JP4206461B2 - Quantitative analysis method and specimen holding tool used therefor - Google Patents

Description

[0001]
[Technical field to which the invention belongs]
The present invention relates to a quantitative analysis method for determining the amount of a component present in an unknown amount of sample and a sample holding tool used therefor.
[0002]
[Prior art]
Conventionally, patients have to go to a medical institution to collect blood and urine for examination and treatment of various diseases. However, these test results cannot be obtained unless waiting until the next visit or waiting for a long time, and there is a problem that it is very time-consuming for patients and medical institutions.
[0003]
In recent years, in order to avoid such a problem, a sample collection card formed of filter paper or the like has been proposed. For example, Japanese Patent Application Laid-Open No. 10-104226 discloses a blood collection card. The patient, for example, impregnates and dries the blood collected by himself on this blood collection card and mails it to a medical institution. Upon receiving this, the medical institution cuts out the blood-impregnated portion of the blood collection card, extracts blood therefrom, and conducts an inspection for each inspection item. And when a patient comes to a medical institution, treatment and diagnosis are performed based on the test result.
[0004]
When using such a blood collection card, for example, since the patient himself collects blood as described above, the amount of blood impregnated in the blood collection card is an unknown amount, and it is not possible to accurately quantify the amount of components in the blood. It was difficult. Therefore, for example, using a filter paper that holds a certain amount of blood in a certain area and cutting out the blood-impregnated portion of the filter paper by a certain area, a method for securing a certain amount of blood sample, or a certain amount that holds a certain amount of blood A method of securing a certain amount of blood sample by using a filter paper of an area and supplying a saturated retention amount of blood to the filter paper has been proposed.
[0005]
[Problems to be solved by the invention]
However, the filter paper as described above has the following problems. For example, when the former filter paper is used, it is necessary to impregnate the entire cut out filter paper with blood, so that it is difficult to select a cut-out portion and to perform the cut-out operation. In addition, when the latter filter paper is impregnated with a saturated retention amount of blood, it is actually necessary to supply blood with a saturation retention amount or more to sufficiently impregnate the blood, which takes time and increases the burden on the patient. . Furthermore, in order to improve the quantitativeness of these quantitative filter papers, the production itself becomes very difficult and complicated, and the cost increases.
[0006]
Accordingly, an object of the present invention is to provide a quantitative analysis method capable of accurately measuring the amount of a component in a sample even when an unknown amount of sample is used, and a sample holding tool used therefor.
[0007]
[Means for Solving the Problems]
In order to achieve the above object, the quantitative analysis method of the present invention is a quantitative analysis method in which a specimen is held in a porous material, the specimen is collected from the porous material, and the amount of components in the specimen is measured. When the specimen is held in the porous material, the detectable substance corresponding to the quantity of the specimen is also held, the detectable substance is also collected when the specimen is collected, and the collected detectable substance is collected. In this method, the amount of the sample is determined from the measured value, and the amount of the component in the sample is determined based on this value.
[0008]
In the quantitative analysis method of the present invention, not only the sample is held in the porous material, but also a detectable substance corresponding to the amount of the sample is held, and this amount is also measured. In this way, even if the amount of the sample retained in the porous material is an unknown amount, by measuring the retained detectable substance, the measured value can be used as a reference for the amount of the sample. Can do. For example, if the ratio between the amount of the retained sample and the amount of the detectable substance to be collected is obtained in advance, the amount of the sample can be obtained from the measured value of the detectable substance as described above. For this reason, according to the quantitative analysis method of the present invention, the amount of the sample can be obtained with excellent accuracy, and the quantitative property of the component amount in the sample is improved. Furthermore, since the present invention does not require the use of a special porous material or the like, for example, in order to improve the quantitativeness, the cost can be reduced. Further, the quantitative operation is simple, and is useful for various tests in clinical medicine, for example.
[0009]
In the quantitative analysis method of the present invention, a porous material that holds a detectable substance (hereinafter referred to as “detectable substance-holding porous material”) is disposed on the specimen-holding porous material as the porous material. Preferably, the detectable substance is also transferred to the specimen-holding porous material by using a composite porous material and transferring the specimen to the specimen-holding porous material through the detectable substance-holding porous material. Thus, if the detectable substance is transferred by the transfer of the specimen, the detectable substance can be transferred according to the amount of the specimen.
