Method of determining fungicidal activity of chemical preparations
SUBSTANCE: in agar plate which is in a Petri dish and inoculated with one of the fungal species of the genus Fusarium, circular holes are cut with the diameter of 8 mm, 0.05 ml of the working solution of the test preparation is applied into them, and placed in a thermostat at a temperature of 24.5-25.0°C, after 2-5 days depending on the type of fungus the diameter of the zone of growth inhibition of fungal mycelium is measured and the activity of the preparations is calculated according to the formula , and it is considered that when A=1 the fungicide is ineffective - the zone of growth inhibition is not formed (D=d), when A=2-3 the preparation activity is low, when A=4 it is average, and when A≥5 it is high.
EFFECT: invention provides accuracy of determining the activity of seed disinfectants and fungicides used in agricultural production.
The invention relates to agriculture and can be used to determine the fungicidal activity of chemical drugs against fungi causing plant diseases.
As a prototype the selected method used in medical research when studying the activity of the antibiotic agent is introduced into the wells on a nutrient medium, planted the studied species of bacteria or fungi. In the case of the presence of antibiotic action around the point of application of the drug is formed a rounded area of stunted growth of the microorganism. Largest diameter and evaluate the effectiveness of the test tools. This method allows to determine the activity of antibiotic substances against many types of microorganisms that cause disease in humans and animals (Birger M. O. and others Handbook of microbiological and virological research methods. - M: Medicine. - 1982; beat V. I., Methods of experimental Mycology: a Handbook. - Kiev. "Naukova Dumka". - 1982; Egorov N. With. Basic knowledge about antibiotics: a Training manual. - M.: Higher school. - 1979).
The main disadvantage of this method is that in medicine to evaluate the activity of the drug is used to the average diameter of the zone of restricted growth". The proposed method is based on determining the activity of a fungicide on a specially designed �matematicheskoi the formula and the assessment of this indicator is designed for scale, allowing you to more objectively evaluate the effectiveness of disinfectant.
Technical problem - ensuring the accuracy of the new method using mathematical formulas and scoring scales for determining the activity of seed disinfectants and fungicides used in agricultural production.
To achieve the technical objectives of the special experiments, the mathematical formula and is composed of the evaluation scale to determine the activity of fungicides.
The experiments were conducted as follows: in the agar plate in a Petri dish and sown with one species of fungi of the genus Fusarium, cut round holes with a diameter of 8 mm. they were added 0.05 ml of the working solution of the test drug. The working solution of fungicide were prepared the same concentration as in the production environment. Petri dishes with a culture of the fungus caused by the fungicide was placed in a thermostat at a temperature of 24.5-25,0°C. in 2-5 days depending on the type of fungus, carried out measurements of the diameter of the zone of stunted growth of fungal mycelium and the expected activity of the preparations according to the formula:
where A is the activity of the fungicide (in arbitrary units);
D - diameter of the zone of stunted growth of fungal mycelium (mm);
d - diameter of the application site fungicide solution (mm) equal to 8 mm.
Grading scale consists of the following values: if A=1 fungicide ineffective area of stunting is not formed (D=d); if A=2-3 activity of the drug is low; when A=average 4 and A≥5 high. Formula (1) shows how many times the diameter of the zone of stunting greater than the seat diameter of the preparation (the diameter of the hole).
The results of application of the method for determining the activity of fungicides is presented in the table.
