JP5172467B2 - Quantitative spectroscopic analysis of blood components - Google Patents

Description

  The present invention relates to a quantitative spectroscopic analysis method for components in blood.

  Quantitative analysis of target substances such as sugars, amino acids, and proteins in blood samples such as serum, plasma, blood cells, and clots separated from collected blood is widely used because it provides useful information for medical care and treatment of diseases Has been.

As a quantitative analysis method of a target substance in a solution, for example, an electromagnetic wave having a frequency resonating with energy of weak interaction such as hydrogen bond, van der Waals bond, π electron interaction, electrostatic interaction (for example, terahertz wave) A terahertz time domain spectroscopy using an absorption characteristic spectrum of the above is known (Non-Patent Document 1).
Terahertz time-domain spectroscopy is a method for directly performing qualitative quantitative analysis by obtaining an absorption characteristic spectrum derived from energy specific to a target substance.
Y. Ueno, R.M. Rungsavan, I.D. Tomita, and K.K. Ajito, Anal. Chem. 2006, 78, 5424-5428.

However, the spectroscopic method as described in Non-Patent Document 1 has a low limit of quantification of about several tens of percent because of the low absorption intensity inherent in the target substance, and cannot cope with highly sensitive quantitative analysis. Therefore, in order to quantify the concentration of the target substance in the sample blood using the electromagnetic wave that resonates with the energy of the weak interaction as described above, it is desired to improve the sensitivity.
Therefore, an object of the present invention is to provide a quantitative spectroscopic analysis method for components in blood that can quantitate the concentration of a target substance in a sample blood with high sensitivity using an electromagnetic wave that resonates with weak interaction energy.

  The quantitative spectroscopic analysis of blood components according to the present invention is a method for quantifying the concentration of a target substance contained in a sample blood of serum, plasma, blood cells, or blood clot, wherein the sample blood is treated with the sample blood. Is frozen as it is or diluted and an inorganic salt is added to obtain a frozen sample, and then the frozen sample is mixed with an inorganic salt component previously contained in the sample blood, and the added inorganic salt Or a frequency that resonates with the energy of a weak interaction formed between an ion generated from the added inorganic salt and the target substance, or between the ion, the target substance, and water molecules around them. In this method, the concentration of the target substance is quantified from an absorption characteristic spectrum obtained by irradiating the electromagnetic wave containing the electromagnetic wave and measuring the electromagnetic wave transmitted through the frozen sample.

The quantitative spectroscopic analysis of blood components according to the present invention is such that the added inorganic salt is zinc, potassium, calcium, chromium, selenium, iron, copper, sodium, magnesium, manganese, phosphorus in the sample blood. , Vanadium, strontium or ammonium cation, or a salt that produces any one of fluorine, chlorine, bromine, iodine, hydroxy acid, sulfurous acid, carbonic acid, nitric acid, phosphoric acid or boric acid anion .
Moreover, it is preferable that the target substance that forms the interaction is a substance mainly composed of any of sugar, organic acid, organic acid salt, amino acid, amino acid salt, polypeptide, and protein.

  The quantitative spectroscopic analysis method for blood components of the present invention can quantitate the concentration of a target substance in sample blood with high sensitivity using electromagnetic waves that resonate with weak interaction energy.

  The quantitative spectroscopic analysis of blood components according to the present invention is a method for quantifying the concentration of a target substance in sample blood from the absorption characteristic spectrum of a frozen sample obtained by freezing sample blood. Hereinafter, an embodiment of the quantitative spectroscopic analysis method of the present invention will be described in detail.

