Method of quantitative evaluation of blood acetic, propionic, isobutyric, butyric, valeric, isocapronic and capronic acids by gas chromatography analysis

FIELD: medicine.

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

 

The invention relates to medical Toxicological research, in particular to sanitary toxicology, and can be used for the quantitative determination of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids in the blood for diagnostic purposes gastroduodenitis.

The method for determination of volatile fatty acids (acetic, propionic, butyric, ISO-butyric, ISO-valerianic, valerianic, nylon and ISO-Caproic) and malondialdehyde in biological environments on a gas chromatograph with a flame ionization detector with temperature programming mode from 60 to 220°C at a rate of 10°C in 1 min analysis Time 45 min (ed. St-in the USSR №1725095 from 29.06.89). According to a known method to produce samples of exudate in the amount of 1-2 ml, the sample is placed in a special sealed vial filled with oxygen-free gas (a mixture of nitrogen and 85%, hydrogen 10% carbon dioxide 5%). Then analyze the equilibrium vapor phase. To do this in a bottle with this breakdown type crystalline potassium bisulfate, close the vial with a rubber stopper with PTFE gasket and put it in a special cartridge for sealing. Hold the temperature at 90°C for 10 min and Then through the metering needle pneumatic dispenser test centuries what is in the column of the chromatograph. The peaks in the chromatograms of the samples of exudate are identified by comparing the time of their release over time of the output peaks of the investigated acids and malondialdehyde in a similar study of the standard sample.

Also there is a method of quantitative determination of organic acids and anhydrides with their joint presence using the method of gas chromatography (ed. St-in the USSR №347656 from 11.03.1969,) consists in the fact that the analyzed sample before chromatographytandem sequentially treated with an ethereal solution of diazomethane, and then a mixture of ethyl alcohol, diethylsulfate and sulfuric acid when heated. The acid under the action of diazomethane formed methyl esters, and anhydrides under the action of ethyl alcohol and diethylsulfate - ethyl esters. The resulting mixture chromatographic.

A known method of determination of acetic, propionic and butyric acids in the saliva for the diagnosis of chronic gastroduodenitis and functional dyspepsia in children (RF patent No. 2270610 04.08.2004,). According to this method prepares a sample of saliva by acidification with 1 ml of saliva one drop of 10% sulfuric acid. Gas chromatographic separation of the analyzed acids carried out in a glass column of 1 m length, 3 mm in diameter, filled Porapak Q (USA) coated repostory acid, under isothermal conditions at a temperature in the column 200°C, the chromatograph MOSH - model 3700. The detector is a flame ionization, carrier gas - helium. The analysis of saliva in the examined children, the authors found that in healthy children, the concentration of acetic acid in saliva was 0.60 mg/DM3, propionic acid - 0,296 mg/DM3, butyric acid - 0,528 mg/DM3. In children with functional dyspepsia content of acetic acid in saliva was 54,64 mg/DM3, propionic acid is 11.11 mg/DM3, butyric acid - 1,14 mg/DM3. In children with chronic gastroduodenitis acetic acid in saliva was detected at a concentration of 474,3 mg/DM3, propionic acid of 49.6 mg/DM3, butyric acid of 50.2 mg/DM3.

In the abstract of SOIC. academic degree Dr. med. Sciences I. Bykov Molecular mechanisms of pathogenesis and the role of metabolic L-carnitine. Novosibirsk, 2006, it is proposed to determine the concentration of short-chain fatty acids in tissues and body fluids by gas chromatography after extraction with diethyl ether n-valerianic acid as internal standard. The range of detectable concentrations of propionic acid in the tissues and blood plasma is 5,03-6,07 mg/DM3ISO-butyric acid - 1,41-2,03 mg/DM3.

Source of information: DOS is N. R., Elliot D., Jones, K., Paper and thin layer chromatography in biochemistry. Methods of separation. Reference biochemist. M.: Mir, 1991, s use paper chromatography to separate formic, acetic, propionic, butyric, valerianic and Caproic acids. Chromatography is carried out in a solvent containing a volatile base, for example MN3the ethylamine. In the case of a mixed solvent of ethanol: 3 M NH3=95:6 ratio retention is 0.31 for formic acid, 0,33 - acetic acid, 0,44 - propionic acid, 0,54 - butyric acid, 0.60 - for valerianic acid and 0.68 - Caproic acid. Detection acids perform a pH-indicator methods using a 0.04%solution of bromocresol green in ethanol or water, or 0.04%-aqueous solution of bromocresol purple in a mixture of formalin: ethanol = 1:5 pH 5, or 0.1%aqueous solution of thymol blue. The sensitivity of detection of 5 mg. The disadvantage of this method is semi-quantitative determination of acids and low detection sensitivity.

