JP2000121562A - Device for quantitatively determining coloring substance and storage medium for quantitatively determining coloring substance - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable the implementation of a simple and highly accurate method for quantitatively determining a coloring substance to be realized. SOLUTION: The device for quantitatively determining a coloring substance is provided with a light source part 11, a testing device supporting base 21, a part 13 for measuring the rate of relative intensity, an information processing part 14, an input part 15, a displaying part 16, and printing part 17 and irradiates a coloring substance in a solid-phase support with light and to measure the transmitted light. In the part 13 for measuring the rate of relative intensity, the above-mentioned transmitted light is dispersed into a plurality of light to measure the quantity of the spectrum of the transmitted light. The information processing part 14 handles the above- mentioned transmitted light by dividing it on the basis of its properties into a plurality of constituents. The rate of relative intensity of a plurality of light of the above- mentioned transmitted light dispersed at the part 13 for measuring a rate of relative intensity is computed to use each constituent of the transmitted light. By this, only the constituents of light scattered and diffused at a coloring substance to be detected and quantitatively determined are handled by separating them from other constituents of light in the transmitted light to be measured.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for quantifying a colored substance for detecting and quantifying a specific component in a sample by using a color reaction, and a display storing a program used for quantifying such a colored substance. The present invention relates to a color medium storage medium.

[0002]

2. Description of the Related Art Conventionally, detection and detection of specific components contained in a sample, for example, glucose, cholesterol or uric acid contained in blood, or components in a biological sample such as glucose, uric acid or protein contained in urine, and the like. At the time of quantification, a test device in which a reagent which reacts with a specific component to be quantified and develops a color on a flat solid support, for example, a test paper which is entirely or partially impregnated with the reagent is widely used.

In such a test device, the degree of coloration (coloring intensity) of a color product formed by a color reaction between a component to be detected / quantified and a reagent depends on the amount of a specific component contained in a sample. As a result, the specific component contained in the sample can be detected and quantified by measuring the degree of coloration. As a method of measuring the degree of coloration, there is a colorimetric method by visual observation, or an optical measurement method of optically measuring the degree of coloration using an analyzer, but in recent years, measurement accuracy, measurement speed, labor saving, etc. Optical measurement methods that are superior in this respect are widely used.

The general procedure of the optical measurement method is as follows.
A sample containing a component to be quantified is obtained by impregnating a solid support, which is a reagent holding portion of a test paper, with a solid support to form a color product, and then irradiating the color product in the solid support with light. Measuring the intensity of light (monochromatic light) of one type of maximum wavelength (specific absorption wavelength of the colored substance) corresponding to the color tone of the solid support containing the colored substance, which is included in the reflected light or transmitted light; Next, the measured intensity is compared with a calibration curve prepared in advance using a sample containing a known concentration of the component to be detected, and the specific component in the sample is quantified (ranked).

Accordingly, optical measurement methods are roughly classified into two types, a reflection method, which is a measurement method using reflected light, and a transmission method, a measurement method using transmitted light, depending on the type of light used for the measurement. Is done.

[0006] The reflection method corresponds to a reflectance or a reflectance obtained by irradiating a colored substance in a solid support with light having a specific absorption wavelength of the colored substance and measuring the amount of reflected light. This is a method of quantifying a colored substance based on parameters. Currently, in general, the quantification of a colored substance is performed by a method of directly measuring the reflectance of light having a specific absorption wavelength of the colored substance, but in this method, contamination of a sample or solid support is measured. It has been pointed out that it is easily affected by surface conditions and the like.

That is, the amount of reflected light measured by the reflection method for directly quantifying a color object from reflected light is the amount of specular reflected light, the amount of white diffuse reflected light (the spectral distribution of which is the same as the amount of diffusely reflected light). , And (other) the sum of the three types of diffuse reflected light, of which the diffuse reflected light is proportional to the content of the coloring matter, but the specular reflected light is the surface of the solid support. It is known that the amount of white diffuse reflection light varies depending on the state, particularly the degree of dryness and humidity, and the amount of white diffuse reflected light also varies depending on the density of the material of the solid support. Therefore, in such a reflection method, the measurement is also performed on the reference solid support in substantially the same state as the solid support for measurement, and the effects of the specular reflected light quantity and the white diffused light quantity fluctuation on the reflected light quantity are removed. Since it is necessary to use a uniform material for the manufacture of the solid support, and control the manufacture and storage of the solid support so that the state at the time of measurement is uniform, The impact must be minimized.

[0008] Therefore, in recent years, a parameter corresponding to the reflectance is calculated to quantify the color product for the purpose of suppressing the influence of the surface state of the solid support and eliminating the use of the reference solid support. The reflection method has been proposed. In those proposals, the spectral intensity obtained by spectrally reflecting the reflected light is adopted as a parameter corresponding to the reflectance, and the sum, difference or ratio of various spectral intensities of the measured reflected light and the reference reflected light is used. In many cases, the effect of the surface state of the solid support is suppressed, such as a wavelength at which the amount of change in the optical characteristics of the reflectance due to the degree of coloration is large as light for measuring the degree of coloration. Select light of a wavelength in the vicinity, and select light of a wavelength with a very small change in optical characteristics due to the degree of coloration as reference light, and determine the difference or ratio of signals obtained from those lights. Proposal for performing arithmetic processing (JP-B-59-779, JP-B-61-12814)
And Japanese Patent Application Laid-Open No. Hei 3 (1994), which measures the spectral intensities of red light, green light and blue light contained in reflected light.
Japanese Patent Application Laid-Open No. 22035/220) or a proposal to represent reflected light by tristimulus values in the XYZ color system of the International Commission on Illumination (CIE) (see Japanese Patent Application Laid-Open No. 7-35744). However, even if the spectral intensity of the reflected light is used in this way, the spectral intensity measured for each spectrum necessarily includes all of the three types of reflected light amounts described above, and the solid-phase support is included in the measurement results. We must accept that the effects of body surface conditions are included to some extent.

On the other hand, in the transmission method, which is another optical measurement method, light having a specific absorption wavelength or light having a specific absorption wavelength of a specific absorption wavelength of a colorant is applied to a colorant in a solid support. This is a method of irradiating and quantifying the color product based on the transmittance obtained by measuring the amount of transmitted light or a parameter corresponding to the transmittance.

In the transmission method, by using transmitted light instead of reflected light, fluctuations that are affected by the surface state of the solid support, such as fluctuations in accordance with the amount of specular reflection or the amount of white diffused light in the reflection method, are eliminated. It is known that it can be removed (see, for example, JP-A-61-41947).

Further, the transmitted light is spectrally measured to measure the spectral intensity, and the spectral intensity is used as a parameter corresponding to the transmittance to quantify the color product based on a plurality of sets of spectral intensity ratios. It is possible to analyze a color object with higher precision than a method of directly quantifying a color object from light, that is, in a method of directly quantifying a color object from reflected light or transmitted light, the degree of coloration is reduced. Although one-dimensionally correlated as the intensity of reflected light or transmitted light of a single wavelength (region), a plurality of spectral intensity ratios of a predetermined combination are obtained from the intensity of reflected light or transmitted light of different wavelengths (region). By associating these data with a distribution pattern of two or more dimensions, it is possible to enhance the mutual analysis ability of the data and improve the measurement accuracy. For example, protein, bilirubin or urobilinogen, In particular the reliability of the analysis results are also known to improve (e.g., JP-A in the analysis of components of the rate of change is less about the color former with respect to reduction 5-209836
Reference).

[0012]

However, in any of the transmission methods, when the irradiated light is transmitted through the solid support, a part of the irradiated light is scattered by the colorant and / or the solid support. The transmitted light is diffused and transmitted, and the rest is parallel transmitted light that passes through the solid support without contacting anything. Therefore, when measuring the amount of transmitted light, the amount of diffuse transmitted light and (the same spectral distribution as the irradiation light) It is desirable to improve the measurement accuracy by separating the parallel transmitted light amount (which is the transmitted light spectral amount), but this point is not considered in the conventional method. In addition, since the degree of scattering of transmitted light that determines the amount of diffuse transmitted light greatly varies depending on the material of the solid support, it is necessary to determine the optical path length of the transmitted light in order to apply Lambert-Beer's law. Have difficulty.

That is, it is generally known that the reciprocal of the logarithm of the amount of light is proportional to the density (Beer's law).
When it can be considered that a color object exists in a medium that is optically transparent and has a constant optical path length at the measurement wavelength (region), the relationship between the transmitted light amount (R) and the density (C) is expressed by the following equation: α (R) = 10- ^C
It is supposed that a straight line should be obtained by plotting the density and the amount of transmitted light on a graph in which the concentration is expressed on a logarithmic scale and the amount of transmitted light is equally spaced, but in the conventional method, a solid phase support containing a colored substance should be obtained. Since the amount of transmitted light of all transmitted light that has passed through the body is used, a plurality of transmitted light components having an indeterminate optical path length that have undergone various reflections and absorptions are included, and the relationship between the transmitted light amount and the density of the coloring matter is included. However, it is difficult to find a certain law, which limits the improvement of accuracy.

As a unique modification of the permeation method which can solve the above problems, a sample solution is brought into contact with a sample carrier having one net, a liquid film of the sample solution is formed on the net of the net, and the liquid film is dried. After being dried, the dried film is irradiated with a light beam beam perpendicularly to the dried film to detect the transmitted light transmitted therethrough, and the detected transmitted light is spectrally analyzed, and the effective volume of the dried film is determined and included in the sample solution. A method for calculating the concentration of one or several analytes has been proposed (JP-A-8-271).
No. 452).

However, this proposed method is disadvantageous in terms of simplicity and portability, because it does not use a solid support for holding the reagent, and requires time and effort to prepare the reagent during analysis. Furthermore, it is a great disadvantage that commercially available test devices such as test papers impregnated with reagents cannot be used.

In general, since the solid support is not optically transparent and has a small amount of transmitted light, it is necessary to irradiate relatively strong irradiation light and / or use a highly sensitive detector. It is desirable to improve the measurement accuracy by increasing the amount of detected transmitted light or taking measures such as eliminating the influence of the transmitted light using the ratio of the spectral intensity of the transmitted light (a parameter independent of the light). However, many of the conventionally proposed transmission methods are disadvantageous because such measures are not taken.

In the case where the solid support is optically translucent, the opal glass method (written by Kazuo Shibata,
Spectral Measurement and Spectrophotometer, p. 18, 1984)
It has been known. However, this method is not intended for research purposes, because it involves unresolved problems, increases the cost of producing suitable measuring instruments, or limits the solid support that can be handled. It seems that the measurement is hardly used, especially in practical simple measuring methods.

Furthermore, regardless of whether the method is a reflection method or a transmission method, in the conventionally proposed optical measurement method, a contaminant having a specific absorption wavelength similar to that of the color substance to be quantified (coexistence) Color) coexists in the sample, for example, when there is a coexisting color product such as hemoglobin or bilirubin contained when whole blood or serum is used as a sample,
Correction for eliminating the effects of the coexisting color products is necessary, but it is hard to say that the conventional optical measurement method has sufficiently corrected the effects of the coexisting color products.

That is, in the conventional method for quantifying a colored substance using a solid support, particularly when a biological material is used as a sample, it is not possible to simply and accurately quantify a colored substance to be detected and quantified. It was very difficult.

The present invention has been made in view of the above problems, and can realize a method for quantitatively determining a colored substance that can be simply and highly accurately determined using a solid-phase support. It is an object of the present invention to provide a color object quantifying device and a storage medium for color object quantification storing a program for such color object quantification.

[0021]

Means for Solving the Problems In order to solve the above-mentioned problems, the present invention provides a colorimetric substance quantification apparatus comprising: a sample; and a solid support for holding a reagent which causes a color reaction with a specific component in the sample. A color object to be quantified in the solid support by optically measuring the color intensity of the color object to be quantified, which is formed in the solid support by contact with a test device having A color quantification device for determining the concentration of light, light irradiation means for irradiating the solid support with light, transmitted light measurement means for receiving light transmitted from the solid support and measuring the amount of light A density calculating means for calculating the density of the color object to be quantified using the measured amount of transmitted light and a reference value acquired in advance, wherein the reference value is used to calculate various types of transmitted light. Depending on its properties, the scattered transmitted light component from the colored material to be quantified and other plural components Is a relative intensity ratio of each spectrum obtained by calculating from the amount of spectrally transmitted light obtained by spectrally separating each transmitted light component into a plurality of lights having a predetermined specific spectral distribution. In the means, the transmitted light from the solid support is separated into a plurality of lights equal to the reference value to measure the amount of spectrally transmitted light, and the density calculation means converts the transmitted light into a plurality of components equal to the reference value. The light intensity of the scattered transmitted light only by the color object to be quantified is calculated by using the obtained relative intensity ratio and the reference value to calculate the relative intensity ratio of the plurality of lights that have been separated and spectrally separated for each transmitted light component. Alternatively, the concentration of the color substance to be quantified is calculated and obtained using the light quantity of the spectrum having the specific absorption wavelength of the color substance to be quantified contained in the light quantity as a quantitative index. With such a configuration, the color product quantification apparatus of the present invention can easily and accurately quantify a color product to be detected and quantified.

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance according to the present invention is a test medium having a solid support which holds a sample and a reagent which undergoes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, A reference value acquisition for obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectral transmission light obtained by spectrally separating each transmitted light component into a plurality of lights having a predetermined specific spectral distribution. Irradiating the solid support with light A transmitted light measuring step of measuring the amount of spectrally transmitted light by separating the transmitted light into a plurality of lights same as the reference value obtaining step, and a separating step of separating the transmitted light into a plurality of components same as the reference value obtaining step Calculating a light amount of only the scattered transmitted light component from the color object to be quantified based on the spectral transmitted light amount and the reference value; and scattering from the color object to be quantified. A density calculating step of obtaining the density of the color object to be quantified from the light amount of only the transmitted light component. With such a configuration, the storage medium for color object quantification of the present invention can provide a color object quantification method that can easily and accurately quantify a color object to be detected and quantified by such a configuration. .

