JP2011059068A - Method and device for analyzing biocomponent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method reduced in limitation for analyzing a biocomponent capable of wholly grasping a state as a living body of an object to be inspected and analyzing a surrounding environment of the object, and a device therefor. <P>SOLUTION: The method for analyzing the component of the object 1 of a living body includes steps of irradiating the object 1 frozen with an electromagnetic wave W1 of 0.1-10 THz, detecting an electromagnetic wave W2 transmitting the object 1 or reflected by the object 1, obtaining an absorption characteristic spectrum, and determining the distribution of the component of the object 1 based on the absorption characteristic spectrum. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method and apparatus for analyzing components of a living body such as marine products and processed fishery products.

As a method for analyzing components contained in foods made from living organisms such as marine products and processed fishery products, there is a method in which components are extracted from a specimen by heating or an organic solvent and analyzed through pretreatment (for example, patents). References 1 and 2).
However, this method enables high-precision analysis, but the process is complicated and time-consuming, and it is difficult to obtain information on the distribution of components in the living tissue.
As a method of analyzing components in a living tissue, there is an analysis method using a fluorescence microscope (for example, see Non-Patent Document 1). According to this method, the distribution of the fluorescently labeled component in the living tissue can be confirmed.

Japanese Patent Laid-Open No. 11-155597 Japanese Patent Laid-Open No. 6-253897

New Chemistry Experiment Course 6, Biomembrane and Transport Membrane (bottom), Tokyo Chemical Doujin, p. 861

However, in an analysis method using a fluorescence microscope, there is a case where a biological tissue cannot be labeled with a fluorescent probe depending on a component to be analyzed.
In addition, since pretreatment such as the introduction of a fluorescent probe to a subject takes time and the biological tissue is affected physiologically by introduction, the state of the living body may change during measurement. It was difficult to accurately grasp the state of the living body placed below.
In addition, since it is difficult to label a substance outside the subject with a fluorescent probe, the components in the environment around the subject cannot be analyzed.
The present invention has been made in view of the above circumstances, has few restrictions on target components, can accurately grasp the state of a subject as a living body, and can also analyze the environment around the subject. An object of the present invention is to provide a biological component analysis method and apparatus.

The biological component analysis method of the present invention is a method for analyzing a component of a living subject, and the frozen subject is irradiated with an electromagnetic wave of 0.1 to 10 THz and transmitted or reflected by the subject. The component distribution of the subject is obtained based on an absorption characteristic spectrum obtained by detecting electromagnetic waves.
In the biological component analysis method of the present invention, the component distribution of the subject can be obtained by detecting the electromagnetic wave at a plurality of points at different positions in the subject.
In the biological component analysis method of the present invention, the distribution of the components of the subject can be measured over time.
In the biological component analysis method of the present invention, the subject is frozen together with the sodium chloride aqueous solution in a state where at least a part is immersed in the sodium chloride aqueous solution, and the electromagnetic wave is detected with respect to the subject. It is preferable to detect the electromagnetic wave of the surrounding aqueous sodium chloride solution to obtain the distribution of the components.
The subject preferably has a thickness in the irradiation direction of the electromagnetic wave of 0.1 to 15 mm.
The subject preferably has a maximum dimension in a plane that intersects the irradiation direction of the electromagnetic wave of 3 to 100 mm.
In freezing the subject, it is preferable that the cooling rate is −0.1 to −5 ° C./second and the ultimate cooling temperature is −200 to 0 ° C.

The biological component analyzer of the present invention is a device for analyzing a component of a subject that is a living body, and is placed on a placement unit on which the frozen subject is placed, and a test subject on the placement unit. An irradiation unit that irradiates an electromagnetic wave of 1 to 10 THz, a detection unit that detects an electromagnetic wave transmitted or reflected by the subject, and an absorption characteristic spectrum obtained by the electromagnetic wave detected by the detection unit. And a calculation / control unit for obtaining a distribution of components.
The biological component analyzer of the present invention includes a movement mechanism that moves the placement unit relative to the irradiation unit and the detection unit, and the movement mechanism moves the placement unit according to the position of the subject. Can be arbitrarily set.
It is preferable that the moving mechanism is capable of moving the mounting portion in the irradiation direction of the electromagnetic wave and an in-plane direction intersecting with the irradiation direction.

