SU1273800A1 - Method of determining ceruloplasmin in blood serum - Google Patents

Abstract

The invention relates to biochemistry, in particular to clinical laboratory research. The purpose of the invention is to improve the accuracy of the method during long-term storage of samples by adding to the sample a solution of etoni in acetate buffer. To do this, two test tubes and one control tube were added with 2 MP acetate buffer, 0.1 ml of the studied biological fluid, and 0.1 ml of a 0.4–1.2% etonium solution in acetate buffer. Samples shaken, closed with rubber stoppers and placed in storage in the refrigerator at. In the aftermath of 120 days, similarly prepared samples are accumulated in the refrigerator. Then, no later than the 120th day, the activity of ceruloplasmin in all samples is determined, which takes significantly less working time. 2 tab. (L

Description

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The invention relates to biochemistry, in particular to clinical laboratory research.

The aim of the invention is to improve the accuracy of the method during long-term storage of samples by adding to the sample a solution of etonium in acetate buffer.

The method is carried out as follows.

Two test tubes and one control tube were filled with 2 ml of acetate buffer, 0.1 ml of the biological fluid studied and 0.1 ml of a 0.4-1.2% etonium solution in acetate buffer. Samples shaken, closed with rubber stoppers and placed in storage in a refrigerator at a temperature of 5-10 ° C. In the next 120 days, similarly prepared samples are accumulated in the refrigerator. On the 120th day or at any convenient day for the scientist, but not later than the 120th day, cerulochtmin activity in all samples is determined, which takes much less working time.

Example. Two experienced and one

control tubes introduced 2 MP

acetate buffer, 0.01 ml of the studied biological fluid and 0.1 ml of etoni solution. The samples are shaken, covered with rubber stoppers and placed in storage in a refrigerator at 5-10 ° C. In the next 120 days, similarly prepared samples are accumulated in the refrigerator. On the 120th or any day convenient for the scientist, but not later than the 120th day, the activity of ceruloplasmin in all accumulated samples is determined, for which 1 MP sodium sulfite solution is added to the control tubes and their contents are agitated. All MP tubes were p-phenylenediamine 1 MP each, shaken and placed in a water ultra-thermostat with a temperature of 37 ° C for 60 min, shaking them every 2-5 min. In this case, ceruloplasmin interacts with p-phenylenediamine and turns it into a blue product. After 1 hour, 7 mp of n-butyl alcohol is poured into all the tubes, which results in the enzymatic reaction in the test tubes being stopped. The tubes are capped and shaken vigorously.

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30 seconds, placed in an ice bath for 10 minutes, then centrifuged at a speed of 1500 rpm for 10 minutes, the upper butanol phase is aspirated, transferred to a cell with a working layer thickness of 10 mm and colorized on a FEC with a light length waves 582 nm (filter number 7). To express the activity of ceruloplasmin in

For arbitrary units, the average arithmetic magnitude of extinction obtained by colorimetry of two parallel test samples against a control is multiplied by 100 and divided by the amount per milligram of protein contained in 0.1 ml of biological fluid. The protein content is determined by one of the known methods.

In tab. Figure 1 shows the extinction of samples in determining the activity of ceruloplasmin by a known method (serum No. 4) on the periods of storage of samples consisting of 2 ml of acetate buffer and 0 ml of human serum in a refrigerator at 5-10 ° C.

In tab. Figure 2 shows the extinctions obtained by the proposed method at different periods of storage, samples consisting of 30 ml of 2 ml acetate buffer, 0.1 ml of serum and 0.4 ml of etoni.

The experiments performed during the tests of the proposed method confirm its economic effect, which is expressed in a significant reduction in the cost of a single assay for ceruloplasmin activity. The use of the proposed method significantly saves the working time of the researcher, as well as the cost of electricity and facilitates the implementation

scientific work.

Claims (1)

Invention Formula The method for determining ceruloplasmin in serum by adding a p-phenylenediamine solution to the sample, extracting the reaction product with n-butanol followed by photometric measurement of the extract, characterized in that, in order to improve the accuracy of the method during long-term storage of samples, 0.4 -1,5% solution of etoni in acetate buffer.