[Spectroscopic analyzer]
FIG. 1 is a schematic diagram showing an example of an analyzer that can be used in the quantitative spectroscopic analysis method of the present invention. As shown in FIG. 1, the spectroscopic analyzer 10 according to the present embodiment generates a light source 12 that generates excitation light 40 that excites an electromagnetic wave in a frequency range of 0.1 to 10 THz, and an electromagnetic wave 42 using the excitation light 40. A cryogen 14 having a pulse generator 22 for irradiating the frozen sample 30 and a detector 24 for detecting the electromagnetic wave transmitted through the frozen sample 30, and a sample blood to which an inorganic salt is added is frozen to obtain a frozen sample 30 at a low temperature. Control and calculation to obtain the absorption characteristic spectrum by controlling the chamber 16, the thickness measuring means 18 for measuring the thickness of the frozen sample, the spectroscope 14, the low temperature chamber 16 and the thickness measuring means 18 and performing spectrum calculation. Means 20.
The frozen sample 30 is a sample obtained by freezing the sample blood in a state where the sample blood is directly or diluted and an inorganic salt is added.

  The light source 12 is not particularly limited as long as it can generate the excitation light 40 that excites electromagnetic waves (terahertz waves) in the frequency range of 0.1 to 10 THz. For example, a pulse laser or titanium sapphire that generates femtosecond laser excitation light. A laser etc. are mentioned.

The spectroscope 14 includes a pulse generator 22 that generates an electromagnetic wave 42 by the excitation light 40 and irradiates the frozen sample 30, and a detector 24 that detects the electromagnetic wave transmitted through the frozen sample 30.
The pulse generator 22 is not particularly limited as long as it can receive the excitation light 40 from the light source 12 and generate the electromagnetic wave 42 (terahertz wave). For example, the non-linear optical crystal, photoconductive antenna, semiconductor, quantum Wells, high-temperature conductive thin films, and the like.
The detector 24 is not particularly limited as long as it can detect the electromagnetic wave that has passed through the frozen sample 30, and examples thereof include a photoconductive antenna.

  The low temperature chamber 16 is not particularly limited as long as it has a freezing function, and it is preferable that the temperature in the chamber can be adjusted and maintained at an arbitrary temperature in the range of −190 ° C. to room temperature. In addition, the freezing function is such that the freezing rate is preferably 10 ° C./min or more, and more preferably 30 ° C./min or more from the viewpoint that it is easy to suppress the precipitation of inorganic salts. An example of such a low temperature chamber 16 is a chamber manufactured by Nippon Thermal Co., Ltd.

  The thickness measuring means 18 is not particularly limited as long as it can measure the thickness of the frozen sample 30, and examples thereof include Mitutoyo calipers.

The control / calculation means 20 is a means for controlling the spectroscope 14, the low temperature chamber 16, and the thickness measurement means 18, and performing spectrum calculation to obtain an absorption characteristic spectrum.
As the control in the control / calculation means 20, for example, the cryogenic chamber 16 is controlled to control the temperature during preparation and measurement of the frozen sample 30, and the measurement is controlled by the control of the spectrometer 14 and the thickness measuring means 18. Etc. Further, as the calculation in the control / calculation means 20, there is a method in which spectrum calculation is performed from the detection value in the detector 24 to obtain an absorption characteristic spectrum. The spectrum calculation includes obtaining a continuous spectrum from the detected value by Fourier transform and normalizing the continuous spectrum per unit thickness based on the thickness information of the frozen sample 30 sent from the thickness measuring means 18. Can be mentioned.
The control / calculation means 20 is not particularly limited as long as such control and calculation can be performed.

[Quantitative spectroscopy]
Hereinafter, a method for quantifying the concentration of a target substance using the above-described spectroscopic analyzer 10 will be described as an example of an embodiment of a quantitative spectroscopic analysis method of the present invention.
The sample blood in the quantitative spectroscopic analysis method of the present invention is a sample to be subjected to a blood test of serum, plasma, blood cells, or blood clot.
The target substance in the present invention is a substance that is contained in the sample blood and forms a weak interaction in the sample blood. For example, sugar (glucose, etc.), organic acid (succinic acid, malic acid, citric acid) Etc.), salts of organic acids, amino acids (methionine, alanine, valine, leucine, isoleucine, proline, phenylalanine, glycine, serine, threonine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, tryptophan, cysteine ), A substance mainly comprising any one of an amino acid salt, a polypeptide and a protein. Here, the weak interaction is an interaction such as a hydrogen bond, van der Waals bond, π-electron interaction, or electrostatic interaction.
In addition, the sample blood contains in advance inorganic salt components such as sodium, zinc, magnesium, iron, potassium, and calcium.

