JPS62123336A - Measurement for hydrogen peroxide - Google Patents

Abstract

PURPOSE:To improve the detection element of hydrogen peroxide, by using reagent solutions, one being a carbonic acid buffer solution with the luminol concentration range of 10<-6>-10<-7>mol/l.0.2mol and the other an aqueous solution with the red blood salt concentration range of 10<-2.5>-10<-2>mol/l. CONSTITUTION:A sample hydrogen peroxide aqueous solution is sampled into a test tube 1 and set on a liminometer. Here, the concentration of 0.2mol carbonic acid solution in a reagent luminol is represented by X and the concentration of a reagent red blood salt aqueous solution Y, and then a reagent solution is prepared within the range of 10<-6>-10<-7> as the concentration of X and 10<-2.5>-10<-2> as concentration of Y. Then, when the luminometer is switched on, the luminol with the concentration X and the red blood salt solution with the concentration Y previously set in a reagent chamber 2 is automatically distributed into a test tube 1 through a solenoid valve 3 and the pump 4. Upon the end of the distribution, a chemical emission by luminol is generated, the quantity of light is converted 5 into a secondly electron quantity. The results are inputted into a miroprocessor 8 via an amplifier 6 and an interface 7 and the data processing is done automatically. Finally, the number of light emissions is printed out 9 to finish a series of measurements.

Description

DETAILED DESCRIPTION OF THE INVENTION A. Industrial Application Field The present invention relates to a method for quantitatively analyzing hydrogen peroxide, and in particular to a method for quantitatively analyzing hydrogen peroxide using luminol-hydrogen peroxide-red blood salt system chemiluminescence. It concerns analytical methods.

B0 Summary of the Invention In this invention, an analytical method for measuring hydrogen peroxide concentration using chemiluminescence of luminol, hydrogen peroxide, and potassium ferricyanide (red blood salt) is developed.
f1 degree 10-6 to 1.0-'mol//-0, 2m
The detection limit of hydrogen peroxide was successfully increased by applying the method of using an ol charcoal fi12 buffer solution and a potassium ferricyanide 10-2.5 to 10-"mol/l" aqueous solution to a highly sensitive luminometer device. .

C9 Conventional technology Luminol is known as a heavy reagent for hydrogen peroxide, and the technology for quantitative analysis of hydrogen peroxide using the chemiluminescence of the luminol-hydrogen peroxide-red blood salt (potassium ferricyanide) system has already been established. It is what was done.

The minimum detection limit of hydrogen peroxide by the above luminol oxidation luminescence method is described in the literature (Sachiko Kamei, Clinical Pathology).

P.358. (1983), the concentration is 8.8 x 101 mol/l. The reagent concentration used in this document is 4X for Lumi and Nor.
10-' mol/l (0, 2 mol/l carbonic acid 871
1) and potassium ferricyanide (red blood salt) are 6×
10-' mol/l (aqueous solution), and the above minimum detection limit value for hydrogen peroxide was obtained under these conditions.

I) 0 Problems to be Solved by the Invention In the cited examples of the above-mentioned documents, attempts are made to increase the luminescence intensity by varying the concentrations of luminol and potassium 7-erythyanide, but this does not apply to the analysis of hydrogen peroxide. Increasing sensitivity should be inconsistent. In addition, as the concentration of hydrogen peroxide in the sample decreases, the luminescence becomes weaker, so the problem seems to be that the luminescence measurement device was unable to measure weak light, making it impossible to increase detection sensitivity. Let's go.

This invention was made to solve this problem, and the detection limit for hydrogen peroxide quantification was
The purpose of this project is to establish a method for analyzing red blood salts using a device that can measure weak light, based on the idea that the chemiluminescence method of red blood salts could be used to improve the results.

Means for Solving Problem E3 The method for measuring filtrated hydrogen oxide concentration according to the present invention is a hydrogen peroxide-luminol-red blood salt chemiluminescence method, in which the luminol concentration m ranges from 10-' to 10-7 mol. /l! -0
, 2 mol carbonate [H] solution, and the concentration range of red blood salt is 10-'' to 10-2 mol/j' aqueous solution.

F0 effect In this invention, since a device capable of measuring weak light was used, the concentrations of luminol and red blood salt were determined.
Each measurable detection limit can be explored over a wide range of combinations of concentrations of each of these reagents.

G. Example Fig. 3 shows a device capable of measuring weak light and a luminometer (manufactured by Meiden Toryo Co., Ltd.) used in 17 on the wood flow side.