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance according to the present invention has a sample and a solid support for holding a reagent that undergoes a color reaction with a specific component in the sample. And optically measuring the color intensity of the color object to be quantified, which is formed in the solid support by contacting with a test device having a test device identification portion composed of an optically readable code. A storage medium storing a program used for a colorant quantification method for determining the concentration of a colorant to be quantified in the solid support, and obtaining a decoding table of the code of the test device identification unit in advance. And, various transmitted light is separated into a scattered transmitted light component from the color object to be quantified and a plurality of other components by its properties, and each transmitted light component has a predetermined specific spectral distribution. Calculation based on the amount of spectrally transmitted light A reference value obtaining step of obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum, and transmitting light obtained by irradiating the solid support with light, a plurality of light beams identical to the reference value obtaining step. A transmitted light measuring step of measuring the amount of spectrally transmitted light by dispersing the light into a plurality of components; a separating step of separating the transmitted light into the same plurality of components as the reference value obtaining step; A code decoding step of classifying the code, collating the code with a code decoding table of the test tool identification unit, and decoding the code; and determining a type and a reference value of a subsequent step used for an operation based on the code decoding step. And a density calculating step of obtaining the density of the color object to be quantified using the spectral transmitted light amount and the reference value according to the determination in the determining step. With such a configuration, the storage medium for quantitatively determining a colored substance of the present invention can provide a method for quantitatively determining a colored substance that can easily and accurately quantify a colored substance to be detected and quantified.

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance according to the present invention comprises a test medium having a sample and a solid support for holding a reagent that undergoes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, Each transmitted light component is calculated from the amount of spectrally transmitted light obtained by dispersing the transmitted light component into light having a specific absorption wavelength of the color product to be quantified and other light having a predetermined specific spectral distribution. Determine the relative intensity ratio and set a reference value for reference. A reference value obtaining step to obtain, and a transmitted light measuring step of measuring the amount of spectrally transmitted light by separating the transmitted light obtained by irradiating the solid support with light, into the same plurality of lights as the reference value obtaining step. Separating the transmitted light into the same plurality of components as the reference value obtaining step, and using only the scattered transmitted light component from the color object to be quantified in the spectral transmitted light amount using the reference value. A component light amount ratio calculating step of calculating a light amount ratio,
Component spectroscopy for calculating the light amount of light having a specific absorption wavelength of the color object to be quantified contained in the scattered transmitted light component from the color object to be quantified based on the ratio obtained in the component light amount ratio calculation step. It is characterized by including a light quantity calculating step and a density calculating step of obtaining the density of the color substance to be quantified from the light quantity of light having the specific absorption wavelength of the color substance to be quantified. With such a configuration, the storage medium for color object quantification of the present invention can provide a color object quantification method that can easily and accurately quantify a color object to be detected and quantified by such a configuration. .

Preferably, in the second to fourth aspects, in the density calculating step, the density of the color substance to be quantified is calculated using a relative intensity ratio diagram. With such a configuration, it is possible to provide a method for quantitatively determining a color product that more easily and accurately quantifies a color product to be detected and quantified.

Preferably, the sample according to any one of claims 2 to 5, wherein the sample contains a coexisting color product in addition to the color product to be quantified, and the solid-phase support has a wavelength of light used for measurement. A solid support that is optically transparent in the region, and always includes, as the reference value, a relative intensity ratio of the spectrum of light corresponding to the scattered transmitted light by the coexisting color product. With such a configuration, it is possible to provide a method for quantitatively determining a colored substance that can easily determine the colored substance to be detected and quantified, particularly in a biological sample.

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance of the present invention comprises a test medium having a sample and a solid-phase support for holding a reagent that undergoes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, A reference value acquisition for obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectral transmission light obtained by spectrally separating each transmitted light component into a plurality of lights having a predetermined specific spectral distribution. Irradiating the solid support with light A transmitted light measuring step of measuring the amount of spectrally transmitted light by separating the transmitted light into a plurality of lights same as the reference value obtaining step, and a separating step of separating the transmitted light into a plurality of components same as the reference value obtaining step A relative intensity ratio diagram creating step of creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value, and only the scattered transmitted light component from the color object to be quantified based on the relative intensity ratio diagram And a density calculating step of calculating the density of the color object to be quantified from the light intensity of only the scattered transmitted light component from the color object to be quantified. . The color medium quantitative storage medium of the present invention having such a configuration can provide a color object quantitative method that can easily and accurately quantify a color object to be detected and quantified particularly in a biological sample.

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance according to the present invention comprises a test medium having a sample and a solid support for holding a reagent which undergoes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, Each transmitted light component is calculated from the amount of spectrally transmitted light obtained by dispersing the transmitted light component into light having a specific absorption wavelength of the color product to be quantified and other light having a predetermined specific spectral distribution. Determine the relative intensity ratio and set a reference value for reference. A reference value obtaining step to obtain, and a transmitted light measuring step of measuring the amount of spectrally transmitted light by separating the transmitted light obtained by irradiating the solid support with light, into the same plurality of lights as the reference value obtaining step. A sorting step of sorting the transmitted light into the same plurality of components as the reference value obtaining step, and a relative intensity ratio diagram creating step of creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value,
A component light amount ratio calculating step of calculating a ratio of a light amount of only the scattered transmitted light component from the color object to be quantified to the spectral transmitted light amount based on the relative intensity ratio diagram, and a component light amount ratio calculating step. A component spectral light amount calculating step of calculating an amount of light having a specific absorption wavelength of the color object to be quantified contained in the scattered transmitted light component from the color object to be quantified based on the ratio. A density calculating step of calculating the density of the color product to be quantified from the amount of light having a specific absorption wavelength of the color product to be quantified. With such a configuration, it is possible to provide a method for quantitatively determining a color product that more easily and accurately quantifies a color product to be detected and quantified.

Further, in order to solve the above-mentioned problems, a storage medium for quantitatively determining a colored substance according to the present invention comprises a test medium having a solid support which holds a sample and a reagent which undergoes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, A reference value acquisition for obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectral transmission light obtained by spectrally separating each transmitted light component into a plurality of lights having a predetermined specific spectral distribution. And contacting the sample with the test device. As soon as possible, the transmitted light obtained by irradiating the solid support with light is separated into the same plurality of lights as in the reference value obtaining step, and the amount of spectrally transmitted light is measured. A transmitted light measurement step repeatedly performed a plurality of times until the measured value of the spectral transmitted light amount is stabilized at a time interval, and a separation step of separating the transmitted light into a plurality of components same as the reference value acquisition step, A relative intensity ratio diagram creating step of creating a relative intensity ratio diagram from the spectral transmitted light quantity and the reference value, and a solid phase support before the formation of the color object to be quantified at the contact time based on the relative intensity ratio diagram A determining step of determining a spectral transmitted light amount and a relative intensity ratio of transmitted light from the body; and a scattered transmission from the color object to be quantified based on the relative intensity ratio and the determined value obtained in the determining step. Light component And a density calculating step of calculating the density of the color object to be quantified from the light intensity of only the scattered transmitted light component from the color object to be quantified. . With such a configuration, the storage medium for quantitatively determining a colored substance of the present invention eliminates the influence of the background, particularly in a biological sample, and accurately measures the colored substance to be detected and quantified. Can be provided.

Further, in order to solve the above-mentioned problems, a storage medium for the determination of a colored substance of the present invention comprises a sample and a solid support which holds a reagent which causes a color reaction with a specific component in the sample. Concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified, which is formed in the solid support by contacting with a tool. Is a storage medium storing a program used for the colorant quantitative method for determining, the transmitted light is separated into the scattered transmitted light component from the colorant to be quantified and other plural components by its properties, Each transmitted light component is calculated from the amount of spectrally transmitted light obtained by dispersing the transmitted light component into light having a specific absorption wavelength of the color product to be quantified and other light having a predetermined specific spectral distribution. Determine the relative intensity ratio and set a reference value for reference. A reference value acquisition step of Tokusuru quickly as possible after contact with the sample and the test device,
The transmitted light obtained by irradiating the solid-phase support with light is divided into a plurality of lights in the same manner as in the reference value obtaining step to start measuring the amount of spectrally transmitted light. A transmitted light measurement step that is repeatedly performed until the measured value of the light amount is stabilized, a separation step of separating the transmitted light into the same plurality of components as the reference value acquisition step, and the spectral transmitted light amount and the A relative intensity ratio diagram creating step of creating a relative intensity ratio diagram from a reference value, and spectroscopy of transmitted light from the solid support before forming the color substance to be quantified at the time of the contact based on the relative intensity ratio diagram A determining step of determining a transmitted light amount and a relative intensity ratio, and scattered transmission from the color object to be quantified in the spectral transmitted light amount based on the relative intensity ratio and the determined value obtained in the determining step. Light component A component light amount ratio calculating step of calculating the ratio of the only light amount, and the coloration to be quantified contained in the scattered transmitted light component from the colorant to be quantified based on the ratio obtained in the component light amount ratio calculating step A component spectral light amount calculating step of calculating the light amount of light having a specific absorption wavelength of the object, and a density calculation of calculating the concentration of the color object to be quantified from the light amount of the light having the specific absorption wavelength of the color object to be quantified And a step. With such a configuration, it is possible to provide a colorant quantification method for eliminating the influence of the background, particularly in a biological sample, and quantifying the colorant to be detected and quantified with higher accuracy.

Preferably, in claim 2 to claim 10, in the reference value obtaining step, the transmitted light is:
Parallel transmitted light without scattering, scattered transmitted light by solid support,
It is characterized in that it is separated into the transmitted light components of the scattered and transmitted light by the color material to be quantified and the scattered and transmitted light by the coexisting color material when present. With such a configuration, it is possible to provide a color substance quantification method for efficiently and accurately quantifying a color substance to be detected and quantified.

[0032] Preferably, in claims 2 to 11, the plurality of lights obtained in the reference value obtaining step are three types of spectroscopy. With such a configuration, it is possible to provide a method for quantitatively determining a color product that efficiently and accurately quantifies a color product to be detected and quantified with a small amount of calculation.

Preferably, in claim 12,
The three types of spectroscopy are primary stimuli of the RGB color system (CIE color system) defined by the International Commission on Illumination.
With such a configuration, a color product to be detected and quantified can be simply and accurately determined using a commercially available device.

Preferably, in claim 12,
The three types of spectroscopy are the primary stimuli of the XYZ color system (CIE1931 standard color system) defined by the International Commission on Illumination. With such a configuration, a color product to be detected and quantified can be simply and accurately determined using a commercially available device.

Preferably, in claim 12,
The three types of spectroscopy are primary stimuli of the UCS color system defined by the International Commission on Illumination. With such a configuration, a color product to be detected and quantified using a commercially available device can be easily and accurately quantified.

Preferably, in the second to fifteenth aspects, Grassmann's law of additive color mixture is used in the calculation process for calculating the concentration of the color substance to be quantified. With such a configuration, it is possible to simply and accurately execute the calculation required for quantifying the color product to be detected and quantified.

It is also preferable that, in the density calculation step, the density of the color substance to be quantified is calculated by the following formula: Log (C) = α (A) ² + β (A) + Γ (where C is the concentration of the color substance to be quantified, A is the amount of scattered transmitted light from the color substance to be quantified or the amount of light having a specific absorption wavelength of the color substance to be quantified contained therein. , And α, β, and γ each represent a constant). With such a configuration, the color substance to be detected and quantified can be quantified with necessary and sufficient accuracy.

Further, in order to solve the above-mentioned problems, the present invention provides a colorimetric substance quantifying apparatus, comprising: a test device having a sample; and a solid support for holding a reagent which performs a color reaction with a specific component in the sample. To determine the concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified formed in the solid support. A colorant quantitative device, a light irradiating means for irradiating the solid support with light, a transmitted light measuring means for receiving transmitted light from the solid support and measuring the amount of light, 18. Density calculating means for calculating the density of the color object to be quantified using the amount of transmitted light and a previously acquired reference value, wherein the calculating means is any one of claims 2 to 17. The present invention is characterized by including the storage medium for quantitatively determining a colored substance described in the section. With such a configuration, the color product quantification apparatus of the present invention can efficiently and accurately quantify the color product to be detected and quantified.

[0039]

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a block diagram showing a color substance quantifying apparatus according to the present invention.

FIG. 1 is a block diagram schematically showing an example of an embodiment of a coloring matter quantifying apparatus according to the present invention.

According to FIG. 1, the apparatus of the present invention comprises a light source unit 11 as a light irradiating unit, a test tool support 21, a relative intensity ratio measuring unit 13 as a transmitted light measuring unit, and an information processing unit as a density calculating unit. Unit 14, input unit 15, display unit 16, and printing unit 1
7. Then, the light source unit 11 emits light, irradiates the test tool mounted on the test tool support 21 and having the solid support, and is provided to face the light source unit 11 with the test tool support 21 interposed therebetween. After the relative intensity ratio measuring unit 13 spectrally receives the transmitted light transmitted through the test device and receives the light, a signal corresponding to the received light amount is generated, and the information processing unit 14 receives the signal from the relative intensity ratio measuring unit 13 While performing various calculations and controlling the operation of the entire apparatus, the input unit 15 accepts instructions from the operator as necessary and outputs information to the display unit 16 and the printing unit 17.

The light source unit 11 includes one or a plurality of light sources 22. The light source 22 is not particularly limited as long as it can output light having a predetermined spectral distribution. Light sources commonly used in the industry, such as halogen lamps, tungsten lamps, and multiple LEDs, such as deuterium discharge tubes and xenon discharge tubes when measuring with ultraviolet light, and Nernst lamps when measuring with infrared light. There can be. The optical path from the light source 22 to the test tool on the test tool support 21 can be configured by using, for example, an optical fiber, a mirror, a lens, or the like, or can be directly irradiated.

The test tool support 21 is a support on which the test tool is mounted at the time of measurement. The entire test tool or a predetermined area of the test tool mounted on the test tool support 21 is evenly irradiated with light from the light source 22. It is arranged to be.

