JP2018031663A - Metabolite analyzing method and metabolite analyzing device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a metabolite analyzing method and a metabolite analyzing device capable of easily determining property of metabolites in a cell or property of body fluid using a transmission method in infrared spectroscopy.SOLUTION: A cell 2 which comprises a storage part 20 having window parts 21 and 22 through which infrared light can transmit and storing analysis object fluid such as solution of a cell and body fluid, with 0.5-50 μm of an optical path length of infrared light in infrared spectroscopy, is used for measuring absorbance of metabolites in the cell in the solution or absorbance of the body fluid using a transmission method in the infrared spectroscopy, so that a metabolite density value of the body fluid or the cell is measured and if the ratio (N1/NN) of a first measurement metabolite density value N1 to a metabolite density value NN of a normal cell or body fluid previously measured at a first wave number H1 is a predetermined ratio R1 or higher, property of the metabolites in the cell or the body fluid subjected to the measurement is determined, in a first determination step.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a metabolite analysis method and a metabolite analyzer that measure the absorbance of a cellular metabolite or body fluid using a transmission method in infrared spectroscopy to determine the properties of the cellular metabolite or body fluid.

2. Description of the Related Art Conventionally, an analysis method for measuring the infrared absorption spectrum of a dried cell using infrared spectroscopy and analyzing the properties of the dried cell is known (see, for example, Patent Document 1).
In contrast, in order to analyze cells alive and non-invasively, the applicant accommodates living cells soaked in a cell culture medium in a cell with a short optical path length having a transparent window. Thus, the infrared absorption spectrum is measured by using the transmission method in the infrared spectroscopy while keeping living cells. When the applicant compares the infrared absorption spectrum of cancer cells and lactic acid, which is a metabolite of the cell, using infrared spectroscopy, there is a wave number region in which the infrared absorption spectrum is similar. I found out.

Japanese National Patent Publication No. 11-502935

The applicant has found that there exists a wave number region in which the infrared absorption spectrum is similar between cancer cells and lactic acid which is a metabolite of the cells. Therefore, by using the infrared absorption spectrum of cell metabolites using the transmission method in infrared spectroscopy, the properties of the cell metabolites in the cell culture medium (medium) can be determined as they are living cells. It is desirable to easily determine and analyze.
Moreover, it is desired that not only cells but also body fluids such as saliva, sweat, urine, blood, plasma, etc. are easily determined by analyzing the properties of body fluids using the transmission method in infrared spectroscopy.

  An object of the present invention is to provide a metabolite analysis method and a metabolite analyzer that can easily determine the properties of cellular metabolites or body fluids using a transmission method in infrared spectroscopy.

  The present invention is a housing part having a window part capable of transmitting infrared light, which contains a liquid to be analyzed such as an aqueous solution of a cell, a body fluid and the like, and an optical path length of infrared light in infrared spectroscopy is 0.5. By measuring the absorbance of the metabolite of the cell in the aqueous solution or the absorbance of the bodily fluid in the aqueous solution by a transmission method in infrared spectroscopy using a cell having a container with a predetermined length of ˜50 μm, the metabolism of the cell or bodily fluid The product concentration value is measured, and the first measured metabolite concentration value measured at a predetermined first wave number belonging to the stretching vibration is measured for a normal cell or body fluid previously measured at the first wave number. The present invention relates to a metabolite analysis method comprising a first determination step of determining a property of a measured cell metabolite or body fluid when a ratio of the first measurement metabolite concentration value to a metabolite concentration value is a predetermined first ratio or more. .

In addition, the present invention is a container having a window that can transmit infrared light, and stores an analysis target liquid such as an aqueous solution of a cell or a body fluid, and the optical path length of infrared light in infrared spectroscopy is 0. Cell or body fluid is measured by measuring the absorbance of the metabolite of the cell in the aqueous solution or the absorbance of the body fluid by the transmission method in infrared spectroscopy using a cell having a container with a predetermined length between 5 and 50 μm. The metabolite concentration value of the first measured metabolite concentration value measured at a predetermined first wave number between 1020 and 1080 cm ⁻¹ is measured normally in the first wave number. When the ratio of the first measured metabolite concentration value to the metabolite concentration value of the cell or body fluid is a predetermined first ratio or more, a first determination step of determining that the cancer is false positive for the measured cell or body fluid is provided. metabolism About the things analytical methods.

Further, in the second determination step performed before the first determination step, the second measured metabolite concentration value measured at a predetermined second wave number between 1680 and 1780 cm ⁻¹ , When the predetermined peak waveform of the infrared absorption spectrum is not obtained at the second wave number, the process proceeds to the first determination step, and when the peak waveform is obtained, the measured cell or body fluid is positive for cancer. It is preferable to provide the 2nd determination process determined to exist.

Further, in the third determination step that is executed after it is determined in the first determination step that the cancer is false positive, the first measurement metabolite concentration value measured at the predetermined first wave number is measured. When the ratio of the first measurement metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is less than or equal to a predetermined second ratio, the cell or body fluid It is preferable to include a third determination step for determining that the cancer is false positive.

Further, in the fourth determination step, which is executed after it is determined in the first determination step that the cancer is not false positive, the first measured metabolite concentration value measured at the predetermined first wave number is measured. When the ratio of the first measurement metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is less than or equal to a predetermined third ratio, the cell or body fluid It is preferable to provide the 4th determination process which determines with cancer being negative.

  The metabolite is preferably lactic acid.

In addition, the present invention is a container having a window that can transmit infrared light, and stores an analysis target liquid such as an aqueous solution of a cell or a body fluid, and the optical path length of infrared light in infrared spectroscopy is 0. A cell including a container having a predetermined length of 5 to 50 μm, a light source unit that irradiates infrared light to a cell or a body fluid stored in the cell, A light detection unit for detecting infrared light transmitted through the body fluid or cells contained in the cell, and the absorbance or body fluid of the metabolite of the cell in the aqueous solution based on the infrared light detected by the light detection unit A concentration measurement unit for measuring the metabolite concentration value of a cell or body fluid by measuring the absorbance of the sample by a transmission method in infrared spectroscopy, and measurement at a predetermined first wave number between 1020 and 1080 cm ⁻¹ The first measured metabolite concentration value When the ratio of the first measured metabolite concentration value to the normal cell or bodily fluid metabolite concentration value measured in advance at the first wave number is greater than or equal to a predetermined first ratio, And a first determination unit that executes a first determination step of determining that is false positive.

Further, the second determination unit that performs a determination operation before the determination operation by the first determination step of the first determination unit, wherein the measurement wave number is measured at a predetermined second wave number between 1680 and 1780 cm ^−1. When the predetermined peak waveform of the infrared absorption spectrum is not obtained at the second wave number for the second measured metabolite concentration value, the process proceeds to the first determination step executed by the first determination unit, It is preferable to further include a second determination unit that executes a second determination step of determining that the measured cell or body fluid is positive for the cancer when the peak waveform is obtained.