Also known a method of determining volatile fatty acids (AGV) using gas-liquid chromatography, as described in the book the authors cottage of wattle L.N., Ilina NI, Kondratova O.A., Stevulova A.M. "Evaluation of disorders in acute intestinal microbiocenosis of infecti the x in children and their correction. Hard. Patient." 2004; 2(9): 11-6. To determine the AGV is proposed to use the infiltration tests. Obtaining infiltration takes place according to the following scheme: in a disposable plastic tube is weighed 2-3 grams content of the sample on an analytical balance accurate to 3 digits, the sample is poured 1 ml of 0.02 N hydrochloric acid, 1 ml of distilled water and 1 ml of a standard substance. The tube is closed with a glass stopper and homogenize the mixture by vigorous shaking. If you want to determine the concentration of AGV in the blood, the mixture was added as a 0.5 ml of 0.01 N perchlorate acid. Tube with a homogeneous mixture is centrifuged for 10 minutes at 6000 rpm the resulting infiltration should be transparent, have an acid reaction. The chromatography was carried out at a temperature of thermostat 140°C, evaporator 240°C. Disposable glass capillary selected microcaecilia supernatant samples. The capillary tube sealed on one side and is attached to a special device input samples by which the capillary is placed in the evaporator and run the program read chromatogram.

Also there is a method of gas-liquid chromatography for quantitative determination of acetic, propionic and butyric acids in the blood of patients with viral hepatitis (Akishin AS, Zolotarev D.V., Akishina M.A., Balykin SCI "short-chain W is rnie acid in the blood of children with viral hepatitis", Ivanovo state medical. Academy of health Ministry of Russia, Ivanovo, 2005). According to a known method the research was conducted on the chromatograph MESH model 3700 with a flame ionization detector. Chromatographic separation of the acids was performed on a glass column of 1 m length, 3 mm in diameter, filled with Porapak Q (USA) coated with a phosphoric acid. The column temperature 200°C, evaporator - 250°C. the Speed of the carrier gas (helium) 25 ml/min In healthy children acetic acid in the blood were found in concentrations 0,040-0,222 mg/DM3, propionic - 0,0148-0,362, oil - 0,024-0,077 mg/DM3. In patients with viral hepatitis acid content in the blood was increased.

The disadvantage of this method is the low sensitivity of acetic acid in the blood 0.04 mg/DM3, propionic - 0.015 mg/DM3oil - 0.024 mg/DM3. Determination of butyric acid interferes with ISO-butyric acid.

The technical result achieved by the invention, is to improve the sensitivity and accuracy of the method of determination of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids at their joint presence in the blood.

The specified technical result is achieved by the proposed method for quantitative determination of acetic, propionic, and what about the oil, oil, valerianic, ISO-Caproic and Caproic acids in the blood by gas chromatography analysis, according to which the blood sample is acidified with 1%sulfuric acid to pH 2-3, carry out the extraction of the designated acid isobutyl alcohol, the volume of which correlates with the volume of the blood sample as 1:1, conduct Centrafricaine for separating proteins, add 2-3 drops of 0.4%aqueous alkali solution and the extract is evaporated to dryness, then to dry the precipitate successively added 1%solution of sulfuric acid and isobutyl alcohol and exercise gas chromatographic separation of the mixture of acids on a capillary column with a flame ionization detector, and the amount of each acid set via a calibration curve.

Add to the dry residue of 1%aqueous solution of sulfuric acid produced in the volume of 1:40, and isobutyl alcohol is 1:20 to the volume of blood samples, respectively.

