阅读说明：本技术 成分分析方法以及成分分析装置 (Component analysis method and component analysis device ) 是由 吉田迅 于 2019-08-21 设计创作，主要内容包括：本发明涉及成分分析方法以及成分分析装置。在使用了连续试样导入的成分分析系统中,即使在微分波形中难以明确确定两个峰之间的边界的情况下也明确确定其边界。包括：测定工序,在流路内分离被连续导入到流路的试样溶液,并随时间对位于流路测定部的试样溶液进行光学测定而得到光学测定值；分析工序,基于光学测定值,分析试样中所含的多个成分,分析工序包括：原始波形获取工序,在二维平面上沿时间轴绘制光学测定值而获取原始波形；测定值微分工序,获取作为沿与时间轴正交的光学测定值的轴将对原始波形进行微分而得到的波形的测定值微分波形；测定值边界决定工序,将与该测定值微分波形的峰顶对应的光学测定值作为多个成分彼此之间的分离边界。(The present invention relates to a component analysis method and a component analysis apparatus. In a component analysis system using continuous sample introduction, even when it is difficult to clearly specify the boundary between two peaks in a differential waveform, the boundary is clearly specified. The method comprises the following steps: a measurement step of separating the sample solution continuously introduced into the channel in the channel and optically measuring the sample solution at the channel measurement unit over time to obtain an optical measurement value; an analysis step of analyzing a plurality of components contained in a sample based on the optical measurement value, the analysis step including: an original waveform acquisition step of obtaining an original waveform by plotting an optical measurement value on a two-dimensional plane along a time axis; a measurement value differentiating step of acquiring a measurement value differential waveform which is a waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and a measurement value boundary determining step of defining an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.)

1. A method of component analysis, comprising:

a measurement step of separating a sample solution continuously introduced into a flow path into a plurality of components in the flow path, and optically measuring the sample solution at a measurement position of the flow path over time to obtain an optically measured value; and

an analysis step of analyzing the plurality of components contained in the sample based on the optical measurement value,

the analysis process comprises:

an original waveform acquisition step of obtaining an original waveform by plotting the optical measurement values along a time axis on a two-dimensional plane;

a measurement value differentiating step of acquiring a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and

and a measurement value boundary determining step of setting an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.

2. The composition analyzing method according to claim 1,

the analyzing process further includes:

a time differentiation step of acquiring a time-differentiated waveform obtained by differentiating the original waveform along the time axis; and

and an integration quantitative step of integrating the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, with a time point corresponding to the separation boundary being an integration boundary, to obtain a value, and calculating the value as a relative content of the component corresponding to the integration section in the sample.

3. The composition analyzing method according to claim 1,

the analyzing process further includes:

and a displacement quantifying step of calculating a distance between sections having the separation boundaries adjacent to each other along the axis of the optically measured value as a relative content of the component corresponding to the section in the sample.

4. A method of component analysis, comprising:

a measurement step of separating a sample solution continuously introduced into a flow path into a plurality of components in the flow path, and optically measuring the sample solution at a measurement position of the flow path over time to obtain an optically measured value; and

an analysis step of analyzing the plurality of components contained in the sample based on the optical measurement value,

the analysis process comprises:

an original waveform acquisition step of obtaining an original waveform by plotting the optical measurement values along a time axis on a two-dimensional plane;

a time differentiation step of acquiring a time-differentiated waveform obtained by differentiating the original waveform along the time axis;

an inverse differentiation step of acquiring an inverse differential waveform in which the reciprocal of the time differential waveform is plotted along the time axis; and

and a time boundary determining step of setting a time point corresponding to a peak top of the inverse differential waveform as a separation boundary between the plurality of components.

5. The composition analyzing method according to claim 4,

the analyzing process further includes:

and an integration quantitative step of integrating the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, with a time point corresponding to the separation boundary being an integration boundary, to obtain a value, and calculating the value as a relative content of the component corresponding to the integration section in the sample.

6. The composition analyzing method according to any one of claims 1 to 5,

the optical measurement value is obtained by a capillary electrophoresis method in which the sample solution is separated by applying a voltage to the sample solution continuously introduced into a capillary as the flow path.

7. A composition analyzing apparatus comprising:

a flow path for continuously introducing a sample solution;

a measurement unit that optically measures the sample solution separated into a plurality of components in the flow path at a measurement position of the flow path over time to obtain an optical measurement value; and

an analysis unit for analyzing the plurality of components contained in the sample based on the optical measurement value,

the analysis section includes:

an original waveform acquiring unit that acquires an original waveform by plotting the optical measurement value along a time axis on a two-dimensional plane;

a measurement value differentiating unit configured to acquire a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and

and a measurement value boundary determination unit configured to determine an optical measurement value corresponding to a peak top of the differential waveform of the measurement value as a separation boundary between the plurality of components.

8. The composition analyzing apparatus according to claim 7,

the analysis section further includes:

a time differentiation unit that acquires a time-differentiated waveform that is a waveform obtained by differentiating the original waveform along the time axis; and

and an integration quantifying unit configured to integrate the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, using a time point corresponding to the separation boundary as an integration boundary, and calculate a value of the integration boundary as a relative content of the component corresponding to the integration section in the sample.

9. The composition analyzing apparatus according to claim 7,

the analysis section further includes:

and a displacement determining unit that calculates a distance between sections having the separation boundaries adjacent to each other along the axis of the optically measured value as a relative content of the component corresponding to the section in the sample.

10. A composition analyzing apparatus comprising:

a flow path for continuously introducing a sample solution;

a measurement unit that optically measures the sample solution separated into a plurality of components in the flow path at a measurement position of the flow path over time to obtain an optical measurement value; and

an analysis unit for analyzing the plurality of components contained in the sample based on the optical measurement value,

the analysis section includes:

an original waveform acquiring unit that acquires an original waveform by plotting the optical measurement value along a time axis on a two-dimensional plane;

a time differentiation unit that acquires a time-differentiated waveform that is a waveform obtained by differentiating the original waveform along the time axis;

an inverse differential unit that acquires an inverse differential waveform in which an inverse of the time differential waveform is plotted along the time axis; and

and a time boundary determining unit configured to determine a time point corresponding to a peak of the inverse differential waveform as a separation boundary between the plurality of components.