A first determination unit that performs a determination operation after the first determination step of the first determination unit determines that the cancer is false-positive, the first measurement being performed at a predetermined first wave number; Regarding the measured metabolite concentration value, a ratio of the first measured metabolite concentration value to a measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is a predetermined second ratio or less. In this case, it is preferable to include a third determination unit that executes a third determination step of determining that the cell or body fluid is false positive for cancer.

A first determination unit configured to perform a determination operation after it is determined in the first determination step of the first determination unit that the cancer is not false positive, the first measurement being performed at a predetermined first wave number; Regarding the measured metabolite concentration value, a ratio of the first measured metabolite concentration value to a measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is a predetermined third ratio or less. In this case, it is preferable to include a fourth determination unit that executes a fourth determination step of determining that the cell or body fluid is negative for cancer.

  The metabolite is preferably lactic acid.

  ADVANTAGE OF THE INVENTION According to this invention, the metabolite analysis method and metabolite analyzer which can determine easily the property of the metabolite of a bodily fluid or a cell using the transmission method in infrared spectroscopy can be provided.

It is sectional drawing which shows the whole structure of the metabolite analyzer 1 which concerns on one Embodiment of this invention. It is a graph which shows the waveform of an infrared absorption spectrum, (A) is a normal cell, (B) is a dense normal cell, (C) is a cancer cell, (D) shows the waveform of the infrared absorption spectrum of lactic acid. It is a graph. It is a graph which shows the waveform of the infrared absorption spectrum of lactic acid, Comprising: It is a graph at the time of changing pH value. It is a graph which shows the relationship (linearity) of the lactic acid density | concentration value and the light absorbency in an infrared absorption spectrum. It is a graph which shows the waveform of the infrared absorption spectrum of the metabolite of a cell, (A) is a cancer cell just before necrosis, (B) is a cancer cell, (C) is a dense normal cell, (D) is a normal cell. It is a graph which shows the waveform. It is a flowchart for determining the property of the metabolite of a cell.

  Hereinafter, embodiments of the present invention will be described with reference to FIGS. FIG. 1 is a cross-sectional view showing the overall configuration of a metabolite analyzer 1 according to an embodiment of the present invention. FIG. 2 is a graph showing a waveform of an infrared absorption spectrum, where (A) is a normal cell, (B) is a dense normal cell, (C) is a cancer cell, and (D) is an infrared absorption spectrum of lactic acid. It is a graph which shows the waveform. FIG. 3 is a graph showing the waveform of the infrared absorption spectrum of lactic acid when the pH value is changed. FIG. 4 is a graph showing the relationship (linearity) between the lactic acid concentration value and the absorbance in the infrared absorption spectrum. FIG. 5 is a graph showing a waveform of an infrared absorption spectrum of a metabolite of a cell, where (A) is a cancer cell just before necrosis, (B) is a cancer cell, (C) is a dense normal cell, (D ) Is a graph showing the waveform of normal cells.

  The metabolite analyzer 1 of the present embodiment uses the transmission method in infrared spectroscopy to determine the properties of metabolites of the cells 11 in the culture solution (aqueous solution, analysis target solution) 10 accommodated in the optical cell 2. It is a device that measures and analyzes living cells without damaging them (non-invasive). In the present embodiment, the metabolite of the cell 11 is, for example, lactic acid. Lactic acid exists in the form of a mixture of non-ionized and ionized lactic acid ions.

  As shown in FIG. 1, the metabolite analyzer 1 includes an optical cell 2 (cell), a holding plate 3 that holds the optical cell 2, a light source unit 4, a light detector 5 (light detection unit), and a control. Part 6. The optical cell 2 is held by the holding plate 3.

  The optical cell 2 includes an upper window plate 21 (window portion), a lower window plate 22 (window portion) disposed below the upper window plate 21 and spaced from the lower surface of the upper window plate 21, and an upper window plate. 21 and a spacer member 23 disposed between the upper window plate 21 and the lower window plate 22 at the edge of the lower window plate 22.

The upper window plate 21 and the lower window plate 22 are formed in a plate shape and are arranged in parallel to each other.
The upper window plate 21 and the lower window plate 22 are formed of a material that can transmit infrared light. For example, the upper window plate 21 and the lower window plate 22 may be any material that transmits infrared light in a wave number region of 1000 cm ⁻¹ or more. For example, in the present embodiment, the material of the upper window plate 21 and the lower window plate 22 is calcium fluoride. In addition, the material of the material of the upper window board 21 and the lower window board 22 is not limited to calcium fluoride, For example, diamond, zinc selenide (ZnSe), zinc sulfide (ZnS), etc. may be sufficient.

  A portion (space) surrounded by the upper window plate 21, the lower window plate 22 and the spacer member 23 constitutes the accommodating portion 20. The storage unit 20 forms a sealed space and stores the cells 11 and the culture solution 10 that are immersed in the culture solution 10.

The amount of oxygen and the amount of carbon dioxide for adjusting the pH value and the water temperature of the culture solution 10 stored in the storage unit 20 of the optical cell 2 are adjusted so that the living cells 11 can be cultured in a living state. It is a culture solution. The culture solution 10 is mainly composed of, for example, a medium (culture solution) in which 5 vol% concentration of CO ₂ gas is dissolved. Thus, the living cells 11 are cultured in a living state in the culture solution 10 inside the container 20.

  The optical cell 2 is irradiated with infrared light from the light source unit 4 (described later) from above to below. Infrared light penetrates the upper window plate 21 and the lower window plate 22 of the accommodating portion 20 in the vertical direction. Therefore, the distance between the lower surface of the upper window plate 21 and the upper surface of the lower window plate 22 in the housing portion 20 is the optical path length L of infrared light in infrared spectroscopy.

  The optical path length L is formed with a short length that is less affected by absorption of infrared spectroscopy into the culture medium 10 and allows measurement of the transmission method in infrared spectroscopy. In the present embodiment, the optical path length L is, for example, 12 μm. The optical path length L is not limited to 12 μm, and for example, a predetermined length between 0.5 and 50 μm is adopted. The optical path length L is set by, for example, the intensity of light from the light source unit 4.

The light source unit 4 is disposed above the optical cell 2. The light source unit 4 irradiates the cells 11 stored in the storage unit 20 with infrared light. The intensity of the light from the light source unit 4 is an intensity that can pass through the culture medium 10 and the cells 11 and does not affect the cells 11 such as damage.
The photodetector 5 is disposed below the optical cell 2. The photodetector 5 detects infrared light that has been irradiated from the light source unit 4 and transmitted through the cells 11 accommodated in the optical cell 2.
Infrared light emitted from the light source unit 4 passes through the upper window plate 21, the culture solution 10, the cells 11, and the lower window plate 22 and is detected by the photodetector 5. An infrared absorption spectrum signal by infrared light detected by the photodetector 5 is transmitted to the control unit 6.

