Method of determining procaine in blood plasma

FIELD: chemistry.

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

 

The invention relates to biology, toxicology and analytical chemistry, and in particular to methods for the determination of procaine in plasma, and can be used in the practice of chemical-Toxicological, forensic and clinical laboratories. The method refers to the mass number.

The known method of determination of procaine in biological fluids (urine), which consists in the fact that the analyzed sample is treated with sodium bicarbonate to bring the pH to 8.4 and 8.6, extracted for 5 minutes with an organic extractant composed of a mixture of chloroform and Isobutanol, taken in the ratio of 6:1, when the volume ratio of aqueous and organic phases of 0.3:1, the extract was filtered through anhydrous sodium sulfate, the sodium sulfate was washed with chloroform, the filtrate and wash liquid are combined in the combined solution is injected internal standard, which is cyclized, the solvent from the resulting solution is evaporated to a dry residue, the residue is dissolved in acetylides the reagent, which is a mixture of acetic anhydride and pyridine, taken in the ratio 3:2 by volume, the resulting reaction mixture is heated for 20 minutes at a temperature of 80°C, the excess acetylides reagent is evaporated in a stream of air at 40°C, the residue is dissolved in ethyl acetate and determine the number of analiziruemoi by gas chromatography with mass-selective detection (Melentiev A. B. Screening of medicinal, narcotic substances and their metabolites by gas chromatography with mass-selective detector // problems of expertise in medicine. - So 2, No. 4. - S. 15-21).

The method has a relatively low degree of extraction and insufficiently high accuracy.

The known method of determination of procaine in plasma, consisting in the fact that the analyzed sample is added sodium fluoride to create a concentration of 7.5 mg/ml, adjusting the pH of the mixture to pH 11 using bicarbonate buffer, extracted with dichloromethane at a ratio of volumes of aqueous and organic phases 1:1,2, the extract is separated, evaporated to a dry residue, the residue is dissolved in deionized water, and determine the number of the analyzed compounds in the chromatographic peak area method of high performance liquid chromatography in combination with mass-selective detection (Zientek K. D., Anderson D. F., K. Wegner, Cole C. Quantitatio of Procaine in Equine Plasma by Liquid Chromatography - Linear Ion Trap Mass Spectrometry // Journal of Analytical Toxycology. - 2007. - Vol.31. - P. 87-92).

The method differs insufficiently high accuracy and relatively low degree of extraction.

The closest is the method of determination of procaine in biological fluids, consisting in the fact that in the analyzed sample is injected sodium fluoride to create a concentration of 5 mg/ml, up to ablaut internal standard, which is used as tetracaine, the resulting solution is alkalinized borate buffer solution to 9.5, extracted twice with portions of the organic extractant, which is used as dichloroethane, at a ratio of aqueous and organic phases 0.6 to 1:1 by volume), the organic extracts are separated, combined, the solvent is evaporated to the dry residue in a stream of nitrogen under conditions of heating at 60°C, the residue is dissolved in methanol, chromatographic by HPLC using obremenitve sorbent Radial Pak C18"polar mobile phase acetonitrile - 0,0165 M solution of triethylamine in the ratio of 85:15 by volume and UV detector, registrerat optical density at a wavelength of 288 nm and calculate the number of analyzed compounds in the chromatographic peak area (P. Beaumier, Timmings, S., Fenwick, J. D., Fodi F., Young L. M., Yeow T., Weber, M., Stevenson, A. J. Procaine Determination after Administering Various Procaine Formulations to Standard-breds // Proceedings of the 6 th International Conference of Racing Analysis and Veterinarians. Hong Kong, 1985. - P. 209-216).

The method is characterized not high enough sensitivity.

The technical result of the present invention is to increase the detection sensitivity.

The technical result is achieved by the fact that in the analyzed sample is injected sodium fluoride to create a concentration of 10 mg/ml, the mixture was treated with acetone, the extract is separated is from the fall of the precipitate by filtration, the acetone from the filtrate evaporated in a stream of air at room temperature, the aqueous residue is diluted by adding water, the resulting solution is treated with ammonium sulfate, alkalinized ammonium buffer solution to pH of 9.0 to 9.5, extracted twice with portions of the organic extractant, which is 30% solution of camphor in acetate, with the ratio of aqueous and organic phases 1:1 by volume), the organic extracts are separated, the combined solvent from the combined extracts evaporated in a stream of air at room temperature, the remainder chromatographic in a thin layer of silica gel CTX-1A on the plates "Sorbfil" PTSH-AF-And-UV, using the mobile phase dichloromethane-ethanol in a ratio of 6:4 by volume, the chromatogram shown in UV-light, elute an analyte from the sorbent mixture of acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume, chromatographic by HPLC using obremenitve sorbent Nucleosil C18", polar mobile phase acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume and UV detector, registrerat optical density at a wavelength of 298 nm and calculate the number of analyzed compounds in the chromatographic peak area.