11. The composition analyzing apparatus according to claim 10,

the analysis section further includes:

and an integration quantifying unit configured to integrate the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, using a time point corresponding to the separation boundary as an integration boundary, and calculate a value of the integration boundary as a relative content of the component corresponding to the integration section in the sample.

Technical Field

The present invention relates to a component analysis method and a component analysis apparatus using continuous sample introduction.

Background

The method comprises the following steps: in a component analysis system using continuous sample introduction such as capillary electrophoresis, a curve is obtained with detection data such as absorbance obtained by a detector as a vertical axis and time as a horizontal axis, the curve is created as an original waveform, and component analysis is performed using a differential waveform such as an electropherogram obtained by differentiating the original waveform with respect to time.

Each peak appearing in the differential waveform corresponds to each component contained in the introduced sample. Further, the composition can be determined by the difference in time at which the top of each peak is observed. Furthermore, the area of each peak in the differential waveform is an index of the content of the component in the sample. For example, a differential waveform of a hemoglobin measurement system that introduces blood as a continuous sample of a sample has a shape shown in patent document 1 below.

Disclosure of Invention

Problems to be solved by the invention

In a component analysis system based on continuous sample introduction such as capillary electrophoresis, as described above, a component is identified by a peak recognized in a differential waveform obtained by differentiating a absorbance curve along a time axis, and relative quantification of the component is performed based on an area occupied by the peak in the differential waveform. In this case, a valley portion (referred to as a "valley bottom") occurring between peaks is often defined as a boundary between the peaks, but the valley bottom is often unclear. In particular, when two peaks are fused, the lower of the two peaks is absorbed by the higher peak, and it is sometimes difficult to identify the two peaks, in which case it is difficult or impossible to determine the bottom of the valley.

An object of an embodiment of the present invention is to enable a boundary between two peaks to be clearly specified even when it is difficult to clearly specify the boundary in a differential waveform in a component analysis system using continuous sample introduction.

Means for solving the problems

In a first aspect of the present disclosure, a method of analyzing a component includes: a measurement step of separating a sample solution continuously introduced into a flow path into a plurality of components in the flow path, and optically measuring the sample solution at a measurement position of the flow path over time to obtain an optically measured value; and an analyzing step of analyzing the plurality of components contained in the sample based on the optical measurement value, the analyzing step including: an original waveform acquisition step of obtaining an original waveform by plotting the optical measurement values along a time axis on a two-dimensional plane; a measurement value differentiating step of acquiring a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and a measurement value boundary determining step of setting an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.

In a second aspect of the present disclosure, in the first aspect, the analyzing step further includes: a time differentiation step of acquiring a time-differentiated waveform obtained by differentiating the original waveform along the time axis; and an integration quantification step of integrating the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, using a time point corresponding to the separation boundary as an integration boundary, to obtain a value, and calculating the value as a relative content of the component corresponding to the integration section in the sample.

In a third aspect of the present disclosure, in the first aspect, the analyzing step further includes: and a displacement quantifying step of calculating a distance between sections having the separation boundaries adjacent to each other along the axis of the optically measured value as a relative content of the component corresponding to the section in the sample.

In a fourth aspect of the present disclosure, a method of analyzing a component includes: a measurement step of separating a sample solution continuously introduced into a flow path into a plurality of components in the flow path, and optically measuring the sample solution at a measurement position of the flow path over time to obtain an optically measured value; and an analyzing step of analyzing the plurality of components contained in the sample based on the optical measurement value, the analyzing step including: an original waveform acquisition step of obtaining an original waveform by plotting the optical measurement values along a time axis on a two-dimensional plane; a time differentiation step of acquiring a time-differentiated waveform obtained by differentiating the original waveform along the time axis; an inverse differentiation step of acquiring an inverse differential waveform in which the reciprocal of the time differential waveform is plotted along the time axis; and a time boundary determining step of setting a time point corresponding to a peak top of the inverse differential waveform as a separation boundary between the plurality of components.

In a fifth aspect of the present disclosure, in the fourth aspect, the analyzing step further includes: and an integration quantitative step of integrating the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, with a time point corresponding to the separation boundary being an integration boundary, to obtain a value, and calculating the value as a relative content of the component corresponding to the integration section in the sample.

In a sixth aspect of the present disclosure, in any one of the first to fifth aspects, the optical measurement value is obtained by a capillary electrophoresis method in which the sample solution is separated by applying a voltage to the sample solution continuously introduced into a capillary as the flow path.

In a seventh aspect of the present disclosure, a component analysis apparatus includes: a flow path for continuously introducing a sample solution; a measurement unit that optically measures the sample solution separated into a plurality of components in the flow path at a measurement position of the flow path over time to obtain an optical measurement value; and an analysis unit that analyzes the plurality of components contained in the sample based on the optical measurement value, the analysis unit including: an original waveform acquiring unit that acquires an original waveform by plotting the optical measurement value along a time axis on a two-dimensional plane; a measurement value differentiating unit configured to acquire a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and a measurement value boundary determination unit configured to determine an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.

In an eighth aspect of the present disclosure, in the seventh aspect, the analysis unit further includes: a time differentiation unit that acquires a time-differentiated waveform that is a waveform obtained by differentiating the original waveform along the time axis; and an integration quantifying unit configured to integrate the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, using a time point corresponding to the separation boundary as an integration boundary, and calculate a value of the integration boundary as a relative content of the component corresponding to the integration section in the sample.

In a ninth aspect of the present disclosure, in the seventh aspect, the analysis unit further includes: and a displacement determining unit that calculates a distance between sections having the separation boundaries adjacent to each other along the axis of the optically measured value as a relative content of the component corresponding to the section in the sample.

In a tenth aspect of the present disclosure, a component analysis apparatus includes: a flow path for continuously introducing a sample solution; a measurement unit that optically measures the sample solution separated into a plurality of components in the flow path at a measurement position of the flow path over time to obtain an optical measurement value; and an analysis unit that analyzes the plurality of components contained in the sample based on the optical measurement value, the analysis unit including: an original waveform acquiring unit that acquires an original waveform by plotting the optical measurement value along a time axis on a two-dimensional plane; a time differentiation unit that acquires a time-differentiated waveform that is a waveform obtained by differentiating the original waveform along the time axis; an inverse differential unit that acquires an inverse differential waveform in which an inverse of the time differential waveform is plotted along the time axis; and a time boundary determining unit configured to determine a time point corresponding to a peak top of the inverse differential waveform as a separation boundary between the plurality of components.

