Diagnostic technique for blood serum glucose, total protein and electrolytes by multi-frequency impedance analysis

FIELD: medicine.

SUBSTANCE: method provides blood serum analysis by a bipolar method for multi-frequency impedancemetry to determine impedance modulus (|Z|) and phase angle (φ) at AC low-intensity frequencies 20, 98, 1000, 5000, 10,000, and 20,000 Hz by means of hardware-software complex provided with BIA-lab Compositum software; measurements are taken in a microchamber of 50 mcl; the software automatically calculate the blood serum concentrations of total protein, glucose, chlorides and bivalent ions on the ground of solution of assembled mathematical equations; the result is displayed and can be printed.

EFFECT: higher diagnostic effectiveness by eliminating the need for using chemical agents, reducing the time for analysis, cutting the cost and extending the indications for using the technique.

3 ex

 

The invention relates to medicine, namely to laboratory diagnostics, and is intended to improve the efficiency of the study of glucose and total protein in serum.

The closest analogue is a method of biochemical analysis based on photometric measurement of the intensity of the color of the chemical reagent solution of phenolphthalein in determining the concentration of glucose and protein complexes with copper sulfate in the determination of the concentration of total protein (Laboratory techniques: a Handbook / edited by Menshikov V. V. - M.: Medicine, 1987. - S. 175, 232-233, 261-265). Method of biochemical analysis is performed as follows. After protein precipitation with trichloroacetic acid pH protein-free solution with sodium phosphate adjusted to a point close to the optimal action of glucose oxidase, and carry out the enzymatic reaction. The formed hydrogen peroxide in the presence of copper ions oxidize phenolphthalein to phenolphthalein, which in the neutral colorless area, and in the alkaline region is colored in red. The result is recorded by a spectrophotometric method. To determine the total protein concentration to 0.1 ml of serum was added to 5.0 ml of the working solution biuret reagent (4.5 g sagatavoj salt dissolved in 40 ml of 0.2 mol/l sodium hydroxide solution, is added 1.5 g of copper sulfate and 0.5 g of potassium iodide and races�varaut; made up to 100 ml of 0.2 mol/l sodium hydroxide solution; 20 ml of biuret reagent was mixed with 80 ml of 0.5% solution of potassium iodide) and mixed, avoiding formation of foam. After 30-60 minutes to complete the measurement of the optical density of the solution in the photometer in a cell with a layer thickness of 1 cm at a wavelength of 500-560 nm against blank sample. Calculating the concentration of total protein lead via a calibration curve. The concentration of sodium and potassium define a unified method of flame photometry. For this purpose, diluted with water, the serum is sprayed in the form of tiny droplets with a stream of air enters the flame of a gas burner that potassium gives a weak red-violet and sodium bright yellow staining. The color intensity is measured by a photocell. To calculate the concentrations of potassium and sodium used calibration solution. The results calculated by a calibration curve. The concentration of calcium ions is determined using a unified method of colour reaction with cresolphthalein. This reagent reacts with calcium in alkaline medium complex red-violet color, the intensity of which is proportional to the calcium concentration. The reaction mixture was added 8-okshihinolin that binds metals, preventing the determination of the concentration of calcium.

The disadvantages of the prototype: the existing method of biochemical �analysis is not effective enough, requires the use of chemical reagents and laboratory equipment, specially trained medical personnel (a technician) that has a duration of at least 1 hour, is suitable for use in medical institutions and insufficiently cost-effective.

The task of the invention is to simplify the process and expansion of indications for the use of the method.

The problem is solved with the features specified in the claims, such as a method of diagnosis of glucose concentration, total protein and electrolytes in serum, wherein the human serum used bipolar method policestate elektroindustrie with the definition of modular values of impedance (|Z|) and phase angle (φ) at frequencies 20, 98, 1000, 5000, 10000, and 20000 Hz alternating electric current of low power by using a hardware-software complex, equipped with the computer program "BIA-lab Compositum", carry out the measurement in the microcamera volume of 50 μl the program automatically calculates the concentration of total protein, glucose, chloride and divalent ions in the serum based on the solution of a system of mathematical equations, and the result is displayed and can be printed on the printer.

The technical result of the above precisionautofinance essential features - improving the effectiveness of the method by eliminating the need to use chemicals, reducing the run-time studies, cost reduction and the expansion of indications for the use of the method.

