JPS58153167A - Color reagent for determining phosphoric acid - Google Patents

Abstract

PURPOSE:To prepare a reagent which can determine phosphoric acid simply, rapidly, and reliably, and has an excellent storage stability, by mixing an aqueous solution containing sulfuric acid, molybdate, antimonyl tartrate, and a nonionic surface-active agent with powdery ascorbic acid just before colorimetry. CONSTITUTION:The first agent comprises an aqueous solution of 3-5N sulfuric acid, 500-1,000mg/l (in terms of molybdate ion) of molybdic salt, 50-100mg/l (in terms of antimony) of potassium antimonyl tartrate and 0.15-0.3% polyoxyethylene alkyl ether. Powdery L-ascorbic acid is added as the second agent to the first agent fust before analysis. The mixed reagent is used in determination of phosphoric acid in serum or in soil or water. The both agents can be separately preserved for a long period of time, so that it is not necessary to weight and prepare a reagent each time a measurement takes place, and besides, the determination can be performed without being interfered by protein or the like contained in a solution to be measured.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a coloring reagent for quantitatively analyzing phosphoric acid in serum, for example. has become an important factor to analyze.

By the way, this kind of blood component analysis is mostly carried out by general clinical laboratory technicians, so it is easy to operate, the instruments and reagents are compact and there are few types, and the reagents and reagents are compact. 1IIv must be easy to manufacture and stable for a long period of time.

Conventionally, in this blood component analysis, phosphorus tl (P) is quantified by adding a coloring reagent to a certain amount of serum to develop a color and quantifying it using a spectrophotometer. On the other hand, it is also possible to apply the analysis of soil, water quality, etc. to serum analysis. (Dissolve 1Jl in water and filter.) ■ 0.1M Ascorbine 12 [(Dissolve 164JL of L-ascorbic acid in water to make 1Oil.)■
- Potassium antimonyl phosphate solution (WGo- is prepared by dissolving antimonyl potassium α2te4jIi in water.

Prepare a 4s type solution of IWlg-b/l) in advance, and during color comparison, add 301 ml of solution ■ to the enemy zoom above and mix.
To this, 60 - of solution (2) and 101 - of solution (2) are added and mixed well. For this reason, p4111 of the coloring reagent
! When performing analysis, the operation is complicated, it requires a tatami mat, and therefore requires a long time, and there are also disadvantages such as the above-mentioned liquid has poor shelf life and must be prepared every day. Unfortunately, I can't do that...

There was a point. In addition, with this coloring reagent, if a certain amount or more of a substance such as protein is present in the test liquid, it becomes cloudy and analysis becomes impossible.

This departure F! #11 A coloring reagent for phosphorus identification in serum that was created in view of the above circumstances, has easy coloring operations, good storage stability, and enables accurate quantification of phosphoric acid even if you are not an interlabial technician. The purpose is to provide the following.

The coloring reagent of the present invention consists of a first agent which is an aqueous solution containing sulfuric acid, molybdate, antimonylic acid and a nonionic surfactant, and a second agent which is powdered ascorbic acid.

First, the first agent will be specifically explained by giving examples of its preparation. Distilled Honzuri 700- was mixed with concentrated sulfuric acid All m while cooling, and 0.9 I of ammonium molybdate was added to this.
Add antimonyl potassium acid, 29 and 2 g of polyoxyethylene alkyl ether and dissolve well,
After the amount, distilled water is added to make 1000114, which is used as #Il agent. In the first agent of this preparation example, sulfuric acid 4N1 molybdate ion was 769 μ/l.