In a tenth aspect of the component analysis device according to the eleventh aspect of the present disclosure, the analysis unit further includes: and an integration quantifying unit configured to integrate the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, using a time point corresponding to the separation boundary as an integration boundary, and calculate a value of the integration boundary as a relative content of the component corresponding to the integration section in the sample.

Effects of the invention

In the embodiment of the present invention, in the component analysis system using continuous sample introduction, even when it is difficult to clearly specify the boundary between two peaks in the differential waveform, the boundary can be clearly specified.

Drawings

Fig. 1 is a system schematic diagram showing a component analysis system that can be used to execute the component analysis method of the present embodiment;

FIG. 2 is a top view showing an analysis chip used in the analysis system of FIG. 1;

FIG. 3 is a sectional view taken along line III-III of FIG. 2;

fig. 4 is a block diagram showing a hardware configuration of the control section;

FIG. 5 is a block diagram showing a functional configuration of a component analysis apparatus;

fig. 6 is a block diagram showing a functional configuration of an analysis unit in the first embodiment;

fig. 7 is a flowchart showing an outline of a component analysis method in the first embodiment;

FIG. 8 shows an example of an original waveform with a solid line;

FIG. 9 shows, in dashed lines, a time-differentiated waveform relative to the original waveform of FIG. 8;

fig. 10 is a graph in which the measured value differential waveform with respect to the original waveform is added by a dotted line in fig. 9;

fig. 11 is a block diagram showing a functional configuration of an analysis unit in the second embodiment;

fig. 12 is a flowchart showing an outline of a component analysis method in the second embodiment;

fig. 13 is a block diagram showing a functional configuration of an analysis unit according to a third embodiment;

fig. 14 is a flowchart showing an outline of a component analysis method in the third embodiment;

FIG. 15 shows in dotted lines a measured value differential waveform relative to the original waveform of FIG. 8;

fig. 16 is a block diagram showing a functional configuration of an analysis unit according to a fourth embodiment;

fig. 17 is a flowchart showing an outline of a component analysis method in the fourth embodiment;

fig. 18 is a diagram showing an inverse differential waveform with respect to the time differential waveform in fig. 9 added by a dotted line;

fig. 19 is a block diagram showing a functional configuration of an analysis unit in the fifth embodiment;

fig. 20 is a flowchart showing an outline of a component analysis method in the fifth embodiment;

fig. 21 is a diagram showing the influence of the ambient temperature and the concentration of the specimen during measurement on the time-differentiated waveform obtained in the time-differentiating step, where fig. 21 (a) shows a case where the ambient temperature is 23 ℃ and the sample has a low concentration, fig. 21 (B) shows a case where the ambient temperature is 23 ℃ and the sample has a high concentration, fig. 21 (C) shows a case where the ambient temperature is 8 ℃ and the sample has a low concentration, and fig. 21 (D) shows a case where the ambient temperature is 8 ℃ and the sample has a high concentration;

fig. 22 is a diagram showing the influence of the ambient temperature and the concentration of the specimen during measurement on the time-differentiated waveform obtained in the time-differentiating step, where fig. 22 (a) shows the case of a low-concentration sample at an ambient temperature of 23 ℃, fig. 22 (B) shows the case of a high-concentration sample at an ambient temperature of 23 ℃, fig. 22 (C) shows the case of a low-concentration sample at an ambient temperature of 8 ℃, and fig. 22 (D) shows the case of a high-concentration sample at an ambient temperature of 8 ℃.

Detailed Description

Hereinafter, embodiments of the present disclosure will be described with reference to appropriate drawings.

< ingredient analysis System >

Fig. 1 shows a schematic configuration of a component analysis system a1 that can be used to execute the component analysis method according to the present embodiment. The component analysis system a1 is configured to include a component analyzer 1 and an analysis chip 2. The component analysis system a1 is a system for executing an analysis method by capillary electrophoresis for the sample Sa. The form of the sample Sa is not limited as long as it contains a component that can be analyzed by capillary electrophoresis and is soluble in such a solvent, but in the present embodiment, blood collected from a human body will be described as an example.

Examples of the components included in the sample Sa include hemoglobin (Hb), albumin (Alb), globulin (α, α, β, γ globulin), fibrinogen, and the like, and examples of the hemoglobin include various hemoglobin species such as normal hemoglobin (HbA), variant hemoglobin (HbA1c, HbC, HbD, HbE, HbS, and the like), and fetal hemoglobin (HbF).

Depending on the component to be analyzed, the sample may be previously treated with an appropriate reagent, or a preliminary separation process may be performed in advance by another method (for example, chromatography).

The analysis chip 2 holds the sample Sa and provides an analysis site for the sample Sa in a state of being loaded in the component analysis apparatus 1. In the present embodiment, the analysis chip 2 is configured as a disposable analysis chip which is intended to be discarded after completion of a single analysis. As shown in fig. 2 and 3, the analysis chip 2 includes a main body 21, a mixing tank 22, an introduction tank 23, a filter 24, a discharge tank 25, an electrode tank 26, a flow path 27, and a communication flow path 28. Fig. 2 is a plan view of the analysis chip 2, and fig. 3 is a cross-sectional view taken along line III-III of fig. 2. The analysis chip 2 is not limited to a disposable analysis chip, and may be an analysis chip for a plurality of analyses. The component analysis system according to the present embodiment is not limited to a configuration including the independent analysis chip 2 loaded in the component analysis device 1, and may be a configuration in which a functional portion that performs the same function as the analysis chip 2 is incorporated in the component analysis device 1.

The main body 21 is a base of the analysis chip 2, and the material thereof is not particularly limited, and examples thereof include glass, fused silica, and plastic. In the present embodiment, the main body 21 is configured such that the upper portion 2A and the lower portion 2B in fig. 3 are formed independently and they are joined to each other. Further, the present invention is not limited to this, and the main body 21 may be integrally formed, for example.

