SU983541A1 - Method of determination of sugar in blood - Google Patents

Description

(54) METHOD FOR DETERMINING BLOOD SUGAR

This invention relates to biochemistry and may be used clinically. There is a known method for determining sugar in a cro by precipitating glycogen with ethanol and treating it with sulfuric acid. The resulting glucose is phosphorylated by ATP with the enzyme hexokinase, followed by treatment with ADP. The amount of glucose is determined by the change in the content of NAD-H fl 3-

However, the known method is laborious and does not provide high accuracy (i15-20% errors are possible).

Closest to the present invention is a method for determining blood sugar by obtaining it without a bleach filtrate, treating the latter with an anthrone reagent and photometry of the resulting colored solution 12 3.

However, the known method does not provide high accuracy, time-consuming and requires a large consumption of scarce reagents.

The aim of the invention is to improve the accuracy and acceleration of the method.

The goal is achieved by following the method of determining the blood sisgar by obtaining its protein-free filtrate, treating the latter with an anthrone reagent and photometry of the resulting colored solution, before introducing the anthron-free protein-free | 1 liters of blood is treated with sulfuric acid.

Example. A 0.9 ml of 5% aqueous solution of trichloroacetic acid (TCA) is poured into the centrifuge tube and 0.1 MP of the capillary of blood under test is added here, mixed and centrifuged at 1500 rpm for 10 minutes. The supernatant in the amount of 0.5 ml (corresponds to 0.05 ml of blood) is transferred into an ordinary test tube, here 1 ml of concentrated sulfuric acid and 2 ml of anthron indicator (the reagent is prepared by mixing 34 ml of distilled vodts with 66 ml of concentrated sulfuric acid).

20 acids and dissolve 50 mg anthrone and 1 g thiourea in this mixture (the reagent is stored in a refrigerator). The contents of the test tube were mixed and boiled for 10 minutes.

25 bath, then cooled to room temperature and colorized on a FEC with a red light filter in cuvettes with a working width of 5 mm. Instead of compensating fluid take control

30 solution that is prepared just like

an experimental sample, only 0.5 MP of a 5% solution is taken instead of blood.

 The calculation of the concentration of sugar in the test blood is carried out according to a standard glucose solution, which is treated in the same way as an experimental sample.

An example of building a fault table.

A standard solution is prepared from pure and dried to constant weight glucose, which contains 0.35 mg of glucose in 1 ml.

Five ordinary test tubes are taken, 0.1 ml of standard glucose solution is introduced into the first, 0.2 ml into the second, 0.3 ml into the third, 0.4 ml into the fourth, and 0.5 ml into the fifth. All tubes are filled up to 0.5 ml with distilled water. After that, add 1 ml of concentrated sulfuric acid and 2 ml of anthrone reagent to all tubes. In the test tube for the control solution, i.e. solution for

0.1

Og4 0.2 0.3 0.2 0.3 0.4 0.1 0.5

Payment. To calculate the concentration of sugar in the test blood, the weight amounts of glucose used for calibration should be expressed for each standard point of the table in blood glucose concentrations for the test conditions that occurred in this blood sugar determination. Do this according to the formula where KX C

filling the compensation cuvettes, add 0.5 MP of distilled water and process it in the same way as standard samples. The contents are mixed and the tubes are placed for 10 min in a boiled water bath, after cooling at room temperature or in cold water, the contents of the tubes are colorimetric.

When colorimetrication on a photoelectric colorimeter, they use a red light filter and cuvettes, with a working width of 5 mm. As a compensation liquid, they drink a control

a solution that also acquires some color. The readout of the optical density, i.e., the extrusion, is carried out on the red scale of the right drum. Finally, a calibration table is prepared for the absorbance of standard solutions depending on the amount of glucose in them and the calculation of sugar accumulation in the blood under study is carried out.

0.10 0.20 0.30 0.40 0.50

K - the amount of glucose in the standard according to the calibration table. The proposed method allows reducing the time for conducting one study by 2-3 times, reducing the volume of the studied substrate and reagents by 4-5 times, significantly increasing the accuracy of the method and 2-3 times increasing its sensitivity. KX-C; -KI blood sugar concentration; the volume of blood in which the third method is used to determine the concentration, by obtaining protein-free filtration of the sugar; that is, by processing the latter anthropic volume of the blood, followed by experience, with 65 reagent and photometry to form the formula

5 9 "35416

Shegoz colored solution, about tl and -Idtochnki information,

considering that in order to be taken into account during the examination

improve accuracy and speed-up sposo-1. Biochemical methods of research, prior to the introduction of anthrone reactivation in the clinic. M., 1969, p. 232.

protein-free blood filtrate processed-2. Petrunkina A.M. Practical

wate with sulfuric acid. biochemistry L., 1961, p. 208.

Claims (1)

A method for determining blood sugar; by obtaining a protein-free filtrate of it, processing the last anthrono, 65th reagent and photometry _, where K _x is the concentration of sugar in the blood; C is the volume of blood in which it is required to determine the concentration of sugar; With _l - the amount of blood that went into the experiment 9B3541 colored staining solution, which consists in the fact that, in order to increase the accuracy and speed up the method, a protein-free blood filtrate is treated with sulfuric acid prior to the introduction of an anthron reagent.