RU2649812C1 - Method of extraction from solid vegetable raw material of chemical compounds for pressing zoopathogenic bacteria composition - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: invention relates to pharmaceutical industry, namely to method of obtaining oak bark essence that suppresses zoopathogenic bacteria. Method of obtaining oak bark essence that suppresses zoopathogenic bacteria, comprising treating pre-crushed oak bark with an electrochemically activated cathodic aqueous solution having pH of 7–8 Eh = -300 … -450 mV, with an optimal feed-to-solution ratio of 1:8–1:15, with constant stirring for at least 600 seconds, and subsequent processing of the prepared suspension by ultrasound with a frequency of 22 kHz and the intensity of ultrasound exposure within 50–70 W/cm² within 420–600 seconds.

EFFECT: above described method makes it possible to obtain oak bark essence, which effectively suppresses zoopathogenic bacteria.

1 cl, 3 tbl

Description

The invention relates to medicine, pharmaceutical and food industries and can be used in animal husbandry and other industries.

The proposed extraction method is carried out in several stages: the first is the processing of pre-crushed to fine fractions of solid plant materials with an electrochemically activated (ECA) cathode aqueous solution with the following parameters of pH 7-8 and Eh = -300 ... -450 mV for 600 s at a constant stirring the second - the impact on the prepared suspension by ultrasound with a frequency of 15-22 kHz and an intensity of ultrasonic exposure of 1-70 W / cm ² for 420-600 s.

Known methods for cold water extraction of flavonoids from medicinal plant materials by applying an electric field [1] and by placing the raw material in the cathode space of the cell [2], i.e. using the quality of ECA water.

Significantly accelerate the extraction process and the quality of the extracted substance allows the use of ultrasound.

Known methods of ultrasonic extraction: including a set of ultrasonic effects in the modes of broadband and resonant exposure [3]; including an ultrasonic waveguide [4]; including the use of a conical nozzle for an ultrasonic emitter, the parameters of exposure to ultrasound are determined [5].

However, these methods do not provide a sufficiently complete selection of biologically active substances from plant materials, and the disadvantages of the devices are the complexity of manufacture and installation.

The task of the invention is the intensification and acceleration of the extraction process while increasing the degree of extraction of substances.

The problem is solved, and the technological result is achieved by processing pre-ground solid vegetable raw materials with a particle size distribution of 70 to 2000 μm with a moisture content of not more than 20% [5] ECA by a cathode aqueous solution (pH 7-8 and Eh = -300 ... -450 mV) at the optimal ratio of raw material solution 1: 8-1: 15 [6] with constant stirring for at least 600 s [5] and subsequent ultrasonic extraction of the resulting suspension in the frequency range 15-22 kHz and the intensity of the impact of 50-70 W / cm ² for 420-600 s, after which the resulting ex Rakta strained through a filter.

In our experiment, we used oak bark (Quercus cortex) as plant raw materials.

Based on numerous studies, it was found that the composition of the oak bark extract includes molecular compositions of substances that have antibacterial activity by suppressing the collective bacterial “quorum sensing” [7, 8, 9], and oak protistocidal substances can completely kill protozoa [10 ].

Bacterial “sense of quorum” refers to the ability of microorganisms to communicate and coordinate their behavior through the secretion of molecular signals.

The proposed technical solution is an extraction method for obtaining from a suspension of oak bark a composition of chemical compounds that inhibit the vital activity of pathogenic microorganisms.

The effectiveness of practical applicability was carried out by us in an experiment in the cultivation of broiler chickens.

An aqueous solution of oak bark extract obtained by the proposed method includes six chemical compounds: 4- (3-hydroxy-1-propenyl) -2-methoxyphenol - 0.9 mg / ml;

3,4,5-trimethylhydroxyphenol - 0.4 mg / ml;

4-n-propyl-1,3-benzenediol - 0.3 mg / ml;

4-hydroxy-3-methoxybenzaldehyde - 0.1 mg / ml;

7-hydroxy-6-methoxy-2H-1-benzopyran-2-one - 0.1 mg / ml;

2H-1-benzopyranon-2 - 0.06 mg / ml, respectively having the usual trivial names: coniferyl alcohol, antiarol, propylresorcinol, vanillin, coumarin and scopoletin, identified using chromatography-mass spectrometry and oriented to suppress cavity-dependent communication zoopathogenic bacteria.

The study of the molecular composition of chemical compounds in the C. violaceum CV026 bioassay allowed reproducing the anti-QS effect of the initial plant extract.

The studies were conducted in the conditions of the experimental-biological clinic (vivarium) of the Federal State Budget Educational Institution of Higher Professional Education “Orenburg State University” on broilers of cross-country “Smena-7”. For the experiment, 48 heads of 11-day-old broiler chickens were selected, which were divided into 4 groups by the method of analogues (n = 12). During the experiment, the whole bird was in the same conditions.

The formation of general diets (RR) for the experimental bird during the studies was carried out taking into account the recommendations of VNITIP [11].

The microclimate in the room met the requirements of VNITIP. Experienced poultry was fed 2 times a day, and eatability was recorded daily. Watering was carried out ad libitum.

Upon reaching 15 days of age, an aqueous solution of the composition was additionally included in the diets of the I, II, and III experimental groups. The optimal dosage of the composition in groups was established experimentally.