According to the present invention, the component distribution is obtained based on the absorption characteristic spectrum obtained by detecting the electromagnetic wave transmitted through the subject. Therefore, the method for extracting the component from the subject using heating or an organic solvent, Compared with the method of performing (labeling), a complicated process is unnecessary and the operation is easy.
In addition, since it is not necessary to take out the target component from the subject, the component can be analyzed in the state of being present in the subject, and the distribution of the component can be grasped in relation to the shape of the subject.
Compared with the method of introducing a labeling agent (fluorescent probe, etc.), it is possible to analyze a living subject in a normal physiological state and accurately grasp the state of the living body in the environment. be able to.
Further, there are few restrictions on the components to be analyzed, and a wide range of analysis targets can be selected.
Furthermore, in the conventional method, there is a concern that the state of the living body changes during time-consuming preprocessing, etc., but since the pretreatment is unnecessary in the present invention, the state of the living body placed in the environment is accurately determined. Can grasp.
In addition, since component analysis is possible including the surrounding environment of the subject, the exchange of substances between the subject and the surrounding environment can be observed.

It is a block diagram which shows one Embodiment of the biological component analyzer of this invention. It is an enlarged view which shows the principal part of the biological component analyzer shown in a previous figure. It is a block diagram which shows other embodiment of the biological component analyzer of this invention. It is a figure which shows the time-dependent change of the component distribution of the subject obtained in the Example. It is a figure which shows the time-dependent change of the component distribution of the subject obtained in the Example.

An example of the biological component analysis method and apparatus of the present invention will be described with reference to FIGS.
FIG. 1 is a configuration diagram showing a biological component analyzer 10 (hereinafter simply referred to as analyzer 10) according to an embodiment of the present invention. FIG. 2 is an enlarged view showing a main part of the analyzer 10.
The analysis apparatus 10 includes a placement unit 2 that places a chamber 11 that accommodates the subject 1, an irradiation unit 3 that irradiates the subject 1 with electromagnetic waves, and a detection unit 4 that detects the electromagnetic waves transmitted through the subject 1. The calculation / control unit 5 that obtains the distribution of the components of the subject 1 based on the electromagnetic waves detected by the detection unit 4, the moving mechanism 6 that allows the placement unit 2 to move, and the cooling unit that cools the subject 1 7.

The placement unit 2 is a stage arranged substantially horizontally. The placement portion 2 is sized to allow the chamber 11 to be placed.
The irradiation unit 3 includes a light source 8 and a pulse generator 9.
The light source 8 is not particularly limited as long as it can excite an electromagnetic wave of 0.1 to 10 THz (hereinafter sometimes referred to as a terahertz wave). For example, a pulse laser or a titanium sapphire laser that is femtosecond laser excitation light or the like Is mentioned.
The pulse generator 9 is not particularly limited as long as it can receive excitation light from the light source 8 and generate a terahertz wave. For example, a nonlinear optical crystal, a photoconductive antenna, a semiconductor, a quantum well, a high-temperature conductive thin film, or the like. Is mentioned.

The detection unit 4 only needs to be able to detect the terahertz wave that has passed through the subject 1, and includes a photoconductive antenna, for example.
The moving mechanism 6 moves the mounting unit 2 in the electromagnetic wave irradiation direction (vertical direction in FIG. 1) with respect to the subject 1 and in an in-plane direction intersecting this (in-plane direction perpendicular to the vertical direction in FIG. 1). It is configured to be possible.
In the example of illustration, it has the in-plane moving mechanism 14 which enables the movement of the mounting part 2 in the said in-plane direction, and the raising / lowering mechanism 15 which enables the mounting part 2 to move in the height direction.

The in-plane moving mechanism 14 is connected to the mounting unit 2 and can arbitrarily determine the position of the mounting unit 2 in the X-axis direction (left and right direction in FIG. 1) by extending and contracting by a driving unit (not shown). In addition, the position of the mounting portion 2 in the Y-axis direction (direction perpendicular to the paper surface in FIG. 1) can be arbitrarily determined by moving the paper portion toward the front side and the back side by a drive unit (not shown). Therefore, the in-plane direction position of the placement unit 2 can be arbitrarily determined.
The lifting mechanism 15 moves up and down by the drive unit 16 to arbitrarily determine the position (height position) in the Z-axis direction (vertical direction in FIG. 1) of the mounting unit 2 via the in-plane moving mechanism 14. Can do.

