Method of preparing sample for gas-chromatographic identification of pesticides in biomaterial
SUBSTANCE: method includes selection, and crushing of biomaterial, two-stage extraction of pesticides with n-hexane, purification of biomaterial from coextractive substances with concentrated sulphuric acid, formation of concentrate of n-hexane extract of pesticides, drying, sample formation by dissolution in 0.5-1 ml of n-hexane and carrying out gaschromatographic identification.
EFFECT: invention is characterised by higher effectiveness and accuracy of research and can be used in biology, ecology, medicine for gaschromatographic identification of organochlorine pesticides, namely α-HCCH, β-HCCH, γ-HCCH, DDT, DDD, DDE, in various biomaterials, such as lipids of internal organs and tissues, blood, milk, bird feathers.
2 cl, 1 dwg
The invention relates to methods for sample preparation and can be used in biology, ecology, medicine for gas chromatographic determination of organochlorine pesticides, namely α-HCH, β-HCH, γ-HCH, DDT, DDD, DDE in various biomaterial containing lipids, internal organs and tissues, blood, milk. In addition, as a biomaterial, you can use the feathers of birds.
The known method of sample preparation for gas chromatographic determination of organochlorine pesticides in blood, including extraction of pesticides from blood n-hexane, purified from coextruding substances concentrated sulfuric acid, the concentration of n-hexane extract (see RF patent №2414711, IPC G01N 33/50, G01N 33/48, publication date 20.03.2011).
The lack of technical solutions is a limited scope due to the inability of studies of internal organs and tissues.
As the closest analogue of the adopted method of sample preparation for gas chromatographic determination of organochlorine pesticides in the biomaterial, including screening, grinding and subsequent extraction of pesticides n-hexane, after which the resulting material is purified from coextruding substances concentrated sulfuric acid, to form a concentrate of n-hexane extract of pesticides (see Klimenko M. A. Methods for the determination of microco is icest pesticides. - M.: Kolos. - 1977. - S. 15, 19).
The disadvantage of the closest analogue is the low efficiency and accuracy of the study because of incomplete extraction with n-hexane-related lipids pesticides, as well as the duration of the analysis due to multiple stages of extraction and purification of the extract with sulfuric acid.
The problem to which the invention is directed, is to develop an efficient method of sample preparation for gas chromatographic determination of pesticides in various biomaterial.
Technical result achieved when solving a task, expressed in increasing the efficiency and accuracy of research by conducting a more complete extraction with n-hexane-related lipids of pesticides and reduce the number of stages of extraction and purification from coextruding substances concentrated sulfuric acid.
The problem is solved in that in the method of sample preparation for gas chromatographic determination of pesticides in the biomaterial, including the selection, grinding and subsequent extraction of pesticides n-hexane, after which the resulting material is purified from coextruding substances concentrated sulfuric acid, to form a concentrate of n-hexane extract of pesticides, use crushed the biomaterial in the representative volume containing lipids to the th is subjected to at least twice the extraction of lipids by treatment of biomaterial n-hexane with the combined filtrates, received at various stages of extraction, and for the first time extracted at least 20 minutes, then for at least 10 minutes, then combined filtrate is evaporated at a temperature 69-70°, the resulting fat is weighed and dissolved in n-hexane, after which the named material is poured concentrated sulfuric acid to the destruction of coextruding substances and select hexanamide layer, then the material hexenstieg layer selected and washed from concentrated sulfuric acid to achieve pH=6, then evaporated n-hexane at a temperature lower temperature of its boiling point, and then form the sample, dissolving the resulting material in a fixed volume of n-hexane, preferably 0.5 to 1 ml.
In addition, as a biomaterial use internal organs, mainly the liver, weighing not less than 10 g, and/or tissue, for example muscle mass not less than 10 g, and/or blood, milk, not less than 20 ml in Addition, as a biomaterial, you can use the feathers of birds weighing not less than 5,
Comparative analysis of the essential features of the proposed solution with essential features analogues demonstrates its compliance with the criterion of "novelty".
This distinctive features in the claims solves the following functional tasks.
The sign "use crushed b is material in the representative volume, containing lipids provides research and allows us to expand the scope of application due to the use as a biomaterial is not only blood, but also of organs, tissues. In addition, as a biomaterial, you can use the feathers of birds.