The extraction of the designated acid isobutyl alcohol produced within 5-10 minutes

Centrifugation is carried out at 6000 rpm for 10 minutes

Gas chromatographic separation of the mixture of acids on a capillary column with a flame ionization detector is performed with the use of a hardware-software complex based on gas chromatograph "Crystal 2000" detector-ionization in the flame at the capillary to which the PMC HP-FFAP - 50 m*0,32 mm*0,5 µm at temperatures: column - 70°C-160°C-180°C-280°C; evaporator - 250°C; detector, 280°C; flow rate of carrier gas 1 (nitrogen) - 40 cm3/min; flow rate of carrier gas 2 (nitrogen) - 14-30 cm3/min

Experimentally detected that the specified technical result is exactly the set of proposed signs, the implementation of which achieved high sensitivity and accuracy of these short-chain fatty acids in their joint presence.

In the laboratory has conducted a number of experiments to establish the materiality of signs that form the basis of the proposed method.

Example. According to the claimed method took 2 cm3blood containing acetic, propionic, ISO-butyric, butyric, valeric, ISO-Caproic and Caproic acid, acidified its 1%solution of sulfuric acid to pH 2-3, extracted with 2 cm3isobutyl alcohol for 5-10 minutes To denature the protein extract was centrifuged at 6000 rpm for 10 min, then added 2-3 drops (0.1 cm3) 0,4%-aqueous solution of alkali (sodium or potassium), was evaporated to dryness, the residue was acidified to 0.05 cm31%sulfuric acid solution was added 0.1 cm3ISO-butyl alcohol. Next performed quantitative determination of acids in the prepared sample is x by gas chromatography on a calibration curve using a hardware-software complex based on gas chromatograph "Crystal 2000" detector-ionization in the flame. The completeness of the separation of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids were achieved on a capillary column HP-FFAP - 50 m*0,32 mm*0,5 µm at temperatures: column - 70°C-160°C-180°C-280°C; evaporator - 250°C; detector, 280°C; flow rate of carrier gas 1 (nitrogen) - 40 cm3/min; flow rate of carrier gas 2 (nitrogen) - 14-30 cm3/min

To construct the calibration graph used the following method. In a measuring tube of 10 cm3containing ISO-butyl alcohol in an amount of 5 cm3enter 15 mm3(μl-μl) acetic acid, 10 mm propionic, ISO-butyric and butyric acid and 5 mm3valerianic, ISO-Caproic and Caproic acids. The content in the original certified solution is acetic acid - 3120 mg/cm3, propionic - 1996 g/cm3ISO-butyric acid - 1916 µg/cm3, butyric acid - 1918 µg/cm3, valerianic acid - 930 ág/cm3ISO-Caproic acid - 923 mg/cm3, Caproic acid - 923 mg/cm3. Shelf life certified solution was 12 hours.

Calibration characteristic was determined by the method of absolute calibration. It expresses the dependence of peak area on the chromatogram (mm2) from the ground (mcg) and is based on 5 series of working standard solutions. Each series consisting of 5 standard is bound solutions cook in a measuring tubes of 10 cm3. For this purpose, in each test tube add acetic acid, propionic acid, ISO-butyric acid, butyric acid, ISO-Caproic acid, Caproic acid, valerianic acid in accordance with table 1 and adjusted ISO-butyl alcohol to 2 cm3. The obtained extracts - standard solutions chromatographic on the chromatograph detector ionization in the flame. Optimization of gas chromatographic parameters for the definition of these acids in the blood was performed using a hardware-software complex based on gas chromatograph "Crystal 2000" detector-ionization in the flame. The completeness of the separation of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids were achieved on a capillary column HP-FFAP - 50 m*0,32 mm*0,5 µm at temperatures: column - 70°C-160°C-180°C-280°C; evaporator - 250°C; detector, 280°C; flow rate of carrier gas 1 (nitrogen) - 40 cm3/min; flow rate of carrier gas 2 (nitrogen) - 14-30 cm3/min

Studies on the choice of solvent (organic solvent) to extract the volatile fatty acids from the blood were tested following solvents: benzene, ethyl alcohol, diethyl ether, hexane, ISO-butyl alcohol, and inorganic acids such as hydrochloric, phosphoric, sulfuric.

Poppy is emalina the degree of extraction of the studied acids from blood samples was achieved using sulfuric acid, which was used in further studies.

In the course of the experimental work on the choice of extractant used a number of organic solvents and studied the dependence of the degree of extraction from the type of inorganic acids and nature of organic solvents results are shown in table 2.