The control group is OP; I experimental - OP + 1 ml of an aqueous solution of the composition per kg of live weight (m); II experimental - OP + 2 ml of an aqueous solution of the composition per kg of oil; III experimental - OR + 3 ml of an aqueous solution of the composition per kg g.m. Feeding and watering the birds was carried out by the group method.

The solution of the composition of chemicals was set individually, daily for 6 weeks (table. 1).

After a 6-week period, bacterial contamination of broiler chickens was evaluated, according to which no pathogenic and conditionally pathogenic microflora were found in birds of experimental groups II and III (Table 2).

According to the results of the experiment, the proposed method for extracting oak bark is effective both for suppressing pathogenic microflora and for increasing the gain in live weight (Table 3).

After a 5-week feeding period, poultry of the experimental group III exceeded the analogues of the control group by 20.3% in terms of live weight and 22.5% in average daily growth (Table 3).

Thus, the method of extraction of oak bark, including primary processing of pre-crushed raw materials to a particle size distribution of 70 to 2000 μm with a moisture content of not more than 20% ECA by a cathode aqueous solution with a pH of 7-8 and Eh = -300 ... -450 mV with an optimal ratio of raw materials a solution of 1: 8-1: 15 with constant stirring for at least 600 s and subsequent processing of the prepared suspension with ultrasound at a frequency of 22 kHz with an ultrasonic intensity of 70 W / cm ² for 600 s allows you to get a composition containing six chemical soy distinctions: 4- (3-hydroxy-1-propenyl) -2-methoxyphenol at a dose of 0.9 mg / ml; 3,4,5-trimethylhydroxyphenol - 0.4 mg / ml; 4-n-propyl-1,3-benzenediol - 0.3 mg / ml; 4-hydroxy-3-methoxybenzaldehyde - 0.1 mg / ml; 7-hydroxy-6-methoxy-2H-1-benzopyran-2-one - 0.1 mg / ml; 2H-1-benzopyranon-2 - 0.06 mg / ml, which helps to reduce the development of opportunistic microflora in the body of farm birds and increase its productivity.

Information sources

1. Patent for invention RU No. 2453322. The method of cold water extraction of flavonoids from medicinal plant materials. Published on June 20, 2012.

2. Patent for invention RU No. 2146938. A method of preparing aqueous plant extracts. Published 03/27/2000.

3. Patent for invention RU No. 2390364. The method of extraction of biologically active substances from plant materials. Published on May 27th, 2010.

4. Patent for invention RU No. 2053006. Extractor for the solid-liquid system. Published on January 27th, 1996.

5. Patent for the invention RU No. 2104733. The method of extraction from solid plant materials. Published on February 20, 1998.

6. Hakobyan VB, Ershov Yu.A. The basics of the interaction of ultrasound with biological objects. - M .: Publishing house of MGTU im. Bauman. 2005.222 s.

7. Md. Akil Hossain, Seung-Jin Lee, Ji-Yong Park, Md. Ahsanur Reza, Tae-Hwan Kim, Ki-Ja Lee, Joo-Won Suh, Seung-Chun Park. Modulation of quorum sensing-controlled virulence factors by Nymphaea tetragona (water lily) extract // Journal of Ethnopharmacology 174 (2015). 482-491.

8. Deryabin D.G., Tolmacheva A.A. Antibacterial and anti-quorum sensing molecular composition derived from quvercus cortex (oak bark) extract // Molecules. 2015. T. 20. No. 9. C. 17093-17108.

9. Patent for invention RU No. 2542464. Composition based on phytoextracts that suppresses the sense of quorum in bacteria. Posted 02/20/2015.

10. Tokin B.P. Healing plant poisons. - L .: Publishing house Leningr. University, 1980 .-- S. 25.

11. Fisinin V.I., Imangulov Sh.A., Egorov I.A., Okolelova T.M. and other recommendations for feeding poultry. Sergiev Posad, 2000 .-- 67.

Claims (2)

1. A method of obtaining an extract from oak bark that suppresses zoopathogenic bacteria, comprising treating pre-crushed oak bark to a particle size distribution of 70 to 2000 μm with a moisture content of not more than 20% electrochemically activated cathode aqueous solution with a pH of 7-8 Eh = -300 ... -450 mV, with an optimal ratio of raw material-solution 1: 8-1: 15, with constant stirring for at least 600 seconds, and subsequent processing of the prepared suspension with ultrasound at a frequency of 22 kHz and an intensity of ultrasonic exposure for 50-70 W / cm ² those ix 420-600 seconds. 2. The method according to p. 1, in which the extract from oak bark is focused on the suppression of cavity-dependent communication of zoopathogenic bacteria and contains six chemical compounds: 4- (3-hydroxy-1-propenyl) -2-methoxyphenol at a dose of 0.9 mg / ml; 3,4,5-trimethylhydroxyphenol - 0.4 mg / ml; 4-n-propyl-1,3-benzenediol - 0.3 mg / ml; 4-hydroxy-3 methoxybenzaldehyde - 0.1 mg / ml; 7-hydroxy-6-methoxy-2H-1-benzene-pyran-2-one - 0.1 mg / ml; 2H-1-benzopyranon-2 - 0.06 mg / ml, which helps to reduce the development of opportunistic microflora in the body of farm birds and increase its productivity.