JP2013057560A - Reaction speed analysis device using nuclear magnetic resonance - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a reaction speed analysis device utilizing nuclear magnetic resonance capable of analyzing even resonance speed of a reaction with high temperature variation responsiveness to be induced by variation of temperature by NMR measurement.SOLUTION: A reaction speed analysis device utilizing nuclear magnetic resonance is the device for analyzing speed of a reaction to be induced by variation of temperature by nuclear magnetic resonance measurement, and consists of: 1) an NMR magnet which provides sample solution with a magnetic field; 2) a sample solution circulatory system with: a holding part which is installed in a bore in the NMR magnet, and in which the sample solution receives the magnetic field; annular piping consecutively connected to the holding part, a pump which performs circulatory supply of the sample solution in the piping, a first temperature adjustment part which adjusts temperature of the sample solution outside the NMR magnet, and a second temperature adjustment part which adjusts the temperature of the sample solution in the NMR magnet; 3) a detection/control part with a detection coil which is arranged in the outer peripherals of the holding part to detect an NMR signal of the sample solution, and a spectrometer which receives the NMR signal to be converted into an NMR spectrum or an MRI image picture.

Description

The present invention relates to a method for analyzing the rate of reaction induced by a change in temperature by measurement (NMR measurement) using nuclear magnetic resonance phenomena such as NMR spectrum measurement and MRI measurement, and a temperature variable that makes it possible. The present invention relates to a device with a function.

The change in the state and structure of a substance due to a change in temperature is a property of a very wide range of substances, as represented by the phase transition of water. Proteins that make up living organisms form a unique three-dimensional structure at room temperature, but denature and lose at high temperatures. Some proteins may be denatured by heating, but will rewind and regain their inherent conformation when cooled again.

Spectral measurement using nuclear magnetic resonance (hereinafter referred to as NMR spectrum measurement), which detects changes in the three-dimensional structure and chemical structure of molecules, is a useful measurement method with a large amount of information. Can be detected with high sensitivity. Akasaka et al. Succeeded in obtaining the rate constant of the protein structural change reaction by observing the unwinding process of the protein by NMR spectrum measurement and analyzing the change in the obtained spectrum (hereinafter referred to as NMR spectrum). (Non-Patent Documents 1 and 2).

In the techniques of Non-Patent Documents 1 and 2, the sample tube containing the protein solution installed in the detection part of the NMR magnet is denatured by blowing high-temperature air, and then switched to low-temperature air for rewinding. Guide and perform multiple NMR measurements in succession. Then, the rate constant is quantified by analyzing the NMR spectrum change as an exponential function change. The rate constant obtained in Non-Patent Documents 1 and 2 was about 0.03 s ^{−1 to} 0.04 s ⁻¹ .

However, with the techniques of Non-Patent Documents 1 and 2, it is considered difficult to analyze a reaction that is an order of magnitude faster (rate constant 0.5 s ⁻¹ or so). The reason is that it takes about 10 seconds for the temperature to reach the target value and stabilize after switching the air to be blown. This becomes a dead time, and during this time, the amplitude of the early reaction is significantly attenuated to 1/170, which makes analysis difficult.

As a method for inducing protein denaturation and rewinding, in addition to the above-mentioned air blowing, the sample in which the protein is dissolved is circulated in the circulation flow system by the force of the liquid feed pump, and the heating unit and the cooling unit are included therein. Are inserted to induce denaturation and rewinding, respectively (Non-patent Document 3).

In the method of Non-Patent Document 3, since the liquid feed pump is always in an operating state, the protein sample solution denatured through the heating unit starts the rewinding reaction through the cooling unit and reaches the detection coil for NMR measurement. Only one NMR spectrum was observed at, and no rate constant was obtained.

Moreover, since the cooling part for inducing protein rewinding exists outside the NMR magnet, there is a distance from the detection coil, and the time to reach the detection coil after the rewinding reaction is about 15 seconds. Therefore, even if the liquid feeding pump is stopped and the NMR spectrum change can be observed, the reaction at the level of the rate constant 0.5 s ⁻¹ cannot be analyzed.