From the viewpoint that lactic acid and cancer cells have the same tendency peak at a predetermined wave number in the infrared absorption spectrum, the present invention detects the absorbance in the infrared absorption spectrum at the predetermined wave number, and determines the cell properties. Judgment. In the present embodiment, the lactic acid that is a metabolite of the cell 11 accommodated in the accommodating portion 20 of the optical cell 2 is measured by measuring the absorbance of the lactic acid in the cell 11 using a transmission method in infrared spectroscopy. 11 properties are determined.
Hereinafter, the fact that the lactic acid concentration value of the cell can be used for the determination of the present invention will be described in detail.

Specifically, as a result of measuring the infrared absorption spectrum of a living cell (mouse: NIH3T3 strain, MCA205 strain) using an infrared spectrometer, as shown in FIG. 2, “(A) normal cell” , “(B) dense normal cells” (normal cells with high cell density) and “(C) cancer cells” have an absorption peak of Amide I (C═O stretching vibration) in the vicinity of 1660 cm ⁻¹ , and 1550 cm ^{− In the} vicinity of ¹ , there is an absorption peak of Amide II (N-H bending vibration). Dense normal cells are used as samples assuming false positives for cancer. The reason is as follows. Dense normal cells become oxygen deficient when the cells become dense, and lactic acid is observed as the metabolism changes from aerobic glycolysis to anaerobic glycolysis. Although it is essentially different from carcinogenesis, the absorption peak of the infrared absorption spectrum of dense normal cells means that the cells are constantly undergoing anaerobic glycolysis in the sense that the cells are growing abnormally. Gives the same result. Therefore, dense normal cells are used as samples assuming false positives for cancer.
Further, as shown in FIG. 2, in “(B) dense normal cells” and “(C) cancer cells”, there is a difference in the fine structure of the spectrum at a low wave number around 1040 to 1050 cm ^−1. It was confirmed that there was a peak attributed to -O stretching vibration.

Here, as a biological substance common to “(C) cancer cells” and “(B) dense normal cells”, which is a molecule corresponding to CO stretching vibrations in the vicinity of 1040 to 1050 cm ⁻¹ , “ (D) Lactic acid ”was listed as a candidate.
Usually, the characteristic C═O stretching vibration (wave number 1730 cm ⁻¹ ) used in the quantification of lactic acid by infrared absorption spectrum is changed to lactate ions in which protons are dissociated in a pH state close to that of a living body. Therefore, lactic acid should have reduced C = O absorption and CO ² -asymmetric stretching vibration (wave number 1580 cm ^-1 ). However, this absorption peak at a wave number of 1580 cm ⁻¹ is difficult to quantify because Amide II overlaps. However, it has been confirmed that C = O stretching vibration (wave number: 1730 cm ⁻¹ ) of lactic acid is observed when the pH is extremely lowered due to cell necrosis or the like. It has been demonstrated that it can be detected. Therefore, an attempt was made to determine the amount of lactic acid by the absorption peak in place of C═O, CO ²⁻ .

Main absorption peak of lactic acid as defined in JIS K 8726 (wave ^{number: 3000cm -1, 1740cm -1, 1460cm} -1, 1220cm -1, 1130cm -1 and 1040 cm ^-1) was added to all the absorption peaks observed Was verified in detail using computer simulation and actual measurement of molecular vibrations. As a result, as shown in FIG. 2, in “(D) lactic acid”, absorption at wave numbers 1040 to 1050 cm ⁻¹ does not overlap with absorption of biological substances (proteins, amino acids, nucleic acids, phosphates, etc.) and is independent. It was shown that it can be observed as an absorption peak.

  When the pH value of lactic acid is changed, an absorption peak appears at a predetermined wave number as shown in FIG. For convenience, the absorption peak shown in FIG. 1-No. The number of 12 is attached. In FIG. 3, the waveforms of a plurality of infrared absorption spectra are shown in one drawing for convenience so that the tendency of the absorption peak can be easily understood visually. The origin (0) of the waveform at each pH value is shown separately, and “0” (origin) is shown in the vicinity of the bottom value of each waveform in FIG.

As shown in FIG. 3, when the pH value of lactic acid is changed, the absorption near the wave number of 1050 cm ⁻¹ (peak No. 12 in FIG. 3) is in the pH region (pH value: 5 to 10) close to the living body. Since the absorption intensity does not depend on pH, it is suitable for use as a marker for lactic acid. When the pH value becomes 5 or less (tissue acidification due to cancer), the intensity of absorption near the wave number of 1050 cm ⁻¹ is relatively reduced. However, in this state, as shown in FIG. 3, in the vicinity of a wave number of 1730 cm ⁻¹ (peak No. 1 in FIG. 3), particularly in the vicinity of a pH value of 3.94 to 2.44, it is not dissociated into lactate ions. It has been experimentally verified that the C═O stretching vibration of the lactic acid molecule is manifested from the Amide I shoulder and appears as an absorption peak.

Here, the relationship between cancer cells and the amount of lactic acid will be supplementarily described.
When cells break down glucose to obtain ATP (adenosine triphosphate), it has two metabolic mechanisms, aerobic glycolysis and anaerobic glycolysis, and usually aerobic glycolysis is preferentially performed. Therefore, almost no lactic acid is produced. However, since cancer cells consume a large amount of energy and continue to proliferate, anaerobic glycolysis always increases, producing a large amount of lactic acid, and the pH value of the cells / tissues is lowered. This is called the Warburg effect and is known to be one of the characteristics of cancer cells / tissues. Therefore, conventionally, it is known that the lactic acid concentration of a predetermined cell / tissue or the pH value caused by the lactic acid concentration of a predetermined cell / tissue can be used as an indicator of whether or not the cancer has occurred. .
Applicants confirmed that MCA205, Colo26, and B16 mouse fibroblastoma cell lines, colon cancer cell lines, and melanoma cell lines confirmed the production of lactic acid, increased the cell density, and became normal cells that had become oxygen deficient ( In NIH3T3), the production of lactic acid by anaerobic glycolysis was also confirmed.

  As described above, it is found that there is a relationship between the cancer cells and the lactic acid concentration of lactic acid. Therefore, the applicant decided to use lactic acid as a metabolite of the cell 11 as a biological substance common to cancer cells and dense normal cells as a molecule corresponding to C—O stretching vibration. As a result, the infrared absorption spectrum of lactic acid was measured using the transmission method in infrared spectroscopy, and based on the absorbance, as shown below, in the control unit 6, cancer positive, cancer false positive, cancer Can be determined as negative.

  The control unit 6 controls the entire metabolite analyzer 1. The control unit 6 includes a memory. In the memory of the control unit 6, for example, a normal cell lactic acid concentration value NN (normal cell metabolite concentration value) measured in advance at a first wave number H1, which will be described later, and a determination threshold value in each determination step. Each ratio and various programs related to the operation of the metabolite analyzer 1 are stored. The control unit 6 includes a concentration measurement unit 61, a first determination unit 62, a second determination unit 63, a third determination unit 64, and a fourth determination unit 65.