|Activity of fungicides - seed disinfectants against fungi of the genus Fusarium|
|№ p/p||Drug consumption norm||The activity of the drug (A)|
|the fungal species|
|F. avenaceum||F. culmorum||F. graminearum||F. poae|
|1||Vincit SC, 2 l/t||3,9||4,9||7,6||5,0|
|2||Maxim COP, 2 l/ttd align="left"> 5,6||6,0||6,4||4,0|
|3||Maxim old COP, 1.5 l/t||5,8||5,1||8,2||3,9|
|4||Kolfage super COP, 2 l/t||2,4||4,2||6,9||3,5|
|5||Vitaros FAC, 3 l/t||2,6||3,9||2,0||2,5|
|6||Raxil COP, 0.5 l/t||2,0||1,4||2,9||2,5|
|7||Dividend the old CS,1 l/t||1,9||2,2||1,0||2,2|
|8||Premis COP, 2 l/t||1,0||2,8||1,0||1,0|
|9||Control (sterile water)||1,0||1,0||1,0||1,0|
As the study results show that the tested drugs can be divided into several groups. The first high and close to the average activity (3,9-8,2) includes Vincit IC, Maxim COP and the old Maxim of the COP. Second is Kolfage super COP. Its activity against species of Fusarium graminearum and F. culmorum was high and average (6,9 and 4.2), F. avenaceum and F. roue - low (2.4 and 3.5). The third group includes the fungicides Vitaros VSC and Raxil the COP. An indicator of the activity of the first drug against F. culmorum was close to the average of (3.9) with respect to other types vusalevich mushrooms these funds were at a low level (from 1.4 to 2.6). The latter group includes the Dividend of the old COP and Premis the COP. The activity of these drugs was low (A=1,9-2,8) or absent (A=1).
Thus, the use of the method for determining the activity of fungicides against fungi of the genus Fusarium with application of the formula and the new scale provides a more accurate assessment of the test drugs and recommend them for further study or use in agricultural production.
The effectiveness of this method is expressed in a more precise determination of the fungicidal activity of chemical drugs compared to the diameter of the zone delay� growth."
Method of determining the fungicidal activity of chemical preparations, characterized by the fact that in agar plate in a Petri dish and sown with one species of fungi of the genus Fusarium, cut round holes with a diameter of 8 mm, making of 0.05 ml of the working solution of the test drug and placed in a thermostat at a temperature of 24.5-25.0°C, 2-5 days depending on the type of fungus measurements of the diameter of the zone of stunted growth of fungal mycelium and calculate the activity of the preparations according to the formula:
where: a - active fungicide (in arbitrary units);
D - diameter of the zone of stunted growth of fungal mycelium (mm);
d - diameter of the application site fungicide solution (mm) equal to 8 mm,
and consider that if A=1 fungicide ineffective area of stunting is not formed (D=d), if A=2-3 activity of the drug is low, A=average 4 and A≥5 high.
SUBSTANCE: invention represents a method for preclinical study of cardiotropic antiarrhythmic drugs, involving determining the bioelectric parameters in isolated multicellular perfused preparations and measuring an action potential duration, differing by the fact that the isolated multicellular perfused preparations are presented by rat's pulmonary vein myocardium; the parameters are measured in three operation modes of the multicellular preparations; a resting potential is additionally measured; varying APD 90%, related APD 50%/APD 90%, a spontaneous shear velocity of the resting potential, the most positive membrane potential in the resting preparation, a spontaneous activity train repetition rate, spontaneous action potential train repetition and variability frequency, post-depolarisation number and intensity, as well as a shear membrane potential corresponding to the beginning of train activity are used to evaluate the signs of antiarrhythmic and arrhythmogenic action.
EFFECT: more reliable prediction of the antiarrhythmic action of the potential pharmacological agents and reduction of experimental phase time.
SUBSTANCE: invention relates to method of determining platelet resistance to acetylsalicylic acid (ASA) by impedance analysis of aggregation function of platelets in vivo, in which aggregation activity is analyses after incubation of biological material sample with ASA with application of aggregation inducer, and aggregation of platelets is induced by collagen in optimal concentration 2 mg/ml, and simultaneously with measurement of impedance determination of dynamics of platelet granules release is performed by luminescent method, where before carrying out aggregation samples are calibrated by means of adenosine triphosphate (ATP) standard, value of aggregation amplitude are determined in Ohms by obtained aggregatograms and obtained values are given points: values ≤6 correspond to 0 points, values 7-9 correspond to 1 point, values 10-12 correspond to 2 points, values >12 correspond to 3 points; after that, intensity of ATP release from platelet granules is determined in nmoles and obtained values are given points: values <0.5 correspond to 0 points, values 0.5-1.0 correspond to 1 point, values 1.0-1.5 correspond to 2 points, values 1.5 correspond to 3 points, and then resistance index (RI) is calculated by formula, value of IR parameter points to presence of aspirin-resistance of platelets.
EFFECT: carrying out fast complex diagnostics of platelet resistance to ASA with estimation of functional state of platelet granules before beginning treatment and possibility of preventing development of undesirable thrombotic or hemorrhagic complications.