The cooling unit 7 is configured to cool the inside of the chamber 11 by sending a refrigerant such as air into the chamber 11 from the path 7 a and to freeze the subject 1.
It is preferable that the cooling unit 7 has a cooling performance that can arbitrarily set the temperature of the chamber 11 in the range of −200 to 0 ° C., for example.

In the chamber 11, an upper part facing the irradiation unit 3 and a lower part facing the detection unit 4 are optical windows 11 a through which the terahertz wave can be transmitted.
Reference numerals 12 and 13 in FIG. 1 denote lenses.
As shown in FIG. 2, a cylindrical storage portion 17 capable of storing a sodium chloride aqueous solution 18 (environmental water) is provided at the bottom of the chamber 11.

Next, an example of a method for performing component analysis of the subject 1 using the analyzer 10 will be described.
In the present invention, the subject 1 is a marine product, an agricultural product, a livestock product, a food made of a living body such as a processed product thereof, or the like.
Examples of the target component include inorganic salts, amino acids, organic acids, enzymes, vitamins, carbohydrates, lipids, fatty acids and the like. For example, it is possible to target biological nutrients and umami components.
As the inorganic salt, desirably, in the living body and the environment, any one of zinc, potassium, calcium, chromium, selenium, iron, copper, sodium, magnesium, manganese, phosphorus, vanadium, strontium, ammonium cation, or fluorine , Chlorine, bromine, iodine, hydroxy acid, sulfuric acid, sulfurous acid, carbonic acid, nitric acid, nitrous acid, phosphoric acid, and boric acid.
As amino acids, aspartic acid, glutamic acid, lysine, arginine, histidine, glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, proline, phenylalanine, tyrosine, tryptophan, ornithine, taurine, glucosamine And at least one of these salts.
Examples of the organic acid include inosinic acid, guanylic acid, adenylic acid, xanthylic acid, alginic acid, acetic acid, citric acid, lactic acid, succinic acid, malic acid, and salts thereof.
Examples of the enzyme include at least one of amylase, protease, lipase, cellulase, papain, promeline, pancreatin, superoxide dismutase, catalase, and glutathione peroxidase.
As vitamins, vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, niacin, pantothenic acid, folic acid, biotin, vitamin F, vitamin P, vitamin Q, vitamin And at least one of U, choline, isitol, and paraaminobenzoic acid.
As saccharides, dihydroxyacetone, glyceraldehyde, erythrulose, erythrose, threose, ribulose, xylulose, ribose, arabinose, xylose, lyxose, deoxyribose, psicose, fructose, sorbose, tagatose, allose, altrose, glucose, mannose, Growth, idose, galactose, talose, fucose, fucose, rhamnose, cedoheptulose, sucrose, lactose, maltose, trehalose, turanose, cellobiose, raffinose, melezitose, maltotriose, acarbose, stachyose, fructooligosaccharide, galactooligosaccharide, xylooligosaccharide, milk Fruit oligosaccharide, soybean oligosaccharide, isomaltoligosaccharide, chitooligosaccharide, palatinose, cyclic At least one sugar oligo can be mentioned.
As fatty acids, oleic acid, elaidic acid, erucic acid, nervonic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, EPA (eicosapentaenoic acid), DHA (docosahexaenoic acid), stearidonic acid, arachidonic acid, butyric acid, valeric acid , Caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid.

The subject 1 preferably has a thickness in the electromagnetic wave irradiation direction (vertical direction in FIG. 1) of 0.1 to 15 mm (desirably 0.5 to 2 mm).
If the subject 1 is too thin, the amount of the target component decreases, so that the detection accuracy is lowered. If the subject 1 is too thick, the terahertz wave is difficult to transmit. On the other hand, by setting the thickness within the above range, the detection accuracy can be improved and the terahertz wave can be transmitted with certainty. In addition, multiple reflection and interference are less likely to occur. Therefore, highly accurate analysis is possible.