  Based on the infrared absorption spectrum of the infrared light detected by the photodetector 5, the concentration measuring unit 61 absorbs the lactic acid of the cells 11 in the culture solution 10 contained in the optical cell 2 under the cell culture environment. Is measured by the transmission method in the infrared spectroscopy, thereby measuring the metabolite concentration value of the cell 11. In the present embodiment, the concentration measuring unit 61 measures the lactic acid concentration value as the metabolite concentration value of the metabolite of the cell 11.

Here, the relationship between the absorbance of infrared light measured by infrared spectroscopy and the lactic acid concentration value will be described.
As shown in the graph of FIG. 4, it has been confirmed by experiments that the infrared light absorbance and the lactic acid concentration value have linearity in the range where the lactic acid concentration value is 1000 mmol / L or less. For example, as shown in FIG. In the wave numbers where “1”, “10”, “9”, “8”, “12”, “5”, “3” exist, it is possible to determine whether there is a metabolic abnormality typified by lactic acidosis 5 In the range between ˜50 mmol / L, it has linearity and is quantifiable. In the present invention, the range between 5 and 50 mmol / L is the main determination range.
Therefore, in the present invention, the concentration measuring unit 61 can measure the absorbance of the infrared absorption spectrum and associate the absorbance with the lactic acid concentration value to measure the lactic acid concentration value of the cellular metabolite.

  The 2nd determination part 63 performs the positive determination process (2nd determination process) which determines positive of cancer. The second determination unit 63 performs the determination operation before the determination operation by the first false positive determination step (first determination step) (described later) in the first determination unit 62 (described later). In the positive determination step (second determination step) by the second determination unit 63, when a predetermined peak waveform is obtained at the second wave number H2 of the waveform of “(A) necrotic cancer cell” shown in FIG. Determine if cancer is positive. Specifically, the second determination unit 63 determines a predetermined infrared absorption spectrum at the second wave number H2 for the second measured lactate concentration value (second measured metabolite concentration value) measured at the second wave number H2. When the peak waveform is obtained, it is determined that the measured cell is positive for cancer, and when the predetermined peak waveform of the infrared absorption spectrum is not obtained at the second wave number H2, the first determination unit 62 It transfers to the 1st false positive determination process (1st determination process) to perform.

In the present embodiment, the second wave number H2 is a wave number attributed to stretching vibration. The second wave number H2 is a predetermined wave number between 1680 and 1780 cm ⁻¹ , preferably a predetermined wave number between 1710 and 1750 cm ⁻¹ , more preferably a predetermined wave number between 1715 and 1745 cm ^−1. Is the wave number. In the present embodiment, the second wave number H2 does not mean the wave number of the entire region between 1680 and 1780 cm ⁻¹ , but a predetermined specific one wave number such as 1730 cm ^−1. For example, it is a predetermined specific wave number at or near the peak of the absorption peak of the infrared absorption spectrum.

  Further, the second determination unit 63, for the determination of the peak waveform, for example, in the first derivative of the absorption spectrum of the determination region of the second wave number H2, when there is no point from positive to negative or from negative to positive, It is determined that there is no peak waveform. If there is a point from positive to negative or from negative to positive, it is determined that a peak waveform exists. In this case, when the peak waveform has a shoulder shape, it is determined that no peak waveform exists. Note that the method of determining the peak waveform is an example, and is not limited to this method.

  The 1st determination part 62 performs the 1st false positive determination process (1st determination process) which determines the false positive of cancer. The first false positive determination step in the first determination unit 62 is a step of rough (simple) determination of false positive of cancer. In the present embodiment, the first false positive determination step (first determination step) is a determination to perform screening (sieving) before determining in detail in a later determination step whether there is a possibility of cancer. It is a process.

  Specifically, the first determination unit 62 measures in advance the first measured lactic acid concentration value N1 (first measured metabolite concentration value) measured at the predetermined first wave number H1 at the first wave number H1. The ratio (N1 / NN) of the first measured lactic acid concentration value N1 (first measured metabolite concentration value) to the lactic acid concentration value NN (normal cell metabolite concentration value) of the normal cells is a predetermined first ratio. In the case of R1 or more, it is determined that the measured cell is a false positive cancer. The normal cell lactic acid concentration value NN and the predetermined first ratio R1 measured in advance are stored in a memory (not shown) of the control unit 6. The predetermined first ratio R1 is set to a lower limit ratio at which the cancer can be determined to be false positive for the measured cells.

In the present embodiment, the first wave number H1 is a wave number attributed to stretching vibration. The first wave number H1 is a predetermined wave number between 1020 and 1080 cm ⁻¹ , preferably a predetermined wave number between 1030 and 1070 cm ⁻¹ , more preferably a predetermined wave number between 1035 and 1065 cm ^−1. Is the wave number. In the present embodiment, the first wave number H1 does not mean the wave number of all regions between 1020 and 1080 cm ⁻¹ , but a predetermined specific one wave number such as 1050 cm ^−1. For example, it is a predetermined specific wave number at or near the peak of the absorption peak of the infrared absorption spectrum.

  The third determination unit 64 performs the second false positive determination step (third step) for determining whether the cancer is false positive after the first determination unit 62 determines that the cancer is false positive in the first determination step (first determination step). (Determination step) is executed. The second false positive determination step (third determination step) of the third determination unit 64 is the first false positive determination step (first determination) when it is determined to be false positive in the first false positive determination step (first determination step). This is a step of determining in detail the false positive of cancer after the step).

  The third determination unit 64 assumes that the cancer is determined to be false positive in the first false positive determination step (first determination step), and the first measured lactic acid concentration value measured at a predetermined first wave number H1. For N1 (first measured metabolite concentration value), the ratio of the first measured lactic acid concentration value N1 (first measured metabolite concentration value) to the measured reference concentration value NK measured at a predetermined reference value measured wavenumber HK. When (N1 / NK) is equal to or less than a predetermined second ratio R2, it is determined that the cancer is false positive for the cell. The predetermined second ratio R2 is stored in a memory (not shown) of the control unit 6. The predetermined second ratio R2 is set to an upper limit ratio at which it can be determined that the cancer is false positive for the measured cells.

  The fourth determination unit 65 performs a third false positive determination step (fourth determination) for determining whether the cancer is false positive after the first determination unit 62 determines that the cancer is not false positive in the first determination step (first determination step). (Determination step) is executed. The third false positive determination step (fourth determination step) of the fourth determination unit 65 is the first false positive determination step (first determination) when it is determined that the first false positive determination step (first determination step) is not false positive. This is a step of determining in detail the false positive of cancer after the step).