The mixing tank 22 is a portion where the sample Sa and the diluent Ld are mixed. The mixing tank 22 is formed as a recess opening upward through a through hole formed in the upper portion 2A of the main body 21, for example. The introduction tank 23 is a tank for introducing a sample solution obtained by mixing the sample Sa and the diluent Ld in the mixing tank 22. The introduction groove 23 is formed as a recess opening upward through a through hole formed in the upper portion 2A of the main body 21, for example.

The filter 24 is provided in an opening of the introduction tank 23 as an example of an introduction path to the introduction tank 23. The specific structure of the filter 24 is not limited, and a preferable example thereof is a cellulose acetate membrane filter (manufactured by ADVANTEC, Inc., pore size 0.45 μm).

The discharge groove 25 is a groove located on the downstream side of the flow path 27. The discharge groove 25 is formed as a recess opening upward through a through hole formed in the upper portion 2A of the main body 21, for example. The electrode well 26 is a well into which the electrode 31 is inserted by capillary electrophoresis. The electrode groove 26 is formed as a recess opening upward through a through hole formed in the upper portion 2A of the main body 21, for example. The communication passage 28 connects the introduction groove 23 and the electrode groove 26, and constitutes a conduction path between the introduction groove 23 and the electrode groove 26.

The flow path 27 is a capillary tube connecting the introduction tank 23 and the discharge tank 25, and is a place where an electroosmotic flow (EOF) occurs in the electrophoresis method. The flow path 27 is configured as a groove formed in the lower portion 2B of the main body 21, for example. Further, the main body 21 may be formed with a concave portion or the like for facilitating irradiation of light to the flow path 27 and emission of light transmitted through the flow path 27 as appropriate. The dimension of the flow channel 27 is not particularly limited, but, for example, the width is 25 to 100 μm, the depth is 25 to 100 μm, and the length is 5 to 150 mm. The size of the entire analysis chip 2 is appropriately set in accordance with the size of the flow path 27, and the size and arrangement of the mixing tank 22, the introduction tank 23, the discharge tank 25, and the electrode tank 26.

The analysis chip 2 having the above-described configuration is an example, and an analysis chip having a configuration capable of performing component analysis by capillary electrophoresis can be suitably used.

The component analysis apparatus 1 performs an analysis process for the sample Sa in a state where the analysis chip 2 on which the sample Sa is spotted is loaded. The component analysis device 1 includes an electrode 31, an electrode 32, a light source 41, an optical filter 42, a lens 43, a slit 44, a detector 45, a dispenser 6, a pump 61, a diluent tank 71, a migration liquid tank 72, and a control unit 8. The light source 41, the optical filter 42, the lens 43, and the detector 45 constitute a so-called measuring unit 40 in the present embodiment.

The electrodes 31 and 32 are used to apply a predetermined voltage to the flow path 27 in the capillary electrophoresis method. The electrode 31 is inserted into the electrode groove 26 of the analysis chip 2, and the electrode 32 is inserted into the discharge groove 25 of the analysis chip 2. The voltage applied to the electrodes 31 and 32 is not particularly limited, and is, for example, 0.5kV to 20 kV.

The light source 41 is a portion that emits light used for measuring absorbance as an optically measured value in the capillary electrophoresis method. The light source 41 includes, for example, an LED (light emitting diode) chip that emits light in a predetermined wavelength range. The optical filter 42 attenuates a predetermined wavelength of light from the light source 41 and transmits the remaining wavelengths of light. The lens 43 is used to focus the light transmitted through the optical filter 42 on the analysis site of the flow path 27 of the analysis chip 2. The slit 44 is used to remove unnecessary light that can cause scattering or the like, among the light collected by the lens 43.

The detector 45 receives the light from the light source 41 transmitted through the flow path 27 of the analysis chip 2, and is configured to include, for example, a photodiode, an Integrated Circuit (IC), or the like.

Thus, the path of the light emitted from the light source 41 reaching the detector 45 is an optical path. Then, at a measurement position 27A where the optical path intersects the flow path 27, the solution (i.e., either one of the sample solution and the electrophoretic solution or a mixed solution thereof) flowing through the flow path 27 is measured for the optical measurement value. That is, at the measurement position 27A in the channel 27, the measurement unit 40 measures the optical measurement value of the sample solution. The optical measurement value includes, for example, absorbance. The absorbance indicates the degree of absorption of light in the optical path by the solution flowing in the flow path 27, and indicates the absolute value of the common logarithm of the ratio of the intensity of incident light to the intensity of transmitted light. In this case, a general spectrophotometer may be used as the detector 45. In addition, even if absorbance is not used, it can be used in the present invention as long as it is an optical measurement value such as a simple transmitted light intensity value itself. In the following, a case where absorbance is used as an optical measurement value will be described as an example.

The dispenser 6 dispenses a desired amount of the diluent Ld, the migration solution Lm, and the sample solution, and includes, for example, a nozzle. The dispenser 6 is movable by a drive mechanism, not shown, at a plurality of predetermined positions in the component analyzer 1. The pump 61 is a suction source and a discharge source to the dispenser 6. The pump 61 may be used as a suction source and a discharge source of a port, not shown, provided in the component analysis apparatus 1. These ports are used for filling the migration liquid Lm and the like. Further, a dedicated pump different from the pump 61 may be provided.

The diluent tank 71 is a tank for storing the diluent Ld. The diluent tank 71 may be a tank permanently provided in the component analysis apparatus 1, or may be a tank in which a predetermined amount of the diluent Ld is enclosed and loaded in the component analysis apparatus 1. The migration liquid tank 72 is a tank for storing the migration liquid Lm. The migration liquid tank 72 may be a tank permanently installed in the component analysis apparatus 1, or may be a tank in which a predetermined amount of migration liquid Lm is filled in the component analysis apparatus 1.

The diluent Ld is used to generate a sample solution by mixing with the sample Sa. The main agent of the diluent Ld is not particularly limited, and water and physiological saline may be mentioned, and a preferable example thereof is a liquid having a component similar to that of the migration fluid Lm described later. In addition, as for the diluent Ld, additives may be added as needed in addition to the above-described main agent.