  The subject 1 preferably has a maximum dimension of 3 to 100 mm in a plane intersecting the electromagnetic wave irradiation direction (the in-plane direction). If the maximum dimension is too large, the measurement time becomes long when measuring the entire subject 1 while moving the mounting portion 2 in the in-plane direction, and there is a possibility that the amount of components or the like may change during the measurement. . If the number of measurement points is reduced to avoid this, the resolution is lowered. On the other hand, the detection accuracy can be increased by setting the maximum dimension within the above range.

As shown in FIG. 2, a sodium chloride aqueous solution 18 (environmental water) is stored in the storage unit 17. The sodium chloride concentration of the sodium chloride aqueous solution 18 is preferably 0.3 to 20% by mass (desirably 0.3 to 1% by mass).
The subject 1 is placed in the reservoir 17 so that at least a part thereof is immersed in the sodium chloride aqueous solution 18.
The subject 1 is preferably arranged at a position where a sodium chloride aqueous solution 18 exists around the subject 1. For example, if the subject 1 is arranged at the center of the cylindrical storage portion 17, the sodium chloride aqueous solution 18 exists around the entire circumference.
By using the sodium chloride aqueous solution 18, sodium chloride or ions derived therefrom (sodium ions or chlorine ions) show a weak interaction with components or water molecules in the subject 1 in the subject 1. It becomes like this.
Since terahertz waves resonate with weak interactions such as hydrogen bonds, van der Waals bonds, π-electron interactions, and electrostatic interactions, changes in the resonance frequency and absorption intensity of the energy of the above-described interactions are described later. The used component analysis becomes possible.

In accordance with the temperature in the chamber 11 detected by the temperature sensor 19, the calculation / control unit 5 drives the cooling unit 7 to cool the inside of the chamber 11.
The cooling rate is preferably −0.1 to −5 ° C./second (desirably −1 to −1.5 ° C./second). By setting the cooling rate within this range, sodium chloride is dispersed and taken into the subject 1 without precipitating outside the subject 1, so that sodium chloride or ions derived therefrom (sodium) in the subject 1 Ions or chlorine ions) exhibit the above interaction with the components and water molecules in the subject 1.
The ultimate cooling temperature is preferably −200 to 0 ° C. (desirably −200 to −150 ° C.). By setting the cooling temperature within this range, the above interaction can be enhanced.

As a result, the subject 1 is frozen. In this example, the subject 1 is frozen together with the sodium chloride aqueous solution 18 while being immersed in the sodium chloride aqueous solution 18.
Next, the pulse generator 9 is irradiated with excitation light from the light source 8. The pulse generator 9 generates a terahertz wave W1, is focused by the lens 12, and is irradiated on the subject 1.
When the subject 1 is irradiated with the terahertz wave W1, it resonates with energy of weak interaction such as hydrogen bond, van der Waals bond, π-electron interaction, electrostatic interaction and the like.

The terahertz wave W2 that has passed through the subject 1 is converted into parallel light by the lens 13 and reaches the detection unit 4.
The detection value of the terahertz wave W2 detected by the detector 4 is sent to the calculation / control unit 5, where the calculation / control unit 5 converts the detection value into a function of a frequency of 0.1 to 10 THz by Fourier transform.
Based on this, an absorption characteristic spectrum normalized per unit thickness of the subject 1 and unit concentration of the target component is obtained.
Since the resonance frequency and the absorption intensity of the energy of the interaction depend on the concentration of the component, this absorption characteristic spectrum can be used as an index of the component concentration.

The terahertz wave W2 is preferably detected at a plurality of points where the relative positions of the subject 1 with respect to the irradiation unit 3 and the detection unit 4 are different.
For example, the terahertz wave W <b> 2 can be detected at a plurality of points while moving the mounting unit 2 by the moving mechanism 6. The moving direction is not limited, and may be the in-plane direction or the vertical direction.
As a result, an absorption characteristic spectrum serving as an index of the component concentration is obtained at a plurality of points at different positions in the subject 1, and thus a distribution of the component concentration is obtained.

For example, by detecting the terahertz wave W2 while scanning the subject 1 in the in-plane direction by the in-plane moving mechanism 14, the distribution of the component concentration in the in-plane direction can be obtained.
By detecting the terahertz wave W2 while scanning the subject 1 in the vertical direction by the lifting mechanism 15, the distribution of the component concentration in the thickness direction (the terahertz wave irradiation direction) can be obtained.
In addition, since the absorption characteristic spectrum may change depending on the temperature at the time of analysis, measurement under a plurality of temperature conditions can be performed as necessary. Specifically, the reliability can be confirmed by performing measurement under 2 to 8 kinds of temperature conditions and comparing the absorption characteristic spectra with each other.