  The fourth determination unit 65 is executed after it is determined that the cancer is not false positive in the first false positive determination step (first determination step) of the first determination unit 62. The fourth determination unit 65 assumes that the cancer is determined not to be false positive in the first false positive determination step (first determination step), and the first measured lactic acid concentration value measured at a predetermined first wave number H1. For N1 (first measured metabolite concentration value), the ratio of the first measured lactic acid concentration value N1 (first measured metabolite concentration value) to the measured reference concentration value NK measured at a predetermined reference value measured wavenumber HK. When (N1 / NK) is equal to or less than a predetermined third ratio R3, it is determined that the cell is negative for cancer. The predetermined third ratio R3 is stored in a memory (not shown) of the control unit 6. As the predetermined third ratio R3, an upper limit ratio at which cancer can be determined to be negative for the measured cells is set.

In the present embodiment, the predetermined reference value measurement wave number HK used in the third determination unit 64 and the fourth determination unit 65 is a predetermined wave number between 1060 and 1120 cm ⁻¹ , preferably 1050 to 1110 cm ^−. A predetermined wave number between ^{1 and 1} , more preferably a predetermined wave number between 1045 and 1105 cm ⁻¹ . In the present embodiment, the reference value measurement wavenumber HK does not mean the number of waves of all areas between the 1060～1120Cm ^-1, it means such as a predetermined one particular wave number, for example, 1090 cm ^-1 For example, it is a predetermined specific wave number at or near the apex of the lactic acid concentration value of the absorption peak of the infrared absorption spectrum.

Here, the reason why the absorption peak at the reference value measurement wave number HK is adopted as the reference value in the determination by the third determination unit 64 and the fourth determination unit 65 is as follows.
The absorption peak at the reference value measurement wave number HK is the peak No. shown in FIG. 11 is an absorption peak at a wave number where 11 exists. The peak No. shown in FIG. 11 is also present as lactic acid alone. However, peak no. 11, when the absorption spectrum of a cell is measured, absorption due to C—O stretching vibration of other biomolecules present in large quantities is superimposed. Therefore, peak no. 11 makes it difficult to distinguish between normal cells and abnormal cells.
The applicant applied the peak No. shown in FIG. It was found that normal cells and abnormal cells can be distinguished by using 11 as a criterion for whether the amount of lactic acid is large or small. Therefore, in the determination of the third determination unit 64 and the fourth determination unit 65, the peak No. with less fluctuation when the pH value of lactic acid is changed. The lactic acid concentration value of the absorption peak at the wave number where 11 is present (reference value measurement wave number HK) is adopted as the reference value.

  Next, operation | movement of the metabolite analyzer 1 is demonstrated using the flowchart of FIG. FIG. 6 is a flow chart for determining the properties of cellular metabolites.

First, in step S1 shown in FIG. 6, the second determination unit 63 executes a positive determination process (second determination process). In the positive determination step, the presence or absence of an absorption peak waveform of the infrared absorption spectrum is determined at the second wave number H2. The second wave number H2 is a predetermined wave number between 1680 and 1780 cm ⁻¹ . For example, in the first derivative of the absorption spectrum in the determination region of the second wave number H2, the second determination unit 63 determines the peak when there is no point from positive to negative or from negative to positive. It is determined that the waveform does not exist. If there is a point from positive to negative or from negative to positive, it is determined that a peak waveform exists. In this case, when the peak waveform has a shoulder shape, it is determined that no peak waveform exists.

  If it is determined that the absorption peak waveform of the infrared absorption spectrum exists at the second wave number H2, the process proceeds to step S2, and the metabolite of the cell is determined to be “positive” in step S2, and the process ends. To do. On the other hand, when it is determined that there is no absorption peak waveform of the infrared absorption spectrum at the second wave number H2, the process proceeds to step S3.

In this embodiment, for example, for “(A) necrotic cancer cells” shown in FIG. 5, it is determined that an absorption peak waveform exists at the second wave number H2 (1740 cm ⁻¹ ), and the process proceeds to step S2. In step S2, the metabolite of the cell is determined as “positive”, and the process ends. On the other hand, there is no absorption peak waveform at the second wave number H2 (1740 cm ⁻¹ ) for “(B) cancer cells”, “(C) dense normal cells”, and “(D) normal cells” shown in FIG. Is determined, and the process proceeds to step S3.

In step S3, the concentration measuring unit 61 measures the first measured lactic acid concentration value N1 at the first wave number H1. The first wave number H1 is a predetermined wave number between 1020 and 1080 cm ⁻¹ . The concentration measuring unit 61 converts the absorbance to the lactic acid concentration value by measuring the absorbance in the infrared spectroscopy at the first wave number H1.

In step S4, the concentration measuring unit 61 measures the measurement reference concentration value NK at the reference value measurement wave number HK. The reference value measurement wave number HK is a predetermined wave number between 1060 and 1120 cm ⁻¹ . The concentration measuring unit 61 converts the absorbance into the lactic acid concentration value by measuring the absorbance in the infrared spectroscopy at the reference value measurement wavenumber HK.

  In step S5, the first determination unit 62 executes a first false positive determination step (first determination step). The first false positive determination step by the first determination unit 62 is a step of roughly (simplely) determining false positive of cancer. Whether the ratio (N1 / NN) of the first measured lactic acid concentration value N1 to the lactic acid concentration value NN of normal cells measured in advance at the first wave number H1 is equal to or greater than a predetermined first ratio R1. Determine whether or not. The predetermined first ratio R1 is set to a lower limit ratio at which the cancer can be determined to be false positive for the measured cells. If the ratio (N1 / NN) is greater than or equal to the first ratio R1 (YES), the process proceeds to step S6. When the ratio (N1 / NN) is less than the first ratio R1 (NO), the process proceeds to step S8.

  In the present embodiment, for “(B) cancer cells” shown in FIG. 5, the ratio (N1b / NNb) is equal to or higher than the first ratio R1 ((N1b / NNb) ≧ R1) (YES). The process proceeds to step S6. On the other hand, for “(C) dense normal cells” shown in FIG. 5, the ratio (N1c / NNc) is less than the first ratio R1 ((N1c / NNc) <R1) (NO), and the process proceeds to step S8. . For “(D) normal cells”, the ratio (N1d / NNd) is less than the first ratio R1 ((N1d / NNd) <R1) (NO), and the process proceeds to step S8.

  In step S6 when the ratio (N1 / NN) is equal to or higher than the first ratio R1 (YES) in step S5, the third determination unit 64 performs a second false positive determination (third determination step). The second false positive determination step (third determination step) of the third determination unit 64 is the first false positive when it is determined to be false positive in the first false positive determination step (first determination step) (YES in step S5). After the determination step (first determination step), it is a step of determining in detail whether a cancer is false positive.

  The third determination unit 64 determines whether or not the ratio (N1 / NK) of the first measured lactic acid concentration value N1 to the measurement reference concentration value NK measured at the reference value measurement wavenumber HK is equal to or less than a predetermined second ratio R2. judge. The predetermined second ratio R2 is set to an upper limit ratio at which it can be determined that the cancer is false positive for the measured cells. When the ratio (N1 / NK) is equal to or less than the second ratio R2 (YES), the process proceeds to step S7, and “suspected positive” for cancer is determined in step S7, and the process ends. When the ratio (N1 / NK) is greater than the second ratio R2 (NO), the process proceeds to step S7, where “positive” for cancer is determined in step S7, and the process ends.