JP 2007-315826 A

Akasaka et al. “Construction and performance of a temperature-jump NMR apparatus” Rev. Sci. Instrum. 61, 66-68, 1990 Akasaka et al. “Temperature-jump NMR study of protein folding: Ribonuclease A at low pH” J. Biomol. NMR1, 65-70, 1991 Alder et al. “Structural studies of a folding intermediate of bovine pancreatic ribonuclease A by continuous recycled flow” Biochemistry 27, 2471-2480, 1988 Kitagawa et al. “A new titration system of anovel split-type superconducting magnet NMR spectrometer” Rev. Sci. Instrum. 79, 123109, 2008

In the prior art disclosed in Non-Patent Documents 1 to 3, the time until the NMR spectrum change can be detected after the start of the protein structural change accompanying the temperature change is about 10 seconds or more, and the rate constant is 0.5 s ⁻¹ . It is very difficult to analyze the reaction.

Therefore, the present invention provides a reaction rate analyzer using nuclear magnetic resonance, which can be analyzed using NMR measurement even if the reaction rate of the reaction induced by temperature change is high in temperature change responsiveness. Specifically, the sample solution containing the sample reaches the holding part in the NMR magnet within 1 second after the temperature change starts, and the dead time including the waiting time thereafter is shortened to 2 seconds or less. An object of the present invention is to provide a reaction rate analyzer using nuclear magnetic resonance capable of quantifying the rate constant of a fast reaction of about 0.5 s ⁻¹ using NMR measurement.

In order to solve the above problems, the present invention
(1)
An apparatus for analyzing the rate of reaction induced by a change in temperature by nuclear magnetic resonance measurement,
1) NMR magnet for NMR spectrum measurement or MRI measurement that gives a magnetic field to the sample solution, and 2) a holding unit that is installed in a bore in the NMR magnet to receive the magnetic field, and a ring that is connected to the holding unit. A pipe for circulating the sample solution in the pipe, a first temperature adjusting unit for adjusting the temperature of the sample solution outside the NMR magnet, and a second temperature for adjusting the temperature of the sample solution in the NMR magnet A sample solution circulation system provided with a control unit; 3) a detection coil arranged on the outer periphery of the holding unit for detecting an NMR signal of the sample solution; and a spectrometer for receiving the NMR signal and converting it into an NMR spectrum or an MRI image image. The reaction rate analyzing apparatus using nuclear magnetic resonance is characterized by comprising a detection / control unit.
(2)
The first temperature control unit is a heating unit that heats the sample solution, and the second temperature control unit is a cooling unit that cools the sample solution, or the second temperature control unit is a cooling unit that cools the sample solution. Thus, the second temperature control unit is a heating unit that heats the sample solution, and the reaction rate analyzer using nuclear magnetic resonance according to (1) is configured.
(3)
The nuclear magnetism according to (1) or (2), wherein the spectrometer generates an ON / OFF signal for controlling the driving of the pump, transmits the signal to the pump, and controls the driving of the pump. It was set as the structure of the reaction rate analyzer using resonance.
(4)
In the case of NMR spectrum measurement, the holding unit, the detection coil, and the second temperature control unit constitute a nuclear magnetic resonance probe, and the probe can be attached to and detached from the bore. It was set as the structure of the reaction rate analyzer using the nuclear magnetic resonance in any one of (3).
(5)
In the reaction rate analysis apparatus using nuclear magnetic resonance according to any one of (1) to (4), the sample solution that has been operated by the pump and changed in temperature by the second temperature adjusting unit is fed to the holding unit. The pump is stopped, and NMR measurement of the sample solution in the holding unit is performed every predetermined time. Further, the pump operation, stop, and NMR measurement of the sample solution are repeated several times to obtain an NMR signal. The reaction rate analysis apparatus using nuclear magnetic resonance is characterized in that a plurality of obtained equivalent NMR spectra or MRI image images are integrated to analyze the reaction rate.
(6)
In the reaction rate analyzer using nuclear magnetic resonance according to any one of (1) to (5), the time for feeding the sample solution from the second temperature control unit to the holding unit is within one second. The reaction rate analyzer using nuclear magnetic resonance is characterized.

  Since this invention is the said structure, it has the following effect. Since the second temperature adjusting part (cooling part or heating part) of the sample solution is arranged in the NMR magnet, the reaction state of the sample immediately after the temperature change can be detected. In the case of NMR spectrum measurement, the probe can be easily attached to and detached from the bore by forming a nuclear magnetic resonance probe with the holding unit, the detection coil, and the second temperature adjusting unit.