[0010]
In the quantitative analysis method of the present invention, it is preferable that the specimen contains an aqueous solvent, and the detectable substance can be dissolved in the aqueous solvent. According to this, the detectable substance can be dissolved in the sample by contacting the sample to be transferred, and can be transferred together with the sample. The detectable substance is not particularly limited to a substance that can be dissolved in the aqueous solvent. In addition, for example, the detectable substance is physically released from the detectable substance-holding porous material by migration of the specimen. It may be a thing.
[0011]
In the quantitative analysis method of the present invention, the detectable substance is preferably a water-soluble dye, and examples of the water-soluble dye include indocyanine green (hereinafter referred to as “ICG”), indigo carmine, phenol sulfonephthalein. (Hereinafter referred to as “PSP”). Among these, for example, when various analysis target components are detected by an optical method, the ICG has an absorption region at a near infrared wavelength different from a general detection wavelength and does not prevent detection of the analysis target component. preferable.
[0012]
In the quantitative analysis method of the present invention, the specimen is preferably a living body-derived aqueous liquid specimen, and examples thereof include blood, urine, saliva, lymph fluid, spinal fluid, and interstitial fluid. If the present invention is applied to such biological sample quantification, for example, various diagnoses in clinical medicine can be performed with high accuracy.
[0013]
Next, in the specimen holding tool of the present invention, a detectable substance-holding porous material is disposed on a specimen-holding porous material, and the specimen is passed through the detectable substance-holding porous material, and the specimen-holding porous material. It has the structure of making it hold | maintain. By using this specimen holding tool, the above-described quantitative analysis method of the present invention can be easily performed with excellent accuracy. In order to retain the detectable substance in the porous material, for example, the detectable substance is dissolved or dispersed in water or the like, and this liquid is dropped into the porous material, or the liquid is impregnated with the porous material. Then, it only needs to be dried and can be prepared very easily and at low cost. The specimen holding tool of the present invention is useful for, for example, clinical medical examinations. As described above, the specimen collected by the patient itself is impregnated and dried in the specimen holding tool and transported to a medical institution. Since the quantitative analysis of the components in the sample can be easily performed, both the patient and the medical institution can be saved.
[0014]
In the sample holding device of the present invention, when the analysis target is a component in plasma or serum, it is preferable that a blood cell separation material is disposed on the porous material for sample holding, and more preferably blood cell separation ability. Therefore, a blood cell separator is disposed on the detectable substance-holding porous member. Moreover, it is not limited to such a structure, For example, it is also preferable that the said detectable substance holding porous material has a blood cell separation function. According to this configuration, since the blood cell separation material and the detectable substance-holding porous material are in the same layer, the cost can be further reduced. If the specimen holding device of the present invention having blood cell separation ability is used as described above, for example, blood cells can be separated simply by adding whole blood, and the operation can be simplified.
[0015]
In the specimen holding device of the present invention, the detectable substance is preferably a pigment such as ICG, indigo carmine, PSP, and more preferably ICG, as in the quantitative analysis method.
[0016]
DETAILED DESCRIPTION OF THE INVENTION
FIG. 1 shows an example of the specimen holding tool of the present invention. As shown in the drawing, in this specimen holding tool, a detectable substance-holding porous material 2 is disposed on a specimen-holding porous material 1.
[0017]
The size of the specimen-holding porous material 1 can be appropriately determined according to, for example, the quantity of specimen to be held. For example, when the specimen quantity to be held is about 10 to 100 μl, the length is 1 to 100 mm and the width is 1 The size of the detectable substance-holding porous material 2 is, for example, in the range of 1 to 100 mm in length, in the range of 1 to 100 mm in width, and in the range of 10 to 1000 μm in thickness. is there.
[0018]
The material of the specimen holding porous material 1 is not particularly limited. For example, a filter paper, a glass filter, a resin porous membrane, and the like can be used. Among these, a filter paper is preferable from the viewpoints of cost and ease of handling. . The average pore diameter of the porous material 1 for holding a sample is not particularly limited as long as the sample penetrates and is held, but is, for example, in the range of 0.1 to 100 μm.
[0019]
In addition, the porous material 1 for holding a specimen is used, for example, to maintain a stable component in the specimen to be held, for example, sugars such as sucrose, trehalose, lactose, glucose, salts such as glycine, sodium chloride, potassium chloride, phosphoric acid, etc. You may contain stabilizers, such as buffers, such as a buffer, a citrate buffer, and a Good buffer. The content of the stabilizer can be appropriately determined depending on the type and the like, and is, for example, in the range of 0.01 to 10 mg per 1 cm ^{3 of the} specimen-holding porous material 1.