The relative intensity ratio measuring section 13 is roughly composed of a lens system 23 and an imaging section 24. The lens system 23 is
It comprises one or more optical lenses, and adjusts the optical path length so as to refract the transmitted light emitted from the light source 22 and transmitted through the test device to form an image on the light receiving surface. The imaging unit 24 includes the spectral unit 13
a, the photodetector 13b and the imaging control circuit 42. The spectral unit 13a includes a plurality of spectral filters or dispersive elements for separating transmitted light into a plurality of spectral components having a predetermined specific wavelength (band). In the present embodiment, as shown in FIG. 2, there are three types of spectral filter groups 44 of R, G, and B, and each of the spectral filters is provided immediately above the light receiving surface of the light receiving element 43 of the light detection unit 13b. ,
The three types of spectral filters are provided so that the distribution densities are uniform. The light detection unit 13b includes a light detection element that outputs an electric signal corresponding to the amount of received light, such as a CCD solid-state imaging device 41 that has a light reception element 43 that receives light separated by the light separation unit 13a. Then, the imaging control circuit 42
Controls the light detection elements such as the CCD solid-state imaging element 41, processes signals from the light detection elements, and outputs the processed signals to the information processing unit 14 and the display unit 16 as required.
The output signal when outputting to the display unit 16 is NTS
C signal and various control signals. That is, it can be said that the relative intensity ratio measurement unit 13 including the lens system 23 and the imaging unit 24 has the same function as the imaging unit of the color camera or the color video camera. Therefore, the sampling of the amount of transmitted light in the relative intensity ratio measurement unit 13 can be performed in units of the amount of received light per fixed time, that is, the number of frames.

The light source unit 11, the test tool support 21 and the relative intensity ratio measuring unit 13 described above are all stored in the same case 25 to prevent the influence of stray light originating from the external environment. desirable. At this time, all the devices are fixed in the case and the optical path used for measurement is fixed length, and means for adjusting the positional relationship, for example, a structure that can be provided with an adjusting screw to change the relative position, is used for transportation and so on. It is also possible to adjust the deviation of the optical path length over time or change the optical path length depending on the measurement conditions.

The information processing section 14 has a schematic configuration in which a storage section 14a, an operation section 14b, and an input / output control section 14c are connected to a bus 38. The storage unit 14a includes a read only memory (ROM), a random access memory (RAM), a magnetic storage medium such as a hard disk, and a magneto-optical storage medium such as an MO disk. Store. In this example, an R program in which a program such as a coloring matter quantification program according to the present invention is stored in advance.
An OM 33, a RAM 35 for temporarily storing image data and control signals, and the like, and a magnetic storage medium 34 for storing image data and measurement results and calculation results including reference values for a long term are used. The calculation unit 14b includes a central processing unit (CPU) 37
In accordance with various programs stored in the storage section 14a, information (signal) from each section of the apparatus is processed to control the entire apparatus. The input / output control unit 14c includes an A / D conversion unit 31,
It comprises an image data acquisition unit 32, an input control unit 39a, a display control unit 39b, and a print control unit 39c.
The A / D conversion unit 31 converts the analog signal received from the relative intensity ratio measurement unit 13 into a digital signal and passes the digital signal to the image data acquisition unit 32. The image data obtaining unit 32 calculates the image data and the communication signal obtained by processing the received signal,
Send to. Input control unit 39a, display control unit 39
b and the print control unit 39c are composed of the calculation unit 14b and the input unit 1
5. Provide an interface that enables communication through transmission and reception of signals to and from the display unit 16 and the printing unit 17. For example, the display control unit 39 b
It outputs the B signal and various control signals. The input unit 15
It comprises one or more input means such as a keyboard, a mouse, a touch panel, a bar code reader and / or a pen tablet, and accepts instructions from the operator. The display unit 16 is a display device (display) using a CRT, liquid crystal, or the like.
Alternatively, it receives an NTSC signal from the relative intensity ratio measurement unit 13 and displays a measurement image, a measurement result, a calculation result, and the like individually or simultaneously on a divided screen. Printing unit 17
Is a printing device (printer) of any type such as dot impact, ink jet or laser, and prints information according to an operator's command.

In the color substance quantifying apparatus of the present invention configured as described above, an appropriate test tool is placed on the test tool support 21 and quantification is performed by measuring the transmitted light from the test tool. The concentration of the color product to be obtained is determined and quantified. The operation of the present apparatus at that time will be described.

The light emitted from the light source 22 is applied to the test fixture support table 21.
A predetermined region of the test device placed on the upper surface is irradiated and transmitted. The transmitted light that has passed through the test tool is refracted by the lens system 23 of the relative intensity ratio measurement unit 13, is separated by the spectral filter 44, and forms an image on the light receiving surface of the light receiving element 43 of the CCD solid-state imaging device 41. The light receiving element 43 accumulates an amount of electric charge according to the amount of received spectral light. The imaging control circuit 42 transmits a control signal for outputting the received light quantity of each light receiving element 43, that is, the charge amount information, to the CCD solid-state imaging element 41 at an appropriate timing, and receives the information output from the CCD solid-state imaging element 41. . The imaging control circuit 42 adds various timing control signals to the received information,
A signal is generated, and the information processing unit 14 and / or the display unit 1
6 and transmits the generated NTSC signal. The display unit 16
An image created from the received NTSC signal is displayed. In the information processing unit 14, the A / D conversion unit 31 of the input / output control unit 14c receives a signal from the imaging control circuit 42 and converts the received analog signal (NTSC signal) into a digital signal (RGB).
Signal), and transmits the signal to the image data acquisition unit 32. The image data acquisition unit 32 is a central processing unit 3 of the arithmetic unit 14b.
While controlling the A / D conversion unit 31 based on the instruction from 7, the signal from the A / D conversion unit 31 is received and processed. That is, the A / D converter 31 converts the received NTSC signal into RGB signals in units of one frame, obtains the signal of the number of frames instructed by the central processing unit 37, calculates an average value per frame, The obtained average value is stored in the RAM 35 or the magnetic storage medium 34 of the storage unit 14a as test tool image data. The central processing unit 37 temporarily stores the test tool image data in the RAM 35 as necessary,
An RGB signal is transmitted to the display unit 16 via the display control unit 39b to display an image. In addition, the central processing unit 37 performs an operation according to a program stored in the ROM 33 using the acquired test tool image data and a reference value stored in the magnetic storage medium 34 in advance, that is, the operation result, that is, the amount to be quantified. Obtain the density of the color product. Then, the calculation result is stored in the magnetic storage medium 34 and displayed on the display unit 16 in the same manner as the test tool image data, and according to the operator's command obtained from the input unit 15 via the input control unit 39a, An instruction is issued to the printing unit 17 via the printing control unit 39c to print.

Next, a description will be given of a test tool which can be used in the color substance determination apparatus of the present invention.

The test device which can be used in the present apparatus has a solid support which is a site or a region for holding a reagent which undergoes a color reaction with a color substance to be quantified, and which is inactive against components in the sample. The sample support member is made of a material and usually has a plate shape with a thickness of about 20 to 500 μm. The solid support may be integrally formed with the same material as the other parts of the test device, or may be formed as a separate member of a different material and attached to the test device. In addition, the test device may have, in addition to the solid support, a reference solid support that is the same as the solid support except that it does not hold the reagent.

The composition of the reagent held on the solid support is not particularly limited, and it can be appropriately selected according to the components to be quantified according to common knowledge in the art. For example, as a reagent composition used when quantifying glucose in a sample, a reagent composition containing glucose oxidase, peroxidase and o-tolidine as main components can be exemplified.

Examples of the test device usable in the present invention include a test paper partially impregnated with a reagent, a solid support made of a porous material such as a filter paper impregnated with the reagent or a porous polymer,
And a single-item test device, a multi-item test device, a single-item rate assay test device, or a multi-item test device using a test paper, a glass plate, a resin plate, or the like to which one or more types of reference solid supports are adhered as desired. A rate assay test device and the like can be exemplified. In a single measurement, use a single item test device when quantifying only one component in a sample, a multi-item test device when quantifying multiple components, and a single-item or multi-item rate assay if desired. A test device can be used. Test tools that can be used in the present invention are illustrated in FIGS.

The test device shown in FIG. 3 has a solid support (measurement test section) 51 made of a piece of filter paper and a reference solid support (reference test section) 52 stuck at appropriate intervals. Item test paper 45. Various such single-item test papers are commercially available, and those commercially available products can be used. In addition, as shown in FIG.
The identification mark 53 such as a bar code is directly printed at an appropriate position on the surface where the solid support 51 is attached, or a sticker on which the identification code is printed is attached. By providing the test paper, the type of the reagent held by the solid support 51, the size of the test paper 45, or the size or the adhesion position of the test paper such as the size of the solid support 51 and the reference solid support 52. It is also possible to facilitate the identification of the type. The size of the test paper 45, the solid support 51, and the reference solid support 52 in FIG. 3 are 5 mm × 40 mm, respectively.
It is generally about 3 mm × 3 mm and about 3 mm × 3 mm, but the size of each single-item test paper used in the present invention and each part is not limited thereto, and various commercially available single-item test papers are used. Paper can be used. Also,
A test paper having no reference solid support 52 may be used.

In the test device shown in FIG. 4, a solid support (measurement test section) 51a to 51c composed of a piece of filter paper and a reference solid support (reference test section) 52a to 52c are provided at appropriate intervals. This is the multi-item test paper 46 stuck. As with the single item test paper 45, various types of commercially available products can be used as the multi-item test paper 46, and the test paper identification unit 53 can be provided as desired. The test paper 46, the solid supports 51a to 51c and the reference solid support 52a of FIG.
The size of ~ 52c is 10 mm x 20 mm, 2
It is generally about 2 mm x 2 mm and about 2 mm x 2 mm, but the overall size of the test device and each part used in the present invention is not limited thereto, and various commercially available multi-item test papers may be used. Can be used. Further, the test paper may not have the reference solid supports 52a to 52c.

The test device shown in FIG. 5 is a single item rate assay test paper 47 to which a solid support (measurement test section) 54 composed of a piece of filter paper is adhered. As the single item rate assay test paper 47, various commercial products can be used similarly to the single item test paper 45, and the test paper identification unit 5 can be used as required.
3 can be provided. The sizes of the test paper 47 and the solid support 54 in FIG. 5 are 5 mm × 40 mm and 3 mm, respectively.
It is generally about × 3 mm, but the size of the entire test device and each part used in the present invention is not limited thereto, and various commercially available single item rate assay test papers may be used. it can.

The test device shown in FIG. 6 is a multi-item rate assay test paper 48 on which solid supports (measurement test sections) 54a to 54i made of a piece of filter paper are attached at appropriate intervals. As the multi-item rate assay test paper 48, similarly to the single-item test paper 45, various commercially available products can be used, and a test paper identification unit 53 can be provided as desired. The test paper 48 in FIG. 6, the solid supports 54a to 54
The size of i is 10mm x 20mm, 2mm x
Although it is generally about 2 mm, the size of the entire test device and each part used in the present invention is not limited to this, and various commercially available multi-item rate assay test papers can be used. .

A procedure for quantifying a colored substance using the apparatus for quantifying a colored substance of the present invention and the above-described test device will be described below with reference to FIG.

When the power of the apparatus of the present invention is turned on, a message screen for inputting various information is displayed on the display unit 16 in step S101 to request the input, and the process proceeds to step S102.

In step S102, the input method of the information requested in step S101 is determined, and the input of the information is confirmed according to the input method. Of the information to be input in step S102, the operator has to input the atypical information using the keyboard or mouse of the input unit 15, but the test equipment and the components to be quantified that are actually used The standard information (determined by the test device used) such as the information about the data may be input in the same manner as the non-standard information. A reading unit is provided as one of the input units of the input unit 15, and a table or a vote or the like in which predetermined information is recorded using a code such as a barcode is created in advance in the same manner as the test paper identification unit 53 described above. The information can also be input by reading recorded information from a table or vote using the optical reading means. Alternatively, the test strip used in the present invention is provided with the test strip identification section 53 as described above, and at the time of measurement, the transmitted light from the test strip is analyzed and the information recorded in the test strip identification section 53 is read, so that the apparatus can obtain the predetermined information. Can be obtained, and the trouble of inputting standard information at the time of turning on the power can be omitted (this method is hereinafter referred to as a transmitted light recognition method). However, when the transmitted light recognition method is used, input of fixed information is not performed until measurement is performed.

Therefore, in step S102, it is first determined whether or not to use the transmitted light recognition method based on information from the operator. If it is determined that the transmitted light recognition method is to be used, an irregular pattern is used. After confirming that all relevant information has been input, the process proceeds to step S104. If it is determined that all information is not used, the process proceeds to step S103 after confirming that all information has been input.

In step S103, based on the information obtained in step S102 and the information stored in advance in the storage unit 14a, the test equipment to be used and the component to be quantified are recognized, and various set values are determined accordingly. For example,
Recognition of the type and size of the test device, recognition of the type of color product to be determined formed from the reagents used and the components to be determined, and the time required for the color product formation reaction (color reaction), test Of the light-receiving element to measure the amount of received light according to the size of the tool and the solid support, or the light-receiving element that receives the spectrum having the wavelength closest to the specific absorption wavelength of the formed color object to be quantified Is determined. Then, after all determinations have been completed, the process proceeds to step S104.

In step S104, a message prompting input of a measurement start command is displayed on the display unit 16, and the flow advances to the next step S105. After the operator brings the sample into contact with the test device to impregnate the sample into the solid support and optionally the reference solid support, the operator places the test device on the test device support 21 and issues a measurement start instruction. Enter However, the actual measurement is
The operation is started after a lapse of a predetermined time from the input of the measurement start command. This is particularly important when using a rate assay.

In step S105, it is determined whether or not a measurement start command has been input, and after confirming the input, the flow proceeds to step S106.

In step S106, time measurement is started, and the flow advances to step S107.

In step S107, it is determined whether or not the test device to be used uses the transmitted light recognition method based on the recognition in step S102. If the optical recognition method is not used, the process proceeds to step S109.

In step S108, first, an extra test tool image data acquisition operation is performed once before the usual test tool image data acquisition, and the transmitted light from the test tool is analyzed to record the information recorded in the test paper identification section 53. Is read to obtain standard information, and in the same manner as in step S103, the used test tool and the component to be quantified are recognized based on the obtained information and the reference value, and various set values are determined accordingly. Next, it is determined whether or not the test device to be used is a test device using the rate assay method based on the obtained recognition. If it is determined that the rate test method is to be used, step S110 is performed.
If it is determined that the rate assay method is not used, the process proceeds to step S111.

In step S109, based on the recognition in step S103, it is determined whether or not the test device to be used is a test device that uses the rate assay method. If it is determined that the rate assay method is used, the process proceeds to step S110. The process proceeds to step S111 when it is determined that the rate assay method is not used.

