Method of obtaining material with antibacterial properties based on montmorillonite-containing clays

FIELD: chemistry.

SUBSTANCE: inorganic clay, represented by sodium-calcium, and/or calcium and/or ferrous forms of montmorillonite, is modified with a water solution of silver nitrate with a concentration 0.16-9.9 wt % in a weight ratio clay:water solution of silver nitrate 1:5. Modification is carried out with mixing from 3 to 7 hours at a temperature in the interval from 10°C to the temperature of boiling. The obtained material is washed with distilled water to pH ≈6-5, until excess of silver nitrate is removed, stood at room temperature and decanted. The material is dried at a temperature of 20-160°C.

EFFECT: obtaining an efficient antibacterial material for traditional and veterinary medicine.

2 tbl, 5 ex

 

The invention relates to methods for antibacterial materials by means of modification of natural montmorillonite clays to improve their anti-bacterial characteristics, which allows their use in traditional and veterinary medicine when creating complex of antibacterial drugs.

A method of obtaining an antibacterial material (patent RU 2429857, publ. 2011), which offers the first stage bentonite Na-form to activate the sodium chloride solution with the subsequent removal of anions of chlorine in washing and filtering, at the second stage, the resulting intermediate product intercalary metal ions bactericidal action, for example Ag+by processing in aqueous solutions of inorganic salts of these metals. Remove sodium salt when washing the product with deionized water, filtered, dried and pulverized to a dispersion of particles of 20-150 nm, where according to the invention the activation processes and the intercalation of bentonite carried out using ultrasound with a frequency of 20-50 kHz and intensity 10-100 watts/cm2and the clean up process intercalated product from the sodium salt produced in two stages, first the product is decanted, and the second phase is washed with deionized water containing 30 ppm - 100 ppm of the complexing agents of alkali metal ions on snowcrown-esters with a molecular weight of less than 264. While the resulting material contains 2.35 and 2.95 wt.% silver.

The known method of producing antimicrobial drug (patent US 2012093907 (A1), publ. 2012), according to which 1% of the mass. suspension of nanosilicate plates treated with a solution of silver nitrate (AgNO3) (1% wt.) when the ratio Ag+: clay is 7:93. Add 6~8 ml of methanol. Chemical interaction is carried out with the aid of ultrasonic agitation in a water bath at 70~80°C. the Disadvantage of this method is obtained prior nanosilicate plates of the layered clay minerals, which significantly affects the duration of the process. Also use methanol, which is a toxic substance. The silver content in the samples, a certain mass spectrometry (ICP-MS), corresponds to approximately 120 to 190 parts per billion.

The closest is essentially the invention consists in the modification of inorganic mineral montmorillonite inorganic metal salts in a polar solvent and the subsequent exposure of bentonite in the salt solution, removing the products of the modification of bentonite from the solution, followed by drying at a temperature not exceeding 100°C, according to the invention before the modification of bentonite enriched with ions of Na+by processing 3-10 wt.% aqueous solution is aristovo sodium with subsequent washing and filtering the obtained semi-finished product, then modify 10-20 wt.% a solution of inorganic metal salts, which use silver nitrate or copper sulfate, to produce an extract of modified bentonite in these salt solutions within 12-24 hours, and then cleaning premodification bentonite from the sodium salts by washing and filtration. After drying, the resulting preparation is crushed to a dispersion of particles of 20-150 nm, while the processing of inorganic mineral called solutions are produced at a ratio of mass - bentonite:solution 1:(10-40) (A.A. Abrahamian, V.I. Beklemishev, I.I. Makhonin, P.I. Makhonin, VA Solodovnikov. Method of producing antimicrobial drug. [Description of the invention to the patent RU 2330673 from 22.11.2006]).

The disadvantage of the prototype, as analogues of the above, is that they use only the Na-form of montmorillonite, which limits resource base, because more often montmorillonite clay, presents the sodium-calcium and/or calcium, and/or glandular form of montmorillonite. Also the disadvantages of the prototype can be attributed to the processing and use in the modification of high concentrations of silver nitrate.