2 dwg, 1 tbl, 2 ex
SUBSTANCE: claimed is application of fat emulsion for parenteral feeding as solvent for compounds which are poorly soluble in water. Fat emulsion contains in 1 l of solution: 30 g of refined soybean oil, 30 g of triglycerides with the average chain length, 25 g of olive refined oil, 15 g of purified fish oil.
EFFECT: obtaining solvent for compounds, poorly soluble in water, which makes it possible to determine parameters and spectrum of biological activity of novel compounds of chemical nature at the stages of pre-clinical and clinical tests, which does not change basic biological constants and possesses biological inertness.
2 tbl, 2 ex
SUBSTANCE: invention relates to biology, forensic and analytical chemistry and specifically to methods of determining procaine in blood plasma. The method comprises adding sodium fluoride to blood plasma containing procaine to achieve concentration of 10 mg/ml; treating the obtained mixture with acetone; separating the extract from the precipitate by filtering; evaporating acetone from the filtrate in an air current at room temperature; diluting the aqueous residue by adding water; saturating the obtained solution with ammonium sulphate; alkalising with an ammonium buffer solution to pH 9.0-9.5; extracting twice with portions of an organic extraction agent in the form of 30% camphor solution in methyl acetate, with ratio of the aqueous to the organic phase of 1:1 by volume; separating the organic extracts; combining; evaporating the solvent from the combined extract in an air current at room temperature; chromatographing the residue in a thin layer of silica gel STKH-1A on Sorbfil PTSKH-AF-A-UF plates, using a dichloromethane-ethanol mobile phase in ratio of 6:4 by volume; developing the chromatogram in UV light; eluting the analysed substance from the sorbent with a mixture of acetonitrile-methanol-0.025 M potassium dihydrogen phosphate solution with pH 3.0 in ratio of 10:10:90 by volume; chromatographing by HPLC method using a reversed-phase sorbent Nucleosil C18, a polar mobile phase acetonitrile-methanol-0.025 M potassium dihydrogen phosphate solution with pH 3.0 in ratio of 10:10:90 by volume and a UV detector; measuring optical density at wavelength 298 nm and calculating the amount of the analysed compound from the area of the chromatographic peak.
EFFECT: method improves sensitivity of determination.
3 tbl, 2 ex
SUBSTANCE: invention relates to analytical chemistry. The method is characterised by electrochemically concentrating benzoic acid on the surface of a graphite electrode for 90 s at electrolysis potential of (-0.500) V on a background of 0.1 mol/l sodium hydrogen phosphate, recording polarisation curves with linear potential sweep rate of 25 mV/s and determining concentration of benzoic acid from the peak height in potential range of 0.5-1.6 V relative to a silver chloride electrode.
EFFECT: method provides highly sensitive and rapid determination of benzoic acid in medicinal drugs.
3 ex, 6 tbl
SUBSTANCE: sample is applied on a paper filter, and standard calibration solutions of metronidazole in the concentration range of 10-100 mcl are radially applied on the same filter. The filter is processed with a developing agent containing 5% potassium hydroxide and acetone taken in ratio 2:1 and thermostated at 100°C until coloured spots are visible. A colour intensity of the sample spot is compared to the colour of the spot colour of the reference calibration solutions to determine the metronidazole concentration in the analysed sample.
EFFECT: method enables the fast and reliable monitoring of the metronidazole content and the timely correction of the therapeutic process.
SUBSTANCE: invention refers to medicine, namely to studying and analysing medical preparations, and can be used for standardising herbal raw materials. A method for identification and qualitative measurement of chlorophyll, carotinoids and hydroxycinnamic acids in a combination in great nettle leaves involves a 1-hour fractional extraction, 30 min each of the ground raw material having a particle size of 1.0 mm on a water bath at a temperature of 100°C with 70% ethanol in a ratio of the herbal raw material to the extractant of 1:100, the combination of the extracts and reduction to 100 ml with a solvent, dilution of the prepared solution in a ratio of 2:25 in 96% ethanol, measuring an optical density of the solution in relation to 96% ethanol at maximum absorption 328±1 nm, 442±1 nm and 667±1 nm, calculation of the total content of hydroxycinnamic acids equivalent to chlorogenic acid, carotinoids equivalent to violaxanthin and chlorophyll in the percentage equivalent to an absolute dry mass of the raw material by formulas.