By comparing the obtained absorption characteristic spectrum with a standard absorption characteristic spectrum obtained using a standard sample containing the target component at a known concentration, the component can be specified and its concentration can also be known.
Specifically, the distribution of the components in the subject 1 can be known for inorganic salts, amino acids, organic acids, enzymes, vitamins, carbohydrates, lipids, fatty acids, and the like. The concentration of the component can also be grasped.
Further, if the measurement is performed over time, the change in the component concentration in the subject 1 can be grasped.
For example, if a frozen sample is created for a subject 1 at regular time intervals immediately after being placed in a predetermined environment, and component analysis thereof is performed, information on the increase or decrease of the component can be obtained.

When the subject 1 is immersed in the sodium chloride aqueous solution 18 and frozen together with the sodium chloride aqueous solution 18 around the subject 1, the terahertz of not only the subject 1 but also the sodium chloride aqueous solution 18 therearound is present. It is preferable to detect the wave W2 and obtain the distribution of the component.
As a result, it is possible to obtain information on the increase / decrease of the component around the subject 1 as well as within the subject 1. For example, the intake amount of the component into the subject 1 and the discharge amount from the subject 1 can be examined.

According to the analysis method of the present invention, the component distribution is obtained based on the absorption characteristic spectrum obtained by detecting the terahertz wave W2 that has passed through the subject 1, so that the component is removed from the subject using heating or an organic solvent. Compared to the extraction method and the fluorescent labeling (labeling) method, a complicated process is unnecessary and the operation is easy.
In addition, since it is not necessary to take out the target component from the subject 1, the component can be analyzed in the state of being present in the subject 1, and the distribution of the component can be grasped in relation to the shape of the subject 1.
In the conventional method, the influence of the introduction of the labeling agent (fluorescent probe or the like) on the living body may be a problem. However, the analysis method according to the present invention does not have such a problem. Analysis can be performed in a normal physiological state, and the state of a living body placed in the environment can be accurately grasped.
In addition, in the conventional method, depending on the component to be analyzed, it may be difficult to analyze because it is difficult to introduce a labeling agent (fluorescent probe or the like). A wide range of analysis targets can be selected.
Furthermore, in the conventional method, there is a concern that the state of the living body may change during the pretreatment that takes time, but since the pretreatment is unnecessary in the analysis method of the present invention, the living body placed in the environment is not necessary. The state can be grasped accurately.
In addition, in the conventional method, component analysis is difficult for the environment around the subject 1, but in the present invention, component analysis is possible including the environment around the subject 1. You can observe the exchange of substances with.

FIG. 3 is a configuration diagram showing an analyzer 20 according to the second embodiment of the present invention.
In the following description, the same components as those of the analyzer 10 are denoted by the same reference numerals and description thereof is omitted.
The analyzer 20 includes a placement unit 2 for placing the chamber 11, an irradiation unit 3 for irradiating the subject 1 with the terahertz wave W1, a detection unit 4 for detecting the terahertz wave W3, and a target based on the terahertz wave W3. A calculation / control unit 5 that obtains the distribution of components of the specimen 1, a moving mechanism 6 that can move the placement unit 2, a cooling unit 7 that cools the subject 1, and a reflection unit 21 are provided.

  The reflection unit 21 includes reflection mirrors 22a and 22b that reflect the terahertz wave W1 and irradiate the subject 1, and reflection mirrors 23a to 23c that reflect the terahertz wave W3 reflected by the subject 1.

In order to perform component analysis using the analyzer 20, the irradiation unit 3 generates a terahertz wave W1 and irradiates the subject 1 through the reflecting mirrors 22a and 22b.
The terahertz wave W3 reflected by the subject 1 reaches the detection unit 4 via the reflecting mirrors 23a to 23c, and the detection value of the terahertz wave W3 detected by the detector 4 is sent to the calculation / control unit 5 for calculation / control. In part 5, an absorption characteristic spectrum is obtained.
Since the resonance frequency and the absorption intensity of the energy of the interaction depend on the concentration of the component, this absorption characteristic spectrum can be used as an index of the component concentration.
This makes it possible to know the component distribution in the subject 1 such as inorganic salts, amino acids, organic acids, enzymes, vitamins, carbohydrates, lipids, fatty acids and the like. The distribution of the concentration of the component can also be grasped.