The electrophoretic fluid Lm is a medium which is filled in the discharge tank 25 and the flow path 27 in the analysis step S20 of the electrophoresis method and generates an electroosmotic flow in the electrophoresis method. The migration liquid Lm is not particularly limited, and a migration liquid using an acid is preferable. Examples of the acid include citric acid, maleic acid, tartaric acid, succinic acid, fumaric acid, phthalic acid, malonic acid, and malic acid. The migration liquid Lm preferably contains a weak base. Examples of the weak base include arginine, lysine, histidine, and tris (hydroxymethyl) aminomethane (tris). The pH of the migration solution Lm is, for example, in the range of pH4.5 to 6. Examples of the buffer solution of the migration solution Lm include MES, ADA, ACES, BES, MOPS, TES, HEPES and the like. In addition, in the migration liquid Lm, an additive may be added as necessary, as described in the description of the diluent Ld.

The control unit 8 controls each unit in the component analyzer 1. As shown in the hardware configuration of fig. 4, the control Unit 8 includes a CPU (Central Processing Unit) 81, a ROM (Read Only Memory) 82, a RAM (Random Access Memory) 83, and a Memory 84. The respective components are communicatively connected to each other via a bus 89.

The CPU 81 is a central processing unit and executes various programs or controls each unit. That is, the CPU 81 reads out the program from the ROM 82 or the memory 84 and executes the program with the RAM 83 as a work area. The CPU 81 performs control and various arithmetic processes of the above-described configurations in accordance with programs recorded in the ROM 82 or the memory 84.

The ROM 82 stores various programs and various data. The RAM 83 temporarily stores programs or data as a work area. The memory 84 is constituted by an HDD (Hard Disk Drive), an SSD (Solid State Drive), or a flash memory, and stores various programs including an operating system and various data. In the present embodiment, a program and various data for executing the component analysis method according to the present embodiment are stored in the ROM 82 or the memory 84.

The component analysis apparatus 1 implements various functions shown in fig. 5 using the hardware resources and the respective configurations described above when executing the component analysis method according to the present embodiment. These functions include, in addition to the measurement unit 40, an analysis unit 800 that analyzes a plurality of components contained in the sample Sa based on optical measurement values obtained by optically measuring the sample solution with time by the measurement unit 40, which will be described later.

< first mode >

A component analysis method according to a first aspect of the present disclosure includes: a measurement step S10 of separating the sample solution continuously introduced into the channel 27 into a plurality of components in the channel 27, and optically measuring the sample solution at the measurement position 27A of the channel 27 with time to obtain an optical measurement value; and an analyzing step S20 of analyzing a plurality of components contained in the sample Sa based on the optical measurement values, wherein the analyzing step S20 includes: an original waveform acquisition step S21 of obtaining an original waveform by plotting optical measurement values along a time axis on a two-dimensional plane; a measurement value differentiation step S23 of obtaining a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and a measurement value boundary determining step S24 of defining an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.

The component analysis device 1 of the present embodiment includes: a flow path 27 for continuously introducing the sample solution; a measurement unit 40 that optically measures the sample solution separated into a plurality of components in the flow path 27 at a measurement position 27A of the flow path 27 with time to obtain an optical measurement value; and an analysis unit 800 that analyzes a plurality of components contained in the sample Sa based on the optical measurement values, wherein the analysis unit 800 includes: an original waveform acquisition unit 801 that obtains an original waveform by plotting optical measurement values along a time axis on a two-dimensional plane; a measurement value differentiating unit 803 that acquires a measurement value differential waveform obtained by differentiating the original waveform along an axis of the optical measurement value orthogonal to the time axis; and a measurement value boundary determining unit 804 for determining an optical measurement value corresponding to a peak top of a differential waveform of the measurement value as a separation boundary between the plurality of components.

The functional configuration of the analysis unit 800 in this embodiment is shown in fig. 6. Next, a method of analyzing components in the present embodiment will be described with reference to the flowchart of fig. 7.

In the measurement step S10 shown in fig. 7, the sample solution is continuously introduced into the flow path 27, the sample solution is separated in the flow path 27 by applying a voltage, and when the sample solution reaches the measurement position 27A provided in the middle of the flow path 27, an optical measurement value is obtained by the measurement unit 40. Specifically, an electrode 31 and an electrode 32 are provided upstream and downstream of the flow path 27, respectively, and a voltage is applied therebetween to electrophorese the sample solution by a so-called capillary electrophoresis method.

For example, when the component to be analyzed is hemoglobin as described above, since the molecule surface thereof has a negative charge, the component to be analyzed is electrophoresed toward the electrode 32 as the anode by disposing the cathode as the electrode 31 at the upstream side of the channel 27 and disposing the anode as the electrode 32 at the downstream side. At this time, the electrophoresis speed differs depending on the charged state of the molecule surface. That is, the stronger the negative charge on the surface of the molecule, the faster the electrophoresis speed. Therefore, the component having a high electrophoresis rate reaches the measurement position 27A more quickly at the stage when the sample solution is introduced into the flow path 27.

At the time point when the component having the slow electrophoresis rate reaches the measurement position 27A, the component having the fast electrophoresis rate from the sample solution introduced later also reaches the measurement position 27A at the same time. That is, in the flow path 27, if a component having a high electrophoresis rate reaches the measurement position 27A first, the component will continue to reach as long as the sample solution is continuously introduced into the flow path 27. Further, although the component having the slower electrophoresis speed reaches the measurement position 27A with a delay, the component continues to reach the measurement position 27A as long as the sample solution is continuously introduced into the flow path 27 once it reaches the measurement position 27A.

In other words, at the measurement position 27A, the component having the high electrophoresis rate arrives first, and then the component having the low electrophoresis rate arrives cumulatively. Therefore, the absorbance of the sample solution measured by the measurement unit 40 at the measurement position 27A as an optical measurement value monotonously increases with time.

The original waveform acquiring unit 801 in the analyzing unit 800 shown in fig. 6 acquires an original waveform by plotting the absorbance as an optically measured value on a two-dimensional plane in contrast with another time axis (for example, X axis) as one axis (for example, Y axis) in the original waveform acquiring step S21 in the analyzing step S20 shown in fig. 7. This original waveform is represented, for example, as a solid curve as shown in fig. 8. The optical measurement values in the present embodiment may be acquired as data that can be plotted on a two-dimensional plane, and it is not necessary to actually plot a curve based on such data. This is also the same in other ways mentioned below. Here, the original waveform may be a function in which an optical measurement value (for example, absorbance) is plotted with time as a variable.