Topic "... biomaterial is subjected to at least twice the extraction of lipids by treatment of biomaterial n-hexane with the combined filtrates obtained at different stages of extraction, and for the first time extracted at least 20 minutes, and subsequently at least 10 minutes" provides the possibility of separating the lipids from the crushed biomaterial.
The sign of "the combined filtrate is evaporated at a temperature 69-70°" allows you to remove excess n-hexane while maintaining lipids.
The characteristic of the resulting fat is weighed and dissolved in n-hexane produces a portion of fat known weight lipids for subsequent discharge from her pesticides by gas chromatography.
The characteristic material pour concentrated sulfuric acid to the destruction of coextruding substances and select hexanamide layer allows for efficient removal of lipids saving in material of pesticides.
The characteristic material hexenstieg layer selected and washed from concentrated sulfuric acid to achieve pH=6" allows jet to remove the remnants of concentrated sulfuric acid at achieving an optimal level of acidity and provide a more reliable and durable equipment, namely, the chromatographic column.
The characteristic is evaporated n-hexane at a temperature lower temperature of its boiling point" allows you to remove n-hexane while maintaining the material containing pesticides.
The characteristic shape of the sample, dissolving the resulting material in a fixed volume of n-hexane, preferably 0.5 to 1 ml makes it easy to control the amount of sample for analysis.
In Fig.1 shows a chromatogram of a mixture of standard solutions of pesticides.
The method is as follows.
Pre-produce a selection of biomaterial - internal organs, mainly the liver, weighing not less than 10 g, and/or tissue, for example muscle mass not less than 10 g, and/or blood, milk, not less than 20 ml in Addition, as a biomaterial, you can use the feathers of birds weighing not less than 5,
The selected biomaterial pulverized by grinding in a porcelain mortar with anhydrous sodium sulfate (Na2SO4betw.) to remove moisture. Then prepare the first portion, for which the crushed material is transferred into a conical flask, pour 40-50 ml of n-hexane and extracted with not less than 20 minutes After that, the liquid part is filtered in a pear-shaped flask, previously dried and weighed on an analytical balance to the fourth digit. Next, prepare the second portion, for that crushed the biomaterial from the nutrient conical flask pour 20 ml of n-hexane and extracted for 10 min, then the liquid portion connected to the first portion of the filtrate in pear-shaped flask.
Then the combined filtrate is evaporated on a water bath with temperature 69-70°C (the boiling point of n-hexane - 68,742°C), select a portion of the resulting fat and determine its weight.
After weighing the fat pear-shaped flask was dissolved in n-hexane (25 ml), then the resulting material is poured concentrated sulfuric acid (5-7 ml), gently shaken and left for a few hours (2-4 hours). After the destruction of coextruding substances, when the mixture is stratified, select hexanamide layer glass syringe into a separating funnel with a volume of 250 ml.
Next hexanamide layer in a separating funnel cleaned with concentrated sulfuric acid to obtain a colorless layer of sulfuric acid. After that hexanamide layer washed with distilled water until neutral pH=6 by universal paper indicator. Then washed hexanamide layer is dried by filtering through sodium sulfate in sharp-bottom flask with one neck with cut (volume of 25-100 ml). Forth from sharp-bottom flask is evaporated n-hexane at a temperature lower temperature of its boiling point, for example on a rotary evaporator, or leave for a few days until complete evaporation of n-hexane. Then form the sample, dissolving the resulting Mat is real in a fixed volume of n-hexane, preferably 0.5 to 1 ml, and spend gas chromatographic determination of the content of organochlorine pesticides on the equipment of known construction in the standard modes. If the peaks on the chromatogram are not too large, the material of the sample add a controlled amount of n-hexane.
1. The method of sample preparation for gas chromatographic determination of pesticides in the biomaterial, including the selection, grinding and subsequent extraction of pesticides n-hexane, after which the resulting material is purified from coextruding substances concentrated sulfuric acid, to form a concentrate of n-hexane extract of pesticides, characterized in that use crushed the biomaterial in the representative volume containing lipids, which expose at least a twofold extraction of lipids with n-hexane filtered extracts each stage with the consolidation of the obtained filtrate, and the duration of the first extraction 20 minutes, and the next 10 minutes, then combined filtrate is evaporated at a temperature 69-70°, the resulting residue is weighed and dissolved in n-hexane, after which the solution is poured concentrated sulfuric acid to the destruction of coextruding substances and selected material from hexenstieg layer, which is washed from concentrated sulfuric acid up to the achievements pH=6, then evaporated n-hexane at a temperature lower temperature of its boiling point, and then form the sample, dissolving the material obtained in 0.5-1 ml n-hexane.