In the quantitative spectroscopic analysis method of the present invention, the frozen sample 30 is obtained by freezing the sample blood in a state where the sample blood is intact or diluted and an inorganic salt is added. When diluting the sample blood, an inorganic salt may be added before diluting the sample blood, or the sample blood may be diluted after adding the inorganic salt. Alternatively, an aqueous solution in which an inorganic salt is dissolved may be used. Dilution of the sample blood is preferably performed with water, particularly preferably with pure water.
The inorganic salt can be dissolved using, for example, a stirrer.

The inorganic salt added in the present invention is weak in the sample blood between the inorganic salt or an ion generated from the inorganic salt and the target substance, or between the ion and the target substance and water molecules around them. There is no particular limitation as long as it forms an interaction.
As an inorganic salt, zinc, potassium, calcium, chromium, selenium, iron, copper, sodium, magnesium, manganese, phosphorus, vanadium, strontium or ammonium is used in the blood sample because it is excellent in determining the concentration of the target substance. Preference is given to any cation or a salt that produces any anion of fluorine, chlorine, bromine, iodine, hydroxy acid, sulfurous acid, carbonic acid, nitric acid, phosphoric acid or boric acid.
Only one inorganic salt may be used alone, or two or more inorganic salts may be used in combination.

  The amount of the inorganic salt added to the sample blood may be 1% by mass to the saturated concentration, and varies depending on the type of the inorganic salt as shown in FIG. It is preferable that the concentration of the inorganic salt is 1 to 50% by mass with respect to 100% by mass. If the concentration is 20% by mass or more, the absorption characteristic spectrum obtained regardless of the type of inorganic salt becomes remarkable, and the concentration of the target substance can be easily determined. Moreover, if the said density | concentration is 10 mass% or less, it will become easy to dissolve inorganic salt in sample blood irrespective of the kind of inorganic salt.

As a method of obtaining the frozen sample 30 by freezing the sample blood to which the inorganic salt has been added, for example, the sample blood to which the inorganic salt has been added is dropped on a sample holder or the like and placed in the low temperature chamber 16 so that the low temperature is reduced. A method of freezing by lowering the temperature in the chamber 16 may be used.
The rate of freezing the sample blood is not particularly limited as long as the sample blood can be rapidly frozen, and is preferably 10 ° C./min or more, and more preferably 30 ° C./min or more. If the speed at which the sample blood is frozen is 10 ° C./min or more, it is easy to suppress the precipitation of inorganic salts during freezing, so that the concentration of the target substance can be easily determined.

  The thickness of the frozen sample 30 is not particularly limited, and can be determined according to the performance and size of the spectroscope 14 and the thickness measuring means 18. However, if the thickness of the frozen sample 30 is too thin, the amount of the target substance and inorganic salt becomes too small, and the absorption intensity in the absorption characteristic spectrum obtained becomes weak, which may make it difficult to determine the concentration of the target substance. There is. If the frozen sample 30 is too thick, the electromagnetic wave 42 is difficult to pass through the frozen sample 30. In addition, considering the interference between the electromagnetic wave 42 and the thickness of the frozen sample 30, the thickness of the frozen sample 30 is preferably 1 to 3 mm.