In addition, the reaction constant of 0.5 s ⁻¹ or more can be quantified by setting the sample solution feeding time within one second from the temperature control device to the holding unit. Further, the pipe for feeding the sample solution is annular, and by controlling the ON / OFF of the drive of the pump, the NMR measurement of the sample solution can be repeatedly performed in the holding unit, and the NMR signals are integrated. Thus, the signal / noise ratio is improved, and the reaction induced by the temperature change can be accurately detected.

FIG. 1 is a schematic diagram of a velocity analysis apparatus using nuclear magnetic resonance according to the present invention. FIG. 2 is a schematic diagram of another embodiment of a velocity analyzing apparatus using nuclear magnetic resonance according to the present invention. FIG. 3 is an NMR spectrum change graph associated with the protein rewinding reaction. This is a result of obtaining a rate constant of 0.47 s ⁻¹ by exponential function fitting of the time change of the NMR peak intensity (position indicated by the arrow in FIG. 3).

  Hereinafter, the present invention will be described in detail with reference to the drawings. As shown in FIG. 1, the reaction rate analyzer 1 using nuclear magnetic resonance according to the present invention is for NMR spectrum measurement, and includes (1) NMR magnet 4 and (2) sample solution circulation system 2 and (3 ) The detection / control unit 3.

  (1) As the NMR magnet 4, a magnetic magnet that has been conventionally used for NMR spectrum measurement that applies a magnetic field to a sample solution can be employed. For example, the NMR magnet 4 may be a superconducting magnet (Patent Document 1, Non-Patent Document 4) that generates a relatively large space that is easy to access the split gap at the center part of the horizontal magnet. It does not have to be horizontal. Alternatively, an NMR magnet used for medical MRI measurement may be used.

  (2) The sample solution circulation system 2 is installed in a bore in the NMR magnet 4, a holding part 2 e that is a place where the sample solution receives a magnetic field from the NMR magnet 4, an annular pipe 2 a connected to the holding part 2 e, A pump 2b for circulating the sample solution in the pipe 2a, a heating unit 2c (first temperature adjusting unit) for heating the sample solution outside the NMR magnet 4, and a cooling unit 2d for cooling the sample solution in the NMR magnet 4 (Second temperature control unit). The holder 2e can adopt a conventional test tube for NMR spectrum measurement when acquiring an NMR spectrum. In addition, when acquiring an MRI image image by MRI measurement, the thing equivalent to a test tube, such as an artificial heart, is employable.

  By installing the cooling unit 2d in the bore in the NMR magnet 4, the distance from the detection coil 3a can be shortened, and the dead time can be shortened. The heating unit 2c and the cooling unit 2d can be exemplified by a heat exchanger such as a heat pump.

  Specifically, in the heating part 2c, heating is performed by bringing a heat medium into direct or indirect contact with the pipe 2a. Further, in the cooling unit 2d, which is the second temperature adjusting unit in the NMR magnet 4, in order to make direct or indirect contact with the pipe 2a, a refrigerator or a cooler (not shown) installed with the refrigerant outside the NMR magnet 4. For example, a method of circulating through a pipe separately.

  The pump 2b is preferably capable of ON / OFF control by a contact signal or the like. Moreover, as for liquid feeding capability, it is desirable to be able to send liquid at the speed | rate of 10 mL / min or more with respect to the piping 2a of internal diameter 0.5mm, for example.

  Therefore, for the pump 2b and the pipe 2a for circulating the sample solution, a tube made of a chemically stable and heat resistant material of about 100 ° C. such as a pump used in high performance liquid chromatography or polyethylene ethylene ketone (PEEK). Is desirable (Patent Document 1, Non-Patent Document 4).

  Further, if the pipe 2a is such a flexible material, the probe 5 can be easily put in and out of the bore. In FIG. 1, the pipe 2 a is arranged so as to penetrate the NMR magnet 4 vertically, but the pipe 2 a after the holding part 2 e may be connected to the pump 2 b from the lower side of the NMR magnet 4.

  (3) The detection / control unit 3 is arranged on the outer periphery of the holding unit 2e, and includes a detection coil 3a that detects an NMR signal that is a nuclear magnetic resonance signal of the sample solution, and a spectrometer 3b. The spectrometer 3b irradiates the sample solution with the radio wave 3c through the detection coil 3a, and receives the NMR signal 3d through the detection coil 3a. When the NMR spectrum is measured, the NMR spectrum is obtained by Fourier transforming the NMR signal 3d. When performing MRI measurement, the NMR signal 3d is converted into an MRI image. In addition, an ON / OFF signal 3e for controlling the driving of the pump 2b is generated and transmitted.