A solution to this problem is achieved by measuring the electrical impedance of the blood serum, placed in a special microcamera, bipolar method using policestate sensing an alternating electric current of low power. The computer program "BIA-lab Compositum" (Certificate of state registration of computer programs "BIA-lab Compositum" No. 2012613493, April 13, 2012 copyright Holders: Mishlanov Y. V., Mishlanov V. Yu., Mishlanova I. V., Mishlanova S. L. Authors: Mishlanov Y. V., Mishlanov V. Yu., Mishlanova I. V., Mishlanova S. L.) allows you to register two physical characteristics of bioelectrical impedance: a modular value of the impedance (|Z|) and phase angle (φ) in series at 6 frequencies: 20, 98, 1000, 5000, 10000 and 20000 Hz. At each frequency of the probing AC electric current is recorded 576000 measurements of the analyzed parameters, and a computer program calculates the average value of these indicators is based on the equations of conformity. The entire measurement period is 18 seconds.

The method is as follows. Blood studies are taken from �octavos a vein or a finger of the patient. Isolated serum and 50 µl her make the micro camera unit using the pipette. The electrodes of hardware and software complex, for example, "BIA-lab Spiro" (Patent RF №2487662, publ. 20.07.13. IPC A61B 5/08) or other (Patent RF "Device for measuring impedance of biological media" No. 2462185 from 19.07.2011. IPC A61B 5/08, publ. 27.09.12) connected to the contacts of the micro cameras include the instrument in a second-by-second visualization of the results at a frequency of 20000 Hz, then write the results of the study successively at frequencies of 20, 98, 1000, 5000, 10000 and 20000 Hz. Total recording time of 18 seconds (the program automatically changes the modes of registration of bioelectrical impedance 6 frequency bands, recording the results of measurements for 3 seconds at each frequency of the probing AC electric current, which corresponds 576000 measurements of parameters measured). The result is recorded automatically in a folder named after the name of the patient. Total execution time of the study less than 1 minute.

Diagnosis the concentration of total protein, glucose, potassium ions and calcium in the sample of blood serum is carried out in automatic mode with the help of the computer program "BIA-lab Compositum", and calculate the concentration according to formulas that take into account the magnitude of the modular value of the impedance |Z| and f�basic angle φ, obtained by probing serum alternating electric current of low power on 6 frequencies: 20, 98, 1000, 10000 and 20000 Hz.

Used the following equation corresponding to the used measuring chamber. To calculate the concentration of glucose: SGL=1,86*Exp(-0,058*f98), where SGL is the concentration of glucose in mmol/l, f98 - value of the angle φ on the frequency of the probing AC electric current 98 Hz. Calculating the concentration of total protein was carried out using the formula: Sa=171,8529*Exp(-0,85*(f10000/f20000)), where Sa is the total protein concentration in g/l, f10000 - value of the angle φ on the frequency of the probing AC electric current 10000 Hz, f20000 - value of the angle φ on the frequency of the probing AC electric current 20000 Hz. Calculating the concentration of calcium ions was carried out using the formula: CCA=-2,7667+1,4374*Log10(Z5000), where Sa is the total protein concentration in mmol/l, Z5000 - the value of the modular values of |Z| at the frequency of the probing AC electric current of 5000 Hz. Calculating the concentration of potassium ions was carried out using the formula: SC=3,0793-0,009*(Z1000/f1000)-0,00001274*(Z1000/f1000)2, where ck is the concentration of potassium ions in mmol/l, Z1000 - the value of |Z| at the frequency of the probing AC electric current of 1,000 Hz, f1000 - value of the angle φ on the frequency of the probing AC electric current of 1,000 Hz.

Error method to determine the concentration of glucose does not turn into a�exceeds 0.5 mmol/l, to determine the total protein concentration is not more than 10 g/l, to determine the concentration of potassium ions is not more than 0.5 mmol/l, calcium - not more than 0.3 mmol/L.

Examples of specific performance.

Example 1.

Patient B., 51 years old, diagnosed with multiple myeloma, diffuse alopecia form, variant G.

The total protein concentration in serum, a certain method is 93,1 g/l. the Concentration of glucose in serum, determined by, is 6.0 mmol/l. the Concentration of potassium ions is 4.5 mmol/l. the Concentration of calcium ions is 2,36 mmol/L.