[0020]
The material of the detectable substance-holding porous material 2 is not particularly limited as long as the detectable substance can be held, but for example, a resin porous film, a filter paper, a glass filter, a mesh, a cloth, a gel, and the like can be used. Examples of the material for the resin porous membrane include polysulfone, polyester, nylon, nitrocellulose, and polycarbonate. Further, the average pore diameter is not particularly limited as long as the specimen and the detectable substance permeate and permeate, but it is, for example, in the range of 0.1 to 100 μm.
[0021]
As the detectable substance, for example, a water-soluble dye as described above can be used, and it is not particularly limited as long as it does not react with the analysis target component and does not interfere with detection of the analysis target component.
[0022]
For example, when the analysis target component is detected by an optical method, it is preferable that the detectable substance has absorption in a wavelength region different from the detection wavelength region of the analysis target component. Moreover, it is not restrict | limited to the above pigment | dyes etc., For example, the substrate which is not colored (for example, colorless), the substrate which does not have an absorption wavelength as it is, etc. can be used. Such a substrate can be detected in the detection stage by, for example, coloring it by an enzyme reaction or a chemical reaction.
[0023]
The amount of the detectable substance retained in the detectable substance-holding porous material 2 can be appropriately determined depending on the type of the detectable substance, etc., for example, a range of 0.1 to 10,000 μg per 1 cm ^{3 of the} porous material, Preferably, it is the range of 1-1000 micrograms. Specifically, when the detectable substance is ICG, for example, the range is 1 to 1000 μg, preferably 10 to 500 μg, per 1 cm ³ of the volume of the porous material.
[0024]
The detection of the detectable substance in the porous material is performed by, for example, dropping a solution obtained by dissolving or dispersing the detectable substance in water onto the porous material, or immersing the porous material in the liquid and then drying the porous material. You can do it. The liquid is prepared to have a concentration in the range of 1 to 10,000 mg / liter, for example. The drying process may be, for example, natural drying or air drying, and the processing conditions are, for example, a temperature range of 1 to 200 ° C. and a processing time of 10 seconds to 3 days.
[0025]
For example, the specimen holding tool may be formed by simply laminating the detectable substance-holding porous material 2 on the specimen-holding porous material 1, or both of them within a range that does not hinder the migration of the specimen and the detectable substance. , And may be integrated using an adhesive or the like.
[0026]
Next, based on FIG. 1, an example in which quantitative analysis is performed with this sample holding tool using urine as a sample and ICG as a detectable substance will be described.
[0027]
First, urine is dropped on the detectable substance-holding porous material 2. Then, urine contacts ICG while moving inside the detectable substance-holding porous material 2 in the thickness direction. The ICG is dissolved in accordance with the amount of urine that comes into contact with the urine and moves along with the urine in the thickness direction of the detectable substance-holding porous material 2 so that the both reach the specimen-holding porous material 1 and impregnate it. Is done.
[0028]
After the specimen holding tool is dried by air drying or natural drying, the detectable substance-holding porous material 2 is removed from the specimen holding tool, and the specimen-impregnated portion of the specimen-holding porous material 1 is cut or punched. Do. For punching, for example, a punch or the like can be used. The size of punching or the like is not particularly limited, but is, for example, in the range of 1 to 30 mm in diameter. In addition, it is preferable that the portion to be punched out contains a large amount of the sample-impregnated portion. However, according to the present invention, the amount of the sample can be quantified regardless of the portion to be punched and its area. Unlike that, there is no problem in quantitativeness even if a portion not impregnated with the specimen is included.
[0029]
A section obtained by punching or the like is placed in, for example, a tube or the like, and an extraction solution is added to the section, and the urine and ICG are extracted and collected. The extraction solution is not particularly limited as long as it can extract both of the above and does not affect the detection of the component to be analyzed in the sample. For example, a buffer solution, physiological saline, purified water, protein solution, etc. Can be used. Examples of the buffer solution include various buffer solutions containing phosphoric acid, citric acid, hydrochloric acid, acetic acid and the like, and the pH is, for example, in the range of pH 3-9. The amount of the extraction solution added is not particularly limited as long as the amount is known, but can be appropriately determined depending on the size of the slice, for example, and is, for example, in the range of 1 to 1000 times the volume of the slice. In addition, since quantitative property can be further improved, for example, it is preferable to make the addition amount of the extraction solution constant with respect to the size of the section. Further, the extraction processing time is not particularly limited, but is, for example, in the range of 1 to 300 minutes.