When the original waveform is observed along the time axis, a portion having a large slope as shown by a solid arrow in fig. 8 results from an increase in the optical measurement value when a certain component first reaches the measurement position 27A. The portion with a small slope as indicated by the broken-line arrow indicates that the next component has not yet reached the measurement position 27A. That is, the portion of the original waveform in which the slope increases indicates that the component in the sample solution has reached the measurement position 27A. Such a portion of the original waveform where the slope is large is represented as a peak waveform appearing in a waveform obtained by differentiating the original waveform along the time axis (referred to as a "time-differentiated waveform") as shown by a dashed-line curve in fig. 9. These peak waveforms are determined as waveforms from the respective components. On the time axis, it is shown that the electrophoretic velocity of the component corresponding to the peak waveform located on the further left side is higher than the electrophoretic velocity of the component corresponding to the peak waveform located on the further right side.

Here, in the case where a plurality of peak waveforms exist in the time-differentiated waveform, adjacent peak waveforms are divided by valley portions, i.e., valley bottoms B, located therebetween. The peak top T of the peak waveform is a maximum value in the time-differentiated waveform, and corresponds to a portion where the slope becomes maximum corresponding to the maximum value in the original waveform. On the other hand, the bottom B is a minimum value in the time-differentiated waveform, and the slope becomes extremely small in the original waveform. In other words, in the original waveform, a portion where the slope is steep corresponds to the peak top T, and a portion where the slope is gentle corresponds to the bottom B.

On the other hand, when the original waveform shown in fig. 8 is observed from the axis (Y axis) of the optical measurement value, the slope of the portion corresponding to the peak top T becomes gentle, and the slope of the portion corresponding to the bottom B becomes steep.

Focusing on this point, the measured value differentiating step S23 in the analyzing step S20 shown in fig. 7 is performed. That is, in the measured value differentiating step S23, the measured value differentiating unit 803 in the analysis unit 800 shown in fig. 6 differentiates the original waveform along the axis of the optical measured value to obtain a measured value differentiated waveform as shown by the dotted line curve in fig. 10. Here, the measurement value differential waveform may be a function obtained by differentiating the time of the original waveform using the optical measurement value as a variable.

As shown in fig. 10, the peaks T1 'to T6' in the measured-value differential waveform correspond to the valleys B1 to B6 in the time differential waveform, respectively. That is, the measured-value differential waveform is alternately expressed in terms of the peak top and the valley bottom with respect to the time differential waveform. In other words, the bottom portions B1 to B6 in the time differential waveform clearly appear as peak tops T1 'to T6' in the measurement value differential waveform, respectively.

Then, the measurement value boundary determining section 804 in the analysis section 800 shown in fig. 6 determines the optical measurement values S1 to S6 corresponding to the peaks T1 'to T6', respectively, as the separation boundaries of the components in the measurement value boundary determining step S24 in the analysis step S20 shown in fig. 7.

According to the configuration of the first aspect described above, the bottom portions B1 to B6, which are required to be recognized as the both ends of the peak waveform in the time-differentiated waveform, can be recognized as the peak tops T1 to T6 ° in the measured-value differentiated waveform, respectively. The optically measured values s1 to s6 corresponding to the peaks T1 'to T6' may be used as boundaries for separation of components contained in the sample Sa.

Note that, although it is not necessary to refer to the time differential waveform at all in order to obtain the peaks T1 'to T6' in the measurement value differential waveform, the contents mentioned in the above description and the time differential waveforms in fig. 9 and 10 are for explaining the meanings of the peaks T1 'to T6' in the measurement value differential waveform.

< second mode >

The component analysis method according to the second aspect of the present disclosure includes, in addition to the configuration of the component analysis method according to the first aspect, the analysis step S20 including: a time differentiation step S22 of obtaining a time-differentiated waveform obtained by differentiating an original waveform along a time axis; and an integral quantification step S28 of integrating the time-differentiated waveform with an integration interval having adjacent integration boundaries as both ends, with the time points corresponding to the separation boundaries as integration boundaries, to obtain a value, and calculating the value as the relative content of the component corresponding to the integration interval in the sample Sa.

The component analysis device 1 according to the present embodiment is configured such that the analysis unit 800 includes, in addition to the configuration of the component analysis device 1 according to the first embodiment: a time differentiation unit 802 that acquires a time-differentiated waveform obtained by differentiating an original waveform along a time axis; the integral quantitative unit 808 integrates the time-differentiated waveform with respect to the time-differentiated waveform at a time point corresponding to the separation boundary as an integral boundary in an integral section having adjacent integral boundaries as both ends, and calculates the value as the relative content of the component corresponding to the integral section in the sample Sa.

The functional configuration of the analysis unit 800 in this embodiment is shown in fig. 11. Next, a method of analyzing components in the present embodiment will be described with reference to a flowchart of fig. 12.

The measurement step S10 shown in fig. 12 is the same as in the first embodiment.

The original waveform acquiring unit 801 in the analyzing unit 800 shown in fig. 11 acquires an original waveform by plotting absorbance as an optical measurement value on a two-dimensional plane in contrast with another time axis (for example, X axis) as one axis (for example, Y axis) in the original waveform acquiring step S21 in the analyzing step S20 shown in fig. 12. This original waveform is represented, for example, as a solid curve as shown in fig. 8. The original waveform acquisition step S21 is the same as in the first embodiment.

Next, the time differentiation unit 802 in the analysis unit 800 shown in fig. 11 acquires a time-differentiated waveform obtained by differentiating the original waveform along the time axis in the time differentiation step S22 in the analysis step S20 shown in fig. 12. The time-differentiated waveform is shown as a dashed curve in fig. 9 and 10. The relationship between the original waveform and the time-differentiated waveform is the same as described in the first mode.

Next, the measured value differentiating section 803 in the analyzing section 800 shown in fig. 11 differentiates the original waveform along the axis of the optical measured value in the measured value differentiating step S23 in the analyzing step S20 shown in fig. 12, and obtains a measured value differential waveform shown by a dotted curve in fig. 10. The measured differential waveform is also the same as in the first embodiment.

Next, the measurement value boundary determining section 804 in the analysis section 800 shown in fig. 11 determines the optical measurement values S1 to S6 corresponding to the peak tops T1 'to T6' in fig. 10 as the separation boundaries of the components in the measurement value boundary determining step S24 in the analysis step S20 shown in fig. 12.