Method policestate electrical impedance analysis using a hardware-software complex, equipped with the computer program "BIA-lab Compositum", defined concentration: protein and 86.5 mmol/l, glucose 5.6 mmol/l, potassium of 4.7 mmol/l, sulphate of 2.3 mmol/L.

The presented example demonstrates the high accuracy of the application policestate electrical impedance analysis for the diagnosis of the concentration of total protein, glucose, potassium ions and calcium in blood serum in clinical practice.

Example 2.

Patient K., 60 years. Diagnosis: diabetes mellitus, type 2, average flow, uncompensated. Complications: Micro - and macroangiopathy, lipodystrophy liver, diabetic foot syndrome. KHAN 1 tbsp.

The concentration of glucose in serum, ODA�divided method is 12.8 mmol/L. Method policestate electrical impedance analysis using a hardware-software complex, equipped with the computer program "BIA-lab Compositum", defined glucose concentration of 13.0 mmol/L.

The presented example demonstrates the high accuracy of the application policestate electrical impedance analysis for the diagnosis of glucose concentration in the serum of the patient with diabetes mellitus in a state of decompensation of carbohydrate metabolism and excess of the normal range of glucose concentration in the blood.

Example 3.

Patient K., aged 29. Diagnosis: diabetes mellitus, type 2, average flow, uncompensated. Complications: Micro - and macroangiopathy, lipodystrophy liver, retinopathy 1 article Hypoglycemic condition on the background of alcoholic intoxication mild. Competing diagnosis: Pneumonia, focal, in the lower lobes of the right and left, not severe. ONE 1 tbsp.

The concentration of glucose in serum, determined by, is 2.3 mmol/L. Method policestate electrical impedance analysis using a hardware-software complex, equipped with the computer program "BIA-lab Compositum", determined the concentration of glucose was 2.7 mmol/L.

The presented example demonstrates the high accuracy of the application policestate electrical impedance analysis in SL�tea diagnosis of glucose concentration in the serum of the patient with diabetes mellitus in a state of decompensation of carbohydrate metabolism and a decrease in the concentration of glucose in the blood.

The proposed method is characterized by the following features. In the proposed method adopts the definition of electrical impedance of the blood serum in a special microcamera volume of 50 µl in the mode policestate analysis that allows to determine the concentration of total protein, glucose and electrolytes on the basis of mathematical solutions of the system of equations based on the experimental results.

Method for the diagnosis of glucose concentration, total protein, potassium ions and calcium in blood serum, characterized in that for the human serum used bipolar method policestate elektroindustrie with the definition of modular values of impedance (|Z|) and phase angle (φ) at frequencies 98, 1000, 5000, 10000, and 20000 Hz alternating electric current of low power by using a hardware-software complex, equipped with the computer program "BIA-lab Compositum", carry out the measurement in the microcamera volume of 50 µl, calculate the concentration of total protein, glucose, ions of potassium and calcium in blood serum, based on the solution of a system of mathematical equations, wherein the glucose concentration is calculated by the formula: SGL = =l,86*Exp(-0,058*f98), where SGL is the concentration of glucose in mmol/l, f98 - value of the angle φ on the frequency of the probing AC electric current 98 Hz, the calculation �concentratii total protein is carried out using the formula: Sa=171,8529*Exp(-0,85*(f10000/f20000)), where Sa is the total protein concentration in g/l, f10000 - value of the angle φ on the frequency of the probing AC electric current 10000 Hz, f20000 - value of the angle φ on the frequency of the probing AC electric current 20000 Hz, calculate the concentration of calcium ions is carried out using the formula: CCA = =-2,7667+1,4374*Log10(Z5000) where CCA is the concentration of calcium ions in mmol/l, Z5000 - the value of the modular values of |Z| at the frequency of the probing AC electric current of 5000 Hz and calculation of the concentration of potassium ions is carried out by applying the formula: SC=3,0793-0,009*(Z1000/f1000)-0,00001274*(Z1000/f1000)^2, where ck is the concentration of potassium ions in mmol/l, Z1000 - the value of |Z| at the frequency of the probing AC electric current of 1,000 Hz, f1000 - value of the angle φ on the frequency of the probing AC electric current of 1,000 Hz, and the result of calculation of concentrations of biological substances in the serum is displayed and can be printed on the printer.



 

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