Hereinafter, the present invention will be specifically described with reference to examples. In addition, this invention is not limited to an Example.
An analytical test was performed using the analyzer 10 shown in FIG. The analysis device 10 includes a spectroscopic device (manufactured by Hamamatsu Photonics, photoconductive antenna) and a spectroscopic device (manufactured by Advanced Infrared, THz-TDS2004) provided with a pulse generator 9 (manufactured by Hamamatsu Photonics, photoconductive antenna). It was made using. The light source 8 used was Ti-sapphire laser Vitesse (100 femtosecond type, manufactured by Coherent).

As the subject 1, a Yamato swordfish produced from Lake Shinji and having a thickness of 10 mm or less was used. The subject 1 is 40 ° C. at 20 ° C. in artificial seawater having a salinity of 0.5% by mass, which is a relatively low value in the salinity range (0.3 to 1%) of the brackish water area where Yamatojimi is inhabiting. After rearing for hours, it was used for the test.
Depth of artificial seawater (sodium chloride aqueous solution 18) having a salinity concentration of 1%, which is a relatively high value in the salinity range (0.3 to 1%) of the brackish water area, in a cylindrical storage portion 17 having a diameter of 30 mm. The specimen 1 was introduced so as to be 2 mm, and the subject 1 was placed in the center so that at least a part of the specimen 1 was immersed in artificial seawater.
The subject 1 was irradiated with the terahertz wave W1 using the irradiation unit 3, and the terahertz wave W2 was detected by the detection unit 4.

The measurement is performed by scanning a 15 mm square region in the in-plane direction at a position 1 mm in height from the bottom surface of the chamber 11 while moving by 300 μm steps in the in-plane X-axis and Y-axis directions by the moving mechanism 6. It was.
Measurement was performed at 50 points in each of the X-axis and Y-axis directions (total of 2500 points). When the measurement of one point took about 0.5 seconds, the entire measurement took about 20 minutes.
If the maximum dimension in the in-plane direction of the subject 1 is too large, the measurement time becomes long, which may cause a problem of change with time. On the other hand, if the number of points is reduced to avoid this, the resolution is lowered. Then, since the whole measurement time was short and the number of points could be secured sufficiently, it was determined that the measurement conditions in this example were appropriate.
Moreover, although the main component of the shell of the subject 1 is calcium carbonate, the electromagnetic wave of 0.1 to 10.0 THz has good permeability to calcium carbonate, so that the body part inside the shell can be analyzed.
If the subject 1 is too thin, the amount of the target component is reduced and the detection accuracy is reduced. If the subject 1 is too thick, the terahertz wave is difficult to transmit. However, in this example, such a reduction in detection accuracy did not occur. The thickness of specimen 1 was determined to be reasonable.

The main umami component of Yamato Shijimi is an amino acid, of which attention was paid to alanine, glycine, and glutamic acid.
At -195 ° C., alanine crystals are near 2.2 THz, 2.6 THz and 3.0 THz, glycine crystals are 2.8 THz, glutamic acid crystals are 1.2 THz, 2.0 THz, 2.5 THz, and 2. 7 THz and 2.9 THz have their own strong absorption, and it is known that the absorption intensity is almost proportional to the concentration.
Further, it is known that the aqueous sodium chloride solution has a strong absorption around 1.6 THz when rapidly frozen, and the absorption intensity is almost proportional to the concentration.