Next, the absorption characteristic spectrum is obtained by irradiating the frozen sample 30 with electromagnetic waves and measuring the electromagnetic waves transmitted through the frozen sample 30.
The electromagnetic wave irradiated to the frozen sample 30 is an inorganic salt component previously contained in the sample blood, the added inorganic salt or an ion generated from the added inorganic salt and the target substance, or the ion And an electromagnetic wave including a frequency that resonates with the energy of a weak interaction formed between the target substance and water molecules around them. That is, between the inorganic salt component preliminarily contained in the sample blood and the target substance, or between the added inorganic salt and the target substance, or ions generated from the added inorganic salt and the It is an electromagnetic wave including a frequency that resonates with the energy of a weak interaction formed between the target substance or between the ions, the target substance, and water molecules around them.
Examples of weak interactions include interactions such as hydrogen bonds, van der Waals bonds, π-electron interactions, and electrostatic interactions. Specific examples of the electromagnetic wave 42 including a frequency that resonates with the energy of these weak interactions include, for example, an electromagnetic wave (terahertz wave) in a frequency range of 0.1 to 10 THz.

The electromagnetic wave 42 irradiated to the frozen sample 30 is an inorganic salt component, an inorganic salt or an ion generated from the inorganic salt previously contained in the sample blood and the target substance, or the ions, the target substance, and their surroundings. Resonates with the energy of the weak interaction that occurs between water molecules. Then, the electromagnetic wave transmitted through the frozen sample 30 is detected by the detector 24. At the same time, the detector 24 is irradiated with a part of the excitation light 40 for sampling detection.
Then, the detection value obtained by detecting the electromagnetic wave transmitted through the target substance and the detection value obtained by the sampling detection are sent as data from the detector 24 to the control / calculation means 20 and converted into a continuous spectrum of the frequency component of the electromagnetic wave 42 by Fourier transformation. Converted. At the same time, based on the temperature in the cryogenic chamber 16 and the thickness value of the frozen sample 30 sent from the thickness measurement means 18 to the control / calculation means 20, the obtained continuous spectrum is converted into a unit thickness at the analysis execution temperature. Normalization is performed to obtain an absorption characteristic spectrum. The normalization uses a proportional relationship between the thickness of the frozen sample 30 and the absorption intensity of the absorption characteristic spectrum. Specifically, the absorption characteristic spectrum is divided by the thickness of the frozen sample 30 measured by the thickness measuring means 18.
Further, ice prepared by freezing pure water is prepared as a reference sample, and an absorption characteristic spectrum is obtained for the reference sample by performing the same measurement as in the above-described detection step, and is referenced from the normalized absorption characteristic spectrum. The absorption characteristic spectrum of the frozen sample 30 is obtained by subtracting the absorption characteristic spectrum of the sample.

The temperature in the low temperature chamber 16 during analysis is not particularly limited as long as the frozen sample 30 is not thawed. However, the lower the measurement temperature, the less the thermal fluctuation and the sharper spectrum can be obtained. Moreover, since an absorption characteristic spectrum may change according to temperature, it is preferable to obtain an absorption characteristic spectrum on several temperature conditions.
The temperature in the low temperature chamber 16 during analysis is preferably 4 to 300K.

  As described above, an absorption characteristic spectrum is obtained from the electromagnetic wave transmitted through the frozen sample 30. The obtained absorption characteristic spectrum includes an inorganic salt component, an ion generated from an inorganic salt or an inorganic salt, and a target substance, or water around the ion, the target substance, and their surroundings. A peak due to resonance with the energy of the weak interaction formed between the molecules is obtained. By using this peak, the concentration of the target substance in the sample blood can be quantified.

  As a method for quantifying the concentration of a target substance by the quantitative spectroscopic analysis method of the present invention, a sample blood having a known concentration by dissolving the target substance is used as a standard sample, and a calibration curve is obtained from the peak in the absorption characteristic spectrum obtained thereby. (1) and a method for obtaining a standard absorption spectrum from the absorption characteristic spectrum obtained from the standard sample and comparing the standard absorption spectrum with the absorption characteristic spectrum of the target sample blood ( 2).