The method is as follows: in plasmas the blood, containing procaine administered sodium fluoride to create a concentration of 10 mg/ml, the mixture was treated with acetone, the extract is separated from the precipitated precipitate by filtration, the acetone from the filtrate evaporated in a stream of air at room temperature, the aqueous residue is diluted by adding water, the resulting solution is saturated with ammonium sulfate, alkalinized ammonium buffer solution to pH of 9.0 to 9.5, extracted twice with portions of the organic extractant, which is 30% solution of camphor in acetate, with the ratio of aqueous and organic phases 1:1 by volume, organic extracts separate, combine, the solvent from the combined extracts evaporated in a stream of air at room temperature, the residue chromatographic in a thin layer of silica gel CTX-1A on the plates "Sorbfil" PTSH-AF-And-UV, using the mobile phase dichloromethane-ethanol in a ratio of 6:4 by volume, the chromatogram shown in UV-light, elute an analyte from the sorbent mixture of acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume, chromatographic by HPLC using obremenitve sorbent Nucleosil C18", polar mobile phase acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume and UV detector, record the optical system is the density at a wavelength of 298 nm and calculate the number of analyzed compounds in the chromatographic peak area.

The method is illustrated by the following examples.

Example 1

Determination of procaine in the blood plasma of a person

5 cm3the human blood plasma contribute 40 mg of procaine (2-(diethylamino)ethyl-4-aminobenzoate hydrochloride) in the form of 0.1 cm30,04% solution, in the analyzed sample is injected 51 mg of sodium fluoride to create a concentration of 10 mg/ml, mix thoroughly biological fluid with the substance within 2 minutes. The resulting mixture was treated with 10 cm3acetone for 10 minutes under stirring. After this time the liquid extract is separated from the precipitated precipitate by filtration through a paper filter, and the residue is again treated with 10 cm3acetone for 10 minutes under stirring, and filter the extract through filter paper used to filter the first extraction. The advanced filter is washed with 5 cm3of acetone. The filtrate and wash liquid are combined and evaporated acetone in air flow at room temperature.

The aqueous residue is diluted by adding 5 cm3water, the resulting solution is saturated with ammonium sulfate, alkalinized ammonium buffer solution to pH of 9.0 to 9.5, bring the volume of the resulting solution to 10 cm3, extracted with 10 cm330% solution of camphor in ethyl acetate for 10 minutes, the extraction system ostable the t for 3-5 minutes for separation, the organic extract is separated and the extraction process repeated as previously described. Separate extracts are combined into varicellae Cup, evaporated to a dry residue in a stream of air at room temperature.

The residue is dissolved in a small amount of ethanol and quantitatively transferred to the starting line chromatographic plate type "Sorbfil" PTSH-AF-And-UV dimensions of 10×7 cm in the form of a strip with a length of 3-3,5 cm and a width of 0.4-0.5 cm Chromatographic in the presence of substances-witnesses of procaine and camphor in a glass chamber with an internal volume of 600 cm3using eluent dichloromethane-ethanol in a ratio of 6:4 by volume. The obtained chromatogram shown in UV - light. An analyte (procaine) identify the largest Rf matching Rf value substance-standard and component data in terms of 0.33±0,02.

After chromatography was carried out by TLC plot chromatogram with spot analyte cut out, placed in the tube, elute the substance of the sorbent 5 ml of a mixture of acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate (pH 3.0) in the ratio 10:10:90 by volume. 1,0·10-2cm3the resulting solution injected type "milikhrom - 5" with UV detector.

Chromatographic by HPLC. The process of chromatography was carried out carry out in column (80×2 mm, filled obrashennih the new sorbent Nucleosil C18 (particle size 5 μm) using a mobile phase of acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate (pH 3.0 by the addition of H 3PO4) in the ratio 10:10:90 by volume. The feed rate of eluent - 0.1 cm3/min Chromatographic process is carried out at 20°C. the Optical density recorded at a wavelength of 298 nm.