Then, the integrating and quantifying unit 808 of the analysis unit 800 shown in fig. 11 sets the optical measurement values S1 to S6 corresponding to the peak tops T1 'to T6' determined in the measurement value boundary determining unit S24 as the separation boundaries in the integrating and quantifying step S28 of the analysis step S20 shown in fig. 12, and sets the time points T1 to T6 corresponding to the bottom portions B1 to B6 in the time differential waveforms corresponding to these respective separation boundaries. Then, the time-differentiated waveform is integrated for an integration section having adjacent integration boundaries as both ends, and the area occupied by the time-differentiated waveform in the integration section is calculated. The value obtained as the area can be regarded as the relative content of a component corresponding to the integration interval (i.e., a component regarded as a peak waveform in the time-differentiated waveform).

Note that, although the bottom B3 in fig. 10 is not necessarily regarded as a definite minimum in the time-differentiated waveform, the peak top T3' in the corresponding measured-value differentiated waveform can be clearly recognized as a so-called shoulder in the curve. Therefore, the time point t3 corresponding to the bottom B3 can be clearly determined as the integration boundary t3 corresponding to the separation boundary s 3. Then, by integrating the time-differentiated waveform in an integration section having the integration boundary t3 and the adjacent integration boundary t4 as both ends, the relative content can be calculated for the peak waveform P3 which is not necessarily regarded as a clear peak. Of course, with respect to the other peak waveforms P1, P2, P4, and P5, which are considered to be clear peaks, the relative contents of the respective peak waveforms can be calculated by integrating the time-differentiated waveforms in the same manner with t1 to t2, t2 to t3, t4 to t5, and t5 to t6 as integration sections.

< third mode >

In the method for analyzing a component according to the third aspect of the present disclosure, the analyzing step S20 includes, in addition to the configuration of the method for analyzing a component according to the first aspect: the displacement determining step S25 calculates the distance between the sections having the separation boundaries adjacent to each other along the axis of the optically measured value as the relative content of the component corresponding to the section in the sample Sa.

The component analysis device 1 according to the present embodiment is configured such that the analysis unit 800 includes, in addition to the configuration of the component analysis device 1 according to the first embodiment: the displacement quantifying unit 805 calculates the distance between sections having separation boundaries adjacent to each other along the axis of the optically measured value as the relative content of the component corresponding to the section in the sample.

The functional configuration of the analysis unit 800 in this embodiment is shown in fig. 13. Next, a method of analyzing components in the present embodiment will be described with reference to the flowchart of fig. 14.

The measurement step S10 shown in fig. 14 is the same as in the first embodiment.

The original waveform acquiring unit 801 in the analyzing unit 800 shown in fig. 13 acquires an original waveform by plotting the absorbance as an optically measured value on a two-dimensional plane in contrast with another time axis (for example, X axis) as one axis (for example, Y axis) in the original waveform acquiring step S21 in the analyzing step S20 shown in fig. 14. This original waveform is represented, for example, as a solid curve as shown in fig. 8. The original waveform acquisition step S21 is the same as in the first embodiment.

Next, the measured value differentiating section 803 in the analyzing section 800 shown in fig. 13 differentiates the original waveform along the axis of the optical measured value in the measured value differentiating step S23 in the analyzing step S20 shown in fig. 14, and obtains a measured value differential waveform shown by a dotted curve in fig. 15. The measured differential waveform is also the same as in the first embodiment.

Next, the measurement value boundary determining section 804 in the analysis section 800 shown in fig. 13 determines the optical measurement values S1 to S6 corresponding to the peak tops T1 'to T6' in fig. 15 as the separation boundaries of the components in the measurement value boundary determining step S24 in the analysis step S20 shown in fig. 14.

Then, the displacement quantifying unit 805 in the analyzing unit 800 shown in fig. 13 calculates the relative content of the component in the sample Sa as the distances D1 to D5 of the sections divided by the separation boundaries S1 to S6 corresponding to the peak tops T1 'to T6' determined in the first embodiment, respectively, as shown in fig. 15, without acquiring the time-differentiated waveform in the displacement quantifying step S25 in the analyzing step S20 shown in fig. 14. The distances between the sections D1 to D5 shown in fig. 15 correspond to the areas of the peak waveforms P1 to P5 shown in fig. 10, respectively. In the displacement determining step S25, it is not necessary to refer to the time-differentiated waveform at all.

< fourth mode >

A component analysis method according to a fourth aspect of the present disclosure includes: a measurement step S10 of separating the sample solution continuously introduced into the channel 27 into a plurality of components in the channel 27, and measuring the sample solution at the measurement position 27A of the channel 27 with time to obtain an optical measurement value; and an analyzing step S20 of analyzing a plurality of components contained in the sample Sa based on the optical measurement values, wherein the analyzing step S20 includes: an original waveform acquisition step S21 of obtaining an original waveform by plotting optical measurement values along a time axis on a two-dimensional plane; a time differentiation step S22 of obtaining a time-differentiated waveform obtained by differentiating the original waveform along a time axis; an inverse differentiation step S26 of acquiring an inverse differential waveform in which the reciprocal of the time differential waveform is plotted along a time axis; and a time boundary determining step S27 of defining a time point corresponding to the peak top of the inverse differential waveform as a boundary of separation between the plurality of components.

The component analysis apparatus 1 according to the present embodiment includes: a flow path 27 for continuously introducing the sample solution; a measurement unit 40 that optically measures the sample solution separated in the channel 27 at a measurement position 27A of the channel 27 with time to obtain an optical measurement value; and an analysis unit 800 that analyzes a plurality of components contained in the sample Sa based on the optical measurement values, wherein the analysis unit 800 includes: an original waveform acquisition unit 801 that obtains an original waveform by plotting optical measurement values along a time axis on a two-dimensional plane; a time differentiation unit 802 that acquires a time-differentiated waveform obtained by differentiating the original waveform along a time axis; an inverse differential unit 806 that acquires an inverse differential waveform in which the inverse of the time differential waveform is plotted along the time axis; and a time boundary determining unit 807 for defining a time point corresponding to the peak top of the inverse differential waveform as a boundary of separation between the plurality of components.

The functional configuration of the analysis unit 800 in this embodiment is shown in fig. 16. Next, a method of analyzing components in the present embodiment will be described with reference to the flowchart of fig. 17.