In step S110, image data of the test device is acquired with time according to the rate assay method, and the light intensity of transmitted light of the solid support and the relative intensity ratio of each spectrum included in the data are obtained. The density of the coloring matter to be quantified based on the reference value is determined by calculation using the value of (1), and the density is determined. Step S
Details of 110 will be described later.

In step S111, the image data of the test tool is obtained, the light intensity of the transmitted light of the solid support and the relative intensity ratio of each spectrum included in the data are determined, and the values are quantified based on the reference value using these values. The density of the coloring matter to be obtained is calculated by operation and quantified.
Proceed to. Details of step S111 will be described later.

In step S112, the quantitative result is displayed on the display unit 16 and printed by the printing unit 17 as desired. Then, the process proceeds to step S113.

In step S113, the operator is inquired whether to end or continue the measurement operation. If the result of the inquiry is continued, the process proceeds to step S101.

Next, the details of step S110 will be described with reference to FIG.

As shown in FIG. 8, step S110 executed when using the rate assay test device includes substeps S201 to S206.

In sub-step S201, it is determined whether or not a predetermined time has elapsed since the start of the time measurement in step S106, and after elapse of the predetermined time, the process proceeds to sub-step S202.

In sub-step S202, the image data acquisition unit 32 is instructed to acquire the test tool image data, and specifies the number of frames to be used for acquiring the test tool image data. Next, the process proceeds to sub-step S203.

In sub-step S 203, after receiving the test tool image data from the image data acquisition section 32, the received test data is determined based on the received data, various data stored in the magnetic storage medium 34 in advance, and the reference value. The amount of transmitted light from the solid support included in the component image data, the transmitted light component obtained by dividing the transmitted light according to the properties, and the relative intensity ratio of each of these components are obtained by calculation, and the obtained calculation is performed. The result is stored in the magnetic storage medium 34 and / or RA
After storing in M35, the process proceeds to sub-step S204.

In sub-step S204, 1 is added to a counter for recording the order of the test tool image data received in the sub-step S203 after the start of measurement, and the count number and the step S10
The number of pieces of test tool image data to be acquired set in step 3 is compared. If the counted number has not reached the specified number, the process proceeds to sub-step S205. If the counted number has reached the specified number, the process proceeds to sub-step S206.

In sub-step S205, the time elapsed since the data was received in sub-step S203 is changed to the next data acquisition instruction in order to acquire data at the test tool image data acquisition time interval set in step S103. Then, it is determined whether or not the time suitable for transmitting the data has been reached, and after confirming that the elapsed time from the data reception has reached the time, the process proceeds to the sub-step S202.

In sub-step S206, all the test-piece image data obtained in sub-step S203 and the calculation results obtained therefrom, and the reference values stored in the magnetic storage medium 34 in advance, are used to determine the quantity to be determined. The concentration of the color object to be quantified is determined and quantified by using, as a quantitative index, the amount of scattered transmitted light due to only the color object or the amount of spectral light having the specific absorption wavelength of the color object to be quantified included in the light amount. Then, step S110 ends, and the process proceeds to step S112.

Next, details of step S111 will be described with reference to FIG.

Step S111 executed when using a test device other than the rate assay test device includes sub-steps S301 to S304 as shown in FIG.

In sub-step S301, it is determined whether a predetermined time has elapsed since the start of the time measurement in step S106, and after elapse of the predetermined time, the process proceeds to sub-step S302.

In sub-step S302, the image data acquisition unit 32 is instructed to acquire test tool image data, and the number of frames to be used for acquiring test tool image data is specified. Next, the process proceeds to sub-step S303.

In sub-step S 303, after receiving the test tool image data from the image data acquisition unit 32, the received test is performed based on the received data and various data and reference values previously stored in the magnetic storage medium 34. The amount of transmitted light from the solid support (and, if necessary, the reference solid support) included in the component image data, the transmitted light component obtained by dividing the transmitted light into components according to properties, and each of them After calculating the relative intensity ratio of the spectrum and storing the obtained calculation result in the magnetic storage medium 34 and / or the RAM 35, the process proceeds to the sub-step S304.

In the sub-step S304, the test tool image data obtained in the sub-step S303 and the calculation results obtained therefrom, and the magnetic storage medium 34
Using the reference value stored in the, as a quantitative index, the amount of scattered transmitted light only by the color product to be quantified or the amount of spectral light having the specific absorption wavelength of the color product to be quantified included in the light amount, The concentration of the coloring matter to be quantified is determined and quantified. Then, step S111 ends, and step S111 is executed.
Proceed to 112.

[0086] The apparatus for quantitatively determining a color product of the present invention and / or
Alternatively, a method for quantifying a color substance realized by a storage medium for quantifying a color substance will be described.

The outline of the color substance determination method is as follows. The color substance in the solid support is irradiated with light, and the amount of transmitted light is measured. A transmission method is used to quantify the color object based on the set reference value. At this time, the transmitted light is divided into a plurality of components according to its properties, and each transmitted light component is set to a predetermined specific spectral distribution. Is a method for quantifying a colored substance using a solid support, in which the ratio of each spectral intensity to the total transmitted light component intensity (total of the spectral intensity) is used as a parameter for each transmitted light component. .

Here, in the present specification, “solid support”
The term is used to mean a test site or test area that holds a reagent that undergoes a color reaction with a component to be quantified in a test device such as a test paper, but if necessary, has the same configuration except that it does not hold the reagent. The site or region of the test device to be taken may be particularly referred to as a “solid support for reference”.

The expression “separate the transmitted light into a plurality of components depending on the nature” means that the transmitted light is a parallel transmitted light without scattering, a scattered transmitted light by a solid support, and a colorant (to be quantified). This means that the light is separated into respective components such as scattered transmitted light and, in some cases, scattered transmitted light by a coexisting colorant (present in the sample).

The number of components obtained by separating transmitted light into various components is not limited. However, it is preferable to reduce the amount of calculation for quantifying a colored substance by setting the types of the separated components to about three types. When it is necessary to treat light as four or more different components, two components are selected according to the conditions such as the object to be measured and required accuracy, and are converted into one component according to the additive color mixing law. By repeating the procedure, it is preferable to treat the transmitted light as being composed of three types of components and determine the amount of the color product.

In this specification, the above-mentioned “ratio of spectral intensity to total transmitted light component intensity (sum of spectral intensity)” is used.
And a parameter independent of the amount of light is expressed as a relative intensity ratio. At this time, as described above, the spectroscopy is obtained by spectroscopy into a plurality of lights having a specific spectral distribution set in advance for each transmitted light component, and the type (number) is not limited. It is preferable to reduce the amount of calculation for quantifying the color product by setting the types of the spectroscopy to three. If such a plurality of spectral components include a spectral component having a specific absorption wavelength (range) of the color object to be quantified, the amount of transmitted light of such a spectral component is approximately equal to the color of the color object to be quantified. This is advantageous because it fluctuates sharply and fluctuates. In the case of measurement using the visible light region, there are three types of spectroscopy, and the wavelengths of the three types of spectroscopy are RGB (CI) defined by the International Commission on Illumination (CIE).
E) A wavelength corresponding to the primary stimulus of the color system or the XYZ (CIE1931) color system, such that the relative intensity ratio matches the chromaticity coordinates (r, g, b) or (x, y, z). Can be set. Alternatively, the wavelengths of the three types of spectroscopy may be the wavelengths of the primary stimuli of the UCS color system of MacAdams defined by the CIE. When the primary stimulus determined by the CIE is selected as a spectrum, a member for a commercially available optical measuring device that is preset to be measured using the primary stimulus, for example, a light receiving unit component such as a CCD solid-state imaging device can be used. This is advantageous.

First, a method of collecting reference values used in the present method will be described.

In the following description, a plurality of light beams (spectroscopy) having the specific spectral distribution are U, V, and W, respectively.
Assuming three types of light having a wavelength or wavelength range of U, V, and W, the transmitted light amount of each of the spectra is U, V, and W, and the relative intensity ratio of each of the spectra is u, Expressed as v and w. Therefore, u = U / (U + V +
W), v = V / (U + V + W), w = W / (U + V +
W) and u + v + w = 1. Further, which transmitted light component of the plurality of transmitted light components is spectrally represented is indicated by a subscript. For example, the spectral transmittance of the transmitted light component (Z light) _U Z, _{V Z} and _{W Z,} relative intensity ratios express a _u Z, _{v Z} and _{w Z.}

In the present quantification method, as a reference value used for quantification, parallel transmitted light without scattering, scattered transmitted light by the solid support, Spectral transmitted light of three types of spectroscopy of U, V and W for each component of the scattered transmitted light by the color object (to be quantified) and, optionally, the scattered transmitted light by the coexisting color object (present in the sample) Is used to calculate the relative intensity ratio.

[0095] _{U RFW} light is three spectral scattered without parallel transmitted light (RFW _{light), V rf W} light and _{W RFW}
The relative intensity ratios of each light u _rfW , v _rfW and w
_{Since rfW} has the same relative intensity ratio as that of the irradiation light, it can be calculated by measuring the spectral transmission amount of each of the three types of spectrum of the irradiation light.

Light scattered and transmitted by the solid support (rfB light)
U _rfB light, V _rfB light and W
The relative intensity ratios u _rfB , v _rfB and w _rfB of the respective _rfB lights are determined by the transmitted light (rf
BW light) can be obtained by measuring the spectral transmittance of each of the three types of spectral components and calculating their relative intensity ratios. The rfBW light is scattered transmitted light (rf
B light) and parallel transmitted light (rfW light) having no scattering. When the light is represented by three types of spectroscopy and the relative intensity ratio of the spectra is obtained, the sum of the relative intensity ratios of the three spectra is always 1 and when the relative intensity ratio of the two spectra is determined, the remaining one of the spectra is determined. Since the relative intensity ratio is naturally determined, it is known that the original light can be represented by a two-dimensional diagram (relative intensity ratio diagram, chromaticity diagram) in which the relative intensity ratio of the two spectral components is set on the coordinate axis. A relative intensity ratio diagram (chromaticity diagram) in which the light and the rfBW light are plotted is created, and the light amounts of the rfB light and the rfW light included in the rfBW light are calculated based on the additive color mixing law of HG Grassmann. The ratio is calculated, and 3 of the rfBW light is calculated according to the calculated ratio.
R obtained by distributing the amount of spectral transmission of each type of spectrum
From three types of spectral transmission amounts of fB light, the relative intensity ratio u _rfB ,
v _rfB and w _rfB can be determined.

3 of the transmitted light (rfL light) scattered by the colored material
Is a type of spectral _{U RFL light, V r fL} light and W
The relative intensity ratios u _rfL , v _rfL, and w _{rf L} of the respective _rfL lights are 3 with respect to the transmitted light of the aqueous solution containing only the color substance of a certain concentration or the transmitted light of the solid support containing only the color substance of a certain concentration. It can be obtained by measuring the spectral transmission amount of each type of spectrum and calculating their relative intensity ratio.

First, from the transmitted light (rfLW light) of an aqueous solution containing only a color substance at a certain concentration, the relative intensity ratios u _rfL and v
A case where _rfL and w _rfL are obtained will be described.
First, using a conventional absorptiometer, the absorption spectrum of an aqueous solution containing only a colorant at a certain concentration is measured using water as a control, and then the aqueous solution at each wavelength (or wavelength range) of U, V, and W is measured. Of transmitted light U _rfLW , V
_rfLW and W _rfLW were measured and the relative intensity ratio u
_{Determine rfLW} , v _rfLW and w _rfLW . At this time, the measured rfLW light has been corrected for absorption by water, and is regarded as transmitted light including two components of scattered transmitted light (rfL light) and parallel transmitted light (rfW light) without scattering by a colored material. Therefore, a relative intensity ratio diagram (chromaticity diagram) in which the rfW light and the rfLW light are plotted is created, and the ratio of the light amounts of the rfL light and the rfW light included in the rfLW light is calculated based on the principle of additive color mixture. Rf according to the calculated ratio
The relative intensity ratios u _rfL , v _rfL and w _rfL can be determined from the three types of spectral transmission amounts of the rfL light obtained by distributing the three types of spectral transmission amounts of the LW light.

Next, the case where the relative intensity ratios u _rfL , v _rfL, and w _rfL are determined from the transmitted light (rfLBW light) of the solid support containing only the color substance at a certain concentration will be described. First, three types of spectral transmission amounts of rfBLW light are measured, and then a relative intensity ratio is calculated and obtained. The rfLBW light is scattered and transmitted light (r
fL light), transmitted light including three components of scattered transmitted light (rfB light) by the solid support and parallel transmitted light (rfW light) without scattering, and thus rfB light, rfW light, and rfBL.
A relative intensity ratio diagram (chromaticity diagram) in which the W light is plotted is created, and the ratio of the light amounts of the rfB light, the rfL light, and the rfW light included in the rfBLW light is calculated based on the principle of additive color mixture. The relative intensity ratios u _rfL , v _rfL and w are obtained from the three types of spectral transmission amounts of rfL light obtained by distributing the three types of spectral transmission amounts of rfBLW light according to the ratio.
_rfL can be determined. At this time, when the solid support to be used is optically substantially uniform and can be regarded as not absorbing light (that is, optically transparent) at a measurement wavelength (range), the rfLBW light is: Rf
Similar to the LW light, the scattered transmitted light (rfL light) due to the color object
And the transmitted light including two components of parallel transmitted light (rfW light) without scattering. In such a case, rfL
The reference value is obtained using rfLBW light instead of W light,
The labor of preparing an aqueous solution or using an absorptiometer for measuring a liquid sample can be saved.

Relative intensity ratios u _{rfCL of} U _rfCL light, V _rfCL light and W _rfCL light, which are three types of spectroscopy of scattered transmitted light (rfCL light) by coexisting color materials, which are reference values adopted as desired. , V _rfCL and w
_rfCL can be obtained by the same procedure as that for the relative intensity ratios u _rfL , v _rfL, and w _{rfL of the} rfL light spectrum.

By performing the present color substance determination method using the reference values obtained as described above, it is possible to easily and accurately quantify the color substance.

Hereinafter, the first to sixth embodiments of the method for quantifying a colored substance realized by the apparatus and / or the storage medium of the present invention will be described in detail step by step. The sixth form is a form using a rate assay method.

A) First Colored Object Quantitative Method The first form of the colored object quantification method is a method for quantifying a colored substance formed from one type of component of known properties contained in a sample. , Which is the basis of another method for quantitatively determining a colored substance described later. Note that, in order to facilitate understanding, a case will be described in which no coexisting color product exists in the sample.