The peak on the chromatogram with time holding 5,73 min (retention 573 μl) corresponds to procaine.

The quantitative content of procaine determined on the basis of the chromatographic peak area, the equation of the calibration curve and count on a portion of the substance introduced into the plasma of human blood.

The construction of the calibration curve

In a series of volumetric flasks with a capacity of 25 ml make 0,025, 0,25, 1,0, 2,5, 5,0; 10,0 ml of a 0.005% solution of procaine in a solvent mixture of acetonitrile-methanol - 0.025 M solution of potassium dihydrophosphate (pH 3.0 by the addition of H3PO4) in the ratio 10:10:90 by volume and brought to the mark with a mixture of solvents acetonitrile-methanol - 0.025 M solution of potassium dihydrophosphate (pH 3.0 by the addition of H3PO4) in the ratio 10:10:90. 1,0·10-2cm3each of the obtained solutions are injected. The chromatography was carried out is carried out in a column of size 80×2 ml, filled with sorbent "Nucleosil C18 (particle size 5 μm) using a mobile phase of acetonitrile-methanol - 0.025 M solution of potassium dihydrophosphate (pH 3.0 by the addition of H3PO4) in the ratio 10:10:90 by volume and a UV detector. RMSE is ity supply of eluent is 0.1 cm 3/min

The chromatographic process is carried out at 20°C. the Optical density recorded at a wavelength of 298 nm.

According to the results of measurements on the chromatograph build a graph of the dependence of chromatographic peak area against the concentration of the detected substance. Linear in the concentration range 5·10-10-2·10-7he

By the method of least squares to calculate the equation of the calibration curve, which in this case is:

S=295,4721·+0,2759,

where S is the area of the chromatographic peak With the concentration of analyte in khromatograficheskoi sample, ng.

The results of determination of procaine in the blood plasma of a person are shown in table 1.

Example 2

Determination of procaine in the blood plasma of horses

5 g of blood plasma of horses contribute 40 mg of procaine (2-(diethylamino)ethyl-4-aminobenzoate hydrochloride) in the form of 0.1 cm30,04% solution, in the analyzed sample is injected 51 mg of sodium fluoride to create a concentration of 10 mg/ml, mix thoroughly biological fluid with the substance within 2 minutes. The resulting mixture was treated with 10 cm3acetone for 10 minutes under stirring. After this time the liquid extract is separated from the precipitated precipitate by filtration through a paper filter, and the residue is again treated with 10 cm3acetone for 10 minutes PR is stirring and filter the extract through filter paper, used for filtering the first extraction. The advanced filter is washed with 5 cm3of acetone. The filtrate and wash liquid are combined and evaporated acetone in air flow at room temperature.

The aqueous residue is diluted by adding 5 cm3water, the resulting solution is saturated with ammonium sulfate, alkalinized ammonium buffer solution to pH of 9.0 to 9.5, bring the volume of the resulting solution to 10 cm3, extracted with 10 cm330% solution of camphor in ethyl acetate for 10 minutes, the extraction system is left on for 3-5 minutes for separation, the organic extract is separated and the extraction process repeated as previously described. Separate extracts are combined into varicellae Cup, evaporated to a dry residue in a stream of air at room temperature.

The residue is dissolved in a small amount of ethanol and quantitatively transferred to the starting line chromatographic plate type "Sorbfil" PTSH-AF-And-UV dimensions of 10×7 cm in the form of a strip with a length of 3-3,5 cm and a width of 0.4-0.5 cm Chromatographic in the presence of substances-witnesses of procaine and camphor in a glass chamber with an internal volume of 600 cm3using eluent dichloromethane-ethanol in a ratio of 6:4 by volume. The obtained chromatogram shown in UV - light. An analyte (procaine) identify the value of R, coinciding with the magnitude of the Rf substance-standard and component data in terms of 0.33±0,02.

After chromatography was carried out by TLC plot chromatogram with spot analyte cut out, placed in the tube, elute the substance of the sorbent 5 ml of a mixture of acetonitrile-methanol - 0.025 M solution of potassium dihydrophosphate (pH 3.0) in the ratio 10:10:90 by volume. 1,0·10-2cm3the resulting solution injected type "milikhrom-5" with UV detector.

