Method of determination of ethanol and other metabolites content in human blood
SUBSTANCE: invention describes a method of determination of ethanol and other metabolites content in human blood by liquid phase chromatography, including preparation of blood distillates by vapour straight distillation and blood component analyis, characterised by the fact that it is combined with one-stage quantitative determination of ethanol, diethyl ester, acetaldehyde, acetone, methylacetate, ethylacetate, propyl alcohol, isobutyl alcohol, butyl alcohol, isoamyl alcohol with the use of capillary chromatographic columns; the concentration of the determined blood components is calculated by formula: where a is chromatographic study results, mg/dm3; V is a distillate volume, cm3; m is a whole blood weight, g.
EFFECT: method can be used in clinical laboratory diagnostics in studies of metabolic disorders caused by alcohol poisoning, and in judicial medical activity for diagnosing of a degree of intoxication of live persons.
1 ex, 1 tbl, 2 dwg
The invention relates to medicine and can be used in clinical laboratory diagnostics in the study of metabolic disorders in humans caused by alcohol poisoning, and in forensic medical practice for diagnosing the degree of intoxication of living persons.
The generally accepted method for determination of ethyl alcohol in human blood is a method of color reactions. The most popular of the 60-ies of the so-called "nitrite" method and method Widmark. "Nitrite" method is very time-consuming, requires rapid processing of the material researched by double distillation with water vapor, a large number of the investigated material (200-300 g). In addition, the method is not sufficiently specific. The way Widmark is used to determine the degree of intoxication of living persons. this Method is quite time-consuming, research is needed 3-4 portions of the material researched and parallel conducting 3-4 blank experiments, indicating poor reproducibility of the method.
There is a method of quantitative determination of the content of ethyl alcohol in blood and human urine by gas chromatography [Methodological letter dated April 22, 1968, No. 10-95/14-32]. The essence of the method consists in the conversion of alcohols to alkyl nitrites, which are then subjected to chromatographic separation. In osnovatyelyei is the measurement of differences in thermal conductivity of the pure carrier gas, coming in comparative camera detector thermal conductivity - katharometer, and the mixture of the analyte with the carrier gas emerging from the chromatographic column in the measuring chamber of the detector. For the study applies the chromatograph laboratory PI-4.
The concentration of ethanol is determined by calculating the ratio of the peak heights of amylnitrite and isopropylacetate (propylitic) followed by interpolation from the calibration graph. The calibration graphs are built using standard solutions of ethyl alcohol in blood and urine.
A significant drawback of this method is the absence of the possibility of quantitative determination of ethyl alcohol in the simultaneous presence in the studied material ethyl, propyl, and isopropyl alcohol. In addition, the above method does not allow for simultaneous determination of blood ethanol, diethyl ether, acetaldehyde, acetone, methyl acetate, ethyl acetate, propyl alcohol, isobutyl alcohol, butyl alcohol, isoamyl alcohol and other metabolites.
The objective of the invention is to develop a method for simultaneous quantitative determination in human blood ethyl alcohol, diethyl ether, acetaldehyde, acetone, methyl acetate, ethyl acetate, propyl alcohol, isobutyl alcohol, butyl Speer is a, isoamyl alcohol and other metabolites by gas chromatography.
The problem is solved in that in the method of determination of content of ethanol and other metabolites in human blood by gas chromatography, including research blood components, while conducting quantitative determination of ethyl alcohol, diethyl ether, acetaldehyde, acetone, methyl acetate, ethyl acetate, propyl alcohol, isobutyl alcohol, butyl alcohol, isoamyl alcohol and other metabolites in the distillate blood using capillary chromatographic column, calculate the concentration of blood components by the formula:
where and is the result of chromatographic studies, mg/DM3;
V - volume of distillate, cm3;
m is the mass of a sample of whole blood,
Figure 1 shows the chromatogram of distillate whole human blood, in figure 2 - model chromatogram of the mixture.
