0.2 m and a height of 1.2 m with a filter device at the top and bottom (No. 13 and No. 14).

All columns are sealed. An ultrasound device with a power consumption of 5 volts and having a mass of 200 g, which can be connected to an AC electrical network using remote control, is installed in the center of the speakers. Columns No. 1 and No. 3 are equipped at the top and bottom with multi-way opening and closing taps (No. 15). A vacuum hose connected to a vacuum pump (No. 9) is attached to the upper crane of columns No. 1 and No. 3, and the lower crane of columns No. 1 and No. 3 is connected through a hose to a centrifugal pump (No. 10) and the lower taps of columns No. 1 and No. 3 .

From a container (No. 6), where 96 ° concentration alcohol is prepared, a pump (No. 18) is used to fill a column (No. 2) of 35-40 liters of ethyl alcohol, and vacuum vacuum in the columns (No. 1 and No.) is turned on. A vacuum gauge (No. 19) is installed in the upper part of the column (No. 1), which controls the vacuum in the columns (No. 1 and No.).

The ultrasonic device is turned on when the ethanol temperature reaches 20-25 ° C and the vacuum pressure becomes 0.9-0.95 ati, then the column is filled with cooked alcohol from the column (No. 2), opening the bottom crane of the column (No. 1 and No. 2) and through the centrifugal pump using the bottom crane of the column (No. 1), the ethanol supply is manually controlled so that the pressure in the column (No. 1) does not drop below 0.65 bar.

When column No. 1 is filled with alcohol to the upper drain fitting and the extract is drained into column No. 2, the drain rate is reduced and extraction is carried out for 1.5-2 hours at a temperature of + 20-25 ° C in a moving alcohol layer with the device turned on. This leads to an increase in the diffusion of molecules of biologically active compounds from the depths of propolis pieces.

By this we achieve a decrease in the mean free path of the active compounds to the surface of the extract at low temperatures, without losing a large expenditure of energy for heating, thereby eliminating the transfer of wax and other substances to the extract. The ultrasonic device acts on propolis and leads to its powder state, thereby increasing the completeness of extraction of biologically active substances. After 1.5-2 hours of extraction, the vacuum and the ultrasonic device are turned off and, under a pressure of 0.5 atm, through the bottom valve of column No. 1, the alcohol extract of propolis is transferred under pressure to a container No. 4, washed with propolis cake after extraction with 5 liters of ethanol 96% through the top fitting of column No. 1, and then the propolis cake is removed from the column to load with new propolis.

In columns No. 2 and No. 3, the same technological operations are carried out as in column No. 1, the combined alcohol extracts of propolis obtained by extraction in columns No. 1 and No. 3 enter a tank No. 4 equipped with a mixing device. Propolis alcohol extract is treated with multi-functional charcoal sorbent (know-how) to remove heavy metals, pesticides and drug residues from propolis used to treat bees that are toxic.

2 g of a multifunctional sorbent is added per 1 liter of alcohol extract of propolis, periodically stirring for 3 hours, then filtered or separated on a separator. The obtained purified extract from the tank No. 4, freed from impurities, enters the tank No. 5. From the tank No. 5, the extract enters the vacuum evaporation unit No. 7 and evaporated to a vacuum of dry lumps under reduced pressure with a degree of vacuum (0.9-10 ^-1 ). Unloaded from the evaporation tank and dried in a vacuum oven on a baking sheet at 45-50 ° C, adjusted to a moisture content of 9-12%. The finished propolis powder in the amount of 2170.0 g is packed in glass one-liter or two-liter cans of dark glass with hermetically screwed caps or plastic bags and hermetically wrapped, removing air.

The guaranteed shelf life of dry propolis is 4 years from the date of manufacture.

For comparison, propolis was taken from six regions of the Russian Federation and vacuum-ultrasonic extraction and purification with a multifunctional sorbent were performed.

The following table 1 shows the results of the microbiological purity of the dry substance of propolis, which was obtained after vacuum-ultrasonic treatment in the device described above.

Propolis was taken from the Oryol region (No. 1), Krasnodar Territory (No. 2), Pskov Region (No. 3), Mordovia (No. 4), Tula Region (No. 5) and Kursk Region (No. 6), which are indicated in the table under the corresponding by numbers: 1, 2, 3, 4, 5, 6.

Table 1. No. of samples from different regions Bacteria and fungi in total in 1 ml not more than 10 ² Enterobakteraceae and some other gram-negative bacteria in 1 g not more than 10 ^-1 Pseudomonas, Aeruginaosa. Staphylococcus, Freus must be absent one. 36 6 Absent 2. 40 8 - "- 3. 45 four - "- four. 37 6 - "- 5. 40 5 - "- 6. 35 3 - "-

All tested samples of dry purified propolis complies with the requirements of Global Fund XI, issue 2, p. 193 Change No. 3, cat.2.

Table 2. Antimicrobial action of the dry substance of propolis obtained from six different regions of the Russian Federation. No. of samples of regions of the Russian Federation Antimicrobial action Dilution 1: 300 1: 600 1: 1200 The content of heavy metals is not more than 0.001% Note 1: 300 1: 600 1: 1200 one. - + + <0.001 Acc. FS 2. + + <0.001 - "- 3. - + + <0.001 - "- four. - + + <0.001 - "- 5. - + + <0.001 - "- 6. - + + <0.001 - "-

From the data obtained in table 2 it follows that all samples inhibit the growth of the test microorganisms in a dilution of 1: 300

Table 3. The results of the content of the amount of phenolic compounds of dry purified propolis by vacuum-ultrasonic extraction. See Fig.№2 No. of samples of the regions of the Russian Federation The number of propolis dry peeled, taken in g Amount of added alcohol 80% ml Contents the amount of phenolic compounds,% Note: the content of phenolic compounds 2-6% one. 10 up to 1000 5.24 Acc. FS 2. 10 up to 1000 4.26 - "- 3. 10 up to 1000 5.06 - "- four. 10 up to 1000 4.70 - "- 5. 10 up to 1000 4.25 - "- 6. 10 up to 1000 4.93 - "-

Based on the preparation of a dry substance of propolis with purification with multifunctional sorbitol, the studies showed that the biological activity of propolis is interconnected with the purity of the compounds identified in it and, therefore, it allows to obtain a higher quality dry substance and propolis tincture corresponding to regulatory documentation.

Thus, vacuum-ultrasonic technology has been developed in a mobile alcohol extract for producing tincture propolis, which increases the yield of biologically active substances, separates propolis from wax, solids, tar lipid and other low molecular weight and high molecular weight substances, heavy metals, as well as pesticides that get into propolis .

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

The determination of phenolic propolis compounds is presented in samples 1, 2, 3, 4, 5, 6, which indicates the high quality of dry propolis.

Claims (1)

A method of obtaining dry propolis for the preparation of medical preparations containing propolis, which includes grinding raw materials, extraction with ethanol 96% at a temperature of + 20-25 ° C using a vacuum ultrasonic device, filtering, purification of heavy metals and other impurities using a carbon sorbent and subsequent evaporation.

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