An explanatory diagram of Luminometer UPD-8000) is shown. In the figure, (1) is a test tube for analyzing a sample of hydrogen peroxide, (2) is a reagent chamber for preparing and storing luminol and blush salt solutions, and (3) is a test tube for analyzing a sample of hydrogen peroxide.
) is the electromagnetic valve for reagent injection, (4) is the test tube for reagent (
This is a pump for injecting into 1).

In addition, (5) is a photomultiplier that detects light emission from the test tube (1), (6) is an output amplifier for the photomultiplier, (7) is an interface, and (8) is a microcontroller that performs data processing. The processor (9) is a printer for printing data. In addition, ``Shi'' is a shutter controller, fil) is a shutter controller, and @ is a temperature controller for the test tube (1). Note that α3) is a high voltage power supply for the photomultiplier (5). Microprocessor (8)
is equipped with a mode switch α (a) and a memory unit side, and is operated by a keyboard switch αl.

Hereinafter, a measurement procedure using the luminescence filter configured as described above will be explained. First, sample hydrogen peroxide aqueous solution 100μ
Collect the sample into a test tube (1) and transfer it to a luminometer. Here, the concentration of the 0.2 mof/l' carbonic acid solution of reagent luminol is set to X, and the concentration of the reagent red blood salt aqueous solution is set to Y,
X concentration 10-4.5 to 10-7.5, Y concentration 10-
Prepare reagent solutions ranging from ' to 10-. FIG. 1 shows the combinations of X and Y. In addition, the concentration of the hydrogen peroxide sample was 0.10-'.

10-', 1.0-', 10-', 10-',
10-'mo17j' is an aqueous solution of seven types. By switching on the luminometer, luminol with a predetermined concentration
), 250 μe each is automatically dispensed into test tubes (1).

Simultaneously with the end of the partial pressure of these reagent solutions, luminescence occurs due to chemiluminescence of luminol, the amount of light is converted to the amount of secondary electrons by the photomultiplier (5), and the output is output from the amplifier (
6) and input to the microprocessor (8) via the interface 7/8 (7), where the data is automatically processed.
Number of luminescence (CPS: count per 5eeo
nd) is printed by the printer (9), and the series of measurements is completed. - As a measurement example, Fig. 2 shows a luminol concentration of 10
-"+ol/l, red blood salt concentration 10-!mol/l, the relationship curve between the luminescence B (cps) at 15 seconds after dispensing and the hydrogen peroxide concentration. The CV value was always less than 9%. From Figure 2, a concentration was detected that showed a luminescence amount 1.5 times the luminescence amount at a hydrogen peroxide concentration of 0 (i.e., the background value). Defining the limit, the actual value of the detection limit is 5×10-9, which is on the order of 10-'.

Although the results were obtained from the calibration curve in the same manner as in FIG. 2, FIG. 1 shows the detection sensitivity of hydrogen peroxide in logarithmic values for each combination of luminol and red blood salt concentrations X and Y.

In the examples, the case where a luminometer (manufactured by Meiden Kan Co., Ltd., UPD-8000) was used as the measuring device was explained, but the measurement device is not limited to this, and it is possible to use a luminometer having a means for detecting luminol luminescence with a photomultiplier. It goes without saying that a similar effect can be achieved if the method is used for measurement.

H. Effects of the Invention As explained above, this invention uses a device that can measure weak light, so it can trap a wide range of a degree of luminol and red blood salt, and measure measurable hydrogen peroxide and agrochemical degrees. I started thinking about it. As a result, 0.2 mol of luminol
/l! Concentration of carbonic acid slow solution 10-'~10''n+o
l/l, red blood salt aqueous solution eIf10-”' ~10
We were able to increase the detection limit of hydrogen peroxide to 10-s on the order of 10-s.

This value indicates an improvement in sensitivity of approximately one order of magnitude over the conventional value.

[Brief explanation of drawings]

Fig. 1 is an explanatory diagram showing the relationship between filtration reagent concentration and hydrogen peroxide detection sensitivity in one embodiment of the present invention, Fig. 2 is a hydrogen peroxide calibration curve, and Fig. 3 is a block explanatory diagram of the measuring device. be. Agent: Tadashi Sato, Patent Attorney Figure 1 Red Blood Salt>l Takaj7og y moj7/j! I have a few pieces on hand, * 10g (order) x 1-
1 The club is soft and 46 months x 2-2 hits,...! Transparent sunless moon Figure 2

Claims (1)

[Claims] In the hydrogen peroxide-luminol-red blood salt chemiluminescence analysis method, the concentration range of luminol is 10^-^6~10
It is a ^-^7 mol/l·0.2 mol carbonate buffer solution, and the concentration range of the red blood salt is 10^-^2^. ＾5～10
The hydrogen peroxide concentration measuring method uses a reagent solution that is ^-^2 mol/l aqueous solution.