The data in table 2 show that only when using ISO-butyl alcohol along with sulfuric acid, the aggregate of all announced transactions, the proposed method ensures the maximum extraction (the highest degree of extraction) of these acids from blood samples at the selected optimum conditions for gas chromatographic analysis.

The proposed method was investigated by a number of blood samples with different content of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids.

The result of this experiment established that the greatest extraction of the studied acids from the blood occurs in the extraction of ISO-butyl alcohol in an acidic environment (pH 2-3), adding 0.4 percent aqueous solution of alkali (for translation acid salt), centrifugation, evaporation to dryness, the one stripped off acidification of the extract with 1%sulfuric acid solution, in a volume ratio to the blood sample as 1:40, and the introduction of ISO-butyl alcohol in volumetric zootoxin and to the blood sample as 1:20. Moreover, the deviation from these volumetric ratios more or less side led to the increase of error definitions.

The method of measurement using the proposed method provides the results of measurements with an error not exceeding the values given in tables 3 and 4. The data given in tables 3 and 4 show that the detection sensitivity in the analyzed sample volume of blood in the joint presence amounted to: acetic acid is 0.003 ág; propionic acid of 0.002 µg; ISO-butyric acid is 0.001 μg; butyric acid is 0.001 μg; valerianic acid - 0,0005 g; ISO-Caproic acid - 0,0005 g; Caproic acid - 0,0005 mcg.

The error of determination was for acetic acid - 17,66%; propionic acid - 13,93; ISO-butyric acid - 19,06%; butyric acid - 11,21%; valerianic acid - 15,64%; ISO-Caproic acid - 17,56%; Caproic acid - 20,76%, even when their joint presence.

The application of the proposed method can improve the detection sensitivity for acetic, propionic and butyric acids in 2 times in comparison with the known method). It should also be noted that the prior art unknown method of determination of such a circle acids in blood during their joint presence.

The proposed method is available and can be recommended for clinical studies in the laboratory.

Table 1
Standard solutions to establish the calibration graph
The number of mixture12345
Acetic acid, the Volume of the original STD. Solution, cm3235710
The concentration of acetic acid, mg/cm33,12to 4.687,810,92the 15.6
Propionic acid is the Volume of the original STD. Solution, cm3235710
The concentration of propionic acid, mg/cm31,9962,994,996,99998
ISO-butyric acid, the Volume of the original STD. Solution, cm3235710
The concentration of ISO-butyric acid, g/cm30,962,874,79of 6.719,58
Oil acid Volume of the original STD. solution, cm3235710
The concentration of Butyric acid, g/cm30,9592,884,79of 6.719,59
Valeric acid Volume of the original STD. solution, cm3235710
The concentration of Valerianic acid, mg/cm30,471,39 2,333,264,65
ISO-hexanoic acid, the Volume of the original STD. solution, cm3235710
The concentration of ISO-Caproic acid, mg/cm30,461,392,313,23to 4.62
Hexanoic acid, the Volume of the original STD. solution, cm3235710
The concentration of Caproic acid, mg/cm30,461,392,313,23to 4.62

Table 2
The average completeness of the extraction of acetic acid, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids from blood samples by nature organic p is Stroiteley and type of inorganic acids
ComponentOrganic solvent
HexaneISO-butyl alcohol
Type inorganic acid
2% H2SO41% H2SO40,5% H2SO42% H2SO41% H2SO40,5% H2SO4
The degree of extraction, %
1. Acetic acid25±2,05025±2,0020±2,71759±4,05260±4,90555±3,312
2. Propionic acid39±2,13435±2,3530±2,1350±4,2365±5,9945±3,391
3. ISO-butyric acid32±2,2529±1,95 35±2,9980±2,75688±6,8275±6.73 x
4. Butyric acid30±1,29930±2,05225±2,12075±7,4282±7,570±2,251
5. Valeric acid35±2,9233±2,8239±3,0082±6,85of 91.3±8,1389±8,1
6. ISO-hexanoic acid28±2,1229±1,9540±2,9580±7,2195,5±8,6590±8,32
7. Hexanoic acid27±1,9930±2,0535±2,3385±7,0988±7,5484±7.23 percent