First, a reference value for reference is determined in advance as described above. However, in this case, since it is assumed that there is no coexisting color product, the reference value need not be obtained for the scattered transmitted light (rfCL light) by the coexisting color product.

Next, at desired times, the amount of transmitted light of each of the three types of spectra of the transmitted light from the solid support containing the color substance to be quantified was measured, and the relative intensity of each spectrum was determined from the obtained amount of transmitted light. Find the ratio.

Then, the transmitted light (M light) of the solid support containing the color substance to be quantified is converted into the parallel transmitted light (W
Light), scattered transmitted light (B light) by the solid support, and scattered transmitted light (L light) by the color object to be quantified. The density of the color object is calculated based on the intensity ratio and the reference value. At this time, the relative intensity ratio of the spectrum is a parameter independent of the light amount as defined above, and all the light having the same optical tone have the same value. The relative intensity ratio of the spectral components of the light components L and L can be treated as being equal to the relative intensity ratio of the spectral components of the rfW light, the rfB light, and the rfL light obtained as the reference values.

The process of calculating the density of a color product will be specifically described below.

The M light is separated into three components of W light, B light and L light, and each component is divided into three types to be U-spectroscopic and V-spectroscopic.
Spectroscopy and W spectroscopy. If this is expressed by a mathematical expression with respect to the amount of light of each spectrum of M light, the following expressions (1) to (3) are obtained.

[0109]

(Equation 1) Here, U _M , V _M, and W _M , which are the amounts of light of the _M light, are spectrally separated.
Other values are values that cannot be directly measured, but the relative intensity ratio, which is a value related to those values (light quantity), is determined by reference value measurement. As described above, the relationship between each light amount and the relative intensity ratio is represented by the following equations (4) to (9). At this time, of course, the total value of the relative intensity ratios of the three types of spectral components for all the light components is 1, and this is generally represented by the following equation (10) (without subscripts).

[0110]

(Equation 2) Then, L light is a component of the M light, each light amount of the B light and W light _R L, putting the _{R B} and _{R W,} it is possible to represent each of the light amount by the following formula (11) - (13) .

[0111]

(Equation 3) Therefore, the above equations (1) and (2) are replaced by the above equation (4)
To (9), and further transformed by equations (11) to (13).
Are substituted, the following equations (14) to (16) are obtained.

[0112]

(Equation 4) Here, each light component, for _R L (amount _{= R M = R L + R} W + R B of the M light) spectroscopy of the relative intensity ratio as well as the total amount of light, _{R W} and _{R B} each relative ratio _{r L} , R _W
And r _B , r _L , r _W, and r _B are respectively expressed by the following equations (17) to (19), and at this time, the following equation (20) is always satisfied.

[0113]

(Equation 5) Then, dividing both sides of Expression (14) by Expression (16), Expression (1)
7) by deleting the variable _{r W} using equation (20) after assigning to (19), the following equation (21) is obtained.
Similarly, by erasing the variable _{r W} using equation (20) after substituting equation (17) to (19) by dividing both sides of formula (15) in equation (16), the following formula ( 22) is obtained.

[0114]

(Equation 6) Since the two unknowns included in the above two equations (23) and (24) are r _L and r _B , these two unknowns can be obtained by calculation. That is, the following equation (25)
It can be obtained r _L and r _B the matrix equation shown in running.

[0115]

(Equation 7) Further, from Expressions (16) and (17), R _L is expressed as _UM ,
The following formula expressed by V _M and _{W M} (26) is obtained.

[0116]

(Equation 8) By the above calculation, R _L (= U _L + V _L + W _L ), which is the amount of scattered transmitted light (L light) by the color object to be quantified, is
A measurement _U M, _{V M} and _{W M,} as well as pre-measured reference value _{u L, v L, u W} , v W, can be determined from _{u B} and _{v B.}

In this way, in this method, it is possible to obtain _RL , which is the amount of only scattered and transmitted light (L light) by the color object to be quantified, and to obtain other parameters affected by light components. All are erased. Therefore, in the subsequent calculations, the color object to be quantified can be regarded as existing in a medium that is optically transparent and has a constant optical path length at the measurement wavelength (range). Therefore, according to Beer's law, The relationship between the light quantity ( _RL ) and the color density (C) is expressed by the following equation (27).
And plotting the density and the light amount on a graph in which the density is on a logarithmic scale and the light amount is an equally spaced scale gives a line segment representing the relationship between the two.

[0118]

(Equation 9) At this time, the line segment representing the relationship between the color density (C) and the amount of light (R _L ) obtained by creating the above-described graph based on the actual measured values slightly deviates from the equation (27). An approximate expression of a line segment on a graph that is actually obtained according to measurement conditions or desired measurement accuracy can be obtained, and the obtained approximate expression can be used in place of Expression (27). It should be noted that this process can also be executed entirely by calculation by obtaining an approximate expression of a line segment that can virtually draw a graph. From the viewpoint of labor saving and accuracy, it is preferable to execute all processes by calculation. It is advantageous. As the approximation formula obtained in such a manner, the following formula (28), formula (29) and general formula (30) can be exemplified.
It seems practically preferable to use the two-dimensional approximation formula shown by

[0119]

(Equation 10) The constants expressed in Greek letters in the above equations (27) to (30) can be determined from the measured values obtained by performing measurements on samples containing various colored substances having known densities according to a standard method.

When a coexisting color product is present in the sample, the scattered transmitted light (CL light) from the coexisting color product is further separated and the M light is treated as transmitted light composed of four types of components. As a reference value, the scattered transmitted light (rfC
This method is carried out by determining the relative intensity ratio of the spectrum of (L light). At this time, for example, according to the law of additive color mixing, the rfCL light is converted together with the other component light having the closest relative intensity ratio to the relative intensity ratio of the rfCL light and treated as a single component, and It is also possible to simplify the calculation for obtaining the density of the color product.

As described above, in the method for quantifying a colored substance according to the first embodiment, the transmitted light is divided into a plurality of components according to its properties, and the relative intensity ratio of the spectrum obtained by spectrally dividing each component is obtained. Based on the light quantity (R _L ) of only the scattered and transmitted light by the color object, the quantity of the color substance is determined based on the light quantity. It can be minimized.

Therefore, according to this method, the color substance impregnated in the solid support can be quantitatively determined simply and with high precision.

B) Second Colored Object Quantitative Method The second form of the colored object quantitative method is a method for quantifying a colored substance formed from one type of component of known properties contained in a sample. This is one of the modifications of the colorant quantification method of the first embodiment.

In the present method, when selecting the plurality of spectroscopy in the method for quantifying a colored substance of the first embodiment, a wavelength (specific absorption wavelength) at which the colored substance to be quantified specifically absorbs is used. Light is always selected as one of a plurality of spectroscopy, and the color product is quantified based on the spectral transmission amount and the relative intensity ratio of the spectroscopy having the specific absorption wavelength.

In the following, for example, as in the case of the color substance quantification method of the first embodiment, three types of U spectrum, V spectrum, and W spectrum are selected as a plurality of spectrums. Assuming that the wavelength has a specific absorption wavelength W, the present method will be described mainly for the portions different from the colorant determination method of the first embodiment.

In this method, first, the specific absorption wavelength of the color substance to be quantified is determined using a conventional method, and the W spectrum having the wavelength W which is the specific absorption wavelength is determined as one of a plurality of spectra. Adopt and select other spectral components.

Next, after obtaining various reference values in advance in the same manner as in the colorant determination method of the first embodiment, the amount of transmitted light from the solid support containing the sample at a desired time point Is measured.

Then, the transmitted light (M light) of the solid support containing the color substance to be quantified is converted into the parallel transmitted light (W
Light), scattered transmitted light (B light) by the solid support, and scattered transmitted light (L light) by the color object to be quantified. The density of the color object is calculated based on the intensity ratio and the reference value. This calculation process is also generally performed in the same manner as the above-described method for quantifying a colored substance of the first embodiment. However, in this method, after calculating the light amount _RL of only the scattered and transmitted light by the colored substance, the reference value is determined in advance. The relative intensity ratios u _L and v of each of the three types of spectrum of the scattered and transmitted light by the color object, which were obtained as
With _L and w _L, and calculates the amount W _L and W light having a specific absorption wavelength. Then, either of the formula, supra, was used in the coloration colored article Determination of the first form (27) - (30), the concentration of coloration colored article by substituting W _L instead of R _L Calculate and quantify.

As described above, in the color object quantification method of the second embodiment, the light amount R, which is the sum of the spectral light amounts of the scattered and transmitted light by the color object employed in the color object quantification method of the first embodiment, is used. instead _{L, and} is quantified adopted to coloration colored article the amount W _L only spectral (W light) of a specific absorption wavelength of coloration colored article contained in the scattering transmitting light by coloration colored article, the W _L is Since the change in the density of the coloring matter changes with higher sensitivity than that of the _RL , the use of this method enables more accurate quantification.

C) Third Colored Object Quantitative Method The third form of the colored object quantitative method is a method for quantifying a colored substance formed from one type of component of known properties contained in a sample. This is one of the modifications of the colorant quantification method of the first embodiment. This method can also be used in combination with the above-described second embodiment of the method for determining a colored substance.

In this method, while the color substance concentration is calculated only by calculation in the color substance quantification methods of the first and second embodiments, the relative density using a plurality of spectral components described above is used. The density of the color object is calculated using the intensity ratio diagram (chromaticity diagram) together.

Hereinafter, this method is carried out in accordance with the method for quantifying a colored substance according to the first embodiment, and a plurality of spectroscopy, U-spectroscopy, V-spectroscopy, and W-spectral, are performed similarly to the method for quantifying a colored substance of the first embodiment. Assuming that three types of spectroscopy are selected, the present method will be described with respect to portions different from the colorant quantification method of the first embodiment. When the present method is carried out in combination with the colorant quantitative method of the second embodiment, it can be carried out by changing the following description example according to the description of the above B) .

The only difference between this method and the first embodiment is the use of a relative intensity ratio diagram in the process of calculating the concentration of the colored substance, and the other procedures are the same. .

Therefore, after various reference values have been measured in advance in the same manner as in the first embodiment, the amount of transmitted light from the solid support containing the sample at a desired point in time. Is measured.

Then, the transmitted light (M light) of the solid support containing the color substance to be quantified is converted into the parallel transmitted light (W
Light), scattered transmitted light (B light) by the solid support, and scattered transmitted light (L light) by the color object to be quantified. The density of the color object is calculated based on the intensity ratio and the reference value.

In this method, when a certain light is represented by three types of spectral components and the relative intensity ratio of the spectral components is obtained, the sum of the relative intensity ratios of the three spectral components is always 1 and the relative intensity ratio of the two spectral components is determined. Since the relative intensity ratio of the remaining one spectrum is naturally determined, the knowledge that the original light can be represented by a two-dimensional diagram (relative intensity ratio diagram, chromaticity diagram) using the relative intensity ratio of the two spectra as a coordinate axis. Based on this, a relative intensity ratio diagram is created to determine the density of the color product.

FIG. 10 is a graph based on the above findings, where u is the vertical axis,
Using a relative intensity ratio diagram in which v is the abscissa and the coordinates of each light are represented by points with the abbreviations of each light, the transmitted light (M light) of the solid support containing the color substance to be quantified, Scattered transmitted light (L light) due to the color substance to be quantified, scattered transmitted light (B
2 shows the correlation between the light (light) and the parallel transmitted light (W light) without scattering. This FIG. 10 can be created by using the relative intensity ratio obtained from the measured value for the M light, and using the reference values for the L light, B light and W light. FIG.
0, point P is the intersection of the line LB with the line passing through points W and M, point Q is the intersection of the line WL with the line passing through points B and M, and point S Are the points L and M
Respectively, and the intersections of the straight line passing through and the line segment BW.

At this time, the relative ratio R _W : R of the light amount (R _W ) of the W light and the light amount (R _L ) of the L light contained in the M light.
_L is the ratio of R _{L to} the sum of both light quantities (R _W + R _L ), q
w and can be expressed by the following equation (31).

[0139]

[Equation 11] Similarly, the light amount (R _W ) of W light included in M light and B light
The relative ratio _R W between the quantity of light _{(R B): R B} can be expressed by the following equation (32) at the q'w the ratio of _{R B} to the sum of both the amount of light _(R W + _{R B)} .

[0140]

(Equation 12) Furthermore, the relative ratio _R L of the L light amount of light contained in M light _{(R L)} and B light quantity _{(R B): R B} is the sum of both the amount of light (R
It can be expressed by the following equation (33) _L + for _{R B)} at the q''w the ratio of _{R B.}

[0141]

(Equation 13) Further, the amount of M light if _{R M,} the following equation (34) holds.

[0142]

[Equation 14] From the _above, R L, and the specific and the total amount of _{R W} and _{R B} is _determined, it is possible to calculate R L, the respective values _{R W} and _{R B.}

Then, the obtained _{RL is represented} by the above formula (27).
To (30), the density of the color product can be determined and quantified.

As described above, the color substance density calculating process of the present method using the relative intensity ratio diagram can be executed by actually creating the relative intensity ratio diagram and obtaining the length of the line segment. It is also possible to calculate formulas representing various straight lines and line segments obtained by virtually drawing a relative intensity ratio diagram and execute them all by calculation.

As described above, in the colorant determination method of the third embodiment, the transmitted light is divided into a plurality of components according to its properties, and the relative intensity ratio of the spectrum obtained by spectrally separating each component is obtained. The amount of transmitted light (R _L ) of only the scattered and transmitted light by the color object is calculated based on the light amount, and the amount of the color object is determined from the transmitted light amount. Can be minimized, and the color product impregnated in the solid support by the present method can be quantified simply and with high accuracy.

D) Fourth Colored Object Quantitative Method In the fourth form of the colored material quantification method, when a mixture of two or more types of colored substances of known properties is present in a sample, that is, it should be quantified. A method for quantifying a color product formed from one kind of component of known properties and a reagent contained in a sample when a coexisting color product other than the color product is present in the sample.
This is one of the modifications of the method for quantifying a colored substance of the form (1). In addition, this method can be used in combination with the color substance determination method of the second or third embodiment.

This method is a practical and simple method for quantitatively determining a colored substance, which is a modification of the above-described first to third embodiments of the method for quantitatively determining a colored substance, particularly suitable for quantifying a specific component in a biological sample. It is.