The essence of the method is to obtain distillates blood by direct steam distillation, which is then subjected to chromatographic separation. Separated through column chromatography components of the mixture in turn come in a flame ionization detector, the signals which are recorded in the form of a series of chromatogra the practical peaks. The basis of the detection is the measurement of differences in electrical conductivity of a flame of pure carrier gas flowing in comparative camera detector, and a mixture of the analyte with the carrier gas coming from the chromatographic column in the measuring chamber of the detector. For research use gas chromatograph company Hewlett Packard (USA), model 5890, series 2, with a flame ionization detector and electronic regulation of the flow of carrier gas, column chromatographic capillary HP-FFAP (USA) 50 m × 0.32 mm × 0,52 μm, the internal standard 1,2-propylene glycol, the carrier gas is nitrogen, the temperature program of the column thermostat: initial temperature 50 ° C, 10 min isotherm, further programming of the temperature at 15 degrees per minute up to 220 degrees Celsius, 10 min isotherm, the temperature of the injector - 150 degrees Celsius, the temperature of the detector 230 degrees Celsius, the division of the flow - 1:40.
Quantitative determination of ethanol and other metabolites is carried out as follows. Weigh from 10 to 50 g of whole blood in a distillation flask with a capacity of 500 cm3then add 50 cm3of distilled water. The mixture is stirred. The flask with the mixture attached to the reflux condenser with a liquid trap and a fridge with a straight tube. With careful Naga is Ivanyi (rapid expansion) the mixture is subjected to direct distillation and collect exactly 25 cm 3of distilled water in a volumetric flask with a capacity of 25 cm3. The receiving flask should be further cooling. 25 cm3the obtained distillate add 2 mm3internal standard. The mixture was thoroughly stirred and subjected to chromatographic study. Capillary chromatographic column allows to reliably separate the components of the distillate blood and simultaneous determination of ethyl alcohol, diethyl ether, acetaldehyde, acetone, methyl acetate, ethyl acetate, propyl alcohol, isobutyl alcohol, butyl alcohol, isoamyl alcohol and other metabolites.
Chromatograph pre-calibrated by ethyl alcohol and other metabolites, as well as propylene glycol, using at least three of the calibration mixtures. The values of the retention time, the calibration coefficients and the concentration of the internal standard is recorded in the computer memory. The estimated concentration of the internal standard 83,2 mg/DM3.
The study was performed on whole human blood (nine healthy men aged 25 to 39 years).
The results are shown in the table.
|The content of endogenous ethanol and acetaldehyde in human blood|
|№ p/p||The content of ethyl alcohol, 10-3%||The content of acetaldehyde, 10-3%|
The obtained data on the content of endogenous ethanol in blood of the person correspond to the literature, according to which normal, without the introduction of outside, in human blood may contain 0.004% alcohol. In addition, the proposed method allows to reliably diagnose the degree of intoxication of living persons (table, outcome). According to literature data, the alcohol concentration of 0.02 to 0.20% of calls depending on individual sensitivity, the condition and type of nervous activity intoxication varying degrees.
Thus, in the course of the research it is established that the proposed method for determination of content of ethanol and other metabolites in human blood allows to obtain accurate and reproducible results.
Method for determination of content of ethanol and other metabolites in human blood by gas chromatography, including the production of distillates blood by direct steam distillation and research blood components, characterized in that conduct quantitative determination of ethyl alcohol, diethyl ether, acetaldehyde, acetone, methyl acetate, ethyl acetate, propyl alcohol, isobutyl alcohol, butyl alcohol, isoamyl alcohol in one study using capillary chromatographic column, the calculation of the concentration of the detected blood components produced according to the formula:
where and is the result of chromatographic studies, mg/DM3;
V - volume of distillate, cm3;
m is the mass of a sample of whole blood,
SUBSTANCE: patients in their 16 weeks of pregnancy initiated by ART methods are examined for fibrinogen levels, and chorion volume is calculated by formula: V =π/6∙ABC (cm3), where A is a maximum length, B is a maximum width, and C is a maximum thickness of the chorion measured by ultrasonic examination. Further, a prognostic index is calculated: F=1.35*FG-0.02*V-5.75, where FG is fibrinogen levels (g/l), V is chorion volume (cm3), const - 5.75. If the F value is less than 0, favorable prognosis for a foetus is considered, while F more than 0 shows a high probability of intrauterine growth retardation.