2. The method of sample preparation for gas chromatographic determination of pesticides in biomaterial under item 1, characterized in that as a biomaterial using samples taken from the internal organs, mainly the liver, weighing not less than 10 g, and/or tissues, such as muscles, weighing not less than 10 g, and/or blood or milk, not less than 20 ml, and/or sample weighing not less than 5 g is formed from the feathers of birds.
SUBSTANCE: invention represents a method for preclinical study of cardiotropic antiarrhythmic drugs, involving determining the bioelectric parameters in isolated multicellular perfused preparations and measuring an action potential duration, differing by the fact that the isolated multicellular perfused preparations are presented by rat's pulmonary vein myocardium; the parameters are measured in three operation modes of the multicellular preparations; a resting potential is additionally measured; varying APD 90%, related APD 50%/APD 90%, a spontaneous shear velocity of the resting potential, the most positive membrane potential in the resting preparation, a spontaneous activity train repetition rate, spontaneous action potential train repetition and variability frequency, post-depolarisation number and intensity, as well as a shear membrane potential corresponding to the beginning of train activity are used to evaluate the signs of antiarrhythmic and arrhythmogenic action.
EFFECT: more reliable prediction of the antiarrhythmic action of the potential pharmacological agents and reduction of experimental phase time.
FIELD: measurement equipment.
SUBSTANCE: invention comprises a method of determination of nano trace contaminants, supposing the use of emulsion from liquid crystal drops, dispersed in water and capable to change a configuration of liquid crystal drops in presence in the emulsion composition of foreign admixtures, measurement of change of intensity of light scattered by the emulsion, according to which it is possible to evaluate the content and density of required admixtures differing by that for a liquid crystal the compounds capable to trans-cis-transition under the effect of actinic light are selected and before measurement of change of light intensity the emulsion is illuminated additionally by actinic light, thus ensuring change of configuration in liquid crystal drops due to trans-cis-transition in liquid crystal molecules.
EFFECT: increase of sensitivity of method of determination of nano-trace contaminants.
SUBSTANCE: invention refers to a method for identifying living and dead mesozooplankton in seawater samples, which involves taking samples, staining the organisms with suitable colouring material, giving a visual estimation of the colour intensity of the units under the microscope, which is combined with microphotographying the units with an adjustable camera without changing the settings keeping throughout a photographic session of at least one sample; thereafter colour and brightness specifications average for each unit are measured in the formed images with using a painting program, e.g. Adobe Photoshop package, and the units are referred to living or dead by a discriminative analysis of the varied digital values.
EFFECT: improving the method.
SUBSTANCE: identification method of blue pus bacillus Pseudomonas aeruginosa involves inoculation of the test material on a hard substrate, incubation of the inoculum under anaerobic conditions at the temperature of 37°C during 16-18 hours. The obtained bacterial mass in the amount of one bacteriological loop is placed in 300 mcl of a physiological solution and warmed-up at the temperature of 98-99°C during 20-30 minutes and centrifuged at 12000 revolutions per minute during 30 seconds. To a supernatant there added is colouring material for electrophoretic detection in the quantity of 0.5 mcl and 20 mcl is added to a well with a size of 4×1 mm 1.2% agarose gel on TAE-buffer with 10 mcl of 1% ethidium bromide. Besides, the amount of 3 mcl of the solution of standard DNA-marker 1 kb containing DNA fragments in the range of 250-10000 bp is added to the test well. Horizontal electrophoresis is performed during 15-20 minutes, and at detection on the obtained electrophoregramme of the test bacterial mass of three strips radiating in ultraviolet light, one of which corresponds to fragments of standard DNA-marker with the size of 10000 bp, the second one corresponds to fragments of standard DNA-marker with the size of 6000-8000 bp and the third strip at the end of the track in the form of a whisk, which corresponds to fragments of standard DNA-marker with the size of less than 750 bp, there identified is Pseudomonas aeruginosa in the test bacterial mass.