Next, the amount of ICG in the collected liquid is measured, and the amount of urine is obtained from the measured amount. For example, using a standard solution of various known amounts of urine (for example, when the sample is blood, the standard solution is blood) and the sample holding device, the standard solution and ICG are fixed in the same manner. A calibration curve showing the relationship between the standard solution amount and the ICG amount is created by measuring the amount of ICG in the recovery solution and collecting the calibration curve and the ICG in the recovery solution. The amount of urine can be determined from the amount. Examples of ICG detection means include optical means, which can be detected by measuring absorbance in the wavelength range of 700 to 850 nm. The detection means for the detectable substance can be appropriately determined depending on the type of the detectable substance. In addition to the direct optical means as described above, for example, an indirect optical means using a color reaction or the like, Electrochemical means using biosensors, means by freezing point method, etc. can also be employed.
[0031]
Then, the component to be analyzed in urine is measured using the collected liquid, and the amount of component in the urine sample can be determined based on the value of the amount of urine.
[0032]
Next, FIG. 2 shows an example in which the blood cell separation material 3 is arranged on the detectable substance-holding porous material 2 shown in FIG. 1 in the specimen holding tool of the present invention. In FIG. 2, the same parts as those in FIG.
[0033]
The blood cell separation material 3 is an optional component provided when, for example, the analysis target is plasma or serum components in whole blood and it is necessary to remove the blood cells.
[0034]
The size of the blood cell separation material 3 is not particularly limited, but is, for example, a total length range of 1 to 100 mm, a width range of 1 to 100 mm, and a thickness range of 1 to 1000 μm.
[0035]
The material of the blood cell separator 3 is not particularly limited, and for example, a glass filter, a resin porous membrane, and the like can be used. Examples of the resin porous membrane material include polysulfone, polyester, nylon, nitrocellulose, Examples include polycarbonate. The average pore diameter is not particularly limited as long as blood cells can be separated, and is, for example, in the range of 0.1 to 100 μm. Also, use a porous membrane whose pore structure is, for example, a pore structure (an anisotropic pore diameter gradient structure) in which the pore diameter changes continuously or stepwise from one surface to the other surface of the membrane. You can also.
[0036]
As the specimen holding device of the present invention capable of performing blood cell separation in this way, for example, as shown in FIG. 3, a detectable substance-holding porous material 4 having blood cell separation ability is a specimen-holding porous material. The structure arrange | positioned on 1 may be sufficient. The detectable substance-holding porous material 4 having blood cell separation ability can be produced, for example, by holding a detectable substance in the material of the blood cell separating material 3 in the same manner as described above.
[0037]
Next, when the sample is whole blood and a specific component in plasma is an analysis target, ICG is used as a detectable substance, and quantitative analysis is performed using the sample holding tool shown in FIG. 2 or FIG. Will be described. Unless otherwise indicated, the measurement can be performed in the same manner as the quantitative analysis using the specimen holding tool shown in FIG.
[0038]
When the specimen holding tool shown in FIG. 2 is used, when whole blood is dripped onto the blood cell separation material 3, the whole blood moves inside the blood cell separation material 3 in the thickness direction, while the blood cell separation material 3 Blood cells are captured by the pore structure and separated into blood cells and plasma. The separated plasma passes through the blood cell separator 3, reaches the detectable substance-holding porous material 2, and is impregnated therein. Then, the impregnated plasma comes into contact with the ICG while moving the detectable substance-holding porous material 2. The ICG is dissolved in accordance with the amount of plasma in contact with it, moves along with the plasma inside the detectable substance-holding porous material 2, reaches the specimen-holding porous material 1 together with the plasma, and is impregnated therein. . In addition, when using the specimen holding tool shown in FIG. 3, when whole blood is dropped on the detectable substance-holding porous material 4 having blood cell separation ability, the whole blood has a thickness inside the porous material 4. While moving in the direction, it is separated into blood cells and plasma as described above. Further, simultaneously with this separation, the ICG that has come into contact with the plasma component in the whole blood is dissolved in accordance with the amount of plasma as described above, and moves inside the porous material 4 together with the plasma. Then, ICG and plasma reach the specimen holding porous material 1 and are impregnated therein.
[0039]
【Example】
As shown below, a specimen holding tool as shown in FIG. 3 was prepared, and various components in plasma and ICG were measured using this.