EFFECT: use of a method allows higher prediction accuracy of intrauterine growth retardation in the second trimester of pregnancy after auxiliary reproductive technologies.
SUBSTANCE: diagnostic technique for cobalt-induced nephropathy in experimental animals suffering chronic poisoning, involves examination of the animals for erythrocyte and renal tissue malondialdehyde (MDA) concentration and simultaneously, for Na+, K+-ATP-ase activity in renal tissue, and if the MDA values are within the range 4.96±0.03 to 5.32±0.06 nmol/ml of erythrocyte mass and more, and in cortical and medullary substance cells of renal tissue respectively within the range 2.49±0.12 to 2.87±0.06 and 4.56±0.06 to 5.25±0.08 nmol/mg of protein and more, and the Na+, K+-ATP-ase activity values of the cortical and medullary substance of renal tissue within 2.08±0.03 to 1.31±0.14 and 5.28±0.18 to 3.92±0.02 mcmole Rn/mg protein/hour and less respectively, cobalt-induced nephropathy is diagnosed.
EFFECT: technique allows extending a notion of pathogenetic mechanisms of nephropathy development with underlying chronic cobalt-induced intoxication in experimental animals.
5 tbl, 1 ex
SUBSTANCE: bone marrow is sampled from breast; the sample is prepared for analysis and microscopic study in at least 50 fields of vision, in each of which a number of clusters consisting at least of three plasma cells is determined. Further, an evaluation index is described as an arithmetical mean derived from division of all clusters found on a number of the counted fields. If the index is less than 1, an avascular type of the clinical course of multiple myeloma is stated, while the value of this index exceeding 1 enables to state a vascular type of the clinical course of multiple myeloma.
EFFECT: use of the method allows diagnosing a type of the clinical course of multiple myeloma at the earliest stage that shall provide substantially higher clinical effectiveness.
4 dwg, 4 ex
SUBSTANCE: invention relates to medicine, namely to obstetrics, and can be used for predicting syndrome of fetus development retardation (SFDR) in the second trimester in patients with HIV. For this purpose carried out is analysis of screening examination of pregnant women infected with HIV in the second trimester. First determined is viral loading (VL), expressed in number of HIV RNA copies in 1 ml of blood plasma. After that, week of gestation age, at which carrying out of chemical therapy began (BegCT) is fixed and number of anti-viral chemical preparations (NumCP) is estimated in points: in case of monotherapy - 1 point, in case of tritherapy - 3 points. Then, on the basis of data of ultrasound and dopplerometric examination presence of feto-placental failure (FPF) is detected, presence of FPF being estimated as 1 point, absence of FPF - as 0 points. After that, by means of polymerase chain reaction presence in blood of accompanying viral pathology (AVP) in form of hepatitis B virus, hepatitis C virus, cytomegalovirus infection, virus of Herpes simplex is detected, presence of AVP is estimated as 1 point, absence of AVI - as 0 points. In addition, determined are mother's constitution (MC) and father's constitution (FC), hypersthenic type is estimated as 0 points, normosthenic type - as 1 point, hyposthenic type - as 2 points. After that, by formula WEIGHT=2901.2-0.001·VL+24.5·BegCT-37.8·NumCP-23.3·FPF-12.3(AVP)-166.2·MC-21.4·FC predicted value of baby's weight at birth is calculated. If obtained value constitutes less than 2800 g, hypotrophy of newborn baby is predicted, if obtained value is more than 2800 g, it is considered that there is no risk of hypotrophy of newborn baby.
EFFECT: being widely available, method provides possibility of timely prediction of SFDR in pregnant women with HIV, which makes it possible to optimise tactics of pregnancy management in case of HIV infection.
SUBSTANCE: one of the systems is a two-component system and has a reagent component for storing one or more reagents and a processing component for processing one or more reagents during analysis. The reagent component and the processing component can be connected to each other to form a cartridge. Furthermore, the reagent and/or processing component has at least one compartment configured to receive analysis wastes, wherein the reagent component does not participate in reagent processing during analysis, except receiving wastes from the processing component. Another cartridge system is a system having a reagent component for storing one or more reagents, a processing component for processing one or more reagents during analysis and a sensor component, having at least one sensitive element for detecting analyte. The reagent, processing and sensor components are separate components which can be connected to each other to form a cartridge. The invention also discloses a cartridge system having a reagent component for storing one or more reagents, a processing component for processing one or more reagents during analysis and a sample preparation component for sample preparation during analysis. The reagent component and processing component can be connected to each other to form a cartridge.