Therefore, the spectrometer 3b controls the timing of ON / OFF of the drive of the pump 2b, the irradiation of the radio wave 3c, and the detection of the NMR signal 3d, and by repeating this, a plurality of the rate constant analysis for the reaction is performed. The signal / noise ratio is improved by obtaining an equivalent NMR signal 3d and integrating a plurality of NMR spectra (or MRI image images). The ON / OFF control of the pump 2b may be controlled by a timer or the like separately from the spectrometer.

  In the reaction rate analysis apparatus 1 using nuclear magnetic resonance for NMR spectrum measurement, the holding unit 2e, the detection coil 3a, and the cooling unit 2d constitute a nuclear magnetic resonance probe 5. The probe 5 can be attached to and detached from the internal space (bore) of the NMR magnet 4.

  Hereinafter, the sample solution temperature control, liquid feed control, and detection method in the detection coil 3a in the case of the sample solution circulation system 2 in FIG. 1 will be described.

The heating unit 2c heats the sample solution by, for example, placing a commercially available throw-in heater with a thermostat in a metal container filled with 20 L or more of water and heating the water in the container. The cooling unit 2d uses, for example, a commercially available cooling water circulation device installed outside the NMR magnet 4 in a container installed in the vicinity of the detection coil 3a in the NMR magnet 4 to circulate low-temperature water or antifreeze using a silicon tube or the like ( (Not shown).

  The temperature of the sample solution in the heating unit 2c is set to 80 ° C. which is higher than the protein denaturation temperature, and the temperature of the cooling unit 2d is set according to the reaction temperature to be observed in the holding unit 2e (however, below the protein denaturation temperature). To do. For example, when the reaction temperature of the sample solution in the holding part 2e is 40 ° C. and the pipe 2a has an inner diameter of 0.5 mm and the liquid feeding speed is 10 mL / min, the temperature setting of the cooling part 2d is 26 ° C. When the sample solution heated to 80 ° C. is sent by the pump 2b to the holding portion 2e, the temperature is exactly 40 ° C.

Moreover, when the reaction temperature in the holding part 2e is 20 ° C., the same effect can be obtained by setting the temperature of the cooling part 2d to 12 ° C. The coincidence between the target temperature in the holding unit 2e and the set temperature is determined by the coincidence of the water signal position at the time of liquid feed stop equilibrium of the pump 2b and immediately after the pump 2b is operated. At this time, by removing the NMR lock for magnetic field correction, the signal position of water reacts sensitively to temperature changes.

In the heating unit 2c, it is necessary to keep in a denatured state a protein solution having a volume exceeding the sample solution (for example, about 0.5 mL) that is moved by one operation of the pump 2b. In the case where the pipe 2a is bundled and put into the heating unit 2c, the amount is 2 m50 cm if the tube has an inner diameter of 0.5 mm.

On the other hand, the capacity | capacitance containing from the exit of the heating part 2c to the holding | maintenance part 2e needs to be below the capacity | capacitance which moves by one pump 2b action | operation. Thereby, the holding | maintenance part 2e can be satisfy | filled with the sample solution, for example, the protein solution of the reaction process of denaturation and rewinding. When the holding part is 0.18 mL, it is 0.32 mL or less, and 1 m60 cm or less with the above tube.

Moreover, if it is about 20 cm (0.04 mL) with said tube as an area | region through which the sample solution in the cooling part 2d passes, it can cool efficiently. The shorter the distance from the outlet of the cooling unit 2d to the holding unit 2e, the better. However, it is desirable that the distance is 20 cm or less (0.04 mL) with the above tube.

As a result, the flow path capacity from the entrance of the cooling unit 2d at which cooling starts to the center position of the holding unit 2e is within 0.17 mL. If liquid feeding is performed at 10 mL / min, it will reach within 1 second from the inlet of the cooling unit 2d to the holding unit 2e.

  The reaction rate analyzing apparatus 1 using nuclear magnetic resonance thus configured modifies the circulation flow system of Patent Document 1 and constitutes an apparatus for quantifying the rate constant of the reaction induced by the temperature change.

