RU2497539C1 - Antimicrobial activity of robinia seed essence - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, specifically to an agent showing antimicrobial activity.

EFFECT: agent possessing antimicrobial action and representing robinia (Robinia pseudaacacia L) seed essence prepared by collecting the seeds in October and extracting them three times in petroleum benzene under certain conditions.

2 tbl, 1 ex

Description

The invention relates to medicine, the medical industry and can be used, in particular, to the use of essential oil from the seeds of robinia pseudoacacia (Robinia pseudaacacia L.) as an agent with antimicrobial activity.

A known method of producing anti-decubitus oil [RF patent No. 2416425], containing in its composition fatty sea buckthorn oil and additionally fatty oils of tamanu and wheat germ, as well as essential oils of oregano, geranium, cedar, marjoram and other plants.

A known method of obtaining the composition [RF patent No. 23 72096], with fungicidal, antimicrobial and virucidal action, characterized in that it contains as a base oil, water-alcohol extracts and essential oil of monarda monica, propylene-glycolic extracts of pine and poplar, ethereal peppermint oil, bisilic oil, etc. The disadvantages of these methods are that they consist of many components, in addition, the simultaneous extraction of all components of plant materials collected at different times, as they ripen, force allows you to store a number of components in anticipation of others, which leads to the loss of their healing properties.

A known method of producing an extraction of essential oil from coniferous herbs [RF patent No. 2396309], characterized in that the oil exhibits bactericidal properties. The method involves grinding coniferous wood greenery and its extraction with liquefied carbon dioxide at a temperature of 20-22 ° C and a pressure of 6-7 MPa. The disadvantage of this method is the complexity of the technological operations for the extraction of essential oil and the use of expensive equipment.

A known method of producing a biologically active substance [application of the Russian Federation No. 94025595] with antimicrobial, phagocytic, mitotic and antioxidant activity, including the preparation of an ether concentrate by mixing peppermint oil and / or fennel, caraway, anise, dill, coriander, bergamot, lemon, orange eucalyptus, salty, ylang-ylang, clary-sage and other oils and an aqueous extract of wormwood grass, peppermint, ordinary pine buds, licorice and other plants. However, this method has several disadvantages, namely, the low degree of extraction of biologically active substances from plant materials into water, the technological difficulties of the fine filtration of the obtained extract, due to the large volume of the filtered mass, consisting of many components, which may affect the properties of the drug. In addition, the simultaneous extraction of all components of plant materials collected at different times, as they mature, forces the storage of a number of components in anticipation of others, which leads to the loss of their properties.

A known method of producing essential oil of anise lofant [RF application No. 2009143766] by steam extraction method containing methylchavicol, as well as substances with antifungal, bactericidal and disinfectant effects. The main disadvantage of this method is that the anise lofant is a plant introduced to Russia and the technology of cultivating it is quite laborious (regular watering of the plant, fertilizing and presowing cultivation of the land).

A known method of producing essential oil from wormwood [RF patent No. 2356567] by the method of steam distillation, which has antibacterial, disinfectant, antiseptic, anti-inflammatory, wound healing and anti-burn effect. However, these extracts and essential oils cannot be analogues, as they are obtained from other plants.

The problem solved by the invention is the study of the antimicrobial activity of essential oils from plant materials of Robinia pseudoacacia (Robinia pseudaacacia L.). The problem is solved using essential oil from the seeds of pseudoacacia robinia collected in the month of October and obtained by three times extraction with petroleum ether by heating for 45 minutes at a volume ratio of solvent to raw material of 3: 1.

Due to the presence of a complex of biologically active substances in it, essential oil of pseudo-acacia robinia exhibits noticeable antimicrobial activity to a wide range of both gram-negative and gram-positive microorganisms.

The method for studying antimicrobial activity was based on the study of the effect of the active substances of robinia essential oil on the respiratory system of microorganisms. The level of respiration or its suppression was observed according to the degree of glucose consumption with an indicator of methylene blue. Antimicrobial activity was determined by serial dilution in 0.5 ml of nutrient broth [Gerhard F. Methods of General bacteriology. M., 1983. 612 p.]. The strains of hospital infection pathogens were used as test cultures: Escherichia coli CK, Klebsiella 204, B-1919 Bacillus mesentericum-3, Pseudomonas aeruginosa 165, Staphylococcus aureus 209, Proteus vulgaris, Klebsiella pneumoniae, Acinetobusecuscus calcocouscus calcococeus calcococeus calcococecus calcococeus calcococeus calcococecus calcococeus calcococeus calcococeus calcococeus calcococeus calcococecus calcococeus calcococecus calcococeus calcococeus calcococus calcococus The antimicrobial activity of robinia pseudo-acacia essential oil is illustrated by the following example.

Example.

Pure bacterial cultures grown on nutrient agar medium placed in a thermostat at 37 ± 1 ° C were washed off from the medium after 18-20 hours with a 0.89% sodium chloride solution. A suspension was prepared according to the standard turbidity of 1 · 10 ⁹ cells / ml (10 units of turbidity). To obtain the appropriate dilution of microbial suspension, a series of seven-fold serial dilutions were prepared. After determining the "working dose" of the test culture, the essential oil was titrated by double dilution in a volume of 0.5 ml of meat-peptone broth. The last test tube in the control row did not contain essential oil and served as a control of the dihydrogenase activity of the cells of the test culture. As a result, the in vitro microbial load corresponded to the “working dose” of the test culture. Tubes with essential oil and a test culture were exposed for 24 h at 37 ° С, then a methylene blue indicator with glucose and meat-peptone agar was introduced, the contents of the tubes were mixed and incubated for an hour at 37 ± 1 ° С. The result was taken into account by the color of the nutrient medium, if the indicator discolored, then this indicated no suppression, if the color did not change, then this indicated that the respiratory enzymes of the test culture cells were blocked by the active components of the essential oil. Gentamicin sulfate was used as a comparison drug. The results were statistically processed using t-student test.

The results of the study of the antimicrobial activity of the essential oil of robinia pseudoacacia are shown in tables 1 and 2.

Thus, the essential oil from the seeds of Robinia pseudoacacia (Robinia pseudaacacia-L) exhibits a noticeable antimicrobial activity to a wide range of both gram-negative and gram-positive microorganisms.

Application.

Table 1 Antimicrobial activity of essential oil from the seeds of robinia pseudoacacia in relation to strains of gram-negative bacteria Test sample IPC (μg / ml) in relation to E.coli SC Klebsiella 204 Pseudomonas aeruginosa 165 Proteus vulgaris Klebsiella pneumoniae Acinetobacter calcoaceticus Robinia pseudoacacia twenty 5 5 ^* twenty 5 ^* 10 Gentamicin sulfate 2.5 1.25 0.63 ^* 2.5 1.25 1.25 ^* Differences between repetitions are significant at p = 0.95

table 2 Antimicrobial activity of essential oil from the seeds of robinia pseudoacacia in relation to strains of gram-positive bacteria Test sample IPC (μg / ml) in relation to B-1919 Bacillus Mesentericum-3 Staphylococcus aureus Staphylococcus aureus 209 Staphylococcus aureus MRSA Robinia pseudoacacia 5 ^* 0.63 0.63 ^* 5 Gentamicin sulfate 1.25 0.63 0.63 ^* 0.63 ^* Differences between repetitions are significant at p = 0.95

Claims (1)

The antimicrobial agent, which is an essential oil from the seeds of Robinia pseudoacacia (Robinia pseudaacacia L.), obtained by collecting seeds in the month of October, extracting them three times with petroleum ether while heating for 45 minutes at a ratio of 1: 3.