Specifically, in the present embodiment, as shown in FIG. 5, in the case of “(B) cancer cell”, for example, the measurement reference concentration value NKb is measured by setting the reference value measurement wave number HK to 1087 cm ^−1. did. For example, the first measured lactic acid concentration value N1b was measured with the first wave number H1 set to 1045 cm ⁻¹ . In this case, the ratio (N1b / NKb) of the first measured lactic acid concentration value N1b to the measurement reference concentration value NKb is, for example, 1.00. Here, the ratio (N1 / NN) of the first measured lactic acid concentration value N1 to the lactic acid concentration value NN of normal cells measured in advance at the first wave number H1 in the first false positive determination step by the first determination unit 62 is predetermined. In the second false positive determination (third determination step) executed after it is determined that the ratio is equal to or higher than the first ratio R1, it is experimentally verified that the cancer is false positive when the second ratio R2 is equal to or lower than 0.70. ing. Therefore, for example, the threshold value of the second ratio R2 when the cancer is false positive is set to 0.70, and in the second false positive determination (third determination step), the third determination unit 64, for example, 3 False positive of cancer is determined when the ratio R3 is 0.70 or less. In the memory of the control unit 6, for example, 0.70 is stored as the threshold value of the second ratio R2.

  In this embodiment, for example, in the case of “(B) cancer cell” shown in FIG. 5, in step S6, the ratio of the first measured lactic acid concentration value N1b to the measurement reference concentration value NKb ((N1b / NKb) = 1). .00) is larger than the second ratio R2 (0.70) ((N1b / NKb)> 0.7 (R2)) (step S6, NO). As a result, the process proceeds to step S7, and “false positive” of the cancer is determined in step S7 and is terminated.

  In step S8 when the ratio (N1 / NN) is less than the first ratio R1 (NO) in step S5, the fourth determination unit 65 performs a third false positive determination (fourth determination step). The third false positive determination step (fourth determination step) of the fourth determination unit 65 is the first false positive when it is determined that the first false positive determination step (first determination step) is not false positive (NO in step S5). After the determination step (first determination step), it is a step of determining in detail whether a cancer is false positive.

  The fourth determination unit 65 determines whether the ratio (N1 / NK) of the first measured lactic acid concentration value N1 to the measurement reference concentration value NK measured at the reference value measurement wave number HK is equal to or less than the third ratio R3. . As the predetermined third ratio R3, an upper limit ratio at which cancer can be determined to be negative for the measured cells is set. When the ratio (N1 / NK) is equal to or less than the third ratio R3 (YES), the process proceeds to step S9, where “negative” of the cancer is determined in step S9, and the process ends. When the ratio (N1 / NK) is larger than the third ratio R3 (NO), the process proceeds to step S7, and “false positive” of the cancer is determined in step S7, and the process ends.

Specifically, in the present embodiment, as shown in FIG. 5, in the case of “(C) dense normal cells” and “(D) normal cells”, for example, the reference value measurement wave number HK is set to 1087 cm ^{−. As 1} , the measurement standard concentration values NKc and NKb were measured. Further, for example, the first measured lactic acid concentration values N1c and N1b were measured with the first wave number H1 set to 1045 cm ⁻¹ . In this case, in the case of “(C) dense normal cells” shown in FIG. 5, the ratio (N1c / NKc) of the first measured lactic acid concentration value N1c to the measurement reference concentration value NKc is, for example, 0.51. is there. In the case of “(D) normal cell” shown in FIG. 5, the ratio (N1d / NKd) of the first measured lactic acid concentration value N1d to the measurement reference concentration value NKd is 0.46.

  Here, the ratio (N1 / NN) of the first measured lactic acid concentration value N1 to the lactic acid concentration value NN of normal cells measured in advance at the first wave number H1 in the first false positive determination step by the first determination unit 62 is predetermined. In the third false positive determination (fourth determination step) executed after it is determined that the ratio is equal to or higher than the first ratio R1, for example, when the third ratio R3 is equal to or lower than 0.50, it is experimentally determined that the cancer is negative. It has been verified. Therefore, in the third false positive determination (fourth determination step), for example, the fourth determination unit 65 determines that the cancer is negative when the third ratio R3 is 0.50 or less. In the memory of the control unit 6, for example, 0.50 is stored as a threshold value of the third ratio R3.

In the present embodiment, for example, in the case of “(C) dense cells” shown in FIG. 5, the ratio of the first measured lactic acid concentration value N1c to the measurement reference concentration value NKc ((N1c / NKc) = 0.51). ) Is larger than the third ratio R3 (0.50) ((N1c / NKc) = 0.51> 0.50 (R3)) (step S8, NO). As a result, the process proceeds to step S7, and a false positive of cancer is determined in step S7, and the process ends.
In the case of “(D) normal cell” shown in FIG. 5, the ratio of the first measured lactic acid concentration value N1c to the measurement reference concentration value NKc ((N1c / NKc) = 0.46) is the third ratio R3. (0.50) or less ((N1c / NKc) = 0.46 ≦ 0.50 (R3)) (step S8, YES). Thereby, a process transfers to step S9, determines negative of cancer in step S9, and is complete | finished.

According to the metabolite analyzer 1 of the present embodiment, for example, the following effects are exhibited.
The metabolite analyzer 1 of the present embodiment has an upper window plate 21 and a lower window plate 22 that are capable of transmitting infrared light, accommodates a culture solution 10 of cells 11, and an optical path of infrared light in infrared spectroscopy. An optical cell 2 having a container 20 having a predetermined length of 0.5 to 50 μm, a light source unit 4 for irradiating infrared light to a cell 11 housed in the optical cell 2, and a light source Based on the photodetector 5 that detects infrared light irradiated from the unit 4 and transmitted through the cells 11 accommodated inside the optical cell 2, and the culture solution 10 based on the infrared light detected by the photodetector 5 A concentration measurement unit 61 that measures the lactic acid concentration value of the cell 11 by measuring the absorbance of the metabolite of the cell 11 in the infrared spectroscopy, and a predetermined wave number between 1020 and 1080 cm ^−1. First measured lactic acid concentration value N1 measured at the first wave number H1 The measured cells when the ratio (NN / N1) of the first measured lactic acid concentration value N1 to the normal cell metabolite concentration value NN measured in advance at the first wave number H1 is equal to or higher than the predetermined first ratio R1. A first determination unit 62 that executes a first false positive determination step (first determination step) for determining that the cancer is false positive.