The present invention is the creation of a material with antibacterial properties based on non-deficient montmorillonite-containing clay, which mo is no effectively suppress the growth of pathogenic microorganisms.

The technical result of the invention to provide an effective antibacterial material on the basis of non-deficient sodium-calcium and/or calcium and/or iron montmorillonite due to less expensive ingredients used and the duration of the technological process of modifying the silver ions using solutions of AgNO3lower concentrations.

The invention consists in the modification of clays, including inorganic mineral montmorillonite, a silver nitrate solution, washing and subsequent drying, includes the following new features:

- clay, including inorganic mineral montmorillonite, presents the sodium-calcium and/or calcium, and/or glandular form of montmorillonite;

- mass ratio of clay:the modifying agent is 1:5;

- the process of modifying the clay with an aqueous solution of silver nitrate AgNO3with a concentration of 0.16-9.9 wt.% carried out at a temperature in the range from 10°C to the boiling temperature, duration of treatment from 3 to 7 hours;

- obtained modified material was washed with distilled water until and unless you remove the excess silver nitrate to pH≈5-6;

- defend at room temperature and decanted;

- the material is dried at a temperature 20-16°C, in the result that are soft, easily crushed clay material from light brown to dark brown.

The method is implemented as follows.

The mineralogical composition of the original non-deficient sodium-calcium and/or calcium, and/or ferruginous clays:montmorillonite, illite, kaolinite, quartz, Muscovite, calcite, feldspars, where the main sorption material is montmorillonite.

The original clay is poured modifying the silver nitrate solution with a concentration of 0.16-9.9 wt.% the ratio of clay:the modifying agent, equal to 1:5. Stirred for 3 to 7 hours at a temperature from 10°C to the boiling point. Once the product is washed until pH≈5-6 to remove excess silver nitrate and dried at a temperature of 20-160°C. Drying at a temperature less than 20°C occurs in a considerable interval of time and requires the use of cooling equipment that is not economically feasible. The material dried at a temperature over 160°C, have a lower antibacterial action.

Examples

Example 1. The original clay was poured modifying the silver nitrate solution with a concentration of 3.2 wt.% the ratio of clay:the modifying agent, equal to 1:5. Was stirred for 3 hours at a temperature of 20-30°C. Upon completion of the process polucen the th material was washed to remove excess silver nitrate to pH≈5-6, dried at a temperature of 20-40°C.

Example 2. The original clay was poured modifying the silver nitrate solution with a concentration of 3.2 wt.% the ratio of clay:the modifying agent, equal to 1:5. Was stirred for 3 hours at the boiling temperature of the mixture. When the process is complete, the resulting material is washed to remove excess silver nitrate to pH≈5-6, dried at a temperature of 80-105°C.

Example 3. The original clay was poured modifying the silver nitrate solution with a concentration of 0.16 wt.% the ratio of clay:the modifying agent is 1:berelatively for 7 hours at a temperature of 20-30°C. Upon completion of the process, the resulting material is washed to remove excess silver nitrate to pH=5-6, dried at a temperature of 100-160°C.

Example 4. The original clay was poured modifying the silver nitrate solution with a concentration of 0.16 wt.% the ratio of clay:the modifying agent, equal to 1:5. Was stirred for 7 hours at the boiling temperature of the mixture. When the process is complete, the resulting material is washed to remove excess silver nitrate to pH≈5-6, dried at a temperature of 100-160°C.

Example 5. The original clay was poured modifying the silver nitrate solution with a concentration of 9.9 wt.% the ratio of clay:the modifying agent, equal to 1:5. Was stirred for 3 hours at a temperature of 10-15°C. At superseries the resulting material was washed to remove excess silver nitrate to pH≈5-6, dried at a temperature of 100-160°C.

Samples of materials for examples 1-5 was determined by the silver content. Research method was quantitative analysis based on measuring the volume or mass of reagent required for the reaction with the test substance - titrimetrically analysis.