Antimony (sb)? 3~/l, polyoxyethylene alkyl ether 0.2%. However, in the coloring reagent of this invention, sulfur a [) ~ 5M molybdate ion la 0
0~zooo■/l, antimony 0V100■/
I, polyoxyethylene/alkyl ether O, l~O
The coloring effect of XIj can be obtained even in the highest density range. In addition, molybdate salts such as sodium molybdate can be used in addition to ammonium molybdate, and antimonyl tartrate salts such as sodium antimonyl tartrate can also be used. Further, other nonionic surfactants can be used in place of polyoxyethylene alkyl ether, but polyoxyethylene lauryl ether (eg, Br1j 35 Kao Atlas Co., Ltd.) is preferred. The addition of this nonionic surfactant
Interference caused by proteins in the test solution can be prevented.

This is a single L-ascorbic acid powder manufactured by MZ Pharmaceutical Co., Ltd.

Then, during colorimetry, a second agent is dissolved in the @l agent to form a coloring reagent, and a certain amount is added to the test solution to develop color. In the above preparation example, 100111jK 2-dose 4. Dissolve the liquid.

In such a coloring reagent, the preparation of the coloring reagent during color comparison is simply by dissolving the first part K11g2, so the operation can be performed easily and quickly, and there is no need for additional operations.

In addition, molybdenum ions and antimonyl phosphate ions were made into a dilute l12# solution, and ascorbic acid was made into a powdered TT Z agent, so the shelf life of Agents I and II was greatly improved. This further simplifies experimental operations.

Furthermore, since a nonionic surfactant is added, it is possible to remove interference from interfering substances such as proteins coexisting in the test solution.

Next, experiment f was conducted to examine the storage stability of the coloring reagent of this invention.
Ii&show.

[Experiment example]

According to Example III above, the first part and the second part were wrinkled, and the ones after preparation and those stored at room temperature for 2 months were quantitatively added to P, O, 5xo PJN. The storage stability was evaluated by measuring the absorbance of each K.The coloring reagent using the cloth agent immediately after making the Tsumugi rice and #1 showed the absorbance [0,2j(Te1υ- ) With the coloring reagent using the first agent stored at room temperature for 2 months, the absorbance of t' was 0.2 - 8 (7101), and no deterioration of Fi was observed when experimental errors were taken into account.

Next, an experimental example will be shown to demonstrate the effect of adding a nonionic surfactant.

[Experiment example]

0.2 doses of ovalbumin as an interfering substance was added to Tori o, @ standard solution (P, O, 寞 0. OOl So/1) to prepare a test solution. The first part of the above preparation example and the first part obtained by removing polyoxyethylene alkyl ether from this first part.
Prepare an agent (referred to as the first agent) and add 1011 ml of the upper ml test solution.
1 wLt of each was added to 7, color was developed, and the colors were compared. As a result, in the first agent, the P, O, concentration was 0,0 OO95
(2) The result was 1/1, whereas agent 1' caused white turbidity, making colorimetric determination impossible, indicating that the effect was due to ovalbumin.

In addition, this coloring reagent 1IFi can also be used as a coloring reagent for quantifying phosphoric acid in fields such as soil and water quality.

As explained above, the coloring reagent for quantifying phosphoric acid of the present invention is composed of a first agent containing sulfuric acid, a molybdate, an antimoel salt of tartrate, and a nonionic surfactant, and a second agent consisting of powdered ascorbic acid. Therefore,
A color reagent for colorimetry can be prepared easily and quickly, and preparation of mackerel can be prevented.

As a result, colorimetric determination can be performed easily, quickly, and with high precision. In addition, since both the first and second agents have excellent storage stability, they can be prepared in large quantities in advance and separated and used at the time of use. This makes it extremely suitable for field analysis. Furthermore, since the phosphoric acid content can be accurately quantified even when interfering substances such as proteins coexist in the test sample, there is an advantage that the troublesome preliminary removal of these interfering substances is not necessary.

Claims (1)

[Claims] A coloring reagent for quantifying phosphoric acid, comprising an aqueous agent IIl containing sulfuric acid, molybdate, antimonyl tartrate, and a nonionic surfactant, and a second agent consisting of powdered ascorbic acid.