In order to examine the time-dependent changes in the salinity concentrations of alanine, glycine, glutamic acid and artificial seawater, immediately after (1 hour), subject 1 immersed in artificial seawater at 20 ° C., 30 minutes, 1, 2, 3, 6 and After leaving still for 24 hours, it cooled to -195 degreeC with the cooling rate of -1.5 degreeC / sec, and each test object 1 was frozen with artificial seawater and used for the analysis.
FIG. 4 shows the measurement results at 2.6 THz, which is an absorption band specific to alanine. In this figure, the absorption intensity is represented by the color intensity, and the higher the absorption intensity, the darker the color is displayed.
From this result, it was found that the absorption intensity inside the subject 1 increased with time. No increase or decrease in absorption intensity was observed in the sodium chloride aqueous solution 18 around the subject 1.
Similarly, for glycine and glutamic acid, it was found that the absorption intensity inside the subject 1 increased with time.
In addition, when the change in absorption intensity is averaged over the whole body of the subject 1 and the value after 24 hours is compared with the value at 0 hour, alanine is about 5 times, glycine is about 4.5 times, and glutamic acid is about 3 times. It was found to have increased 5 times. The results of this analysis are the results of previous analysis conducted by extracting amino acids from the body of Shijimi (Shimane Prefectural Fisheries Experiment Station Business Report, Vol.1993, Pages.167-175 (1995) Let ’s eat deliciously ”p.6, http://www.sijimi-lab.jp/oisiku.html).

The sodium chloride aqueous solution 18 around the subject 1 was examined for changes in absorption intensity near 1.6 THz, which is an index of salinity.
If any substance flows out from the subject 1, the salinity concentration of the aqueous solution 18 around the subject 1 becomes low, and it is considered that this absorption intensity is observed to be lower than other regions.
As shown in FIG. 5, in the sample after 30 minutes, a region where the intensity decreased in a very small range around the subject 1 was confirmed. In the sample after 1 hour, the area where the intensity decreased was spread so as to diffuse, and the amount of decrease in the intensity increased. In the samples after 2 hours, the decrease in strength was small. The decrease in strength is considered to be because excrement from the subject 1 flowed out to the surroundings (the same effect as sand removal was obtained).

The same Yamato swordfish as in Example 1 was used as the subject 1, and an analysis test was performed using the analyzer 10 shown in FIG.
As environmental water around the subject 1, three kinds of water with different salinity concentrations were used: tap water (saline concentration 0 mass%), artificial seawater with a salinity concentration 0.5 mass% and 1.0 mass%. The environmental water was introduced into the reservoir 17 to a depth of 2 mm, and the subject 1 was placed in the center so that at least a part thereof was immersed.
The specimen 1 immersed in environmental water at 20 ° C. was cooled to −195 ° C. at a cooling rate of −1.5 ° C./sec immediately after (0 hour) and after standing for 24 hours. The whole environment water was frozen.
These samples were subjected to analysis in the same manner as in Example 1. Other test conditions were in accordance with Example 1.

When time-dependent changes in the salinity concentrations of alanine, glycine, glutamic acid and artificial seawater were examined, the absorption strength inherent to alanine, glycine and glutamic acid was obtained using artificial seawater having a salinity of 0.5 mass% and 1.0 mass%. In some cases, it increased within the subject 1 body. The amount of increase increased with increasing salt concentration.
In the surrounding environmental water, these changes in absorption intensity were not observed.
When tap water (salt concentration: 0% by mass) was used, the absorption intensity inherent to alanine, glycine, and glutamic acid decreased inside the subject 1 body. When the change in absorption intensity was averaged over the whole body of the subject 1 and a sample after 24 hours was compared with a sample at 0 hour, alanine and glycine were reduced to about 1/5 and glutamic acid was reduced to about 1/2. all right.
On the other hand, in the surrounding environmental water, these absorption intensity | strength increases were seen, and it was confirmed that the runoff of alanine, glycine, and glutamic acid has occurred from the subject 1.

The same Yamato swordfish as in Example 1 was used as the subject 1, and an analysis test was performed using the analyzer 10 shown in FIG.
As the environmental water around the subject 1, an aqueous solution in which glucose was added to artificial seawater having a salinity concentration of 1% by mass to a concentration of 0.2% by mass was used.
The specimen 1 immersed in environmental water at 20 ° C. was cooled to −195 ° C. at a cooling rate of −1.5 ° C./sec immediately after (0 hour) and after standing for 24 hours. The whole environment water was frozen.
These samples were subjected to analysis in the same manner as in Example 1. Other test conditions were in accordance with Example 1.