The method for obtaining the absorption characteristic spectrum of the standard sample can be the same as the method for obtaining the absorption characteristic spectrum of the sample blood described above, and the preferred embodiment is also the same.
When using the calibration curve of method (1), the calibration curve is calculated from the concentration (known) and absorption intensity of the standard sample at the frequency at which the peak due to resonance with the energy of the interaction occurs in the absorption characteristic spectrum. create. Thereby, the concentration of the target substance can be quantified from the absorption intensity of the same frequency in the absorption characteristic spectrum of the sample blood to be processed.

  In the case of method (2), the standard absorption spectrum is obtained by dividing the absorption characteristic spectrum of the standard sample by the concentration of the target substance in the standard sample. Next, the absorption characteristic spectrum of the target sample blood is compared with the standard absorption spectrum, and the error between the theoretical absorption characteristic spectrum obtained from the product of the standard absorption spectrum and the coefficient and the absorption characteristic spectrum is minimized. By calculating the coefficient, the concentration of the target substance can be quantified.

The comparison using the absorption characteristic spectrum of the target sample blood in the method (1) and the method (2) is performed between a calibration curve based on the absorption characteristic spectrum of the standard sample measured under the same temperature condition or a standard absorption spectrum. The temperature conditions of the absorption characteristic spectrum and the standard absorption spectrum used for comparison are not particularly limited, and may be a single temperature condition. However, since the absorption characteristic spectrum may change depending on the temperature at the time of analysis, it is preferable to perform comparison under a plurality of temperature conditions.
In the method (1), it is preferable to calculate an average value and a standard deviation of the concentration of the target substance obtained at each temperature. In the method (2), it is preferable to calculate an average value and a standard deviation of the coefficients obtained at each temperature.

When targeting multiple target substances in the sample blood, prepare a standard curve or standard absorption spectrum for all the target substances to obtain the concentration of all target substances in the sample blood. Can be quantified.
That is, in the method (1), each target substance in the sample blood is obtained by using a calibration curve obtained from the absorption characteristic spectrum of the standard sample having the same peak for each peak of the absorption characteristic spectrum of the target sample blood. The concentration of can be quantified.
Also in the method (2), by extracting a standard absorption spectrum having the same peak for each peak of the absorption characteristic spectrum of the target sample blood, and calculating a coefficient that minimizes the error between the peaks, The concentration of each target substance in the sample blood can be quantified.

Further, according to the quantitative spectroscopic analysis method of the present invention, the electromagnetic wave 42 irradiated on the frozen sample 30 is analyzed by removing the data of the absorption characteristic spectrum in the frequency region below 0.5 THz and above 2.0 THz for the following reason. You can also. In the frequency region below 0.5 THz, there is a tendency that an error due to the influence of multiple reflection or the like occurring at the interface of the frozen sample 30 becomes large. On the other hand, in the frequency region exceeding 2.0 THz, the noise level tends to increase as the absorption intensity increases toward the high frequency side. Therefore, in some cases, the error in the above-described frequency region in the absorption characteristic spectrum becomes too large, and it may be difficult to quantify the concentration of the target substance.
Therefore, measurement may be performed by removing spectral data in a frequency region that is inappropriate for the quantitative spectroscopic analysis method of the present invention. Here, the frequency region inappropriate for analysis is, for example, in the case of method (2), a region in which the error of the standard absorption spectrum in each concentration condition of the target substance becomes less than 10% as a result of the removal of the frequency region. It is. Specifically, a method of performing measurement using electromagnetic waves in a frequency region of 0.5 to 2.0 THz can be mentioned.

  Further, the baseline of the absorption characteristic spectrum of the sample blood may change due to the output stability of the laser of the apparatus or the thermal action of the optical component. For this reason, it is preferable to perform baseline correction by adding or subtracting a constant or a value having a certain relationship to the frequency in a frequency region with a large error. Thereby, the concentration of the target substance in the sample blood can be quantified more accurately.

