For hyperpolarization 129 Device and method for flow sampling of Xe magnetic resonance molecular probe

Abstract

The present invention belongs to the field of biomedicine technology and particularly relates to a method for 129 A device and method for flow sampling of Xe magnetic resonance molecular probes, wherein the device includes a hyperpolarized Xe gas supply device, an air inlet line, an air return line, a sample tube, a liquid flow device, a liquid inlet line, and a liquid return line; one end of the air inlet line is connected to the hyperpolarized Xe gas supply device, and the other end extends into the sample tube; the sample tube is connected to the hyperpolarized Xe gas supply device through the air return line; one end of the liquid inlet line is connected to the liquid flow device, and the other end extends into the sample tube; the sample tube is connected to the liquid flow device through the liquid return line. The present invention provides a method that does not require complex live animal experiments, saves time and money, and is suitable for hyperpolarized Xe gas flow sampling. 129 Development and research of Xe magnetic resonance molecular probes for hyperpolarization 129 Device and method for flow sampling of Xe magnetic resonance molecular probe.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology and particularly relates to a method for 129 Device and method for flow sampling of Xe magnetic resonance molecular probe. Background Art

[0002] 129 Xe MRI is one of the most popular molecular imaging methods in recent years. 129 The high sensitivity of Xe combined with molecular probes provides a very promising technology development direction for detecting ultra-low concentration chemical molecules or cancer cells. 129 Xe MRI molecular imaging ultimately addresses two challenges: first, the ability to detect ultra-low concentrations of chemical or biological molecules. This means it boasts higher sensitivity than all conventional MRI methods and can detect ultra-low concentrations of specific chemical or biological molecules in the subject. Second, it can spatially locate the location of ultra-low concentrations of chemical molecules or cancer cells.

[0003] Hyperpolarization 129 The acquisition of Xe is mainly achieved through the spin exchange optical pumping technology (SEOP). Circularly polarized laser is used to irradiate a magnetic field containing alkali metal atoms and inert gas. 129 The mixed sample of Xe makes the spin of alkali metal atoms highly polarized. Then, the polarized alkali metal atoms are mixed with the non-polarized 129 Xe undergoes spin exchange collisions, and the polarized alkali metal atoms transfer angular momentum to 129 Xe, compared with the thermal equilibrium polarizability determined by the Boltzmann distribution under the same temperature and magnetic field conditions, the noble gas 129 The spin polarization of Xe nuclei can be enhanced by 10 3 -10 5 times, greatly increased 129 Signal intensity and sensitivity of Xe MRI.

[0004] Using Xe molecular cage to load the above hyperpolarized 129 Xe, constructing hyperpolarized molecules through targeted modification of molecular cages 129 Xe magnetic resonance molecular probes can achieve specificity for target biomolecules 129 Xe MRI. Hyperpolarization 129 The specific mechanism of action of the Xe magnetic resonance molecular probe is: the probe interacts with the target biological molecules, and the chemical shift of the Xe in the cage changes accordingly. Using the chemical shift imaging method, the distribution of the target biological molecules in the body can be obtained, that is, the image, and it can be used to reflect certain specific molecular processes in the body, thereby achieving the purpose of molecular imaging.

[0005] However, researchers developed hyperpolarized129 The detection performance of Xe magnetic resonance molecular probes and experimental probes for target biomolecules is often carried out in liquid NMR tubes. The liquid in the NMR tube is usually static deionized water or organic solvents, which is quite different from the real physiological environment. It is difficult to evaluate the potential of the probe for in vivo or clinical application. Therefore, a series of complex in vivo animal experiments are required, such as in situ collection of the probe's hyperpolarized polarization in animal blood vessels. 129 Xe magnetic resonance spectroscopy is used to evaluate the stability of the probe (stability means that the probe is less affected by the physiological environment of blood, the nuclear magnetic signal of the probe in the blood is less attenuated than that in the nuclear magnetic tube, and it can still detect and image the target biological molecules). This experiment is of great significance: hyperpolarization 129 Xe magnetic resonance molecular probes are generally administered intravenously and reach the area of ​​interest through the blood circulation to detect and image the target biomolecules. If the probe does not perform well in the initial intravascular test (hyperpolarization of the probe in the blood vessel), the probe will be 129 If the Xe magnetic resonance spectrum signal is weak or absent, indicating that the probe is unstable, it is necessary to re-synthesize and optimize it and conduct in vivo experiments again. Repeated synthesis and animal intravascular testing will consume a lot of time, energy and cost for researchers. If there is a device and method that can relatively easily obtain the hyperpolarization of the probe under physiological conditions in the blood, 129 If Xe magnetic resonance spectroscopy data is available, there is no need to conduct multiple complex experiments on living animals, which can save researchers a lot of time, energy and money. Summary of the Invention