EFFECT: method allows quick and complete identification of pigment-shaping and non-pigment strains Pseudomonas aeruginosa in a test bacterial mass.
5 dwg, 2 ex
SUBSTANCE: for analysis of removal with meadow grass of biochemicals the yield fluctuations are accounted depending on the structure of phytocenosis in the form of a ground cover. Conducting statistical data processing of testing the samples of grass from the test plots on the meander, central and terrace near flood plains, and also on the meadows with uneven and tiling location of grass species. And prior to establishment of sample plots the terrain reconnaissance is carried out with the grass cover selected for the measurements, an outline map of location of the components of the grass cover is made, then on each component of the grass cover in the form of a plot at least one temporary test plot is established, the wet and air-dry weight of the sample is determined by weighing on the cut grass sample. At that the tests on biochemical analysis are additionally carried out on the dried samples of grass to determine the concentration of at least three chemical nutrients in the form of mobile nitrogen, potassium and phosphorus oxides, and then by summing the concentrations of these three substances the total summarised removal of substances from the aerial part of grass on all test sites is calculated.
EFFECT: invention enables to determine the productivity of meadows.
9 cl, 12 dwg, 4 tbl, 1 ex
SUBSTANCE: for the purpose of the confirmation of a patient's death caused by ventricular fibrillation accompanying myocardial infarction in the patients died from myocardial infarction in the course of autopsy on the basis of a macroscopic pattern, the pathological process age is determined. This enables selecting examination areas in the heart at the level of necrosis. Absolute values (AV) of the following cell populations are counted in the x600-magnified visual field with using haematoxylin and eosin: necrosis-borderline lymphocytes (Lfb) in myocardial infarction of the age of 1-2 days, necrosis neutrophilic granulocytes (NGn) and fibroblasts (Fbn) in myocardial infarction of the age of 3-5 days. The derived absolute values are compared to threshold values (TV) with the threshold values of Lfb=2, NGn=38, Fbn=1. Each visual field is assigned with a diagnostic coefficient (DC), which is equal to 1.9 at the age of myocardial infarction of 1-2 days, if the absolute values Lfb<2 or -3.9, Lfb>2; if the age of myocardial infarction is 3-5 days, the diagnostic coefficient is -2.1, if AFbn<1, or the diagnostic coefficient is 7.2, if the absolute value of Fbn>1; the diagnostic value of 1.36 is shown by the absolute value of NGn<38; the diagnostic value is -11, if the absolute value is NGn>38. The total value (ΣDC) is calculated by moving between the visual fields within the examined area, and if ΣDC>12.78, the patient's death is stated to have come from ventricular fibrillation; if ΣDC<-12.78, the death is stated to be caused otherwise.
EFFECT: information value and reliability of the examination accompanying forensic medical examination.
1 tbl, 3 ex
SUBSTANCE: invention refers to laboratory diagnostics and can be used for testing biological samples for pathogenic microorganisms. The device for microbiological analysis of the body fluid samples comprises a compartment for incubating sample containers, an analyser for studying the internal atmosphere of the above containers and a sorting system for classifying the above containers in accordance with the carbon dioxide content detected by the above analyser. The sorting system classifies the containers delivered from the above incubation compartment to divide the above containers into the positive containers to be analysed for identifying microorganisms found in the respective samples, and the negative containers wherein the analysis is not required for identifying the microorganisms, and also the unspecified containers to be incubated once again.
EFFECT: invention provides simplifying and accelerating the analysis of the biological samples for the pathogenic microorganisms.
10 cl, 8 dwg
SUBSTANCE: method involves a post-therapeutic cytological examination of smears sampled from septic wounds, a cell composition analysis, including neutrophilic granulocytes and tissue elements - fibroblasts and fibrocytes; the material sampled from the septic wound on the 3rd day from the beginning of therapy is used for the cytological examination to determine accessory signs of therapeutic pathomorphism, namely the morphology of neutrophilic granulocytes, fibroblasts and fibrocytes and the presence of oxyphilic ground substance; observing the nuclear fragmentation and cytoplasm vacuolation in the neutrophilic granulocytes, multinuclearity, nuclear lobulation, karyonuclei in nuclei, basophilia, cytoplasm grain and vacuolation in fibroblasts and fibrocytes, as well as the presence of the oxyphilic ground substance, the treatment is stated to be effective.
