RU2535085C2 - Method of manufacturing antimicrobial preparation from aboveground part of ramson - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the pharmaceutical industry, namely to a method of obtaining a preparation, possessing an antimicrobial activity. The method of obtaining the preparation, possessing he antimicrobial activity, where an aboveground part of freshly harvested bear leek (Allium ursinum L.) or victory onion (Allium victoriale L.) is crushed to a paste-like state, poured with ethyl alcohol, infused tree times, under certain conditions, after each extraction the extracts are poured off, the obtained extracts are combined.

EFFECT: medication, obtained by the claimed method, possesses an expressed antimicrobial activity.

2 cl, 1 tbl, 6 ex

Description

The invention relates to medicine and relates to methods for producing preparations from plant materials - bear onion (Allium ursinum L.) and victorious onion (Allium victoriale L.) (wild garlic), onion family (Alliaceae Juss.), Which can be used as an antimicrobial agent .

Known phytoncide antibacterial agents obtained on the basis of plant materials (plants): alcohol tincture of onions ("Allilchep") [Russian Medicines Register. / Ed. Yu.F. Krylova.- M .: Infarmhim, 1993. - P.164.], Tablets with Alisat garlic powder, Allylsat liquid alcohol extract of garlic, etc. [Mashkovsky MD Medicines: reference edition, 15th ed., Revised., Rev. and add. / Mashkovsky M.D. - M .: New wave, 2006 - S.962]. The closest prototype is a liquid alcohol extract of garlic "Allylsat", used as a fungicidal, antimicrobial, anthelmintic, bacteriostatic agent, etc.

A method of obtaining a liquid alcoholic extract of garlic "Allilsat" is as follows: garlic bulbs are cleaned of scales and chopped in a meat grinder. 34.6 kg of garlic mass is placed in a hermetically sealed percolator, pour 52 l of 90% ethyl alcohol and infuse for 72 hours at room temperature. The resulting extract is drained, a second portion of alcohol in the amount of 24 l is poured into the percolator, and the infusion is repeated again. After 72 hours, the hood is drained and the infusion is repeated a third time with 24 l of extractant. After 72 hours and draining the hood, the pulp is squeezed in the press and the spin is attached to all hoods collected in a common sump. The extract, which has remained for 48 hours, is transferred to a mixer with an anchor mixer, 0.228 kg of essential oils (caraway, dill and mint at 0.3% each) are added, stirring the mixture for an hour with an anchor mixer. After stirring, the allylate is filtered. Determine the content of solids and alcohol in the extract [Muravyov I.A. Technology of drugs: In 2 T. / Muravyov I.A. - M., 1980 .-- T.1. - S.212-213].

The list of hoods for internal use, obtained from plant materials of the Allium species with antibacterial action, is limited. The disadvantages of the extract of garlic "Allylsat" are: the duration of extraction (216 hours), moderate antimicrobial activity. At the same time, the list of drugs with antibacterial action is limited due to the lack of an appropriate plant base. Therefore, the expansion of the arsenal of drugs characterized by antibacterial action is an urgent task.

The technical task of the invention is the development of a method of obtaining funds from the aerial part of freshly harvested wild garlic (bear onion - Allium ursinum l. Or victorious onion - Allium victoriale, onion family - Alliaceae) with a extraction time of 12 hours, characterized by high antibacterial activity.

100 kg of the aerial parts of freshly harvested wild garlic (bear onion - Allium ursinum L. or victorious onion - Allium victoriale L. onion family (Alliaceae)) are minced in a meat grinder and placed in a hermetically sealed percolator. The raw material is poured with ethyl alcohol 40% in the ratio of raw material extractant 1: 1.5 (150 l) and insist at room temperature for 4 hours. Then the resulting hood is drained. A second portion of 40% ethyl alcohol is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 L) and the infusion is repeated again at room temperature for 4 hours. Then the resulting hood is drained. The infusion is repeated for the third time with ethyl alcohol 40% in the ratio of raw material-extractant 1: 0.75 (75 l) and the infusion is repeated again at room temperature for 4 hours. The hood is drained, connected to the first and second hoods. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained. The extract is left to stand for 48 hours and the solids content is checked. Water-alcohol extracts of wild garlic are transparent liquids from light to dark green in color, with a characteristic garlic odor and a burning taste.

The obtained extracts were tested for antimicrobial activity.