[0006] In order to solve the above technical problems existing in the background technology, the present invention provides a method for producing a hyperpolarized 129 Development and research of Xe magnetic resonance molecular probes for hyperpolarization 129 Device and method for flow sampling of Xe magnetic resonance molecular probe.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] One for hyperpolarization 129 The device for Xe magnetic resonance molecular probe flow sampling is characterized in that: the device for hyperpolarization 129The device for flow sampling of Xe magnetic resonance molecular probe includes a hyperpolarized Xe gas supply device, an air inlet pipeline, an air return pipeline, a sample tube, a liquid flow device, a liquid inlet pipeline and a liquid return pipeline; one end of the air inlet pipeline is connected to the hyperpolarized Xe gas supply device, and the other end extends into the sample tube; the sample tube is connected to the hyperpolarized Xe gas supply device through the air return pipeline; one end of the liquid inlet pipeline is connected to the liquid flow device, and the other end extends into the sample tube; the sample tube is connected to the liquid flow device through the liquid return pipeline.

[0009] As an advantage, the sample tube used in the present invention is provided with a hyperpolarized 129 Xe air intake, hyperpolarization 129 Xe gas outlet, liquid inlet and liquid outlet; the gas inlet pipeline is connected by hyperpolarization 129 The Xe gas inlet extends into the sample tube; the sample tube is hyperpolarized 129 The Xe gas outlet and the return gas pipeline are connected to the hyperpolarized Xe gas supply device; the liquid inlet pipeline extends into the sample tube through the liquid inlet; and the sample tube is connected to the liquid flow device through the liquid outlet and the return liquid pipeline.

[0010] As a preference, the hyperpolarized 129 Xe air intake and hyperpolarization 129 The Xe gas outlets are all placed on the top of the sample tube; the gas inlet pipe is connected to the hyperpolarized 129 A sealing ring is provided between the Xe air inlet; the return air line is connected to the hyperpolarized 129 A sealing ring is provided between the Xe air outlets.

[0011] Preferably, the liquid inlet and liquid outlet used in the present invention are both placed in the middle of the sample tube; a sealing ring is provided between the liquid inlet pipeline and the liquid inlet; and a sealing ring is provided between the liquid return pipeline and the liquid outlet.

[0012] Preferably, the hyperpolarized Xe gas supply device used in the present invention comprises a xenon gas bottle and a hyperpolarized Xe gas supply device connected to the xenon gas bottle. 129 Xe generating device; said hyperpolarized 129 The Xe generating device is connected to the sample tube through the air inlet pipe; the sample tube is connected to the hyperpolarized 129 The Xe generating device is connected.

[0013] As an example, the hyperpolarized Xe gas supply device used in the present invention further includes a gas impurity removal device and a gas pressure reducing valve; the xenon gas cylinder is connected to the hyperpolarized Xe gas via the gas pressure reducing valve and the gas impurity removal device. 129 The Xe generating device is connected.

[0014] Preferably, the liquid flow device used in the present invention is a peristaltic pump; both the liquid inlet pipeline and the liquid return pipeline are provided with stop valves.

[0015] A method based on the aforementioned method for hyperpolarization 129 Xe magnetic resonance molecular probe flow sampling device for hyperpolarization 129 The method for Xe magnetic resonance molecular probe flow sampling is characterized in that the method comprises the following steps:

[0016] 1) Set up for hyperpolarization 129 Device for flow sampling of Xe magnetic resonance molecular probe;

[0017] 2) preparing simulated blood for the device in step 1), and perfusing the simulated blood into the device constructed in step 1);

[0018] 3) Preparation of hyperpolarization 129 Xe magnetic resonance molecular probe, the prepared hyperpolarized 129 The Xe magnetic resonance molecular probe is fixedly placed at the bottom of the sample tube of the device constructed in step 1);

[0019] 4) Simulate blood under static conditions or simulate blood under flowing conditions to hyperpolarize the sample tube 129 Xe magnetic resonance molecular probe sampling.

