JPS5861465A - Analytical method for blood - Google Patents

Abstract

PURPOSE:To permit extremely easy measurement of glycosyl hemoglobin, etc. in blood by adding a hemolysis medium prepd. by adsorbing a reagent for hemolysis on a porous material on the sample supply side of a separating medium layer or a reagent layer for inspection of blood of a chromatography or the like. CONSTITUTION:A vinyl resin molding, an inorg. porous plate, etc. having holes of about 1- several mu sizes are used for a porous material to be adsorbed thereon with a reagent for hemolysis. Noionic or ionic surfactants are usable as the reagent for hemolysis, of which polyoxyethylene alkyl allyl ether, salt of sulfuric ester of higher fatty acid, etc. are more perferable in terms of hemolysis characteristic. In this invention, the hemolysis medium is added on the sample supply side of a separating medium layer or a reagent layer for inspection of blood; therefore, the blood analysis by using all the blood samples as they are is made possible without preparing hemolysis samples separately and this method is suitable for analysis of many specimens at one time.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a blood analysis method suitable for measuring glycosylated hemoglobin and the like.

Recently, with the development of clinical testing methods, there has been a need to rapidly analyze a large number of biological components. Particularly when searching for components in blood cells, it is necessary to easily and quickly obtain hemolysis in order to destroy the blood cells and obtain the contents.

However, the usual hemolysis method, in which hemolysis is obtained by mixing and stirring a hemolysis reagent and blood, is complicated because hemolysis of blood cells and blood analysis must be performed in two separate steps. Since it is handled directly, there is a problem of infection, and it is not satisfactory for hemolyzing a large number of specimens at once in clinical tests, etc.

In order to improve this point, the present inventors conducted various studies, found a method suitable for analyzing hemolyzed samples, and arrived at the present invention.

/' In other words, the present invention adds a hemolytic medium in which a hemolytic reagent is adsorbed to a porous material to a separation medium layer such as li-togelafy or to the sample supply side of a blood test reagent layer. A blood analysis method is characterized by the following.

The present invention will be explained below.

First, the hemolysis medium in the present invention is made by adsorbing a hemolysis reagent onto a porous material. The porous material has a diameter l
A material having pores of approximately several microns is used, for example, a molded inorganic porous plate of vinyl resin such as vinyl acetate resin.

The shape of the hemolysis medium is not particularly limited, such as spherical or plate-like, and the size is also not particularly limited.

Usually, a material having a capacity capable of adsorbing a hemolysis reagent of about 5 to t 00 pf is selected.

Examples of the hemolysis reagent to be adsorbed onto this porous material include nonionic or ionic surfactants. for example,
Polyoxyethylene alkyl ether is a nonionic surfactant.

Examples of ionic surfactants include polyoxyethylene sorbitan fatty acid ester, fatty acid glyceride, polyoxyethylene fatty acid ester, polyoxyethylene ester, sorbitan fatty acid ester, sucrose fatty acid ester, and phoroxyethylene alkylamine. Examples include higher fatty acid sulfate ester salts, higher alcohol sulfate ester salts, etc., but phoroxyethylene alkyl allyl ether, higher fatty acid sulfate ester salts, etc. are preferred from the viewpoint of hemolytic properties.

The hemolytic medium in the present invention is prepared by dissolving these hemolytic reagents in an aqueous solvent or an organic solvent such as an alcoholic solvent, and usually contains l to i
About ooμt/me, preferably 10-20μf/ld
The porous material is usually impregnated with this solution for several seconds to several minutes at a temperature around room temperature, and then air-dried or
It is obtained by removing the solvent by heating and drying at a temperature of about IO to ioo°C.

On the other hand, as a separation medium layer for blood components, alumina gel,
Cellulose gel, starch gel, ion exchange resin, etc.
Examples include those normally used as packing materials for column chromatography and liquid chromatography.

Further, the blood test reagent layer is a quantitative reagent for hemoglobin and intrablood cell components. A T P ('adenin
etriphosphate), 2.3-DPG
(,2,,? -diphospho-glycera
Examples include those made of an enzyme reagent for measuring tθ) etc. dissolved in a buffer solution. .

For example, a glass or plastic column is filled with a separation medium, a buffer solution used for elution is added to the upper part of the separation medium layer (on the sample supply side) to a height of approximately 1000 ml, and a hemolytic medium is added to the column. Blood ioμt to be analyzed next
When added to the column and stirred with a tabletop mixer etc. for ~1.0 seconds, hemolysate is formed on the separation medium in the Camum, and by eluting this, blood can be immediately analyzed in the column.

In this case, the hemolytic reagent eluted from the hemolytic medium flows into the separation medium layer or the blood test reagent layer.

Since the concentration is extremely low, it does not interfere with analysis.

In the blood analysis method of the present invention, for example, in order to quantitatively examine abnormalities in glucose metabolism, chromatography method is used.
It is suitably used when measuring (bin).

In this case, a weakly acidic cation exchange resin is used as the separation medium layer. Examples include polymers obtained by crosslinking unsaturated carboxylic acids such as methacrylic acid and acrylic acid with crosslinking agents such as divinylbenzene and divinyltoluene.

The blood analysis method of the present invention allows blood analysis to be performed using a whole blood sample as it is without separately preparing a hemolyzed sample, so it is suitable for analyzing a large number of samples at once, and it is also suitable for analyzing blood directly. Since no operation is required, it has the advantage of preventing infections such as hepatitis viruses.

EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples unless the gist thereof is exceeded.

Example 1 Porous material made of vinyl acetate resin (φ = g mm, thickness 2
m) in an aqueous surfactant solution lθμl/d (triton/00t manufactured by Rohm and Haas), and then dried in an air bath at SO°C for 3 hours.

To measure glycosyl hemoglobin, a plastic column (g=/crn, height 1 lcrn) packed with a weakly acidic ion exchange resin as a separation medium was charged with 50 θμ.
Add t glycosylated hemoglobin elution buffer.

Next, a porous, spherical or disc-shaped hemolysis medium was added to each column, and a weighed/Opi whole blood sample was added to the system, and the column was gently shaken, or 0.5 ~/, When stirred for 0 seconds, the blood cells become hemolyzed, and the applicant for the column: Mitsubishi Chemical Industries, Ltd. and 1 other representative Patent attorney Hase - 1 other person

Claims (1)

[Claims] (1) In a blood analysis method using a blood analysis column having a blood component separation medium layer or a blood test reagent layer,
A method for analyzing blood, which comprises adding a hemolysis medium in which a hemolysis reagent is adsorbed to a porous material to the sample supply side of the separation medium layer or the blood test reagent layer.