Table 3
The range is Sereni, values of accuracy, repeatability, reproducibility when implementing the proposed method
The name of the component to be determined and the measurement range, µg/cm3The measure of repeatability (relative standard deviation repeatability), σr, %The measure of reproducibility (relative standard deviation of reproducibility)%Accuracy rate (bounds the relative error with probability P=0,95), ±δ, %
Acetic acid, from 3 to 16 on.19,114,7117,66
Propionic acid, 2 to 10 Inc.4,485,9913,93
ISO-butyric acid, from 1 to 10 Inc.6,184,4619,06
Butyric acid, from 1 to 10 Inc.5,61a 4.8311,21
Valeric acid, from 0.5 to 5.0 on.5,0215,64
ISO-hexanoic acid, from 0.5 to 5.0 on.3,533,9317,56
Hexanoic acid, from 0.5 to 5.0 on.3,394,2720,76

Table 4
The limits of repeatability and reproducibility when implementing the proposed method (with probability P=0,95)
The name of the component to be determined and the measurement range, µg/cm3The limit of repeatability (relative value allowable discrepancies between the two results of parallel measurements), rn, %Limit intralaboratory reproducibility (relative value allowable discrepancies between the two measurement results obtained in one laboratory, but in different conditions),%
Acetic acid, from 3 to 16 on12,941,31
Propionic acid, 2 to 10 Inc.12,411,66
ISO-butyric acid, from 1 to 10 incl. 17,121,24
Butyric acid, from 1 to 10 Inc.15,53of 1.34
Valeric acid, from 0.5 to 5.0 on.KZT 12.391,39
ISO-hexanoic acid, from 0.5 to 5.0 on.9,771,09
Hexanoic acid, from 0.5 to 5.0 on9,381,18

1. The method of quantitative determination of acetic, propionic, ISO-butyric, butyric, valerianic, ISO-Caproic and Caproic acids in the blood by gas chromatography analysis, characterized in that the blood sample is acidified with 1%sulfuric acid to pH 2-3, carry out the extraction of the designated acid isobutyl alcohol, the volume of which correlates with the volume of the blood sample as 1:1, carry out the centrifugation for the separation of proteins, add 2-3 drops of 0.4%aqueous alkali solution and the extract is evaporated to dryness, then to dry the precipitate sequentially EXT is make 1%solution of sulfuric acid and isobutyl alcohol and exercise gas chromatographic separation of the mixture of acids on a capillary column with a flame ionization detector, and the amount of each acid set via a calibration curve.

2. The method according to claim 1, characterized in that the adding to the dry residue of 1%aqueous solution of sulfuric acid produced in the volume of 1:40, and isobutyl alcohol is 1:20 to the volume of blood samples, respectively.

3. The method according to claim 1, characterized in that the extraction of the designated acid isobutyl alcohol produced within 5-10 minutes

4. The method according to claim 1, characterized in that the centrifugation is carried out at 6000 rpm for 10 minutes

5. The method according to claim 1, characterized in that the gas chromatographic separation of the mixture of acids on a capillary column with a flame ionization detector is performed with the use of a hardware-software complex based on gas chromatograph "Crystal 2000" detector ionization in flames on a capillary column HP-FFAP - 50 m×0.32 mm×0,5 m at temperature: column - 70°C-160°C-180°C-280°C; evaporator - 250°C; detector, 280°C; flow rate of carrier gas 1 (nitrogen) - 40 cm3/min; flow rate of carrier gas 2 (nitrogen) - 14-30 cm3/min.



 

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1 ex, 5 tbl

FIELD: chemistry.

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.

4 tbl

FIELD: chemistry.

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

FIELD: medicine.

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

FIELD: chemistry.

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: medicine.

SUBSTANCE: for an assay, 5-7 cm3 of blood is taken, and before extraction the sample is pre-treated with 2 cm3 of 60% sulphuric acid; the extraction process is executed with 30 cm3 of n-hexane once, and gas chromatography is preceded with single treatment of n-hexane extract with 10 cm3 of concentrated sulphuric acid.

EFFECT: invention provides higher reliability of α-HCCH, γ-HCCH test results and twofold reduced time of sample preparation.

1 ex, 1 tbl

Gas analysis method // 2410679

FIELD: test equipment.