[0040]
(Preparation of specimen holding tool)
ICG (Lot. GG01: manufactured by Tokyo Chemical Industry Co., Ltd.) was dissolved in water to a concentration of 200 mg / 100 ml to prepare an ICG solution. Then, 75 μl of this ICG aqueous solution was dropped onto a polysulfone porous membrane (BTS-sp, Lot. 9043PL2: manufactured by US Filter) having a thickness of 320 μm and 15 × 15 mm using an autopipette, and then at room temperature (about 25 ° C.) The ICG holding porous material 4 having blood cell separation ability was left to air dry at room temperature (about 60%) overnight.
[0041]
Sucrose (manufactured by Nacalai Tesque) was dissolved in a saline-isotonic PBS solution (Phosphate-buffered saline: pH 7.4) to a concentration of 250 g / liter to prepare a sucrose / PBS solution. Then, a filter paper (BFC180: manufactured by Whatman) is immersed in the sucrose / PBS solution, and is left to stand overnight at room temperature (about 25 ° C.) and normal humidity (about 60%) to air-dry it. Porous material 1 was obtained. Then, the ICG holding porous material 4 having the blood cell separation ability was disposed on the plasma holding porous material 1, and this was used as a specimen holding tool. Using this sample holding tool, a plasma sample was prepared as follows.
[0042]
(Preparation of plasma sample)
Whole blood was collected using a heparin-containing blood collection tube (Insepack-E: manufactured by Sekisui Chemical Co., Ltd.), and this was used as a specimen. 150 μl of the specimen was dropped onto the surface of the ICG holding porous material 4 having the blood cell separation ability of the blood holding tool and dried at a temperature of 25 ° C. for 1 hour. Then, the ICG holding porous material 4 having the blood cell separation ability is removed from the blood holding tool, and the plasma-impregnated portion of the plasma holding porous material 1 is randomized to a diameter of 6 mm using a stationery punch. Punched and placed in a 1.5 ml disposable tube. Further, 300 μl of the PBS solution was added as an extraction solution and allowed to stand at room temperature for 20 minutes, and then the supernatant was taken out and used as a plasma sample solution. In the same manner, a total of 20 plasma sample solutions were prepared. The plasma impregnation part was punched so that the plasma concentration varied.
[0043]
Using these plasma sample solutions, the ICG amount and the following various component amounts were measured by the method described below. ICG measured the absorption of the plasma sample solution at a wavelength of 805 nm using an automatic analyzer (BM-8: manufactured by JEOL Ltd.). Glucose (hereinafter referred to as “Glc”) is glucose II-HA Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.), total cholesterol (hereinafter referred to as “TC”) is Determiner L TCII (manufactured by Kyowa Medex), Glutamic- Pyrvic transaminase (hereinafter referred to as “GPT”) is transaminase HR-II (GPT-7070: manufactured by Wako Pure Chemical Industries, Ltd.) and alkaline phosphatase (hereinafter referred to as “ALP”) is ALPII-HA test Wako (7150: Wako Pure Chemical Industries, Ltd.), creatine kinase (hereinafter referred to as “CPK”) is CK E-HA Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.), and amylase (hereinafter referred to as “Amy”) is Amy II-HA test. Wako (manufactured by Wako Pure Chemical Industries, Ltd.) and creatinine (hereinafter referred to as “Cre”) are pure auto CRE-N (Daiichi Chemicals Co., Ltd.) and urea nitrogen (hereinafter referred to as “BUN”) are urea nitrogen II-HA test Wako (7070: manufactured by Wako Pure Chemical Industries, Ltd.) and triglyceride (hereinafter referred to as “TG”). ), Triglyceride E-HA test Wako (manufactured by Wako Pure Chemical Industries, Ltd.) and HDL-cholesterol (hereinafter referred to as “HDL-C”), Choletest HDL (manufactured by Daiichi Chemicals), respectively, It measured according to the usage method. In various measurements, purified water was used as a blank. The results of these various component amounts are shown in FIGS. 4 to 13 together with the results of ICG amounts. Each figure is a graph showing the correlation between the concentration of each component and the ICG concentration in the plasma sample solution. The formula in the figure shows the correlation and R ² shows the correlation coefficient.