Titrimetrically analysis to determine the amount of silver in the materials obtained according to examples 1-5 were carried out using indicators that secures the equivalence point of the titration. When conducting titrimetricheskogo analysis, by definition, in the samples of silver content (wt.%) used as reagents concentrated nitric acid as the titrant solution - ammonium thiocyanate or potassium, an indicator solution salesonline alum. As a result of the research showed that the sample of example 1 contains 3.36 wt.% silver, the study sample according to example 2 contains 3.61 wt.% silver, test specimens according to example 3 and 4 contain silver in the amount of 0.10 and 0.20 wt.% accordingly, the study sample according to example 5 contains 4,35 wt.% silver. In addition, the chemical composition enriched and modified samples of materials according to examples 1-5 were determined by x-ray fluorescence analysis x-ray spectrometer ARL OPTIM'X table 1).

Table 1
Average chemical composition of samples of materials in terms of oxides, wt.%
The material sampleSiO2Al2O3Fe2O3TiO2MgOCaOK2ONa2OAg2OΣ
Enriched clay60,1219,365,270,943.04 from8,872,400,38-100,38
Example 159,3713,404,370,892,058,284,050,266,9999,60
Example 259,6916,794,060,922,346,182,167,5199,65
Example 359,8718,90of 4.380,853,038,703,760,2199,09
Example 460,3118,605,900,823,228,122,300,4299,69
Example 558,78of 13.054.26 deaths0,832,307,783,338,67 100,00

Testing the effectiveness of the bactericidal properties of the material based on montmorillonite clay modified with silver ions was carried out under sterile conditions using sterile equipment and materials. For tests were used sterile Petri dishes containing sterile mastopathy agar (MPA) or blood agar with a pH of 7.2 to 7.4. The thickness of the layer of chilled MPA or blood agar - 2,5-3,0 mm In the nutrient medium, cooled to 45-48°C, made sterile sample material in the range from 1.56 to 100 mg per 1 ml of culture medium and suspended the studied strains of microorganisms. In the control Cup with a nutrient medium made only a suspension of the studied microorganisms. Cultivation of Escherichia coli, Pseudomonas aeruginosa. Salmonella dublin, Salmonella enteritidis, Staphylococcus hyicus, Staphylococcus intennedius, Staphylococcus aureus was performed on MPA, a Proteus vulgaris and Candida albicans on blood agar. Determination of the sensitivity of microorganisms to the materials obtained according to examples 1-5, depending on their concentration in the MPA and blood agar, was performed after culturing in an incubator at 37°C for 16-18 hours. The results are shown in table 2, allowed to set their minimum bacteriostatic concentration. Evidence of the effectiveness of the bactericidal properties of the material based montmorillonite-containing clay modified with silver ions, obtained in examples 1-5 are presented in table 2.

Table 2
The sensitivity of microorganisms to the received materials in examples 1-5
MicroorganismsThe concentration of the material, mg/mlThe number of CFU/ml on McFarland
example 1example 2example 3example 4example 5control **
Salmonella Dublin12,50**
6,25**3·1082·10812·108
3,1252·1081·1089·108 8·108*
1,569·1086·10812·10815·1083·108
Salmonella enteritidis12,50**22·108
6,25**2·1081·108
3,1252·1086·10818·10820·108*
1,5618·10815·10822·10824·1082·108
Staphylococcus hyicus25,00 **36·108
12,503·1081·108
6,25**9·1088·108
3,1256·1089·10833·10830·108*
1,5615·10818·10839·10839·1082·108
Proteus vulgaris12,50**36·10s
6,25 **6·1083·108
3,1252·1083·10830·10830·108*
1,5618·10821·10836·10836·1083·108
Staphylococcus aureus25,00**36·108
12,50**6·1082·108
6,252·1081,5·10833·10830·108*
3,125 21·10815·10839·10836·1081,5·108
Escherichia coli25,00**39·108
12,50**3·1081·108
6,252·1081,5·10836·10830·108*
3,12518·1089·10845·10839·1086·108
Pseudomonas aeruginosa25,00** 39·108
12,50**8·1086·108
6,253·1086·10830·10821·108*
3,12518·10815·10833·10833·1083·108
Staphylococcus intermedius25,00**30·108
12,50**2·1081,5·108
6,252·1083·10821·10818·108 *
3,12524·10818·10833·10830·1083·108
Candida albicans25,00**39·108
12,50**6·1083·108
6,258·1089·10833·10830·108*
3,12515·10815·10845·10839·1086·108
* Minimum bacteriostatic concentration;
** In the control experiment who used sterile mastopathy or blood agar without the introduction of clay.