SUBSTANCE: invention refers to analysis of the quantity of impurities in carbon dioxide during manufacturing and/or cleaning process. Measurement method of concentration of impurities during gas cleaning consists in the fact that, first, gas flow containing impurities passes through gas absorbing device during the time period at ambient temperature or higher so that impurities can be absorbed. Than gas flow movement is stopped. Then, desorption and analysis of impurities in stopped gas flow movement is performed by means of detector. At that, impurities have been chosen from the group consisting of H2S, COS, dimethyl sulphide, benzene, aldehydes, spirits with low length of carbon chain and hydrocarbons. Also, in the proposed method, gas absorbing device includes column with absorbent layer in gas chromatograph, and gas is desorbed from column with absorbent layer through gas-distributing column.

EFFECT: improving accuracy and reducing costs for measurement of concentration of impurities during gas cleaning.

9 cl, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to analytical chemistry. The method is realised as follows: a sample of ground up premix is filled with a hydrochloric acid solution and put into an opaque case which is put into an ultrasonic bath. Extraction is carried out for 15-20 minutes at 38-42°C and centrifuging is then carried out for 10 minutes at 8000 rpm. The mixture is then brought up to the mark in a measurement flask. The obtained solution undergoes chromatographic separation on a column with Purospher sorbent. Chromatography conditions: eluent A - 0.005 M lithium perchlorate solution, pH=2.5; eluent B- acetonitrile; elution gradient mode - from 0 to 26% eluent B for 14 minutes.

EFFECT: high efficiency and accuracy and possibility of detecting a wider range of vitamins independent of the premix base.

2 dwg, 5 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: method for chromatographic analysis of a substance involves exposing the separated mixture of substances carried by a carrier through a chromatographic column to acoustic oscillations. Before chromatographic analysis, a liquid nematic crystal is deposited on the wall of the chromatographic column, where the said crystal is directed across the propagation of sound oscillations.

EFFECT: high efficiency of separating an analysed mixture of substances into components using sound waves.

1 dwg

FIELD: oil and gas industry.

SUBSTANCE: gas chromatograph includes chamber for samples with piston position sensor, which is connected through sample valve to pipeline and through oil pump to reservoir for compensation of hydraulic oil pressure, electrical thermostat with temperature sensor and chromatograph tube located inside thermostat, which is in-series connected on one side through rotating sample injector, zeolite filter, the first return valve and isolating valve of chromatograph with connection line of sample valve and chamber for specimens, in-series connected on the other side to the second return valve, fraction detector, bottle with sample portion and the second pressure sensor. At that, rotating sample injector is in-series connected to pressure reducer, valve for transporting medium, bottle with compressed nitrogen and the first pressure sensor, bypass line with bypass valve is parallel connected to rotating sample injector, chromatograph tube and fraction detector, and circuit of electronic telemetry is connected to output of fraction detector. Method of downhole gas chromatography is proposed as well.

EFFECT: development of device allowing to perform gas chromatography for determining the type of well fluids in well in real time.

3 cl, 5 dwg

FIELD: chemistry.

SUBSTANCE: device for chromatographic separation of substances contains three chromatographic columns connected to each other by crossover channels fitted with switching elements and extra channels fitted with switching elements which are connected to a source of the separated medium, eluent stream and system of receivers for collecting fractions. A controlled flow divider, one or more detectors and an analytical column are fitted at the output of the chromatographic separation system. Also in order to increase output and efficiency of the device for chromatographic separation of substances dissolved in supercritical fluids, the receivers for collecting fractions are fitted with level sensors and their outputs are further fitted with pilot-controlled valves which prevent diffusion of collected substances between receivers. The device also has a collector with a receiver and a flow regulator for the stream of fluids evaporated when pressure falls below the critical value. The device also has a high-pressure pump whose output is also connected to the flow regulator with pressure sensors at the input and output and a flow sensor, which guide part of the stream of formed fluids through the pilot-controlled valve into one or more spherical reactors which have an outer insulating layer and outer and inner heat chambers connected to heat or cold sources, temperature sensors, and the other part of the stream is directed to the analytical column and reactor. The reactor is connected through the pilot-controlled valve to the spherical collector of fluid solutions which is similar to the reactor whose output is connected to liquid batch collection device.

EFFECT: more accurate batching and increased output and efficiency of the disclosed device.

1 dwg

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.

8 dwg

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