  Therefore, the ratio of the first measured lactic acid concentration value N1 to the lactic acid concentration value NN of normal cells previously measured at the first wave number H1 using the optical cell 2 using the transmission method of infrared spectroscopy (NN / It can be determined that the cancer is false positive only by determining that N1) is equal to or greater than the first ratio R1. Thereby, it can be easily determined that the cancer is false positive for the cell. As a result, a false positive of cancer can be easily determined even in an early stage of canceration or an inflammatory state that cannot be said to be cancer.

  In addition, the metabolite analyzer 1 of the present embodiment has an upper window plate 21 and a lower window plate 22 that can transmit infrared light, and an optical path length of infrared light in infrared spectroscopy is 0.5 to 50 μm. The absorbance of the metabolite of the cell 11 in the culture solution 10 is measured by the transmission method in the infrared spectroscopy using the optical cell 2 including the storage unit 20 having a predetermined length between. Therefore, by using the transmission method in the infrared spectroscopy, the absorbance of the metabolite of the cell 11 in the culture solution 10 can be measured quantitatively with high accuracy rather than using the reflection method in the infrared spectroscopy. Can do.

Further, in the metabolite analyzer 1 of the present embodiment, the second determination unit 63 is configured to execute the determination operation before the determination operation by the first false positive determination step (first determination step) of the first determination unit 62. Then, the second determination unit 63 uses the predetermined infrared absorption spectrum at the second wave number H2 for the second measured lactic acid concentration value measured at the predetermined second wave number H2 between 1680 and 1780 cm ⁻¹ . When the peak waveform is not obtained, the process proceeds to the first false positive judgment step (first judgment step) executed by the first judgment unit 62, and when the peak waveform is obtained, the measured cell is positive for cancer. A positive determination step (second determination step) is performed.

  Therefore, when a predetermined peak waveform of the infrared absorption spectrum is obtained at the second wave number H2 before the determination operation by the first false positive determination step (first determination step) is performed, cancer is positive for the cell. It can be easily determined. Therefore, when it is determined that the cancer is positive in the positive determination step (second determination step), the first false positive determination step (first determination step) may not be performed, and the positive determination step (second determination step). The first false positive determination step (first determination step) can be executed only when it is determined that the cancer is not positive. Thereby, since it is not necessary to perform the 1st false positive determination process (1st determination process) about all the cells, determination operation can be simplified.

Further, in the metabolite analyzer 1 of the present embodiment, the third determination unit 64 that is executed after the first determination unit 62 determines that the cancer is false positive in the first false positive determination step (first determination step). The measurement reference concentration value NK measured at a predetermined reference value measurement wave number HK between the measurement wave numbers of 1060 to 1120 cm ⁻¹ for the first measured lactic acid concentration value N1 measured at the predetermined first wave number H1. When the ratio of the first measured lactic acid concentration value N1 to N1 (N1 / NK) is equal to or less than the predetermined second ratio R2, the second false positive determination step (third determination step) for determining that the cancer is false positive for the cell is executed. A third determination unit 64 is provided.
For this reason, in the first false positive determination step (first determination step), the cells determined to be false positive in the cancer are determined in more detail, so that the cancer is false positive or positive in detail with high accuracy. Can be easily determined.

In the metabolite analyzer 1 of the present embodiment, the fourth determination unit 65 is executed after it is determined in the first false positive determination step (first determination step) of the first determination unit 62 that the cancer is not false positive. The measurement reference concentration value NK measured at a predetermined reference value measurement wave number HK between the measurement wave numbers of 1060 to 1120 cm ⁻¹ for the first measured lactic acid concentration value N1 measured at the predetermined first wave number H1. When the ratio (N1 / NK) of the first measured lactic acid concentration value N1 to the predetermined third ratio R3 is less than or equal to the predetermined third ratio R3, a third false positive determination step (fourth determination step) for determining that the cell is negative for cancer is performed A fourth determination unit 65 is provided.
Therefore, in the first false positive determination step (first determination step), by determining in more detail about the cells for which the cancer is determined not to be false positive, the cancer is false positive or negative for the cells with high accuracy. Can be easily determined.

As mentioned above, although preferred embodiment was described, this invention can be implemented with a various form, without being limited to embodiment mentioned above.
For example, in the above-described embodiment, the determination is performed by measuring the concentration of the metabolite of the cell, but the present invention is not limited to this. For example, the determination may be made by measuring the concentration of bodily fluids such as saliva, sweat, urine, blood and plasma. The concentration of the body fluid may be measured by measuring the concentration of the body fluid itself, or by measuring the concentration of the aqueous solution containing the body fluid in a state where the body fluid is thinned.

  Moreover, in the said embodiment, although the metabolite of the cell utilized for determination of this invention was lactic acid, it is not limited to this. The cellular metabolite used in the determination of the present invention may be a substance other than lactic acid. Examples of cellular metabolites include pyruvate, adenosine triphosphate, adenosine diphosphate, and the like.

  In the said embodiment, although the positive of cancer, the false positive, and negative were determined, it is not limited to what performs the determination of canceration. What is necessary is just to judge the property of a cell metabolite or a body fluid.

  In the embodiment, in the metabolite analyzer, the first determination unit 62 determines the peak waveform, and the second determination unit 63 to the fourth determination unit 65 perform the determination based on the ratio of the metabolite concentration values. However, the present invention is not limited to this. Instead of the determination performed by the first determination unit 62 to the fourth determination unit 65 in the metabolite analyzer, a person may perform the determination.

In the embodiment, the reference ratio value used for the determination by the third determination unit 64 and the fourth determination unit 65 is the second ratio R2 based on the measurement reference concentration value NK measured at the reference value measurement wavenumber HK. Alternatively, the determination is performed by calculating the third ratio R3, but the present invention is not limited to this. For example, the reference density value used for the determination of the third determination unit 64 and a fourth judging unit 65, and the absorption peak of AmideI existing near 1660 cm ^-1, such as the absorption peak of AmideII existing near 1550 cm ^-1 A based value can also be used.

In the said embodiment, although the 1st wave number was made into the predetermined wave number between 1020-1080cm < ^-1 >, it is not limited to this. The first wave number may be any wave number belonging to the stretching vibration. In the mid-infrared region measured by infrared spectroscopy, the spectrum intensity due to stretching vibration is stronger than the spectrum intensity due to bending vibration. Therefore, it is important in the present invention to measure the absorbance at the wave number attributed to the stretching vibration.

  In the above embodiment, the culture solution 10 and the cell 11 of the cell 11 are accommodated in the accommodation unit 20 of the optical cell 2 and the absorbance of the metabolite of the cell 11 in the culture solution 10 is measured. However, the present invention is not limited to this. . Only the culture solution 10 from which the cells 11 have been taken out may be stored in the storage unit 20 of the optical cell 2, and the absorbance of the metabolites of the cells 11 in the culture solution 10 may be measured.