Chromatographic by HPLC. The process of chromatography was carried out carry out in column (80×2 mm, filled obremenitve sorbent "Nucleosil C18 (particle size 5 μm) using a mobile phase of acetonitrile-methanol - 0.025 M solution of potassium dihydrophosphate (pH 3.0 by the addition of H3PO4) in the ratio 10:10:90 by volume. The feed rate of eluent - 0.1 cm3/min Chromatographic process is carried out at 20°C. the Optical density recorded at a wavelength of 298 nm.

The peak on the chromatogram with time holding 5,73 min (retention 573 μl) corresponds to procaine.

The quantitative content of procaine determined on the basis of the chromatographic peak area, the equation of the calibration curve and count on a portion of the substance introduced into the blood plasma of horses.

Scheme for the construction of calibration curve for determination of p is ocaina and the equation of this graph is presented above in example 1.

The results of determination of procaine in the blood plasma of horses presented in table 2.

The proposed method is compared with the prototype 2-8 times increases the detection sensitivity and 8-9% increases the degree of extraction of procaine from blood plasma.

Comparative characteristics of the proposed and known methods are presented in table 3.

Table 1
The results of determination of procaine in the plasma of human blood (n=5; P=0.95)
No.Made of procaine, ug per 5 cm3the human blood plasmaFoundMetrological characteristics, %
the peak area on the chromatogram, the river. unitsmcg in khromatograficheskoi sampleμg in terms of the linkage made in biojidkosti% of the paid in biojidkosti sample
14023362,787,906·10-239,53298,83 x=98,21
24022086,347,474·10-237,36893,42S=3,47
34023803,048,055·10-240,276100,69Sx-=1,55
44024197,378,158·10-240,792101,98Δx=4,31
54022744,567,690·10-238,44896,12ε =4,39

Table 2
The results of determination of procaine in the blood plasma of horses (n=5; P=0.95)
No.Made of procaine, mg per 5 cm3plasma horsesFoundMetrological characteristics, %
the peak area on the chromatogram, the river. unitsmcg in khromatograficheskoi sampleμg in terms of the linkage made in biojidkosti% of the paid in biojidkosti sample
14024131,018,166·10-240,832102,08x=98,40
24023303,697,886·10-2 39,42898,57S=3,28
34023909,418,091·10-240,456101,14Sx-=1,47
44022417,277,586·10-237,92894,82Δx=4,08
54025547,287,630·10-238,15295,38ε=4,14

Table 3
Comparative characteristics of razlagaemogo and known methods (for example, studies of blood plasma of horses) (n=5; p=0.95)
Comparative ratesThe proposed methodThe known method
1. Sensitivity (open minimum), g/cm3biomaterial2,5·10-95-20·10-9
2. The degree of extraction98,40±4,14%90±5%

Method for determination of procaine in plasma, consisting in the fact that in the analyzed sample is injected sodium fluoride, the resulting solution is alkalinized buffer solution, extracted twice with portions of the organic extractant, private separate extracts are pooled, the solvent is evaporated to the dry residue, the residue is dissolved, chromatographic by HPLC using obremenitve sorbent, polar mobile phase and UV detector, record the optical density and calculate the number of analyzed compounds in the chromatographic peak area, characterized in that the sodium fluoride in the analyzed sample is injected to create a concentration of 10 mg/ml, after the introduction of sodium fluoride obtained mixture is treated with acetone, the extract is separated from the precipitated precipitate by filtration, the acetone from the filtrate evaporated in a current of air is and at room temperature, the aqueous residue is diluted by adding water, the resulting solution is treated with ammonium sulfate, alkalinized ammonium buffer solution to pH of 9.0 to 9.5, as the organic extractant use 30% solution of camphor in the acetate, the ratio of aqueous and organic phases is 1:1 by volume, after evaporation of the extractant to the dry residue residue chromatographic in a thin layer of silica gel CTX-1A on the plates "Sorbfil" PTSH-AF-And-UV, using the mobile phase dichloromethane-ethanol in a ratio of 6:4 by volume, the chromatogram shown in UV-light, an analyte elute from the sorbent mixture of acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume, when chromatographicaliy by HPLC as obremenitve sorbent used Nucleosil C18, as a polar mobile phase - acetonitrile-methanol-0.025 M solution of potassium dihydrophosphate with pH 3.0 in the ratio 10:10:90 by volume, optical density recorded at a wavelength of 298 nm.



 

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25 cl, 1 dwg

FIELD: chemistry.

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

FIELD: medicine.

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.

3 ex

FIELD: medicine.

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

FIELD: chemistry.

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

FIELD: chemistry.

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.

9 ex

FIELD: chemistry.