  First, a system capable of analyzing the temporal change of the nuclear magnetic resonance spectrum by enabling the ON / OFF of the pump 2b to be controlled by the ON / OFF signal 3e from the spectrometer 3b and stopping the circulation of the sample solution during the NMR measurement. Become.

Next, the cooling unit 2d is incorporated into the NMR magnet 4 and the dead time is reduced to 2 seconds or less by adjusting the temperature so that the sample solution reaches the target temperature immediately when it reaches the holding unit 2e. Is realized.

In this system, the signal / noise ratio can be improved by repeating heating, cooling, and NMR measurement and integrating the obtained equivalent nuclear magnetic resonance spectra. With the above system, it is possible to quantify the rate constant with a high accuracy of about 0.5 s ⁻¹ or more.

The superconducting magnet part for NMR spectrum measurement holds a liquid helium bath in which the superconducting coil is immersed in the NMR magnet 4, and a probe 5 for detecting the NMR signal 3d is placed in a narrow space (bore) in the center part. An electric heater and a gas flow system for adjusting the temperature of the detection coil 3a and the holding part 2e are installed in the probe 5 (not shown).

In addition to this, incorporating the sample solution circulation system 2 and further incorporating the cooling part 2d therefor has not been performed so far.

  In particular, in most commercially available probes 5 for NMR measurement, an electric heater that can be detached from the outside is installed in the detection coil 3a, and there has been no idea of further disposing a cooling part 2d.

  The reaction rate analyzer 11 using nuclear magnetic resonance in FIG. 2 is for NMR spectrum measurement, and exchanges the positions of the heating unit 2c and the cooling unit 2d of the reaction rate analyzer 1 using nuclear magnetic resonance in FIG. The sample solution circulation system 12 has the first temperature adjusting unit as the cooling unit 2d and the second temperature adjusting unit as the heating unit 2c. Depending on the sample solution, temperature control method, and measurement purpose, the reaction rate analyzer 1 using nuclear magnetic resonance and the reaction rate analyzer 11 using nuclear magnetic resonance are used properly.

Next, using the reaction rate analysis apparatus 1 using nuclear magnetic resonance shown in FIG. 1, bovine pancreatic ribonuclease A (EC 3.1.27.5, Sigma R5500) was used as a sample solution at a concentration of 0.3 mM. Explains the results of analyzing the reaction rate constant of the temperature-dependent conformational change of protein using a protein solution dissolved in 99.9% heavy water containing 20 mM deuterated acetic acid (pH 3.1: indicated by pH meter). To do. It should be noted that the reaction rate analysis apparatus using nuclear magnetic resonance according to the present invention is not limited to measuring a reaction of a three-dimensional structure change of a protein, but can be used for reaction rate analysis of a chemical reaction capable of NMR measurement.

In the test, in the reaction rate analyzer 1 using the nuclear magnetic resonance having the configuration shown in FIG. 1, the pipe 2a was a PEEK tube having an inner diameter of 0.5 mm. Further, the temperature of the heating part 2c was adjusted to 80 ° C., the cooling part 2d was adjusted to 26 ° C., and the temperature of the detection coil 3a portion constituting the probe 5 in the NMR magnet 4 was separately adjusted to 40 ° C. by gas flow. Then, the pump 2b is operated for 3.5 seconds to supply liquid, and after the stop, a waiting time of 1.2 seconds is set, and thereafter, NMR measurement is performed 7 times every 2.0 seconds to obtain an NMR signal 3d. Respectively (FIG. 3).

  In the liquid feeding time of 3.5 seconds, the first 1 second or so is a dead time in which the pressure does not increase and the liquid is hardly fed, but the subsequent 2.5 seconds flow at an average speed of 12 mL / min, and 560 μL is flown. Transferred.

  The flow path volume from the inlet of the cooling part 2d of the sample solution circulation system 2 employed in Example 1 to the central part of the holding part 2e is 165 μL, and after the cooling is started by the cooling part 2d, the liquid is sent to the detection coil 3a. The arrival time until the end was about 0.8 seconds. Then, 2.0 seconds obtained by adding 1.2 seconds, which is a waiting time before the NMR measurement after the pump is stopped, to the arrival time is a dead time in the reaction rate analysis apparatus 1 using nuclear magnetic resonance.