  In the said embodiment, although the metabolite concentration value of the living cell 11 in the culture solution 10 was measured, it is not limited to this. Metabolite concentration values of aqueous solutions of cells other than the culture solution may be measured. Examples of the aqueous solution of cells other than the culture solution include an aqueous solution in which dead cells that cannot be cultured are immersed. Further, for example, the concentration of a metabolite in an analysis solution such as a nutrient solution that is not a culture solution before immersing cells or a physiological saline used for calibration of a metabolite analyzer may be measured. At the time of calibration of the metabolite analyzer, the metabolite analyzer can be calibrated by measuring that the metabolite concentration value in the physiological saline is zero and the first ratio being zero.

1 Metabolite Analyzer 2 Optical Cell (Cell)
4 Light source 5 Photodetector (photodetector)
10 Culture solution (aqueous solution, analysis target solution)
11 cells 20 housing part 21 upper window plate (window part)
22 Lower window plate (window)
61 Concentration Measurement Unit 62 First Determination Unit 63 Second Determination Unit 64 Third Determination Unit 65 Fourth Determination Unit H1 First Wave Number HK Reference Value Measurement Wave Number L Optical Path Length N1 First Measurement Lactate Concentration Value (First Measurement Metabolite Concentration Value value)
NN Normal cell lactate concentration value (normal cell metabolite concentration value)
NK Measurement standard concentration value R1 1st ratio R2 2nd ratio R3 3rd coverage

Claims (11)

An accommodation part having a window part capable of transmitting infrared light, containing an analysis target liquid such as an aqueous solution of a cell or body fluid, and an optical path length of infrared light in infrared spectroscopy is between 0.5 and 50 μm By measuring the absorbance of the metabolite of the cell in the aqueous solution or the absorbance of the body fluid by the transmission method in the infrared spectroscopy using the cell having the container of the predetermined length, the metabolite concentration value of the cell or body fluid Make measurements,
For the first measured metabolite concentration value measured at a predetermined first wave number where the measured wave number belongs to the stretching vibration, the first relative to the metabolite concentration value of normal cells or body fluids measured in advance at the first wave number. A metabolite analysis method comprising a first determination step of determining a property of a measured metabolite or body fluid of a cell when a ratio of measured metabolite concentration values is a predetermined first ratio or more. An accommodation part having a window part capable of transmitting infrared light, containing an analysis target liquid such as an aqueous solution of a cell or body fluid, and an optical path length of infrared light in infrared spectroscopy is between 0.5 and 50 μm By measuring the absorbance of the metabolite of the cell in the aqueous solution or the absorbance of the body fluid by the transmission method in the infrared spectroscopy using the cell having the container of the predetermined length, the metabolite concentration value of the cell or body fluid Make measurements,
With respect to the first measured metabolite concentration value measured at a predetermined first wave number between 1020 and 1080 cm ⁻¹ of the measured wave number, the metabolite concentration value of normal cells or body fluids measured in advance at the first wave number A metabolite analysis method comprising a first determination step of determining that a measured cell or body fluid is false positive for a measured cell or body fluid when the ratio of the first measured metabolite concentration value is a predetermined first ratio or more. The second determination step is performed before the first determination step, and the second measurement metabolite concentration value measured at a predetermined second wave number between 1680 and 1780 cm ⁻¹ is the first determination step. When a predetermined peak waveform of an infrared absorption spectrum is not obtained at two wave numbers, the process proceeds to the first determination step, and when the peak waveform is obtained, cancer is positive for the measured cell or body fluid. The metabolite analysis method according to claim 2, further comprising a second determination step of determining. The third determination step is executed after it is determined that the cancer is false positive in the first determination step, and the measured wave number of the first measured metabolite concentration value measured at the predetermined first wave number is When the ratio of the first measured metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is equal to or less than the predetermined second ratio, cancer is detected for the cell or body fluid. The metabolite analysis method according to claim 2 or 3, further comprising a third determination step of determining that it is a false positive. The fourth determination step is executed after it is determined in the first determination step that the cancer is not false positive, and the measured wave number is measured for the first measured metabolite concentration value measured at the predetermined first wave number. When the ratio of the first measured metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is equal to or lower than a predetermined third ratio, cancer is detected for the cell or body fluid. The metabolite analysis method according to claim 2 or 3, further comprising a fourth determination step of determining negative.   The metabolite analysis method according to claim 1, wherein the metabolite is lactic acid. An accommodation part having a window part capable of transmitting infrared light, containing an analysis target liquid such as an aqueous solution of a cell or body fluid, and an optical path length of infrared light in infrared spectroscopy is between 0.5 and 50 μm A cell provided with an accommodating portion of a predetermined length;
A light source unit for irradiating the body fluid or cells contained in the cell with infrared light;
A light detection unit that detects infrared light irradiated from the light source unit and transmitted through cells or body fluids contained in the cell;
Based on the infrared light detected by the light detection unit, the absorbance of the metabolite of the cell or body fluid is measured by measuring the absorbance of the metabolite of the cell in the aqueous solution or the absorbance of the body fluid by the transmission method in infrared spectroscopy. A concentration measuring unit for measuring,
With respect to the first measured metabolite concentration value measured at a predetermined first wave number between 1020 and 1080 cm ⁻¹ of the measured wave number, the metabolite concentration value of normal cells or body fluids measured in advance at the first wave number A first determination unit that executes a first determination step of determining that the measured cell or body fluid is false positive for the measured cell or body fluid when the ratio of the first measured metabolite concentration value is equal to or greater than a predetermined first ratio. Product analyzer. A second determination unit that performs a determination operation before the determination operation by the first determination step of the first determination unit, the measured wave number being measured at a predetermined second wave number between 1680 and 1780 cm ⁻¹ . For the second measured metabolite concentration value, when a predetermined peak waveform of the infrared absorption spectrum is not obtained at the second wave number, the process proceeds to the first determination step executed by the first determination unit, and the peak waveform The metabolite analyzer according to claim 7, further comprising a second determination unit that executes a second determination step of determining that the measured cell or body fluid is positive for cancer when obtained. A third determination unit that executes a determination operation after it is determined that the cancer is false positive in the first determination step of the first determination unit, the first measured metabolism measured at a predetermined first wave number; For the product concentration value, the ratio of the first measured metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is less than or equal to a predetermined second ratio The metabolite analyzer according to claim 7 or 8, further comprising a third determination unit that executes a third determination step of determining that the cancer is false positive for cells or body fluids. The fourth determination unit that performs a determination operation after it is determined that the cancer is not false positive in the first determination step of the first determination unit, the first measurement metabolism measured at a predetermined first wave number For the product concentration value, the ratio of the first measured metabolite concentration value to the measurement reference concentration value measured at a predetermined reference value measurement wave number between 1060 and 1120 cm ⁻¹ is less than or equal to a predetermined third ratio The metabolite analyzer according to claim 7 or 8, further comprising a fourth determination unit that executes a fourth determination step of determining that cancer is negative for a cell or body fluid.   The metabolite analyzer according to any one of claims 7 to 10, wherein the metabolite is lactic acid.

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