In general, a biological sample, such as blood, serum, or urine, has a color tone similar to that of a color product to be detected and quantified, and a coexisting color product that affects a measured value as a so-called background. Often. Therefore, when such a coexisting color product is present in the sample, the transmitted light (M light) of the solid support containing the color product to be quantified is converted into the parallel transmitted light (W light) without scattering. Scattered transmitted light (B light) by the solid support, scattered transmitted light (L
Light) and scattered and transmitted light (CL light) due to coexisting colorants present in the sample.

It is possible to determine and determine the concentration of the color substance to be quantified by separating the M light into the four types of components and handling the same as it is in the color substance quantification methods of the first to third embodiments. However, as the number of chemical species to be separated and measured increases (that is, the number of types of transmitted light components to be separated increases), the amount of calculation increases, and the slight variation in the calculated parameter value affects the quantitative result. Tends to be large.

However, if an optically transparent solid support that does not absorb light in the measurement wavelength range such as the visible region is used as the solid support, it is included in the four types of transmitted light components in the measurement wavelength range. Parallel transmitted light without scattering (W light)
And the spectral distribution of the scattered transmitted light (B light) by the solid support can be regarded as the same, so that the calculation can be omitted by including the W light in the B light, for example. Thereby, the component of the M light only needs to be classified into three types, and the amount of the colored material can be practically and simply quantified without increasing the amount of calculation and the influence of parameter fluctuation.
Such a quantification method is a practical quantification method that can be widely applied, including quantification of components contained in a biological sample.

Accordingly, the colorant determination method of the fourth embodiment is the same as the colorant determination method of the first to third embodiments, except that the solid-state support which is optically transparent in the measurement wavelength range is used. And the parallel transmitted light (W light) without light and the scattered transmitted light (B light) by the solid support can be treated as the same light,
In addition, the scattered transmitted light (CL
This is a method that always uses the relative intensity ratio of a plurality of spectral components of light.

That is, according to this method, the transmitted light (M light) of the solid support containing the color substance to be quantified is calculated according to the color substance quantification method of the first to third embodiments. Using either one of the parallel transmitted light (W light) without scattering or the scattered transmitted light (B light) by the solid support and the scattered transmitted light (CL light) by the coexisting colorant present in the sample, The color is corrected based on the baseline and the background and based on the light amount R _L of the scattered transmitted light (L light) by the color object to be quantified or the light amount of the spectrum having the specific absorption wavelength of the color object in the L light. The concentration of the substance can be determined and quantified.

Hereinafter, the present method is carried out in accordance with the method for quantifying a colored substance according to the first embodiment, and a plurality of spectroscopy, U-spectroscopy, V-spectroscopy and W-spectroscopy, are carried out in the same manner as in the first embodiment. Assuming that the three types described above are selected, the present method will be described with respect to portions different from the colorant quantification method of the first embodiment. When the present method is carried out in combination with the colorant quantification method of the second or third embodiment, the following explanation example may be changed according to the description of the above B) or C) .

This method differs from the first embodiment of the method for quantitatively determining a colored substance in that a solid support having no light absorption in a measurement wavelength range is used, and a parallel transmitted light (rfW) having no scattering in a reference value. In other words, the relative intensity ratio of the spectrum of the scattered and transmitted light (rfCL light) due to the coexisting color product is used without omitting the relative intensity ratio of the spectrum of the light), and the other procedures are the same.

Therefore, after various reference values are measured in advance in the same manner as in the first embodiment, the amount of transmitted light from the solid support containing the sample is determined at a desired time. Is measured. However, as described above, the parallel transmitted light (r
The measurement of the reference value can be omitted for the relative intensity ratio of the spectrum of the fW light), and the scattered and transmitted light (rfC
The relative intensity ratio of the spectrum of (L light) is always acquired.

Next, the transmitted light (M light) of the solid support containing the color substance to be quantified is scattered transmitted light (B light) by the solid support and the scattered transmitted light (B light) by the color substance to be quantified. L light)
And each component of the scattered and transmitted light (CL light) due to the coexisting colorant present in the sample is handled separately, and is displayed based on the measured spectrally transmitted light amount and relative intensity ratio of the M light and the above reference value. Calculate the density of the color object. This calculation process is performed in the same manner as in the above-described first to third embodiments of the method for quantitatively determining a colored substance, in which the parallel transmitted light (W light) having no scattering in the description of the above A) , B) and C) coexists. It can be carried out by replacing it with the scattered and transmitted light (CL light) by the object.

Here, it will be specifically described that the present method can be carried out by using a relative intensity ratio diagram in accordance with the above-described color form quantification method of the third embodiment.

When the sample contains two types of color products, a color product to be quantified and a coexisting color product, the transmitted light (M light) of the solid support containing the color product to be quantified is , Parallel transmitted light without scattering (W light), scattered transmitted light by solid support (B light),
It can be regarded as a system containing each component of the scattered transmitted light (L light) by the color material to be quantified and the scattered transmitted light (CL light) by the coexisting color material. This is as shown in FIG. 11A. In FIG. 11a, the symbols B to L attached to the respective points represent corresponding abbreviations of component light, for example, point B is scattered transmitted light (B light) by the solid support.
Represents the coordinates of. At this time, if the solid support is optically transparent in the measurement wavelength range, the point W and the point B overlap one point and become a point B (W) as shown in FIG. 11C. However, this is a theoretically obtainable state, and in actual measurements, in most cases, points W and B are plotted as separate points in close proximity as seen in FIG. 11b. However, the point W and the point B are located very close to each other, and it is possible to treat the point W and the point B in an approximate manner. The error caused by this approximation is within a range that can be tolerated in general use, and it is determined that the benefit brought by the approximation by suppressing the increase in the amount of calculation and the effect of parameter fluctuation is greater.

By approximation as described above, FIG.
The relative intensity ratio diagram for the M light shown in FIG. 10 is the same as that of FIG. 10 used for describing the colorant quantification method of the third embodiment except that the W light and the CL light are exchanged. According to the same procedure as that of the color substance quantification method of the third mode, the concentration of the color substance to be quantified can be determined and quantified.

As described above, according to this method, by using an optically transparent solid support at the measurement wavelength, parallel transmitted light (W light) without scattering and scattered transmitted light (B light) by the solid support are obtained. Light) can be treated as light of the same component, and the relative intensity ratio of the spectrum of the scattered transmitted light (CL light) due to the coexisting colorant present in the sample is always obtained as a reference value, thereby obtaining the baseline. Since the correction can be simplified and the background correction can be performed, the density of the color substance to be quantified can be easily and simply determined with practical accuracy.

E) Fifth Colored Object Quantitative Method In the fifth form of the colored substance quantitative method, when a mixture composed of two or more types of colored substances of known properties is present in a sample, that is, it should be quantified. This is a method for quantifying a colored substance formed from one kind of component of known properties and a reagent contained in a sample when a coexisting colored substance other than the colored substance is present in the sample. This is one of the modifications of the method for quantifying a colored substance of the form (1).

This method is a practical method for quantitatively determining a colored substance obtained by modifying the above-described method for determining a colored substance of the third embodiment so as to be particularly suitable for quantitatively determining a specific component in a biological sample.

As described above, when the coexisting color product is present in the sample, the transmitted light (M light) of the solid support containing the color product to be quantified is converted into the parallel transmitted light (M) without scattering. W light), scattered transmitted light (B light) by a solid support, scattered transmitted light (L light) by a colorant to be quantified, and scattered transmitted light (CL light) by a coexisting colorant present in the sample. It is necessary to handle each component separately.
Although the M light can be quantified by determining the concentration of the coloring matter to be quantified by separating and handling the four components as it is, as the number of chemical species to be categorized and measured increases, the amount of calculation increases. In addition, there is a tendency that a slight change in the calculated parameter value has a large effect on the quantitative result.

Therefore, in the present method, in the measurement of a biological sample in which correction of the coexisting color product in the background is important, a reagent that forms a color product by color reaction with a component to be quantified in the sample is used. By using a test device including a solid support to be retained and a reference solid support that does not retain the reagent for measuring the coexisting colorant present in the sample, the calculation amount is increased and the background is reduced. Suppress the effects.

That is, in this method, as a test device, for example, FIG.
By using a test paper 54 provided with a solid support 51 and a reference solid support 52 as shown in FIG. 5 and adopting a color substance concentration calculation procedure of the color substance quantitative method of the third embodiment, Facilitates correction for background effects.

In the following, this method is carried out in accordance with the third embodiment of the method for quantifying a colored substance, and similarly to the third embodiment, the U-spectroscopy, V-spectroscopy and W spectroscopy 3
Assuming that the type has been selected, the present method will be described with respect to portions different from the color form quantification method of the third embodiment.

The present method is different from the colorimetric substance determination method of the third embodiment in that, in addition to a solid support containing a sample, a reference solid support containing a sample is measured by measuring the amount of spectrally transmitted light to obtain a relative value. The point at which the intensity ratio is determined and the concentration of the color product is calculated using the relative intensity ratio, and the reference value used in the calculation process are the transmitted light (rfBW) of the sample-free solid support.
This is a point including the relative intensity ratio of the spectral intensity of light (light) and the spectral intensity of the transmitted light (rfLBW light) of the solid support containing only a colorant at a certain concentration.

Therefore, in the same manner as in the third embodiment, the above-mentioned reference value is measured in advance in the same manner as in the third embodiment. The amount of transmitted light from the phase support is measured.

Next, the transmitted light (M light) of the solid support containing the color product to be quantified and the coexisting color product is converted into parallel transmitted light (W light) without scattering and scattered transmitted light by the solid support. (B light) transmitted through the solid support (BW light), scattered transmitted light (L light) by the color object to be quantified, and transmitted light (B light) through the solid support.
W light) and the transmitted light (LBW light) of the color substance to be quantified and the solid support, the scattered transmitted light (CL light) of the coexisting color substance present in the sample, and the transmitted light of the solid support. (BW
Light) and the transmitted light (CLB) of the solid support
W light) is divided into three components, and the transmitted light (rfM light) of the reference solid support containing the color material to be quantified but not containing the color material to be quantified (rfM light) is converted into parallel transmitted light (W light) without scattering. Light) and the transmitted light (BW light) of the solid support composed of the scattered transmitted light (B light) of the solid support, and the scattered transmitted light (CL light) of the coexisting colorant present in the sample and the solid phase The light quantity and the relative intensity ratio of the M light and the rfM light, which are separately handled and treated in two components, a coexisting color product composed of the light transmitted through the support (BW light) and the light transmitted through the solid support (CLBW light) , And the density of the color object is calculated based on the reference value. This calculation process is executed using the relative intensity ratio diagram (chromaticity diagram) as described in the above C) , in the same manner as in the color substance quantification method of the third embodiment described above.

Hereinafter, the color substance density calculation process of the present method will be described in detail with reference to the drawings.

FIG. 12 is a relative intensity ratio diagram in which reference values used in the present method and values obtained by measurement are plotted. In the drawing, the symbols BW to rfM attached to the respective points represent the corresponding abbreviated component lights, for example, the point BW represents the coordinates of the transmitted light (BW light) of the solid support. Also, the point P
Is the intersection of a straight line passing through the points BW and LBW and a straight line passing through the points M and rfM. The light represented by this point P (coordinate P) is referred to as P light.

[0172] At this time, the relative ratio of the quantity of M optical _{(R M)} Included RFM light quantity in _{(R RFM)} and P light quantity _{(R P) R rfM: R} P is the line segment M- M is the length of rfM
M and the length of the line segment MP as MP, the following equation (3)
5).

[0173]

(Equation 15) Accordingly, the light amount _{R P} and its spectral relative intensity ratio _u P of P light from 12 and above equation (35), _{v P} and _{w P}
Can be requested.

Similarly, the light amount of the BW light (R _BW ) and the light amount of the LBW light (R _BW ) included in the light amount of the P light (R _P )
_LBW ) and the relative ratio R _BW : R _LBW is a line segment P-BW
Is the length of PBW, and the length of the line segment P-LBW is PLB
W can be expressed by the following equation (36).

[0175]

(Equation 16) Therefore, it is possible to obtain a light quantity _{R LBW} of LBW light in the light amount _{R P} of the P beam from FIG. 12 and the above equation (36), and its spectral relative intensity ratio _{u LBW, v LBW} and _{w LBW.}

However, since the rfM light does not include the LBW light, the light amount of the LBW light in the P light is nothing but the light amount of the LBW light in the M light. Therefore, among the color substances to be quantified and the transmitted light (LBW light) of the solid support determined above, the scattered transmitted light (B light) and the parallel transmitted light without scattering (W light) by the solid support are: Assuming that the light is substantially constant, the light amount R _{L of the} LBW light
_By substituting _BW into any one of the above equations (27) to (30), it is possible to determine and quantify the density of the color product.

However, since the LBW light contains the B light and the W light in addition to the L light, in order to obtain the density of the color object with higher accuracy, the relative amount of the light amount of the LBW light and its spectrum is required. from the intensity ratio, and the amount R _L of the L light in M light, it calculates its spectral relative intensity ratio u _L, v _L and w _L, it is preferable to determine the concentration of coloration colored article from these values .

That is, the LBW light includes three components: scattered transmitted light (L light) by a colored substance, scattered transmitted light (B light) by a solid support, and parallel transmitted light (W light) without scattering. Since the light is light, a relative intensity ratio diagram (chromaticity diagram) in which the B light, W light and BLW light are plotted is created, and the B light, L light and W light included in the BLW light are formed based on the principle of additive color mixture. The relative light intensity ratios u _L , v _L and w _L are calculated from the three types of spectral transmission amounts of L light obtained by calculating the ratio of the light amounts and distributing the three types of spectral transmission amounts of the BLW light according to the calculated ratios. Can be requested.

Then, the obtained R _L , or, for example, when the W spectrum has the specific absorption wavelength W of the color substance, the light quantity W _L of the W spectrum in the L light is calculated by the above-mentioned formulas (27) to (27). 30) can be substituted for any one of the above to determine and quantify the density of the colored substance.

In this way, the color substance density calculating process of the present method using the relative intensity ratio diagram can be executed by actually creating the relative intensity ratio diagram and obtaining the length of the line segment. It is also possible to calculate formulas representing various straight lines and line segments obtained by virtually drawing a relative intensity ratio diagram and execute them all by calculation.