EFFECT: improved cartridge system.
45 cl, 27 dwg
SUBSTANCE: invention describes method of noninvasive potentiometric determination of oxidant/antioxidant activity of biological tissues, which includes bringing analysed object in contact with electroconductive medium, which containing mediator system and estimation of oxidant/antioxidant activity by change of potential difference on electrodes, introduced into electroconductive medium, and electroconductive medium represents gel, which contains as mediator system a pair of chemical compounds, containing element in different degrees of oxidation, electrodes through gel contact with the analysed object, and oxidant/antioxidant activity is determined by formulae. For realisation of method claimed is device, which includes apparatus for measuring potential difference and filled with electroconductive medium reservoir with working electrode and electrode of comparison, connected with device for measuring potential difference, where reservoir is made open from one side. As electroconductive medium applied is gel, and working electrode is made in form of plate, placed from the side of open part of reservoir and partially covering it.
EFFECT: increase of reliability and accuracy of obtained results.
4 cl, 5 ex, 6 dwg
SUBSTANCE: method includes determining complex of serum cytokines in period of disease beginning. In case of tick-born encephalitis level of interleukin-1 α is determined in interval 298-358 pg/ml, interferon-γ-295-350 pg/ml and interleukin-10-115-150 pg/ml; at ixodid tick-borne borreliosis - interleukin-1 α 178-270 pg/ml, interferon-γ 125-190 pg/ml and interleukin 10-11.5-18 pg/ml; in case of mixed infection, generated by virus of tick-borne encephalitis and borrelia - interleukin-1 α- 30-75 pg/ml, interferon-γ 115-130 pg/ml and interleukin-10 -22-65 pg/ml.
EFFECT: method is simple, makes it possible to determine disease causative agent reliably and accurately in short terms.
1 tbl, 3 ex
SUBSTANCE: method of predicting development of arterial hypertension in pregnant women consists of realisation of DNA separation from peripheral venous blood, carrying out polymerase chain reaction, finding out data about presence of polymorphism of α-adducin 1 gene. If carrying genotype 460WW of gene of α-adducin 1 ADD1 G460W is detected, conclusion about risk of hypertension development in pregnant women is made.
EFFECT: increased efficiency of predicting arterial hypertension development in pregnant women.
1 ex, 1 tbl, 2 dwg
SUBSTANCE: method includes DNA separation from peripheral venous blood, analysis of genotype of locus - 322 VNTR of receptor of tumour necrosis factor of 2 type. In case if genotype 2/2-322 VNTR TNFR2 is determined -little area of myomatous knots is predicted. In case of genotypes 1/1-322 VNTR TNFR2 or 2/1-322 VNTR TNFR2, large area of myomatous knots is predicted.
EFFECT: invention application makes it possible to obtain criteria of estimating myomatous knots area, define tactics and individualize treatment basing on obtained data.
2 tbl, 2 ex
SUBSTANCE: invention describes method of predicting risk of development of therapeutic resistance in patients with bronchial asthma, by analysis of patients blood 3 month after beginning of treatment, DNA, separated from lymphocytes of peripheral venous patients blood, is analysed, values of levels of genes INC2, INC3, GJA8, SV2 expression are determined by microchips Affymetrix and probability of relating individuum to group with high risk of developing therapeutic resistance and low risk of therapeutic resistance development, and, R1 for individuums with high risk of therapeutic resistance development is estimated by formula: R1=-351.966-, 274*INC2+46.608*INC3+129.530*GLA8+105.438*SV2A, where 274; 46.608; 129.530; 105.438 - numeric values are coefficients; (-351.966) is constant for individuums with high risk of BA development; INC2, INC3, GJA8, SV2 are genotytpes, and R2 for individuums with low risk of therapeutic resistance development is calculated by formula: R2=-403.217-64.874*INC2+55.603*INC3+141.648*GJA8+116.342*SV2A, where 64.874; 55.603; 141.648; 116.342 are numeric values which are coefficients; (-403.217) is constant for individuums with high risk of BA and at R1>R2 high risk is predicted , and at R1<R2 low risk of therapeutic resistance is predicted in patient with bronchial asthma.