The results of the experiment are presented in examples 1-6. The use of ethyl alcohol 40%, 70%, 90% as an extractant guarantees maximum extraction of active substances. The temperature regime, the duration of extraction and the extractant were identified experimentally and guarantee the optimality of the process.

Examples of specific performance of the method

Example 1

100 kg of the aerial parts of freshly harvested wild garlic (bear onion - Allium ursinum L.) are minced in a meat grinder and placed in a hermetically sealed percolator, filled with ethyl alcohol 40% in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature. Then the resulting hood is drained. A second portion of 40% ethyl alcohol is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 L) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained.

In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the content of solids (3.26%).

Example 2

100 kg of the aerial parts of freshly harvested wild garlic (bear onion - Allium ursinum L.) are minced in a meat grinder and placed in a hermetically sealed percolator, filled with ethyl alcohol 70% in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature. Then the resulting hood is drained. A second portion of ethyl alcohol 70% is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 l) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained. In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the content of solids (2.83%).

Example 3

100 kg of the aerial parts of freshly harvested wild garlic (bear onion - Allium ursinum L.) are minced in a meat grinder and placed in a hermetically sealed percolator, filled with 90% ethyl alcohol in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature. Then the resulting hood is drained. A second portion of ethyl alcohol 90% is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 l) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained. In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the solids content (2.15%).

Example 4

100 kg of the aerial parts of freshly harvested wild garlic (victorious Allium victoriale L. onion) are minced in a meat grinder and placed in a hermetically sealed percolator, poured with ethyl alcohol 40% in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature . Then the resulting hood is drained. A second portion of 40% ethyl alcohol is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 L) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained.

In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the solids content (2.13%).

Example 5

100 kg of the aerial parts of freshly harvested wild garlic (victorious Allium victoriale L. onion) are minced in a meat grinder and placed in a hermetically sealed percolator, filled with ethyl alcohol 70% in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature . Then the resulting hood is drained. A second portion of ethyl alcohol 70% is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 l) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained. In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the content of solids (2, 25%).

Example 6

100 kg of the aerial parts of freshly harvested wild garlic (victorious Allium victoriale L. onion) are minced in a meat grinder and placed in a hermetically sealed percolator, filled with ethyl alcohol 90% in the ratio of raw material extractant 1: 1.5 (150 l) and insisted for 4 hours at room temperature . Then the resulting hood is drained. A second portion of ethyl alcohol 90% is poured into the percolator in a ratio of raw material-extractant 1: 0.75 (75 l) and the infusion is repeated again. After four hours, the hood is drained. Insisting is repeated for the third time in the ratio of raw material extractant 1: 0.75 (75 l). After four hours, the hood is drained, connected to the first and second hoods in the sump. The pulp is squeezed in the press and the spin is attached to the hoods assembled into a common sump, 385 liters of hood are obtained. In parallel, determine the moisture content of the raw materials. All measurements are carried out with an accuracy of 0.0002 g. The completeness of the extraction is controlled by the content of the amount of oxidizable compounds and the content of solids (2.11%).

When studying the antimicrobial properties of the target product was studied:

1) extracts of wild garlic (bear onion - Allium ursinum L.) liquid alcohol, obtained using 40%, 70%, 90% ethyl alcohol,

2) extracts of wild garlic (victorious onion - Allium victoriale L.) liquid alcohol, obtained using 40%, 70%, 90% ethyl alcohol,

3) garlic extract liquid alcohol "Allylsat".

In the study of the antimicrobial activity of the target product, a method was used based on the ability of drugs to diffuse into agar infected with test cultures of microorganisms and inhibit the growth of the latter [Handbook of Microbiological and Virological Research Methods / Ed. Birgera M.O. - M .: Medicine, 1982. - S.172-177.].