EFFECT: method provides availability, simplicity, efficiency and low error of measurement.
3 dwg, 1 ex
SUBSTANCE: method of determining fat-soluble vitamins A, D2, E and β-carotene which are present at the same time includes separating the fat-soluble vitamins from a substance by extraction with 96% ethanol, separating the alcohol extract of vitamins using a separating funnel, successive chromatography using Sorbfil PTSKH-P-A silica gel plates on a polymer substrate using two eluents with a different range, time of saturating the chamber with eluent vapour of 20 minutes and elution time of 55 min; drying the plates at temperature not lower than 80°C in a temperature-controlled chamber for 3-5 min, treating the plates with a developer - 5% alcohol solution of phosphatomolybdic acid; according to the invention, the eluents used are hexane:chloroform (19:1) and hexane:chloroform (3:1), and detection of the chromatographic zone of β-carotene is carried out before treating the plates with a developer in day light.
EFFECT: simpler and faster process of determining fat-soluble vitamins.
10 dwg, 3 ex
SUBSTANCE: invention relates to medicinal drug quality control and provides a method of authenticating and determining the quantitative content of benzethonium chloride in medicinal drugs, which includes separating components of the drug via high-performance liquid chromatography, where the eluent used is a mixture of acetonitrile, tetrabutylammonium hydrophosphate, disubstituted potassium phosphate and water, with content of tetrabutylammonium hydrophosphate of 2.5 mM, content of disubstituted potassium phosphate of 5 mM, wherein separation of the components of the drug is carried out with column length of 125 mm.
EFFECT: high accuracy and faster analysis since there is no need for special sample preparation.
SUBSTANCE: invention relates to a method of screening a non-teratogenic substance comprising bringing a test substance into contact with cereblon or a fragment of cereblon, evaluating the capacity of the test substance to bind to cereblon or the fragment of cereblon, and selecting a test substance that does not bind to cereblon or the fragment of cereblon or a test substance exhibiting lower capacity to bind to cereblon or the fragment of cereblon compared with thalidomide.
EFFECT: improved method.
8 cl, 19 dwg, 8 ex
FIELD: microbiology, pharmaceutical agents.
SUBSTANCE: invention relates to method for determination of genome response of specific cells to vegetable extract action. Method for drug screening includes 1) treatment of specific cells with vegetable extract; 2) protein or RNA isolation from said treated cells; 3) identification of isolated protein or RNA; 4) determination of compound(s) in said vegetable extract; 5) treatment of said cells with determined compound(s); 6) protein or RNA isolation from said treated cells; and 7) determination of compounds providing expression or suppression of said protein or RNA which have another concentration than in untreated said specific cells.
EFFECT: identification of individual compound(s) for screening of new pharmaceutical agents or new pharmaceutical application of existing drugs.
FIELD: medicine, in particular urology.
SUBSTANCE: method relates to method for determination of pathogenic activity of microorganism staphylococcus epidermidis isolated from sperm. Microorganism samples isolated from donors are incubated with spermine or spermidine in concentration of 0.02-0.06 mg/ml followed by determination of survived microorganism amount by comparison of sample and standard optical density. Increased microorganism amount in sample in contrast with standard denotes the pathogenic activity of analyzed strain.
EFFECT: method with improved selectivity and accuracy.
3 ex, 2 tbl
FIELD: medicine, microbiology.
SUBSTANCE: invention relates to a method for assay of pathogenicity of microorganism Staphylococcus epidermidis. Method involves incubation of experimental samples isolated from donors with lactofferin taken in the concentration 320-580 ng/ml followed by counting amount of revived microorganisms as compared with control value. Advantage of invention involves enhancing sensitivity of method.
EFFECT: improved assay method.
1 tbl, 3 ex
SUBSTANCE: method involves determining quantitative and qualitative composition of short chain fatty acids of C2-C4 fraction in feces and peripheral blood serum to evaluate fatty acids metabolism at various levels (hepatic, vesicular and intestinal).