As described above, according to this method, the background correction can be performed with a small amount of calculation by using a test device provided with a reference solid support in addition to the solid support. The concentration of the color substance to be quantified can be determined easily with high accuracy.

F) Sixth Colored Object Quantitative Method The sixth form of the colored object quantitative method is used when a mixture of a plurality of colored objects is present in a sample, that is, other than the colored object to be quantified. A method for quantifying a color product formed from one kind of component of known properties and a reagent in a sample when a coexisting color product is present in the sample. This is one of the modifications.

The present method is a method for quantifying a colored substance obtained by modifying the method for quantifying a colored substance of the third embodiment described above so as to be suitable for a test device for a rate assay.

In this method, a time-dependent change in the formation of a colored substance caused by a color reaction by bringing a sample into contact with a solid support using a test device for a rate assay, that is, a time-dependent change in the degree of coloration (coloring intensity). The change is measured at a fixed time interval using the amount of transmitted light as an index, and the concentration of the colored substance is determined and quantified based on the difference between the measured transmitted light amount and the relative intensity ratio of the spectrum. Since the measurement is performed at a plurality of time points, it is possible to suppress the fluctuation of the surface state of the solid support and the influence of the coexisting color product.

In the following, this method is carried out in accordance with the third embodiment of the method for quantifying a colored substance, and similarly to the third embodiment, a plurality of spectroscopy, U spectrum, V spectrum and W spectrum. Assuming that the following three types are selected, the present method will be described with respect to portions different from the third embodiment of the method for quantitatively determining a colored substance.

The difference between this method and the third embodiment is that the amount of transmitted light from the solid support containing the sample is measured a plurality of times at fixed time intervals and the relative intensity of the spectrum is measured. And the reference value used in the calculation process is the ratio of the transmitted light (rfBW light) of the solid support that does not contain the sample. This is a point that includes the relative intensity ratio of the spectral intensity and the spectral intensity ratio of the transmitted light (rfLBW light) of the solid-phase support containing only a color substance at a certain concentration.

Therefore, first, the above-mentioned reference value is measured in advance in the same manner as in the third embodiment.

Next, at a desired time, the amount of transmitted light from the solid support containing the sample is measured. In this method, since a test device for a rate assay is used, the amount of transmitted light is measured by solidifying the sample. It is started as soon as possible after contact with the phase support and is repeated an appropriate number of times at appropriate short time intervals. In the following description, measurement of the measured transmitted light and the quantity, referred to as M light and R _M collectively, when handled individually, are numbered according to the measured order, for example, the n-th has been transmitted light Mn light refers to the light amount thereof and _{R Mn.} .

Then, the transmitted light (M light) of the solid support containing the color substance to be quantified is converted into parallel transmitted light (W light) without scattering.
Light transmitted through the solid support (BW light) composed of light transmitted through the solid support and light transmitted through the solid support (BW light); BW light) and light transmitted through the solid support (LBW)
Light), and the coexisting color product and the transmitted light (CLBW light) of the coexisting color product and the transmitted light (BW light) of the solid support, which are scattered and transmitted by the coexisting color product (CL light) present in the sample. 3)
The density of the coloring matter is calculated based on the transmitted light amounts and the relative intensity ratios of the measured plurality of M lights and the above-mentioned reference values. This calculation process is executed using the relative intensity ratio diagram (chromaticity diagram) as described in the above C) , in the same manner as in the color substance quantification method of the third embodiment described above.

Hereinafter, the color substance density calculation process of the present method will be described in detail with reference to the drawings.

FIG. 13A is a relative intensity ratio diagram plotting reference values and measured values used in the present method.
FIG. 13B is an enlarged view showing a region surrounded by a dotted line Z in FIG. 13A. In the drawing, the symbols of BW to Mm attached to the respective points represent the corresponding abbreviations of light, for example, the point BW represents the coordinates of the transmitted light (BW light) of the solid support. The point P is an intersection of a straight line passing through the points BW and LBW and a straight line passing through the points Mm and Mn. This point P (coordinate P)
Is referred to as P light. Similarly, point M0 is
The intersection of a straight line passing through the point BW and the point M1 and a straight line passing through the point Mm and the point Mn. The light represented by the point M0 (coordinate M0) is referred to as M0 light. However, both the Mm light and the Mn light were measured after the color reaction caused by the contact between the sample and the solid support was terminated and the amount of M light became constant,
That is, the M light is such that R _Mm = R _Mn .

At this time, the M0 light is the M light at the moment when the sample comes into contact with the solid support, that is, before the start of the formation of a colored substance by the color reaction, so that the BW light and the CLBW light are emitted.
It can be considered that the light includes only two components of light and does not include LBW light. And the relative intensity ratio u of the spectrum
_M0 , _vM0 and _wM0 can be determined from the relative intensity ratio diagram of FIG. 13a or by calculation. Therefore, when approximated by quantity _{R M1} of M1 light measured the quantity _{R M0} of M0 light in its immediate vicinity, the light quantity _U M0 of the spectral light amount _{R M0} of M0 _{light, V M0} and _{W M0} is obtained.

From the above, it can be seen that the M0 light can be handled in the same manner as the transmitted light (rfM light) of the reference solid support used in the above-described fifth embodiment of the method for determining a colored substance. Therefore,
The rfM light in the description of E) is read as M0 light, and the concentration of the color substance to be quantified can be determined and quantified.

That is, the relative light amount of the _M light (R _M0 ) and the light amount of the _P light (R _P ) included in the light amount (R _Mm ) of the M light after the light amount becomes constant, for example, the M _m light. Ratio R _M0 : R
_P represents the length of the line segment Mm-M0 as MmMO, and the line segment M
The length of m-P can be represented by the following equation (37), where MmP is set.

[0195]

[Equation 17] Accordingly, the light amount _{R P} and its spectral relative intensity ratio _u P of P light from 13 and above equation (37), _{v P} and _{w P}
Can be requested.

Similarly, the light quantity (R _BW ) of the BW light and the light quantity (R _BW ) of the LBW light included in the light quantity (R _P ) of the P light
_LBW ) and the relative ratio R _BW : R _LBW is a line segment P-BW
Is the length of PBW, and the length of the line segment P-LBW is PLB
W can be expressed by the following equation (38).

[0197]

(Equation 18) Therefore, it is possible to obtain a light quantity _{R LBW} of LBW light in the light amount _{R P} of the P beam from FIG. 13 and the above equation (38), and its spectral relative intensity ratio _{u LBW, v LBW} and _{w LBW.}

However, since the M0 light does not include the LBW light, the light amount of the LBW light in the P light is nothing but the light amount of the LBW light in the M light. Therefore, among the color substances to be quantified and the transmitted light (LBW light) of the solid support determined above, the scattered transmitted light (B light) and the parallel transmitted light without scattering (W light) by the solid support are: Assuming that the light amount is substantially constant, the light amount R _{LB of the} LBW light
_By substituting _W into any one of the above formulas (27) to (30), it is possible to determine and quantify the density of the color product.

However, since the LBW light includes the B light and the W light in addition to the L light, in order to obtain the density of the color object with higher accuracy, the relative amount of the light amount of the LBW light and its spectrum is required. from the intensity ratio, and the amount R _L of the L light in M light, it calculates its spectral relative intensity ratio u _L, v _L and w _L, it is preferable to determine the concentration of coloration colored article from these values .

That is, the LBW light includes three components: scattered transmitted light (L light) by a colored substance, scattered transmitted light (B light) by a solid support, and parallel transmitted light (W light) without scattering. Since the light is light, a relative intensity ratio diagram (chromaticity diagram) in which the B light, W light and BLW light are plotted is created, and the B light, L light and W light included in the BLW light are formed based on the principle of additive color mixture. The light intensity ratio is calculated, and the light intensity of the three types of LW light obtained by distributing the light intensity of the three types of light of the BLW light according to the calculated ratio is calculated from the light intensity of the three types of light _L and the relative intensity of the light. Ratio u _L ,
v _L and w _L can be determined.

The obtained R _L or, for example, when the W spectrum has the specific absorption wavelength W of the color product, the light quantity W _L of the W spectrum in the L light is calculated by the above formulas (27) to (30). ) Can be substituted for any one of them to determine and quantify the density of the color product.

In this way, the color substance density calculation process of the present method using the relative intensity ratio diagram can be executed by actually creating the relative intensity ratio diagram and obtaining the length of the line segment. It is also possible to calculate formulas representing various straight lines and line segments obtained by virtually drawing a relative intensity ratio diagram and execute them all by calculation.

As described above, according to this method, background correction can be performed with a small amount of calculation by using a test device for a rate assay. The concentration can be determined.

As described in detail above, the color object quantification methods of the first to sixth embodiments realized by the color object quantification apparatus and the storage medium for color object quantification of the present invention are as follows. The transmitted light is treated as a plurality of components depending on its properties, and the scattered transmitted light (L
By using the light amount R _{L of the} light) and / or the light amount of the spectrum having the specific absorption wavelength of the color substance contained in the L light, to obtain the concentration of the color substance more simply and more accurately than in the past. can do.

The light quantity of the spectroscopic light having the specific absorption wavelength of the color substance contained in the scattered transmitted light (L light) from the color substance to be quantified, which is the quantitative index, depends on the state change of the solid support. It is more clearly shown by the following Test Examples that this is an excellent quantitative index that is not performed.

<Test Example> First, Food Blue No. 1 which is a coloring agent was selected as a coloring matter to be quantified, and a sample containing Food Blue No. 1 at various concentrations within a range of 0 to 500 mg / dl. An aqueous solution was prepared. Next, a test paper that does not hold a reagent is employed as a solid support, and three types of spectroscopy, namely, U spectroscopy at a wavelength of 435 nm, V spectroscopy at a wavelength of 546 nm, and W spectroscopy at a wavelength of 700 nm are selected as spectroscopy used for the measurement. A reference value was obtained using the aqueous solution. Then, the test paper is impregnated with the sample aqueous solution, and immediately thereafter,
The amount of transmitted light was measured after 5 minutes and after 30 minutes.

A spectrometer having a specific absorption wavelength of a color substance to be quantified and included in the total amount of transmitted light (R _M ), which has heretofore been widely used as a quantitative index of a color substance (herein, referred to as W spectrum). the quantity W _M on the vertical axis, the logarithm of coloration colored article concentration Log
FIG. 14 shows a graph in which C is the horizontal axis, and W _L which is a quantitative index used in the present invention, that is, the light amount W _{L of} W spectrum included in the light amount of scattered and transmitted light (L light) by the colorant to be quantified. Are plotted on the horizontal axis, and the logarithmic value LogC of the color substance density is plotted on the horizontal axis in FIG. In FIGS. 14 and 15, △, □, and Δ indicate the results immediately after the test paper was impregnated with the sample aqueous solution (○), 15 minutes after the impregnation (□), and 30 minutes after the impregnation, respectively (△). The measured transmitted light amount is shown.

[0208] As apparent from FIG. 14, as the amount W _M and W spectral contained in the total transmitted light that has been used conventionally quantitative index, a large amount of change with time. It is considered that the fluctuation of the measured value is caused by the evaporation of water in the sample over time and the surface state of the solid support changed. Therefore, when the conventional W _M perform quantification of coloration colored article as a quantitative indicator, it is necessary to manage the exact measurement conditions with respect to time elapsed from the sample impregnation.

On the other hand, the amount of light W _{L of the} W spectrum contained in the scattered transmitted light (L light) by the color object to be quantified, which is a quantitative index used in the present invention, depends on the elapsed time from the sample impregnation. However, a constant value is shown depending on the color density.

From this test example, the quantitative index W used in the present invention was
_It will be understood that _L is an excellent quantitative index that is not affected by changes in the state of the solid support, unlike the conventional quantitative index.

[0211]

As described above, the color substance quantifying apparatus and the color medium quantification storage medium of the present invention provide a novel color substance quantification method. The colored material in the phase support is irradiated with light, and the transmitted light is quantified based on a parameter corresponding to the transmittance obtained by measuring the amount of transmitted light. Depending on its properties, it is divided into a plurality of components, and each transmitted light component is separated into a plurality of lights having a predetermined specific spectral distribution,
Since the ratio of each spectral intensity to the total transmitted light component intensity (sum of the spectral intensities) is used as a parameter for each transmitted light component,
It is possible to treat only the scattered and diffused light components from the colored object to be quantified separately from other light components in the measured transmitted light, which makes the baseline and background correction much easier than before. Even when a sample containing a large amount of co-existing color products such as a biological sample and having conventionally had difficulty in correcting the background is used, the effect of being able to easily and accurately quantify the color products is exerted.

[Brief description of the drawings]

FIG. 1 is a block diagram showing an example of an embodiment of a color product quantification apparatus of the present invention.

FIG. 2 is a schematic diagram illustrating an example of a configuration of an imaging unit of the color object quantification device of the present invention.

FIG. 3 is a schematic diagram showing an example of a single item test device used in the color product quantification device of the present invention.

FIG. 4 is a schematic view showing an example of a multi-item test device used in the color product quantification device of the present invention.

FIG. 5 is a schematic diagram showing one example of a single item rate assay test device used in the color product quantification device of the present invention.

FIG. 6 is a schematic diagram showing one example of a multi-item rate assay test device used in the color product quantification device of the present invention.

FIG. 7 is a flowchart showing an example of an operation flow of the coloring matter determination device of the present invention.

FIG. 8 is a flow chart partially showing an example of an operation flow when a rate assay method is used in the color product quantification apparatus of the present invention.

FIG. 9 is a flowchart partially showing an example of an operation flow when the rate assay method is not used in the color product quantification apparatus of the present invention.

FIG. 10 is a relative intensity diagram used in one embodiment of the method for quantifying a colored substance realized by the present invention.

FIG. 11 is a diagram of a color substance determination method realized by the present invention.
It is a relative intensity figure used in the form different from 0.

FIG. 12 is a diagram of a color substance determination method realized by the present invention.
It is a relative intensity figure used in the form different from 0 and 11.

FIG. 13 is a diagram of a color substance determination method realized by the present invention.
It is a relative intensity figure used in the form different from 0-12.

FIG. 14 is a graph showing the relationship between the total amount of transmitted light, which is a quantitative index of the conventional color substance determination method, and the density of the color substance.

FIG. 15 is a graph showing the relationship between the amount of scattered transmitted light from a color product and the density of the color product, which are quantitative indicators used in the present invention.