EFFECT: increase of accuracy and self-descriptiveness.
4 ex, 3 tbl
SUBSTANCE: invention relates to a method for ion-exchange separation of methionine and glycine and can be used in biochemistry, pharmaceutical and food industry. The method involves separation of methionine and glycine in two steps. At the first step amino acids undergo sorption with enrichment of the sorbent phase with glycine, and the solution at the output enriched with methionine. For this purpose, polyampholyte Purolite S950 in H-form is prepared. The mixture of two aliphatic amino acids undergoes sorption in a countercurrent column with a fixed sorbent layer. For this purpose, a solution containing a mixture of glycine and methionine is fed from below and glycine is undergoes sorption on polyampholyte Purolite S950. Methionine, appears at the output, the aqueous solution of which is sorbed in a receiver at the output of the column and after a certain time - the amino acids. Sorption is stopped. During sorption, samples are collected at defined time intervals. Total concentration of amino acids is controlled using an iodimetric method, and concentration of methionine is controlled using a spectrophotometric method, while glycine concentration is controlled based on concentration difference: between total concentration and methionine concentration. The degree of separation of the initial solution is equal to 60%. At the second step, glycine is eluted with hydrochloric acid solution at pH 1.2 from the sorbent while feeding glycine-containing eluate from the top, and sorbed in the receiver. Concentration of glycine is equal to 70%. After desorption of glycine, the mixture of amino acids undergoes complete desorption. Polyampholyte takes the initial shape and is ready for operation. Samples are collected at defined time intervals and each sample is analysed using iodometric and spectrophotometric methods. For complete separation of glycine from methionine, the two-step process of separating the mixture of amino acids obtained at the output of the column is repeated.
EFFECT: method enables efficient separation of methionine and glycine by combining sorption and desorption processes while excluding the sorbent regeneration step, and reduce the volume of wash water without using considerable amount of auxiliary reactants.
2 dwg, 2 tbl, 1 ex
SUBSTANCE: analysed sample is infused with an organic insulating agent. The obtained extract is purified through chromatography on a column of silica gel L 40/100 µ, while performing elution with a mixture of organic solvents. The analysed substance is determined using a chromatographic technique using a mobile phase which contains hexane, dioxane and propanol-2. The organic insulating agent is toluene. The toluene extract is dehydrated with anhydrous sodium sulphate. During the purification process, hexane is first passed through the column and elution is then carried out with a hexane-dioxane-propanol-2 solvent mixture (8:3:0.6 by volume). Eluate fractions containing the analysed substance are merged. The eluent is evaporated. The residue is dissolved in the hexane-dioxane-propanol-2 solvent mixture (15:5:1 by volume) and detection is carried out using high-performance liquid chromatography (HPLC) in a 64×2 mm column filled with Silasorb-600 sorbent using a hexane-dioxane-propanol-2 mobile phase (15:5:1 by volume) and a UV detector.
EFFECT: high accuracy and sensitivity of analysis.
3 ex, 4 tbl
SUBSTANCE: there is described a method of quantitative cyclosporine A evaluation in patients' blood involving blood protein precipitation by adding an aqueous solution of zinc sulphate and methanol, mixing, centrifuging and sampling a centrifugate; separating the centrifugate ingredients by reverse phase high-yield liquid chromatography, mass-spectrometre detecting cyclosporine A and evaluating the cyclosporine A concentration with plotting a calibration curve; blood protein are precipitated with using whole blood; blood protein precipitation is followed by additional salt impurity precipitation by adding methanol to the centrifugate to the general concentration not less than 90 vol. %, mixing again, centrifuging and sampling the centrifugate; separating the centrifugate ingredients, detecting and evaluating the cyclosporine A concentration.