The above-described quantitative spectroscopic analysis of blood components according to the present invention can quantitate the concentration of a target substance in sample blood with high sensitivity using electromagnetic waves that resonate with weak interaction energy.
This is because, in the quantitative spectroscopic analysis method of the present invention, an inorganic salt is used, and between the inorganic salt or an ion generated from the inorganic salt and the target substance, or between the ion and the target substance and water molecules around them. This is because the absorption intensity of the absorption characteristic spectrum becomes stronger than that of the existing technology that directly measures the intrinsic energy of the target substance.

Hereinafter, the present invention will be described in detail with reference to Examples and Comparative Examples. However, the present invention is not limited by the following description.
Using the spectroscopic analyzer 10 illustrated in FIG. 1, the concentration of the target substance in the sample blood was quantified. The spectroscopic analyzer 10 uses a vitasse (100 femtosecond type, manufactured by Coherent) as a light source 12, a pulse generator 22 (photoconductive antenna, manufactured by Hamamatsu Photonics), and a detector 24 (photoconductive). A spectroscope 14 equipped with an antenna (manufactured by Hamamatsu Photonics) and a low temperature chamber 16 (low temperature chamber with instantaneous cooler, manufactured by Nippon Thermal) and thickness measuring means 18 were used.

[Preparation of frozen sample]
The target substance was glucose in serum. Serum was used as sample blood, which was diluted twice with pure water to obtain a 50% serum aqueous solution, and sodium chloride, an inorganic salt, was added to the diluted sample blood so that the concentration was 20% by mass. In addition, sample blood A was prepared by adding glucose as a target substance so that the concentration was 0.2% by mass. Sample blood B was prepared in the same manner as sample blood A except that glucose was not added. Furthermore, a 20 mass% sodium chloride aqueous solution (sample C) was prepared.

These three types of sample blood A, B and sample C are dropped onto a sample holder having a diameter of 6 mm, cooled from room temperature to −190 ° C. at a rate of −30 ° C. per minute by the low temperature chamber 16, and frozen at a thickness of 2 mm. Samples A to C were obtained.
In addition, ice was prepared using the same amount of pure water and used as a reference sample when measuring the absorption characteristic spectrum.

[Experimental Example 1]
The frozen sample A obtained by the sample preparation described above is placed in the thickness measuring means 18 and measured with an electromagnetic wave in the frequency range of 0.1 to 10.0 THz, and the continuous spectrum obtained by the measurement is obtained per unit sample thickness. Then, the absorption intensity spectrum of the frozen sample A was obtained by subtracting the absorption intensity of the reference sample obtained by the same measurement. For frozen samples B and C, absorption characteristic spectra of the frozen samples were obtained in the same manner as frozen sample A.
The absorption characteristic spectra of the frozen samples A to C are shown in FIG.

  As shown in FIG. 3, a characteristic peak was obtained in the absorption characteristic spectrum of the frozen sample A (particularly around 1.8 THz). This peak was not obtained in the absorption characteristic spectra of the frozen samples B and C. In addition, even if measurement is performed only with an aqueous glucose solution or serum only under the same concentration conditions, an absorption characteristic spectrum like that of the frozen sample A cannot be obtained. Therefore, these peaks (1.8 THz) It is not a peak corresponding to the intrinsic energy of chemical components in serum, but a peak corresponding to the energy of weak interactions formed between glucose or chemical components in serum and ions derived from inorganic salts. I understood.

Subsequently, glucose in serum was quantified.
[Example 1]
Serum is used as sample blood, and this is diluted twice with pure water to obtain a 50% serum aqueous solution, and glucose is added so that the concentrations thereof are 0.05 mass%, 0.1 mass%, and 0.2 mass%, respectively. And three types of blood samples D to F were prepared by adding sodium chloride, which is an inorganic salt, to a concentration of 20% by mass.
Next, with respect to the sample blood D to F, frozen samples D to F were prepared in the same manner as in Experimental Example 1, and the absorption characteristic spectrum of each frozen sample was measured.
The absorption characteristic spectra of the frozen samples D to F are shown in FIG.