[Explanation of symbols]

Reference Signs List 11 light source unit (light irradiation unit) 13 relative intensity ratio measurement unit (transmitted light measurement unit) 13a spectral unit 13b light detection unit 14 information processing unit (concentration calculation unit) 14a storage unit 14b calculation unit 14c input / output control unit 15 input unit Reference Signs List 16 display unit 17 printing unit 21 test fixture support base 22 light source 23 lens system 24 imaging unit 25 case 31 A / D converter 32 image data acquisition unit 33 ROM 34 magnetic storage medium 35 RAM 37 central processing unit (CPU) 38 bus 39a Input control unit 39b Display control unit 39c Print control unit 41 CCD solid-state imaging device 42 Imaging control circuit 43 Light receiving device 44 Spectral filter 45 Single item test paper 46 Multi item test paper 47 Single item rate assay test paper 48 Multi item rate assay test paper 51 Solid support of single-item test paper (measurement test section) 51a-51c Polynomial Solid support for test paper (measurement test section) 52 Solid support for reference of single item test paper (reference test section) 52a to 52c 53 Test paper identification section 54 Solid support for single item rate assay test paper ( Measurement test section) 54a-54i Solid support of multi-item rate assay test paper (measurement test section)

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G045 AA13 AA16 CA25 CB03 DA01 DA31 DA36 DA69 FA13 FB11 FB16 GC10 GC11 GC12 HA10 JA01 JA06 JA07 2G054 AA07 CA23 CA26 CA30 CE01 EA04 EA05 EA06 EB05 FA17 FA18 FA19 FA33 FA03 FB04 GA03 GB10 GE06 JA01 JA05 JA20

Claims (18)

[Claims] 1. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. An apparatus for quantitatively determining a color product, wherein a concentration of a color product to be quantified in the solid support is determined by optically measuring a color intensity of the color product, and the solid support is irradiated with light. A light irradiating means, a transmitted light measuring means for receiving a transmitted light from the solid support and measuring a light quantity, and the quantification using the measured transmitted light quantity and a previously acquired reference value. Density calculating means for calculating and calculating the density of the color product to be obtained, wherein the reference value is a scattered transmitted light component and a plurality of other components from the color product to be quantified by various properties of the transmitted light. And a plurality of transmitted light components having a predetermined specific spectral distribution. The relative intensity ratio of each spectrum obtained by calculating from the amount of spectrally transmitted light obtained by spectrally separating the transmitted light from the solid support into a plurality of lights equal to the reference value in the transmitted light measuring means. The transmitted light is separated into a plurality of components equal to the reference value by the density calculating means, and a relative intensity ratio of the plurality of lights obtained by dispersing each transmitted light component is calculated. Then, using the obtained relative intensity ratio and the reference value, the amount of scattered transmitted light only by the color object to be quantified or the amount of spectral light having a specific absorption wavelength of the color object to be quantified included in the light amount Wherein the density of the color object to be quantified is calculated and obtained using the following as a quantitative index. 2. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component from the color object to be quantified and a plurality of other components according to its properties, and each transmitted light component is separated into a plurality of lights having a predetermined specific spectral distribution. A reference value obtaining step of obtaining a reference intensity by calculating the relative intensity ratio of each spectrum by calculation from the amount of spectrally transmitted light obtained, and transmitting light obtained by irradiating the solid support with light, The light is split into the same plurality of lights as in the reference value obtaining step. A transmitted light measuring step of measuring the amount of transmitted light, a separating step of separating the transmitted light into a plurality of components same as the reference value acquiring step, and the quantitative determination based on the spectral transmitted light amount and the reference value. A component light amount calculating step of calculating only the amount of the scattered transmitted light component from the color object; and a density for obtaining the concentration of the color object to be quantified from the light amount of only the scattered transmitted light component from the color object to be quantified. A color medium quantitative storage medium storing a program used for a color substance quantitative method, comprising a calculating step. 3. A test device comprising: a sample; a solid support for holding a reagent capable of undergoing a color reaction with a specific component in the sample; and a test device including a test device identification portion comprising an optically readable code. To determine the concentration of the color substance to be quantified in the solid support by optically measuring the color intensity of the color substance to be quantified formed in the solid support. A storage medium storing a program used for a color object quantification method, wherein a decoding table of a code of the test device identification unit is obtained in advance, and various transmitted lights from the color object to be quantified by its properties. Separated into scattered transmitted light components and other components,
A reference value acquisition for obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectral transmission light obtained by spectrally separating each transmitted light component into a plurality of lights having a predetermined specific spectral distribution. A transmission light measuring step of measuring the amount of spectrally transmitted light by dispersing transmitted light obtained by irradiating the solid support with light into the same plurality of lights as in the reference value obtaining step; A classification step of classifying the reference component into the same plurality of components as the reference value acquisition step, and classifying the code of the test device identification unit from the amount of spectrally transmitted light, collating with the decoding table of the code of the test device identification unit, A code decoding step of decoding a code; a determination step of determining a type and a reference value of a subsequent step used for an operation based on the code decoding step; and the spectral transmission light according to the determination of the determination step. The amount and by using the reference value, coloration colored article quantified storage medium storing a program used in coloration colored article assay which comprises a density calculation step of determining the concentration of coloration colored article to be the quantified. 4. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component and a plurality of other components from the color product to be quantified by its properties, and each transmitted light component has light having a specific absorption wavelength of the color product to be quantified and A reference value obtaining step of obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectrally transmitted light obtained by spectrally splitting the light into a plurality of lights having a predetermined specific spectral distribution, Obtained by irradiating the solid support with light A transmitted light measuring step of dispersing the transmitted light into the same plurality of lights as in the reference value obtaining step and measuring the amount of spectrally transmitted light; and a separating step of separating the transmitted light into the same plurality of components as the reference value obtaining step And a component light amount ratio calculating step of calculating a ratio of the light amount of only the scattered transmitted light component from the color object to be quantified to the spectral transmitted light amount using the reference value, and the component light amount ratio calculating step. A component spectral light amount calculating step of calculating a light amount of light having a specific absorption wavelength of the color object to be quantified included in the scattered transmitted light component from the color object to be quantified based on the obtained ratio; A density calculation step of obtaining the density of the color product to be quantified from the amount of light having a specific absorption wavelength of the color product to be determined. Coloration colored article Quantitative storage medium storing. 5. The coloration according to claim 2, wherein, in the density calculation step, the density of the color object to be quantified is calculated using a relative intensity ratio diagram. Storage medium for material quantification. 6. The sample according to claim 1, wherein the sample contains a colorant to be quantified in addition to the colorant to be quantified, and the solid support is an optically transparent solid support in a wavelength range of light used for measurement. The present invention according to any one of claims 2 to 5, wherein the reference value includes a relative intensity ratio of the spectrum of light corresponding to the scattered and transmitted light by the coexisting color material, as the reference value. Storage medium for color substance quantification. 7. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component from the color object to be quantified and a plurality of other components according to its properties, and each transmitted light component is separated into a plurality of lights having a predetermined specific spectral distribution. A reference value obtaining step of obtaining a reference intensity by calculating the relative intensity ratio of each spectrum by calculation from the amount of spectrally transmitted light obtained, and transmitting light obtained by irradiating the solid support with light, The light is split into the same plurality of lights as in the reference value obtaining step. A transmitted light measuring step of measuring the amount of transmitted light; a separating step of separating the transmitted light into the same plurality of components as the reference value obtaining step; and creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value. A relative intensity ratio diagram creating step, a component light amount calculating step of calculating only the amount of scattered transmitted light component from the color object to be quantified based on the relative intensity ratio diagram, A density calculating step of calculating the density of the color object to be quantified from the light amount of only the scattered transmitted light component of the color object quantification method. 8. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component and a plurality of other components from the color product to be quantified by its properties, and each transmitted light component has light having a specific absorption wavelength of the color product to be quantified and A reference value obtaining step of obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectrally transmitted light obtained by spectrally splitting the light into a plurality of lights having a predetermined specific spectral distribution, Obtained by irradiating the solid support with light A transmitted light measuring step of dispersing the transmitted light into the same plurality of lights as in the reference value obtaining step and measuring the amount of spectrally transmitted light; and a separating step of separating the transmitted light into the same plurality of components as the reference value obtaining step A relative intensity ratio diagram creating step of creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value; and from the color object to be quantified in the spectral transmitted light amount based on the relative intensity ratio diagram. A component light amount ratio calculating step of calculating a ratio of the light amount of only the scattered transmitted light component, and the quantification included in the scattered transmitted light component from the color object to be quantified based on the ratio obtained in the component light amount ratio calculating step A component spectral light amount calculating step of calculating the light amount of the light having the specific absorption wavelength of the color product to be determined; and the quantification from the light amount of the light having the specific absorption wavelength of the color product to be quantified And a density calculation step for calculating the density of the color object to be performed. 9. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component from the color object to be quantified and a plurality of other components according to its properties, and each transmitted light component is separated into a plurality of lights having a predetermined specific spectral distribution. A reference value obtaining step for obtaining a reference value for reference by calculating a relative intensity ratio of each spectrum by calculation from the amount of spectral transmitted light obtained by the method, as soon as possible after contacting the sample with the test tool, Transmission obtained by irradiating the solid support with light The,
The transmission is started by measuring the amount of spectrally transmitted light by dispersing the light into the same plurality of lights as in the step of acquiring the reference value, and repeating this a plurality of times at fixed time intervals until the measured value of the amount of spectrally transmitted light is stabilized. A light measurement step, a separation step of separating the transmitted light into the same plurality of components as the reference value acquisition step, and a relative intensity ratio diagram creation step of creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value. A determining step of determining a relative intensity ratio and a spectrally transmitted light amount of transmitted light from the solid support before the formation of the color substance to be quantified at the contact time based on the relative intensity ratio diagram; A component light amount calculating step of calculating the light amount of only the scattered transmitted light component from the color object to be quantified based on the relative intensity ratio and the ratio value obtained in the ratio determining step, from the color object to be quantified Coloration colored article Quantitative storage medium storing a program used in coloration colored article assay which comprises a density calculation step of determining the concentration of coloration colored article to be the quantified from the light amount of only the scattered transmitted light component. 10. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. A storage medium storing a program used in a colorant determination method for determining the concentration of a colorant to be quantified in the solid support by optically measuring the color intensity of a colorant, The transmitted light is separated into a scattered transmitted light component and a plurality of other components from the color product to be quantified by its properties, and each transmitted light component has light having a specific absorption wavelength of the color product to be quantified and A reference value obtaining step of obtaining a reference value for reference by obtaining a relative intensity ratio of each spectrum by calculation from the amount of spectrally transmitted light obtained by spectrally splitting the light into a plurality of lights having a predetermined specific spectral distribution, After contact between the sample and the test device As soon ability as far as, the transmitted light obtained by irradiating light to said solid support,
The transmission is started by measuring the amount of spectrally transmitted light by dispersing the light into the same plurality of lights as in the step of acquiring the reference value, and repeating this a plurality of times at fixed time intervals until the measured value of the amount of spectrally transmitted light is stabilized. A light measurement step, a separation step of separating the transmitted light into the same plurality of components as the reference value acquisition step, and a relative intensity ratio diagram creation step of creating a relative intensity ratio diagram from the spectral transmitted light amount and the reference value. A determining step of determining a relative intensity ratio and a spectral transmission light amount of transmitted light from the solid support before the formation of the color product to be quantified at the time of the contact based on the relative intensity ratio diagram; A component light amount ratio calculation unit that calculates a ratio of the light amount of only the scattered transmitted light component from the color object to be quantified to the spectral transmitted light amount based on the intensity ratio and the fixed value obtained in the setting step. And calculating the light amount of light having a specific absorption wavelength of the color object to be quantified included in the scattered transmitted light component from the color object to be quantified based on the ratio obtained in the component light amount ratio calculation step. A component spectral light quantity calculation step, and a density calculation step of calculating the density of the color substance to be quantified from the light quantity of light having a specific absorption wavelength of the color substance to be quantified. A storage medium for quantifying colored substances, which stores a program used for the quantification method. 11. In the reference value acquiring step, the transmitted light is a parallel transmitted light without scattering, a scattered transmitted light by a solid support, a scattered transmitted light by a colorant to be quantified, and if present, The storage medium for quantifying a colored substance according to any one of claims 2 to 10, wherein the storage medium is classified into each transmitted light component of scattered transmitted light by the coexisting colored substance. 12. The storage medium according to claim 2, wherein the plurality of lights obtained in the reference value obtaining step are three types of light. . 13. The storage device for quantifying a colored substance according to claim 12, wherein the three kinds of spectroscopy are primary stimuli of an RGB color system (CIE color system) defined by the International Commission on Illumination. Medium. 14. The method according to claim 12, wherein the three types of spectroscopy are primary stimuli in the XYZ color system (CIE1931 standard color system) defined by the International Commission on Illumination. Storage medium. 15. The storage medium according to claim 12, wherein the three types of spectroscopy are primary stimuli of the UCS color system defined by the International Commission on Illumination. 16. The method according to claim 2, wherein in the calculation process performed to calculate the concentration of the color substance to be quantified, Grassmann's law of additive color mixture is used. Storage medium for quantitative determination of color products. 17. In the density calculating step, the density of the color substance to be quantified is calculated by the following equation: Log (C) = α (A) ² + β (A) + γ (where C is to be quantified) The concentration of the color substance, A is the amount of scattered transmitted light from the color substance to be quantified or the amount of light contained in the color substance having a specific absorption wavelength to be quantified, and α, β and γ are constants. 17. The storage medium according to any one of claims 2 to 16, wherein the storage medium is used to determine the colorant. 18. A sample to be quantified, which is formed in the solid support by bringing a sample into contact with a test device having a solid support holding a reagent that causes a color reaction with a specific component in the sample. What is claimed is: 1. An apparatus for quantitatively determining a color product, wherein a concentration of a color product to be quantified in the solid support is determined by optically measuring a color intensity of the color product, and the solid support is irradiated with light. A light irradiating means, a transmitted light measuring means for receiving a transmitted light from the solid support and measuring a light quantity, and the quantification using the measured transmitted light quantity and a previously acquired reference value. Density calculation means for calculating and calculating the density of a color product to be obtained, wherein the calculation device includes the color medium quantitative storage medium according to any one of claims 2 to 17. Colored object quantitative device.