EFFECT: method allows facilitating analysis simplicity and universality with providing adequate sensitivity and selectivity ensured by the absence of necessity for the internal standard and online extraction and lower requirements to specification of the used mass spectrometre by conducting preliminary impurity precipitation.
SUBSTANCE: proposed method comprises forcing analysed product into chromatograph first circuit to define carbon sulphide at its concentration exceeding 0.1 wt % and, at a time, into second circuit at carbon sulphide concentration lower than 0.1 wt %. First circuit comprises piston-type metering valve and packed columns arranged in heated temperature-controlled cabinet and filled with polymer adsorbent, 0.1-1.5 m-long precolumn and 0.5-5 m-long main column, and heat conductivity detector. Second circuit comprises piston-type metering valve, packed capillary columns arranged in heated temperature-controlled cabinet and filled with polymer adsorbent, 0.1-1.5 m-long precolumn and 15-50 m-long main column with their ID making 0.23-0.32 mm, and sulfur-selective detector. Metering valves are arranged sequentially in both circuits along sample feed direction.
EFFECT: shorter easier process.
5 cl, 1 dwg, 2 tbl, 1 ex
SUBSTANCE: urine is sampled, centrifuged that is followed by solid-phase extraction with Oasis HLB sorbent with using 100% acetonitrile as an extraction fluid for dimethyl terephthalate extraction. Said solid-phase extraction is conducted by consequent passing 100% acetonitrile, distilled water, the urine sample after centrifugation, distilled water, 20% aqueous acetonitrile and 100% acetonitrile as the extraction fluid through the sorbent; then the prepared extract is analysed by liquid chromatography with using as a mobile phase mixed acetonitrile and water in the at the varying ratio 25:75 vol. % to 90:10 vol. % respectively in a gradient mode which is enabled with combining chromatography by supplying at first the mobile phase containing mixed acetonitrile and water in the ratio 25:75 vol. % for 10 minutes. Then increasing the acetonitrile concentration in the mobile phase to 90 vol. % for 5 minutes and passing such mobile phase for another 5 minutes is followed by decreasing a volume amount of acetonitrile to 25 vol. % for 5 minutes and passing such mobile phase through a column for 10 minutes, while an amount of dimethyl terephthalate is determined by a calibration chart.
EFFECT: high sensitivity of the method combined with selectivity and availability for routine analyses.
5 cl, 6 tbl
SUBSTANCE: invention describes a method of quantitative evaluation of blood acetic, propionic, isobutyric, butyric, valeric, isocapronic and capronic acids by gas chromatography analysis wherein a blood sample is acidified with 1 % sulphuric acid to pH 2-3, evaluated acids are extracted with isobutyl alcohol volume of which is related to the blood sample volume as 1:1. The protein separation is enabled by centrifugation. 2-3 drops of 0.4 % alkali is added, and the extract is evaporated dry, further the solid residue is added consistently with 1 % sulphuric acid and isobutyl alcohol that is followed with gas chromatography separation of the mixed acids in a capillary column with a flame ionisation detector, and the amount of each acid is evaluated by a calibration diagram.
EFFECT: higher sensitivity and accuracy of the method of quantitative evaluation of acetic, propionic, isobutyric, butyric, valeric, isocapronic and capronic acids if found in blood together.
5 cl, 1 ex, 4 tbl
SUBSTANCE: disclosed is a method of detecting unknown substances in body fluids of patients taking narcotic or psychoactive substances. The method involves preparation of three body fluid samples - the first through extraction with re-solution, the second through acid hydrolysis and the third through enzymatic hydrolysis. The first sample undergoes GC/MS analysis at temperature gradient of 15°C/min and data are analysed by comparing with a data base from which features of the unknown substance are detected, specifically spectra with m/z values which coincide with basic ions of the narcotic or psychoactive substance or metabolites and content of the unknown substance in the sample. The second sample undergoes GC/MS analysis at temperature gradient of 25°C/min and the third sample undergoes GC/MS analysis also at temperature gradient of 15°C/min and, if content of the unknown substances in the last two samples is higher than the in the first, the narcotic or psychoactive substance undergoes GC/MS analysis for presence of the unknown substance also at temperature gradient of 15°C/min, and if also not present in the basic substance. Presence of the unknown substance in intact body fluid is also checked, for which a sample of the intact body fluid is prepared via acid hydrolysis and undergoes GC/MS analysis at temperature gradient of 15°C/min and 25°C/min, and if the unknown substance is detected in the intact body fluid, the substance is classified as endogenous, and in the absence of features, an aliquot of the first sample is mixed with the sample of intact body fluid. The sample is prepared via acid hydrolysis of the mixture. The sample undergoes GC/MS analysis at temperature gradient of 15°C/min and 25°C/min. Further, content of the unknown substance is determined from results of both analysis modes and then compared with content of the known substance in the first sample. If content values of the unknown substance in the said three samples coincide, the unknown substance is classified as a new, previously unknown product of metabolism of the basic narcotic or psychoactive substance.