EFFECT: high accuracy in measuring metabolism characteristics at various levels; accelerated process.
FIELD: gene engineering.
SUBSTANCE: the present innovation deals with transferring a mutant gene due to homologous recombination into animal embryo. The animal obtained is characterized by the capacity to express mutant protein of presenylin-1 and induction of beta-myeloid protein production that leads to the development of progressing nervous disease in hippocampus or peripheral department of cerebral cortex, It is, also, suggested to apply several plasmids carrying a mutated gene. It is, also, described the way to obtain primary cell culture or subcultivated cell out of obtained mutated animals. Moreover, several methods are, also, suggested for testing the substances for usefulness in therapeutic and/or prophylactic procedures at treating Alzheimer's disease. They deal with introducing a tested substance for mutated animal to evaluate the data obtained. The obtained mutated animals could be applied as model animals while studying Alzheimer's disease nature.
EFFECT: higher efficiency.
25 cl, 8 dwg, 10 ex
FIELD: medicine, clinical pharmacology.
SUBSTANCE: invention relates to a method for identification of antidepressant possessing an anti-itching effect by using the computer program for measurement of models of chemical structures. Method involves visual analysis of all stable conformers of antidepressant molecule and antidepressant possessing anti-itching effect is identified in detection at least one stable conformer comprising structure of two inter-centered aromatic rings wherein their planes are able to orient at angle 120° with respect to one another. Also, invention relates to using antidepressant identified by above described method in treatment of itching. Invention provides identifying anti-itching effect of antidepressants.
EFFECT: improved method for identification.
2 cl, 7 ex
FIELD: analytical methods in medicine.
SUBSTANCE: concentration of miramistin (C26H47N2O) in an aqueous medium is found from spectrophotometrically measured optical density of solution. Spectrophotometric measurement is performed in the region of maximum UV absorption of solution, namely within 261-263 nm wavelength range.
EFFECT: simplified analytical procedure at rather high accuracy level.
SUBSTANCE: method involves scanning pulse with glass plate placed on pulse measurement point. The glass plate bears biological test system containing composition of 0.1% aqueous amino acid solutions like tryptophan, aspartic acid, alanine, treonine, valine, serine, glycine and tyrosine taken in equal proportions, 0.5% aqueous solution of dopamine, 0.5% aqueous solution of histamine, 12% aqueous solution of magnesium sulfate taken in 3:3:13 proportion. The plate is hold on pulse for 1-2 min before and 10-15 min after giving the pharma- or parapharmaceutic under study to a patient. Then, the plate is dried at T=+18-20°C during 2-3 min and another glass plate is covered with biological test system surrounded with wall of the pharma- or parapharmaceutic under study arranged along periphery and dried at T=+18-20°C during 2-3 min. The biological test system structures formed on the plates are compared in polarized light with quartz compensator. Structural similarity being detected, identification of pharma- or parapharmaceutic introduced into human organism is to be recorded.
EFFECT: enhanced effectiveness in identifying biologically active substances introduced into human organism.
FIELD: medicine, medicinal toxicology, microbiology, biology.
SUBSTANCE: invention relates to a method for testing different immune preparations, for example, immunoglobulins. Method involves carrying out the combined culturing test-strain of microorganism with the corresponding immune preparation under standard conditions of the human embryo monolayer of cultured cells. Results are estimated by degree of the adhesion decrease value of test-strain of microorganism as compared with control value. Invention provides elevating rate in evaluation of an anti-infectious activity of immune preparations based on approaching conditions of evaluation to macroorganism conditions.
EFFECT: improved method of evaluation.
2 tbl, 2 ex
FIELD: analytical chemistry.
SUBSTANCE: invention relates to method for quantitative determination of thiotriazoline and pyracetam in complex drugs by high performance chromatography, wherein silicagel with grafted 3-(chlorodimethyl)-propyl-N-dodecylcarbamate having particle size of 5 mum is used as sorbent; and degassed 0.05 M aqueous solution of potassium dihydrophosphate is used as mobile phase. Mobile phase velocity is 1 ml/min, and column temperature is 30°C. Method of present invention makes it possible to determine content of two abovementioned active ingredients simultaneously.
EFFECT: simplified process of sample preparation.
3 ex, 3 tbl