EFFECT: invention provides objective clinical findings of the wound process even on the third post-therapeutic day and can be used for the reliable determination of the clinical effectiveness in septic wounds.
1 ex, 5 dwg
SUBSTANCE: invention refers to medicine. Substance of the early diagnostic technique for chronic renal disease consists in using a dosage range of dopamine of 1 to 3 mcg/kg of body weight and a standard water load of 200 ml. No glomerular filtration rate increase testifies to the presence of early signs of chronic renal disease.
EFFECT: using the declared technique enables standardising the examination as high as possible by precise dosage measurement of the preparation.
2 tbl, 2 ex
SUBSTANCE: carbon nanostructure surface is modified with (2,4,5-triiodophenyl)-methanol; the modified carbon nanostructures are analysed to measure a iodine amount; the prepared modified carbon nanostructures are administered into an experimental animal's body to take organs and tissues thereafter, to homogenise them in a 0.5-2 M NaOH solution, to sample a homogenate, to dilute with water, to expose the diluted sample to ultrasound to a temperature of 40-70°C, to determine a iodine amount in the prepared sample by mass spectrometry with inductively bound plasma and to calculate the carbon nanostructure content in the sample as shown by a difference of the iodine amount in the sample prior to administration of the modified carbon nanostructures and after administration thereof into the body and to re-calculate this iodine amount into the carbon nanostructure content in the sample with the use of the initial iodine content in the modified carbon nanostructure.
EFFECT: method provides monitoring in vivo of the carbon nanostructure distribution in the body.
2 cl, 6 dwg, 3 ex, 2 tbl
SUBSTANCE: method involves examining blood erythrocytes. Blood erythrocyte suspension fluorescence intensity is measured at wavelength of 500 nm polarized along light axis and then at wavelength of 493 nm in perpendicular to the light axis for each polarization direction with following anisotropy value being calculated. Its deviation from norm proves availability of hemolytic disease.
EFFECT: accelerated examination process; reduced risk of traumatic injuries of fetus or newborn.
FIELD: veterinary medicine.
SUBSTANCE: method involves carrying out blood erythrocyte micro electrophoresis in alternating electric field with sign change frequency equal to 0.3-1 Hz. Electric field having current intensity equal to 4.5-5.0 mA is applied. Treatment is applied at 36-38°C. Oscillation amplitude being equal to 3 mm and less, endotoxicosis is detected.
EFFECT: high accuracy and simplicity of the method.
SUBSTANCE: method involves applying integrated diagnostics approach with blood serum examination being applied using infrared spectrometry methods. Sample absorption spectra in the area of 1200-1000 cm-1 are recorded, absorption peak heights having maximum at 1170, 1165, 1140, 1125, 110, 1070, 1025 cm-1 points are measured. Then, mean peak height C and ratio X of peak height at 1125 cm-1 point of maximum to C are calculated. X≥0.7 being the case, light myocardium injury degree is to be diagnosed. 0.7>X≥0.53 being the case, moderate severity degree case is to be diagnosed. X<0.53 indicates grave injury to take place.
EFFECT: high reliability of diagnosis.
FIELD: analytical methods in medicine.
SUBSTANCE: cells of tested organs are subjected to alternating cytological electrophoresis. When cell vibration amplitude rises by at least 20%, drug is regarded to be appropriate for treatment.
EFFECT: increased drug selection efficiency.
FIELD: medicine, hepatology.
SUBSTANCE: the present innovation deals with detecting bilirubin content both before and after curative starvation, moreover, curative starvation one should introduce antipyrine at the dosage of 1 mg/kg, in a day it is necessary to detect antipyrine content in saliva and urine and bilirubin content and at bilirubin content being 30-40 mM/l one should starve at daily intake of 400-450 kcal/d for 3 d, then one should again detect bilirubin level and at its increase by 50-100% one should again introduce antipyrine at the dosage of 1 mg/kg and according to its delayed half-life by 20-50% it is possible to diagnose Gilbert's disease.
EFFECT: higher accuracy of diagnostics.
FIELD: medicine, otorhinolaryngology.