From the data presented in table 2, it is seen that the materials obtained in examples 1, 2 and 5 had a more pronounced bacteriostatic action, than the specimens that were obtained in examples 3 and 4. This table illustrates and explains the different manifestation of the sensitivity of the test organism to various antibacterial materials obtained in examples 1-5, as in these forms respectively contained 3,36; 3,61; 0,10; 0,20 and 4.35 wt.% silver. Investigational antibacterial materials obtained in examples 1 and 2, inhibited the growth and colonization of Salmonella Dublin, Salmonella enteritidis, Staphylococcus hyicus on the surface of MPA and Proteus vulgaris on blood agar already at a concentration of 6.25 mg in 1 ml of culture medium. More effective bacteriostatic effect on these bacteria had a material obtained according to example 5, at a concentration of 3.125 mg/ml of culture medium. At the same time, the minimum bacteriostatic concentration of the studied antibacterial materials obtained in examples 1 and 2, for Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus intermedius on the surface of MPA and Candida albicans on blood agar was 12,50 mg of material per 1 ml of culture medium. Antibacterial material obtained in example 5, inhibited the growth of these microorganisms at a concentration of 6.25 mg/ml of culture among the s. To determine the bactericidal concentration of antibacterial materials obtained in examples 1-5, with swabs of cups, where there was no growth of the test organism, to produce crops on a dense nutrient medium MPA and blood agar, which did not contain the studied material. After cultivation of these crops in thermostat at 37°C for 16-18 hours, there was no growth of microorganisms with swabs MPA and blood agar, in which the minimum bacteriostatic concentration of antibacterial material was 3,125; 6.25 and 12,50 mg/ml of culture medium. The same bacteriostatic action of antibacterial materials obtained in examples 3 and 4 showed against Salmonella Dublin, Salmonella enteritidis and Proteus vulgaris, but at concentrations 12,50 mg/ml, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus intermedius and Candida albicans these materials influenced bakteriostaticheski concentration 25,00 mg/ml At the same time enriched the native form of montmorillonite clay at a concentration of 100 mg/ml MPA and blood agar did not suppress the growth of the test organism, but, on the contrary, strengthened. The number of colony forming units in flush with the surface of the dense nutritional environment experienced Petri dishes were 1.1-1.9 times more than in the control, i.e. not containing native form of clay. In affect, the, the task of creating a material with antibacterial properties based on sodium-calcium and/or calcium, and/or glandular montmorillonitic clays, which can effectively suppress the growth of pathogenic microorganisms, using the modification of solutions of AgNO3lower concentrations compared to the prototype, it was decided.

A method of obtaining a material with antibacterial properties based on montmorillonitic clays, consisting in the modification of clays, including inorganic mineral montmorillonite, a silver nitrate solution, washing and subsequent drying, characterized in that clay, including inorganic mineral montmorillonite, presents the sodium-calcium and/or calcium, and/or glandular form of montmorillonite; the mass ratio of clay:the modifying agent is 1:5, and the concentration of the modifying agent is an aqueous solution of nitrate of silver is 0.16-9.9 wt.%; the inoculation process is conducted under stirring for 3 to 7 hours at a temperature in the range from 10°C to the boiling temperature, washing the obtained modified material is carried out with distilled water until pH≈5-6 until you remove the excess silver nitrate; advocate at room temperature and decanted, then mater what al is dried at a temperature of 20-160°C.



 

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SUBSTANCE: in a complex preparation containing a carrier representing an enterosorbent; the enterosorbent is modified by immobilising high-disperse silver - nanosilver in a concentration of 0.01 - 1.0 wt % on its surface. The enterosorbent represents activated carbon, kaolin, bentonit, or enterodesum, or monocrystalline cellulose. A modifying silver-containing solution - a nanosilver source - is silver clusters in an aqueous solution.

EFFECT: higher specific antimicrobial activity.

2 cl, 3 tbl, 2 ex

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