As shown in FIG. 4, characteristic peaks were obtained in the absorption spectra of the frozen samples D to F (particularly around 1.8 THz). Further, from the comparison of the absorption spectra of the frozen samples D to F, there is a correlation between the absorption intensity at the peak of the absorption characteristic spectrum and the concentration of glucose, and the peak in the absorption characteristic spectrum is an ion generated from glucose and an inorganic salt. It was found that the peak corresponds to the energy of the weak interaction formed between the two.
Moreover, the result of having plotted the relationship between the absorption intensity of the peak of 1.8 THz and the density | concentration of glucose is shown in FIG. As shown in FIG. 5, a good linear relationship was obtained between the absorption intensity at the peak of 1.8 THz and the glucose concentration. Thus, it was shown that glucose can be quantified by using the absorption intensity of this peak.
Moreover, since it was shown that 0.05 mass% glucose can be quantified by this example, the detection limit is at least as compared with the conventional direct qualitative quantitative analysis method that performs quantification from the absorption intensity specific to the target substance. It can be expected that it can be increased by 100 times or more, and it has been found that quantitative analysis can be performed with extremely high sensitivity.

  According to the quantitative spectroscopic analysis method of the present invention, an electromagnetic wave including a frequency resonating with energy of weak interaction such as hydrogen bond, van der Waals bond, π electron interaction, electrostatic interaction, or the like is used. The concentration of the target sample can be quantified with high sensitivity. Therefore, it can be suitably used for quantifying the concentration of a target substance in a sample blood to be subjected to blood tests such as serum, plasma, blood cells, and blood clots.

It is the schematic diagram which showed an example of the spectroscopic analyzer which can be used for the quantitative spectroscopic analysis of this invention. It is the figure which showed the solubility curve of various inorganic salt with respect to 100 g of water. It is the figure which showed the absorption characteristic spectrum of frozen sample AC. It is the figure which showed the absorption characteristic spectrum of frozen samples D-F. It is the figure which plotted the relationship between the absorption intensity of the peak of 1.8 THz of FIG. 4, and glucose concentration.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 10 Spectroscopic analyzer 12 Light source 14 Spectrometer 16 Low temperature chamber 18 Thickness measurement means

Claims (3)

A method for quantifying the concentration of a target substance contained in a blood sample of any one of serum, plasma, blood cells or clots,
The sample blood is frozen with the sample blood as it is or diluted and an inorganic salt is added to obtain a frozen sample,
In the frozen sample, an inorganic salt component previously contained in the sample blood, the added inorganic salt, or an ion generated from the added inorganic salt and the target substance, or the ion and the target From the absorption characteristic spectrum obtained by irradiating an electromagnetic wave containing a frequency resonating with the energy of weak interaction formed between substances and water molecules around them, and measuring the electromagnetic wave transmitted through the frozen sample Quantitative spectroscopic analysis of blood components to quantify the concentration of target substances.   The added inorganic salt is a cation of zinc, potassium, calcium, chromium, selenium, iron, copper, sodium, magnesium, manganese, phosphorus, vanadium, strontium or ammonium, or fluorine in the sample blood. 2. The quantitative spectroscopic analysis of blood components according to claim 1, wherein the salt produces an anion of any one of chlorine, bromine, iodine, hydroxy acid, sulfurous acid, carbonic acid, nitric acid, phosphoric acid or boric acid.   The target substance that forms the interaction is a substance mainly comprising any one of a sugar, an organic acid, an organic acid salt, an amino acid, an amino acid salt, a polypeptide, and a protein. Quantitative spectroscopic analysis of blood components.

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