SUBSTANCE: the present innovation deals with treating diseases of the upper respiratory tract. One should detect disorders of mucociliary transport (MCT) due to measuring the motor parameters of ciliated epithelium cilia, at least, the rate of mucociliary transport that includes material sampling for further testing as ciliary epithelial cells and registration of cilia fluctuations in human respiratory tract due to TV microscopy performed in patient's lifetime. Material for testing should be sampled from mucosal surface of respiratory tract, and therapeutic tactics should be chosen depending upon motor velocity of cilium's tip obtained based upon mathematical modeling followed by calculations by the formula: , where Vc - velocity of cilium's tip [mcm*sec1]; C1, C2, α - calculated as average values being characteristic for every of human body states under investigation: healthy persons, patients with either acute or chronic forms of sinusitis by the developed model of cilia movement; C1 - curvature of cilium's working length, constant being equal to 1/38 mcm-1; C2 - cilium's curvature from its working length up to the tip [mcm-1]; α - cilium's inclination angle against vertical position [degrees]; π = 3.1417 - constant; L, Ts and Te - parameters measured individually in every patient; L - cilium's length [mcm]; Ts - time from the onset of movement till maximal straightening of cilium's tip [sec]; Te - time of efficient impact [sec]. Moreover, the value being Vc = 9.6 - 11.3 mm/min corresponds to normal MCT functioning. For patients with chronic purulent sinusitis beyond exacerbation accompanied with MCT disorder it is necessary to prescribe complex therapy: surgical or conservative therapy with preparations of secretolytic and secretomotory action. For patients with chronic purulent sinusitis beyond exacerbation without any MCT disorder one should prescribe surgical therapy. All patients with acute sinusitis with MCT disorder should be prescribed complex conservative therapy with preparations of secretolytic and secretomotory action. The method enables to specify optimal therapy based upon scientifically proved indications of investigations by taking into account individual peculiarities of every patient.
EFFECT: higher efficiency.
5 dwg, 3 ex, 2 tbl
FIELD: medical engineering.
SUBSTANCE: device has non-absorbing substrate having hydrophilic target region, which is covered with reagent by applying non-impact micro-drop printing method to produce practically uniform reagent layer. The device is in particular usable for measuring blood coagulation time. Preferential invention embodiment involves determining blood coagulation time by carrying out monitoring of light transition through the target region as the blood sample cover interacts with the reagent.
EFFECT: high reliability of analysis results.
20 cl, 9 dwg
SUBSTANCE: one should register the values of pH and Redox potential (Red) of liquid means in bio-objects simultaneously. It is necessary to plot a graph ▵pH/▵Red according to fluctuations of variation values in measured parameters at their temporal diagram to transfer it into a columnar one where the height of every column is proportional to the area of this graph between neighboring measurements, correspondingly. According to the ratio of area sums of positive columns to that of negative ones during a certain period of time ( from a minute- to a year-long ones) one should conclude upon a state in a bio-object. The value ranged 0.5-2 is considered to be a standard. The method enables to provide efficient control for a two-phase systemic process.
EFFECT: higher efficiency and accuracy of diagnostics.
1 cl, 9 dwg, 5 ex, 1 tbl
FIELD: medicine, clinical neurology, neurosurgery.
SUBSTANCE: one should perform crystallographic study if cerebrospinal fluid and at protein concentration in liquor samples being from 0.50 g/l and higher at predominance of crystals as suppressed dendrites upon a crystallographic picture one should diagnose a malignant cerebral tumor, and at protein concentration being below 0.50 g/l and predominance of crystals as branched dendrites - a benign cerebral tumor. The innovation enables to fulfill not only due diagnostics of cerebral neoplasms, but detect the degree of tumor process malignancy at the early stages of its development.
EFFECT: higher accuracy of differential diagnostics.
3 dwg, 2 ex, 1 tbl
FIELD: biology, medicine, in particular histological investigation of shells with natural surface by using light-optical microscope.
SUBSTANCE: claimed method includes surface relief investigation by using plane field microscope in reflected light at one-side falling shadow-forming lighting. Light-reflecting ability of specimen surface is provided by silver impregnation. Said impregnation is carried out by specimen hold in 10 % silver nitrogen solution for 10 min followed by reducing thereof with 1 % ascorbic acid solution for 1 min.
EFFECT: method for investigation of natural shell surface relief of improved quality due to visual representation of surface three-dimension image on tissue level.
3 cl, 4 dwg