[0044]
As shown in FIGS. 4 to 13, a high correlation was observed between the concentrations of various components in the plasma and the ICG concentrations. This is because the ICG amount and the plasma amount are in a proportional relationship, and the large amount of ICG means that the amount of plasma is also large, and the amount of various components increases according to this plasma amount. . As can be seen from these results, if a detectable substance such as ICG is held in a porous material together with the sample and the amount of the sample in the recovered solution is measured, the amount of the sample is determined from a calibration curve prepared in advance, for example. This improves the quantitativeness of the amount of component to be analyzed. The correlation decreases as the ICG concentration increases, but this is eliminated by further diluting and measuring the plasma sample.
[0045]
【The invention's effect】
As described above, according to the quantitative analysis method of the present invention, the amount of the sample can be determined, so that the quantitativeness of the components in the sample is improved. Such a quantitative analysis method of the present invention is useful for diagnosis in clinical medicine, for example.
[Brief description of the drawings]
FIG. 1 is a cross-sectional view showing an embodiment of a specimen holding tool of the present invention.
FIG. 2 is a cross-sectional view showing another embodiment of the specimen holding tool of the present invention.
FIG. 3 is a cross-sectional view showing still another embodiment of the specimen holding tool of the present invention.
FIG. 4 is a graph showing the correlation between ICG concentration and glucose concentration in a plasma sample in one example of the present invention.
FIG. 5 is a graph showing a correlation between ICG concentration and TC concentration in a plasma sample in the Example of the present invention.
FIG. 6 is a graph showing the correlation between ICG concentration and GPT concentration in a plasma sample in the Example of the present invention.
FIG. 7 is a graph showing the correlation between ICG concentration and ALP concentration in a plasma sample in the Example of the present invention.
FIG. 8 is a graph showing a correlation between ICG concentration and CPK concentration in a plasma sample in the Example of the present invention.
FIG. 9 is a graph showing the correlation between ICG concentration and Amy concentration in a plasma sample in the Example of the present invention.
FIG. 10 is a graph showing the correlation between ICG concentration and Cre concentration in a plasma sample in the Example of the present invention.
FIG. 11 is a graph showing a correlation between ICG concentration and BUN concentration in a plasma sample in the Example of the present invention.
FIG. 12 is a graph showing a correlation between ICG concentration and TG concentration in a plasma sample in the Example of the present invention.
FIG. 13 is a graph showing the correlation between ICG concentration and HDL-C concentration in a plasma sample in the Example of the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Porous material for specimen holding 2 Detectable substance holding porous material 3 Blood cell separation material 4 Detectable substance holding porous material having blood cell separation ability

Claims (8)

A quantitative analysis method for holding a specimen in a porous material, collecting the specimen from the porous material, and measuring the amount of a component in the specimen,
As a porous material, using a composite porous material in which a porous material that holds a detectable substance is disposed on a porous material for specimen holding,
The specimen includes an aqueous solvent,
The detectable substance is a water-soluble dye that is soluble in the aqueous solvent,
By holding a detectable substance corresponding to the amount of the specimen when the specimen is held in the porous material, the specimen is transferred to the porous material for specimen holding through the porous material holding the detectable substance. The detectable substance is also transferred to the porous material for holding a specimen, the detectable substance is also collected at the time of collecting the specimen, the amount of the collected detectable substance is measured, and the specimen is calculated from the value. A quantitative analysis method for determining an amount and further determining an amount of a component in the sample based on this value. Water-soluble dye, quantitative analysis according to claim 1, wherein at least one dye selected from the group consisting of indocyanine green, indigo carmine and phenolsulfonphthalein. Specimen, the quantitative analysis according to claim 1 or 2 which is an aqueous liquid specimen of biological origin. The quantitative analysis method according to claim 3 , wherein the specimen is at least one specimen selected from the group consisting of blood, urine, saliva, lymph, cerebrospinal fluid, and interstitial fluid. A specimen holding tool used in the quantitative analysis method according to any one of claims 1 to 4,
A porous material holding a detectable substance is disposed on the porous material for holding the specimen,
The detectable substance is a water-soluble dye that is soluble in an aqueous solvent,
A specimen holding tool for holding a specimen on the specimen-holding porous material through a porous material that holds the detectable substance. The specimen holding device according to claim 5 , wherein the analysis target is a component in plasma or serum, and a blood cell separation material is disposed on a porous material holding a detectable substance. The specimen holding device according to claim 5 , wherein the analysis target is a component in plasma or serum, and the porous material holding a detectable substance has a blood cell separation function. The specimen holding device according to any one of claims 5 to 7 , wherein the detectable substance is at least one dye selected from the group consisting of indocyanine green, indigo carmine, and phenol sulfonephthalein.

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