As for the glucose to be analyzed, a resonance band characteristic in the frequency region of 0.1 to 10.0 THz is not observed in the aqueous solution or the sample frozen thereof, but sodium chloride in the artificial seawater used for the environmental water Or, a weak interaction occurs between ions derived from this (sodium ion or chlorine ion) and glucose, and this resonates in the frequency range, making it possible to analyze using changes in absorption intensity. Become.
In order to cause a weak interaction between sodium chloride or sodium chloride-derived ions and glucose, sodium chloride needs to be dispersed and taken into the subject 1 without being precipitated outside the subject 1. It is considered that this state was realized by adopting the cooling rate and the ultimate cooling temperature.
As a result of the test, an absorption band was observed near 1.6 to 1.8 THz due to the interaction between sodium chloride (or derived ions) and glucose. The intensity of the absorption band increased in the body of the subject 1 with time.
Therefore, the uptake of glucose from the surrounding environment into the subject 1 was confirmed. Further, since the increase or decrease in the absorption intensity was small in the region around the subject 1, it was found that the increase or decrease in the glucose concentration was small.

From the results of Examples 1 to 3, according to the present invention, the chemical substance in the environment immediately before freezing and the biological tissue placed in this environment without introducing or staining the labeling agent to the subject 1 and its It was found that the concentration distribution can be grasped in relation to the shape of the subject 1.
It was also found that components (inorganic salts, etc.) that are difficult to introduce labeling agents (fluorescent probes, etc.) can be analyzed.
Moreover, since pre-processing is unnecessary, it is not necessary to consider the state change in pre-processing, and it was confirmed that the state of the living body placed in the environment can be accurately grasped.
In addition, since component analysis including the surrounding environment of the subject 1 is possible, it was confirmed that the exchange of substances between the subject 1 and the surrounding environment can be observed.

DESCRIPTION OF SYMBOLS 1 ... Subject, 2 ... Mounting part, 3 ... Irradiation part, 4 ... Detection part, 5 ... Calculation / control part, 6 ... Moving mechanism, 7 ... Cooling Part, 10, 20 ... analyzer, 14 ... in-plane moving mechanism, 15 ... elevating mechanism, 18 ... sodium chloride aqueous solution (environmental water).

Claims (10)

A method for analyzing a component of a living subject,
Based on an absorption characteristic spectrum obtained by irradiating the frozen subject with an electromagnetic wave of 0.1 to 10 THz and detecting the electromagnetic wave transmitted or reflected through the subject, a distribution of the components of the subject is obtained. The biological component analysis method characterized by the above-mentioned.   The biological component analysis method according to claim 1, wherein the distribution of the component of the subject is obtained by performing detection of the electromagnetic wave at a plurality of points at different positions in the subject.   The biological component analysis method according to claim 1, wherein the distribution of the component of the subject is measured over time. The subject is frozen with the aqueous sodium chloride solution, with at least a portion immersed in the aqueous sodium chloride solution,
The said electromagnetic wave of the said sodium chloride aqueous solution in the circumference | surroundings is detected with the detection of the said electromagnetic wave about the said test object, The distribution of the said component is obtained, The any one of Claims 1-3 characterized by the above-mentioned. The biological component analysis method.   The biological component analysis method according to claim 1, wherein the subject has a thickness in the irradiation direction of the electromagnetic wave of 0.1 to 15 mm.   The biological component analysis method according to any one of claims 1 to 5, wherein the subject has a maximum dimension in a plane that intersects the irradiation direction of the electromagnetic wave of 3 to 100 mm.   The freezing temperature of the subject is -0.1 to -5 ° C / second, and the ultimate cooling temperature is -200 to 0 ° C. The biological component analysis method according to 1. An apparatus for analyzing a component of a living subject,
A placement section for placing the frozen subject;
An irradiation unit that irradiates a subject on the placement unit with an electromagnetic wave of 0.1 to 10 THz;
A detection unit for detecting electromagnetic waves transmitted or reflected by the subject;
A biological component analyzer comprising: a calculation / control unit that obtains a distribution of components of the subject based on an absorption characteristic spectrum obtained by electromagnetic waves detected by the detection unit. A moving mechanism for moving the placement unit relative to the irradiation unit and the detection unit;
The biological component analyzer according to claim 8, wherein the moving mechanism can arbitrarily set the position of the subject by moving the placement unit.   The biological component analyzer according to claim 8 or 9, wherein the moving mechanism is capable of moving the placement unit in an irradiation direction of the electromagnetic wave and an in-plane direction intersecting with the irradiation direction.

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