The measurement step S10 shown in fig. 17 is the same as in the first embodiment.

The original waveform acquiring unit 801 in the analyzing unit 800 shown in fig. 16 acquires an original waveform by plotting absorbance as an optical measurement value on a two-dimensional plane in contrast with another time axis (for example, X axis) as one axis (for example, Y axis) in the original waveform acquiring step S21 in the analyzing step S20 shown in fig. 17. This original waveform is represented, for example, as a solid curve as shown in fig. 8. The original waveform acquisition step S21 is the same as in the first embodiment. Here, the original waveform may be a function in which an optical measurement value (for example, absorbance) is plotted with time as a variable.

Next, the time differentiation unit 802 in the analysis unit 800 shown in fig. 16 acquires a time-differentiated waveform obtained by differentiating the original waveform along the time axis in the time differentiation step S22 in the analysis step S20 shown in fig. 17. The time-differentiated waveform is shown as a dashed curve in fig. 9. The relationship between the original waveform and the time-differentiated waveform is the same as described in the first mode.

Next, the inverse differentiation unit 806 in the analysis unit 800 shown in fig. 16 plots the reciprocal of the absorbance in the time-differentiated waveform obtained in the time-differentiated step S22 along the time axis in the inverse-differentiated step S26 in the analysis step S20 shown in fig. 17, thereby obtaining an inverse-differentiated waveform shown by a dotted line in fig. 18. When the reciprocal of absorbance in the time-differentiated waveform is plotted on a two-dimensional plane having one axis of the optical measurement value and the other axis of the time axis, the reciprocal does not need to be plotted as an absolute value, but may be plotted as a relative value to the extent that the correspondence with the time-differentiated waveform becomes clear. The reciprocal of absorbance as used herein may be obtained as data that can be plotted on a two-dimensional plane, and is not necessarily actually plotted as a curve based on such data.

Here, the peak top and the bottom in the time-differentiated waveform correspond to the bottom and the top in the inverse-differentiated waveform, respectively, which is the same as the measured-value differentiated waveform described in the first embodiment. Therefore, as shown in fig. 18, the peaks T1 ″ to T6 ″ in the inversely differentiated waveform correspond to the valleys B1 to B6 in the time-differentiated waveform, respectively. In other words, the bottom portions B1 to B6 in the time-differentiated waveform clearly appear as the peak tops T1 "to T6" in the inverse-differentiated waveform, respectively.

That is, the time boundary determining section 807 in the analysis section 800 shown in fig. 16 determines time points T1 to T6 corresponding to the peak tops T1 "to T6" in the inverse differential waveform as the separation boundaries of the components in the time boundary determining step S27 in the analysis step S20 shown in fig. 17.

According to the configuration of the fourth aspect described above, the bottom portions B1 to B6, which are originally required to be recognized as the both ends of the peak waveform in the time-differentiated waveform, can be recognized as the peak tops T1 ″ to T6 ″ in the inversely-differentiated waveform. The time points T1 to T6 corresponding to the peak tops T1 "to T6" may be used as boundaries for separation of components contained in the sample Sa.

< fifth mode >

The component analysis method according to a fifth aspect of the present disclosure includes, in addition to the component analysis method according to the fourth aspect, an integration/quantification step S28 of integrating the time-differentiated waveform at a time point corresponding to the separation boundary with respect to the time-differentiated waveform as an integration boundary in an integration section having adjacent integration boundaries as both ends to obtain a value, and calculating the value as a relative content of the component corresponding to the integration section in the sample Sa.

The component analysis device 1 according to the present embodiment is configured such that the analysis unit 800 includes, in addition to the component analysis device 1 according to the fourth embodiment: the integral quantitative unit 808 integrates the time-differentiated waveform with respect to the time-differentiated waveform in an integration section having adjacent integration boundaries as both ends, with the time points corresponding to the separation boundaries as integration boundaries, and calculates the value as the relative content of the component corresponding to the integration section in the sample Sa.

The functional configuration of the analysis unit 800 in this embodiment is shown in fig. 19. Hereinafter, the component analysis method in the present embodiment will be described with reference to the flowchart of fig. 20. However, the original waveform acquisition step S21, the time differentiation step S22, the inverse differentiation step S26, and the time boundary determination step S27 in the measurement step S10 and the analysis step S20 shown in fig. 20 are the same as those in the fourth embodiment.

In the time boundary determining step S27 shown in fig. 20, in fig. 18, the time points T1 to T6 corresponding to the respective peak tops T1 "to T6" identified in the fourth embodiment are separation boundaries corresponding to the respective valley bottoms B1 to B6, and the time points T1 to T6 as these separation boundaries become integration boundaries. Then, the integral quantifying unit 808 in the analyzing unit 800 shown in fig. 19 integrates the time-differentiated waveform in the integration section having the adjacent integration boundary as both ends in the integral quantifying step S28 in the analyzing step S20 shown in fig. 20, and calculates the area occupied by the time-differentiated waveform in the integration section. The value obtained as the area can be regarded as the relative content of a component corresponding to the integration interval (i.e., a component regarded as a peak waveform in the time-differentiated waveform).

Note that, here, the bottom B3 in fig. 18 is not necessarily regarded as an explicit minimum value in the time-differentiated waveform, but the peak top T3 ″ in the inversely differentiated waveform corresponding thereto can be explicitly identified as a so-called shoulder in the curve. Therefore, the time point t3 corresponding to the bottom B3 can be clearly determined as the integration boundary t 3. Then, by integrating the time-differentiated waveform in an integration section having the integration boundary t3 and the adjacent integration boundary t4 as both ends, the relative content can be calculated for the peak waveform P3 which is not necessarily regarded as a clear peak. Of course, with respect to the other peak waveforms P1, P2, P4, and P5, which are considered to be clear peaks, the relative contents of the respective peak waveforms can be calculated by integrating the time-differentiated waveforms in the same manner with t1 to t2, t2 to t3, t4 to t5, and t5 to t6 as integration sections.

< others >

In another embodiment of the present invention, a component analysis method other than the capillary electrophoresis method may be used, in which a sample solution is separated by some means other than voltage application (for example, chromatography) at the time point of introduction into the flow path, and measurement is performed in a flow path having a width larger than that of the capillary.