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: medicine.

SUBSTANCE: method involves taking blood at 1 ml per a sample into the Vakutainer evacuated test tube with sprayed ethylene diaminetetraacetic acid 2.2 mg; a reference sample 1.0 ml and a sample 1.0 ml with weighted xenobiotics are taken from the total volume into the Eppendorf tubes; 30-minute incubation is followed by using a uniform procedure to prepare blood smears by applying on a dry slide to prepare a smear to be stained according to Leishman; blood corpuscles are analysed by the Zeiss light microscope at a magnification 1500. A microscope field showing echinocytes with styloid processes of the membrane not found in the reference sample is a sign of the xenobiotic membrane toxicity.

EFFECT: invention enables an instant assessment of the membrane toxicity of the substance.

FIELD: chemistry.

SUBSTANCE: lactic acid is transferred from a sample into a solution and voltammetric accumulation of lactic acid in the stirred solution is performed while bubbling with an inert gas for 30 s and with electroaccumulation potential of 1.2-1.4 V relative to a saturated silver chloride electrode on a background electrolyte of 0.1 M Na2HPO4, followed by detection of cathode peaks in differential mode of recording voltamperograms with potential sweep rate of 30-40 mV/s; concentration of lactic acid is determined from peak height in the potential range of 0.25-0.40 V by a standard addition method.

EFFECT: method is simple, does not require a large amount of reactants and labour costs.

2 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: method consists in the application of glutationreductase and catalase enzymes as test-objects for determination of the anti-oxidant activity by the ratio of the rate of enzymatic reaction on the test-object after the substance addition and the rate of enzymatic reaction before the substance addition, which must be larger than 1. Preliminarily, before addition to an incubation medium, the samples of essential oil of Siberian fir are diluted with dimethylsulphoxide in a ratio of 1:1.

EFFECT: increased accuracy of determination.

2 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention relates to a method of early detection of muscular degenerative diseases and to a method of prediction and/or determination of a therapeutic efficiency of a therapeutic preparation and/or a method of disease therapy by measurements of tetranor-PGDM (11,15-dioxo-9α-hydroxy-2,3,4,5-tetranorprostan-1,20-dioic acid) in a subject's urine sample and comparison of its content relative to a sample, separated from a healthy individual. A muscular degenerative disease is identified in a patient if the concentration or content of tetranor-PGDM in a sample, separated in the patient is higher than the concentration or content of tetranor-PGDM in a sample, separated from a healthy individual. An efficiency of the therapeutic preparation and/or the method of therapy of the muscular degenerative disease is determined by comparison of the content of tetranor-PGDM in the sample, separated from the patient with the muscular degenerative disease before and after introduction of the therapeutic preparation. If the measured content of tetranor-PGDM in the sample considerably or inconsiderably decreases after the introduction of the therapeutic preparation, the method of therapy is efficient. The invention also relates to a set for diagnostics of muscular degenerative diseases, which includes an antibody to tetranor-PGDM, labelled tetranor-PGDM and optionally, at least, one compound, selected from a group, consisting of an antibody to immunoglobulin, a diluting solution for the sample, a diluting solution for the antibody and labelled tetranor-PGDM, tetranor-PGDM standard of the known concentration, a substrate for an enzyme immunoassay and a stopping solution for the enzyme immunoassay.

EFFECT: invention is efficient and simple in implementation.

7 cl, 1 tbl, 1 ex, 2 dwg

FIELD: process engineering.

SUBSTANCE: invention relates to fluid treatment. Proposed device comprises phase components feed and discharge means and tubular flow extraction chamber with fluid and carrier gas feed and discharge pipes. Extraction chamber is placed vertically in thermostat. It features hydrophilic inner surface and taper at top end aligned with capillary pipe so that a round slot is formed, or with funnel furnished with hollow cone secured so that a circular slot is formed between funnel surface and cone surface. Gas unions are arranged at acute angle to chamber axis while relative to chamber surface they are fitted tangentially. Method of extraction comprises feed of components into extraction chamber in countercurrent and discharge of carrier gas therefrom enriched in volatile components. Note here that tubular extraction chamber temperature is stabilised. Fluid flow forced into said chamber is converted into coaxial flow flowing over its concave surface in thin film while carrier gas flow is swirled in upward spiral. Mass exchange is executed at countercurrent of phase components or at stationary has phase.

EFFECT: increased extraction and higher sensitivity of analytic systems.

5 cl, 5 dwg

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