With this dead time, an amplitude of 1/3 or more remains even in a reaction with a rate constant of 0.5 s ⁻¹ , and the analysis of the reaction rate is sufficiently possible. Of course, the number of measurements while the pump is stopped for the predetermined time is appropriately changed depending on the reaction rate and type.

  Then, the pump 2b is turned on and off, and the NMR measurement of the sample solution is repeated a plurality of times. A plurality of seven nuclear magnetic resonance spectra with different times since the stop of the pump 2b are acquired and integrated for each time. Increased the ratio.

  In FIG. 3, a set of 448 times was acquired for the NMR spectrum obtained by performing NMR measurement at seven different times at once, and a spectrum integrated every time was displayed. The measurement time of the first NMR spectrum was set to 0, and a total of 7 NMR spectra were displayed in an overlapping manner at intervals of 2 seconds.

  Further, the encircled portion of FIG. 3 is enlarged and displayed to the right. The position of the arrow indicates the peak position used for the exponential function fitting shown in FIG.

  From FIG. 3, it can be observed that the peak derived from the denatured protein decreases while the peak derived from the unwound protein increases.

The change in peak intensity was exponentially fitted to obtain a reaction rate constant (0.47 s ⁻¹ ) of approximately 0.5 s ⁻¹ (FIG. 4). Perhaps even a reaction rate constant as large as about 30% can be sufficiently analyzed.

DESCRIPTION OF SYMBOLS 1 Reaction rate analysis apparatus using nuclear magnetic resonance 2 Sample solution circulation system 2a Pipe 2b Pump 2c Heating part 2d Cooling part 2e Holding part 3 Detection and control part 3a Detection coil
3b spectrometer 3c radio wave 3d NMR signal 3e ON / OFF signal 4 NMR magnet 5 probe 11 reaction rate analyzer 12 using nuclear magnetic resonance sample solution circulation system

Claims (6)

An apparatus for analyzing the rate of reaction induced by a change in temperature by nuclear magnetic resonance measurement,
1) NMR magnet for NMR spectrum measurement or MRI measurement for applying a magnetic field to the sample solution;
2) A holding unit installed in a bore in the NMR magnet for receiving the magnetic field of the sample solution, an annular pipe connected to the holding unit, a pump for circulating the sample solution in the pipe, and the sample solution A sample solution circulation system comprising a first temperature adjusting unit for adjusting the temperature outside the NMR magnet, and a second temperature adjusting unit for adjusting the temperature of the sample solution inside the NMR magnet;
3) a detection coil that is arranged on the outer periphery of the holding unit and detects a NMR signal of the sample solution; and a detection / control unit that includes the spectrometer that receives the NMR signal and converts it into an NMR spectrum or an MRI image image;
A reaction rate analysis apparatus using nuclear magnetic resonance, characterized by comprising: The first temperature control unit is a heating unit that heats the sample solution, and the second temperature control unit is a cooling unit that cools the sample solution, or the second temperature control unit is a cooling unit that cools the sample solution. The reaction rate analysis apparatus using nuclear magnetic resonance according to claim 1, wherein the second temperature control unit is a heating unit that heats the sample solution. The nuclear magnetism according to claim 1 or 2, wherein the spectrometer generates an ON / OFF signal for controlling the driving of the pump, transmits the signal to the pump, and controls the driving of the pump. A reaction rate analyzer using resonance. In the case of NMR spectrum measurement, the holding unit, the detection coil, and the second temperature adjusting unit constitute a nuclear magnetic resonance probe, and the probe is detachable from the bore. A reaction rate analyzer using the nuclear magnetic resonance according to claim 3. The reaction rate analyzer using nuclear magnetic resonance according to any one of claims 1 to 4, wherein the sample solution that has been operated by the pump and changed in temperature by the second temperature adjusting unit is supplied to the holding unit. The pump is stopped, the pump is stopped, and the NMR measurement of the sample solution in the holding unit is performed every predetermined time. Further, the set of the operation of the pump, the stop, and the NMR measurement of the sample solution is repeated a plurality of times. A reaction rate analysis apparatus using nuclear magnetic resonance, wherein a reaction rate is analyzed by integrating a plurality of equivalent NMR spectra or MRI image images obtained as signals. The reaction rate analyzer using nuclear magnetic resonance according to any one of claims 1 to 5, wherein the time for feeding the sample solution from the second temperature control unit to the holding unit is within 1 second. A reaction rate analyzer using nuclear magnetic resonance characterized by the above.

Images