EFFECT: possibility of unique identification of chemical compounds and their fragments in arbitrary combinations while increasing accuracy and rapidness of detection.
SUBSTANCE: sodium fluoride is added to an analysed sample in amount of 10% of the mass of the biological object and infused twice in 45 minutes with portions of ethyl acetate, the mass of each of which is twice higher than the mass of the biological material. Separate extractions are combined, filtered through anhydrous sodium sulphate. The solvent from the filtrate is evaporated at temperature 50-60°C. The residue is dissolved in a mixture of hexane-dioxane-propanol-2 solvents. Chromatography is performed in a column with silica gel L 40/100 µ using a hexane-dioxane-propanol-2 mobile phase. The eluate fractions which contain the analysed substance are merged. The eluate is evaporated. The residue is dissolved in a mixture of hexane-dioxane-propanol-2 solvents and the analysed substance is determined via high performance liquid chromatography (HPLC) in a 64x2 mm column filled with Silasorb 600 sorbent using a hexane-dioxane-propanol-2 mobile phase and a UV detector.
EFFECT: invention shortens the duration of detecting tetraethyl thiuram disulphide in blood and increases its sensitivity.
3 tbl, 2 ex
SUBSTANCE: biological tissue is crushed, processed twice for 30 minutes with portions of ethyl acetate, weight of each twice exceeding weight of a biological object; prepared extractions are combined, filtered through anhydrous sodium sulphate; a solvent is evaporated from the filtrate; the residue is dissolved in acetonitrile; the prepared solution is watered down in the volume ratio 1:4, extracted twice in portions of chloroform, volume of each being equal to volume of a hydrophilic layer; the chloroform extracts are combined, steamed to a dry residue; the residue is dissolved in mixed solvents hexane-dioxane-propanol-2, cleaned in a silica gel column L 40/100µ with using a mobile phase hexane-dioxane-propanol-2; eluate fractions containing an analysed substance are combined; the eluent is evaporated; the residue is dissolved in mixed solvents hexane-dioxane-propanol-2 and analysed by a HELC method in a column of dimensions 64×2 mm filled with the sorbent Silasorb 600 with using a mobile phase hexane-dioxane-propanol-2 and a UV detector.
EFFECT: invention allows higher selectivity, sensitivity and accuracy of biological material analysis for tetramethylthiuramdisulfide.
4 ex, 5 tbl
SUBSTANCE: in the method of detecting phenol in aqueous solution via reverse-phase micro-column high-performance liquid chromatography with a preliminary sample preparation step through liquid-liquid extraction with acetonitrile, extraction is carried out at temperature 263±2 K for 30 minutes with ratio of equilibrium volume of water to the organic phase equal to 1:1.
EFFECT: simple and cheap method, high degree of extracting phenol, low detection limit.
1 ex, 3 tbl
FIELD: chemical engineering; medical engineering.
SUBSTANCE: method involves plotting two chromatograms one of which is based on radioactivity (No 1) and the other one on ultraviolet absorption (No 2) or on radioactivity (No 1) and on fluorescence (No 2) and chromatogram specific relative to ultraviolet absorption (No 3) or relative to fluorescence (No 3). Material quality is estimated to be the more high the more close studied labeled compound peak shape is to trapezoid shape on the third chromatogram.
EFFECT: high accuracy of the method.