The activity of the studied extracts was determined in 3 Petri dishes of the same diameter with a flat bottom. 15 ml of molten nutrient agar was poured into plates mounted horizontally. An agar layer in a Petri dish was poured with 1-2 ml of a suspension of the test microbe in physiological saline. Then the excess suspension was completely removed, the surface of the agar was dried for 30 minutes. Then 6 holes (“wells”) were drilled with a drill (6 mm in diameter) at a distance of 2.5 cm from the center and at the same distance from each other, “wells” were filled with samples of extracts and a solvent in an appropriate concentration (control). The cups were left at room temperature for 30 minutes, after which they were placed in a thermostat at a temperature of 37 ° C, without turning over, strictly horizontally to get round zones. A sterile filter was placed under the lids of the Petri dishes to prevent condensation from entering the wells. Inhibition zones were measured after 16 hours. Evaluation of the results was carried out according to the diameter of the zones of growth inhibition around the "well", including the diameter of the "well" itself:

- the absence of a zone of growth inhibition - the test culture is not sensitive to a given concentration of the drug;

- diameter of the zone of growth inhibition 10 mm - moderate sensitivity of the culture to a given concentration of the drug;

- diameter of the zone of growth inhibition of more than 10 mm - high sensitivity of the test culture to a given concentration of the drug.

The results of the experiment are presented in table 1.

The research results presented in table 1 show that wild garlic extracts (bear onions - Allium ursinum L.) obtained using 40%, 70%, 90% ethyl alcohol, and wild garlic extracts (victorious onion - Allium victoriale L.). ) obtained using 40%, 70%, 90% ethyl alcohol, have: a pronounced antimicrobial effect against gram-positive cocci of the genus Staphylococcus: Staphylococcus aureus (209); Staphylococcus aureus (Type); Staphylococcus epidermidis Wood - 46, Salmonella gallinarum; Escherichia coli 055; spore-forming microorganisms of the genus Bacillus: Bacillus anthracoides - 96; Bacillus anthracoides; Bacillus subtilis, as well as Pseudomonas aeruginosa and Proteus vulgaris (Table 1).

The control, Allilsat liquid garlic extract, showed moderate antimicrobial activity against Staphylococcus aureus (209); Staphylococcus aureus (Type); Staphylococcus epidermidis Wood - 46, Salmonella gallinarum; Escherichia coli 055; Bacillus anthracoides - 96; Bacillus anthracoides; Bacillus subtilis, Pseudomonas aeruginosa and Proteus vulgaris (Table 1).

Table 1 The results of the antibacterial action of wild garlic extracts Extractant The extraction time at each stage of extraction, h The diameter of the zone of growth inhibition of test cultures of microorganisms, mm one 2 3 four 5 6 7 8 9 10 eleven Wild garlic extract (bear onion) 40% four 32 thirty 33 32 28 25 28 29th 32 28 29th 70% four 22 eighteen 12 fourteen 13 12 fifteen fourteen 19 12 fourteen 90% four 10 10 10 10 12 13 17 fifteen 17 eleven 13 Wild garlic extract (victorious onion) 40% four 31 29th 32 33 29th 26 27 28 31 29th thirty 70% four 23 eighteen 12 13 12 12 fourteen 13 eighteen 12 13 90% four eleven eleven eleven eleven 13 fourteen eighteen 16 eighteen 12 fourteen Control (garlic extract "Allylsat") 90% 72 9 9 9 9 9 9 10 10 10 10 10 Test cultures: 1. Staphylococcus aureus (209); 2. Staphylococcus aureus (Type); 3 Staphylococcus epidermidis Wood-46; 4 Escherichia coli 675; 5. Escherichia coli 055; 6. Bacillus subtilis L2; 7. Bacillus anthracoides - 96; 8. Bacillus anthracoides - 2; 9. Salmonella gallinarum; 10. Pseudomonas aeruginosa; 11. Proteus vulgaris.

Claims (2)

1. The method of obtaining funds with antimicrobial activity, characterized in that the aerial part of freshly picked bear onions (Allium ursinum L.) is crushed to a mushy state, poured with ethyl alcohol 40%, or 70%, or 90%, insisted three times, for 4 hours at room temperature, with a ratio of raw material-extractant: during the first extraction 1: 1.5, the second extraction 1: 0.75, the third extraction 1: 0.75, after each extraction, the extracts are drained, the obtained extracts are combined. 2. A method of obtaining funds with antimicrobial activity, characterized in that the aerial part of the freshly picked victorious onion (Allium victoriale L.) is crushed to a mushy state, filled with ethyl alcohol 40%, or 70%, or 90%, insisted three times, for 4 hours at room temperature, with a ratio of raw material-extractant: during the first extraction 1: 1.5, the second extraction 1: 0.75, the third extraction 1: 0.75, after each extraction, the extracts are drained, the obtained extracts are combined.