JPS58137756A - Deproteinization method of blood serum - Google Patents

Abstract

PURPOSE:To remove protein which is a harmful material in analysis thoroughly and easily from a blood serum sample, by adding a water-soluble org. solvent to the serum to produce the precipitate of protein, then adding a water absorbent further thereto. CONSTITUTION:A water-soluble org. solvent is added to a blood serum which is required to be deproteinized in order to be subjected to analysis such as determination of medicines in blood, determination of traces of hormones in blood, etc. to allow protein to precipitate. A water absorbent is then added to a soln. mixture of the serum and the org. solvent, to force the water in the soln. to be absorbed in the absorbent, thereby removing the water in the soln. The protein dissolved in the water is deposited and settled by such removal of the water, and the serum is deproteinized thoroughly; therefore, the decrease in the analytic accuracy by the harmful material of the protein is prevented in the analysis to be executed thereafter.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for deproteinizing serum during serum analysis.

Conventionally, in order to remove proteins that interfere with the measurement of blood drug concentration (in serum), 4Il&solvent or ai
After precipitating the protein by adding diarrhea,
The commonly used method is to separate the precipitate by Rokusaku, but with this method, it is difficult to completely remove the proteins in the serum, and the protein-free serum is then separated using liquid chromatography. During analysis, residual proteins often appear as interfering substances, reducing the accuracy of analysis.

In view of the drawbacks of the conventional color-adding protein removal methods, the present invention has been made with the aim of providing a method that can completely and easily perform protein removal. The present invention provides a serum protein removal method characterized by adding a water-soluble organic solvent to precipitate proteins, and then further adding a water-absorbing agent.

In the present invention, the water-soluble organic solvent refers to an organic solvent that can be wetted with water in any quantitative relationship, and specifically includes, for example, methanol, ethanol, l-propanol, 2-
Propatool, allyl alcohol #O alcohols, ethylene glycol, 1.2-propanediol, 1.3
Examples include polyhydric alcohols such as propanediol, ethers such as ethylene oxide and propylene oxide, ketones such as acetone, acid amides such as formamide and dimethylformamide, nitriles such as acetonitrile, dioxane, and tetrahydrofuran.

Of these, acetonitrile, acetone, dioxane and tetrahydrofuran are particularly preferably used.

In addition, the water-absorbing agent used in this product is one that absorbs water and is insoluble in the organic solvent used.
Barium oxide, aluminum oxide, phosphorus pentoxide, boric anhydride, magnesium oxide, calcium chloride, sodium chloride, calcium chloride, magnesium chloride, magnesium perchlorate, barium perchlorate, calcium sulfate, **
Examples include steel, aluminum sulfate, potassium chloride, and nitrous chloride.

The subject of protein removal according to the present invention is serum, which is generally required to be subjected to protein removal for analysis such as blood drug therapy and blood am hormone measurement. To do this, first, the organic solvent is added to the serum to precipitate the proteins in the serum. Although it depends on the type of water-absorbing agent to be added later, it is preferable to set the volume to 1 volume or more based on the addition amount line of the organic solvent and the blood ml volume.

Next, the water absorbing agent is added to the mixed AI liquid of serum and an organic solvent, and the water in the solution is absorbed by the water absorbing agent Klk, thereby removing the water in the solution. By removing this water, the proteins dissolved in the water precipitate out and the serum is completely deproteinized.Therefore, in the subsequent analysis, the accuracy of the analysis is reduced due to protein interfering substances. That will no longer be the case.

Note that the water-absorbing agent may be used as long as it has enough electricity to absorb the water in serum, and there are no particular restrictions on how it can be used.

Furthermore, in order to take out the serum sample from which protein has been removed according to the present invention for analysis, it may be separated into a dissolved part, a precipitate, and a solid substance in the water-absorbing agent by using an intensive method such as centrifugation. Alternatively, the liquid mixture to which the water-absorbing agent has been added may be allowed to stand to allow the precipitate to settle, and then the upper liquid may be collected.

As mentioned above, the serum protein removal method of the present invention is characterized by adding a water-soluble organic solvent to the serum to be protein-removed to cause protein precipitation, and then further adding a water-absorbing agent. Proteins, which are interfering substances in analysis, are completely and easily removed from the serum sample, resulting in an excellent effect of improving the accuracy of analysis.

Examples of the present invention will be described below.

Examples and Comparative Examples 0.4 d of a water-soluble organic solvent acetonitrile was added to 0.2 d of serum rich in the anti-temperature drugs primidone and ethosuximide to form a precipitate of proteins in the wrinkled serum, and then anhydrous sodium sulfate was added as a water-absorbing agent. 0.22 was added, shaken, and allowed to stand still.

When the supernatant fluid was collected and the amount of water-soluble protein in the cough device fluid was measured using the 7-enol reagent method, the content was found to be 25 mef.
/- or less.

My father, put the above liquid into a 4.61 diameter x 25 (1 m column)
Product name ZORBAX-OD8. Dubo Co., Ltd.) was set on the fast liquid RI4 MADROGRAPH, and 3 was used as the eluent.
Analyzed by UV-UV-210n ultraviolet detection using 5-thiacetonitrile aqueous solution (flow rate, 1.3j/*),
Distinct peaks of primidone and ethosuximide were obtained, and the quantification of these anti-tencan drugs was possible.

On the other hand, for comparison, acetonitrile 0.4 was added to 0.21 serum containing anti-temperature drugs primidone and ethosuximide.
The supernatant solution collected from the serum sample in which sif was added to produce protein was measured using the phenol reagent method, and the result was 11! 500% water-soluble protein in the liquid
It was found that it contained more than F/-. The above solution was analyzed by high-pressure column chromatography in the same manner as in the above example, but water-soluble protein was eluted as an interfering component, and the peak of this component overlapped with the peaks of primidone and ethosuximide. Therefore, it was not possible to quantify these anti-depressant drugs.

patent applicant Sekisui Chemical Co., Ltd. Representative: Mototoshi Numa

Claims (1)

[Claims] 1. A serum protein removal method characterized by adding a water-soluble organic solvent to the protein removal serum to cause protein O precipitation, and then further adding a water-absorbing agent.