[0020] Preferably, in step 4) of the present invention, the hyperpolarization of the sample tube in the simulated blood under static conditions is 129 The specific implementation method of Xe magnetic resonance molecular probe sampling is:

[0021] The simulated blood is perfused into the sample tube through the liquid inlet and liquid inlet. When the simulated blood overflows from the liquid outlet to the liquid return line, the power of the peristaltic pump is turned off, and the stop valves on the liquid inlet and liquid return lines are closed; the xenon cylinder is opened, and the gas in the xenon cylinder is depressurized by the gas pressure reducing valve, and then purified by the gas impurity removal device and enters the hyperpolarized 129 Xe generation device, through hyperpolarization 129 Hyperpolarization generated by Xe generator 129 Xe gas passes through the intake pipe and hyperpolarized 129 Xe enters the sample tube through the air inlet. After a period of ventilation, the ventilation in the sample tube is stopped, and then the hyperpolarized Xe in the sample tube is immediately 129 Xe magnetic resonance molecular probe is used to sample and collect 129 Xe spectrum of Xe magnetic resonance molecular probe, completing the hyperpolarization of blood in the sample tube under static conditions 129 Xe magnetic resonance molecular probe for sampling;

[0022] In the step 4), the hyperpolarization of the sample tube in the simulated blood flow condition is 129 The specific implementation method of Xe magnetic resonance molecular probe sampling is:

[0023] The peristaltic pump injects simulated blood into the sample tube through the liquid inlet and liquid inlet, and waits for the simulated blood to overflow from the liquid outlet to the return liquid pipeline and flow to the peristaltic pump to form a closed liquid circuit; turn on the power of the peristaltic pump, open the shut-off valves on the closed liquid inlet and return liquid pipelines; the simulated blood in the liquid circuit begins to circulate at a certain flow rate under the action of the peristaltic pump; turn on the xenon bottle, and the gas in the xenon bottle is depressurized by the gas pressure reducing valve, and then purified by the gas impurity removal device before entering the hyperpolarized 129 Xe generation device, through hyperpolarization 129 Hyperpolarization generated by Xe generator 129 Xe gas passes through the intake pipe and hyperpolarized 129 Xe enters the sample tube through the air inlet. After a period of ventilation, the ventilation in the sample tube is stopped, and then the hyperpolarized Xe in the sample tube is immediately 129 Xe magnetic resonance molecular probe is used to sample and collect 129 Xe spectrum of Xe magnetic resonance molecular probe, completing the hyperpolarization of the sample tube under simulated blood flow conditions 129 Xe magnetic resonance molecular probe sampling.

[0024] Preferably, the simulated blood in step 2) of the present invention is prepared by mixing glycerol and physiological saline in a volume ratio of 4:6, and the mixture is added with a final concentration of 3×10 -3 M of calcium chloride, the viscosity of the simulated blood at 37° C. is 3 mPa·s;

[0025] In step 3), hyperpolarization 129 The preparation method of Xe magnetic resonance molecular probe is as follows: placing a PTFE tube in a Tris-HCl buffer solution with a concentration of 10mM and a pH of 8.5, adding dopamine hydrochloride with a final concentration of 2g / L and ZnCl2 with a final concentration of 4g / L, stirring at room temperature at a stirring speed of 550rpm-650rpm for 24h; after removing the PTFE tube, washing it with deionized water, drying it in a 60℃ oven for 5-8 hours, and then placing it in a 1.5M 2-methylimidazole aqueous solution for activation at room temperature for 5h; finally, placing the PTFE tube in a reactor containing ZIF-8 membrane synthesis solution and reacting it at 120℃ for 4h, taking it out after the reaction is completed, washing it with deionized water, and drying it in a 60℃ oven for 5-8 hours to obtain hyperpolarized 129 Xe magnetic resonance molecular probe, the hyperpolarized 129The Xe magnetic resonance molecular probe is a ZIF-8 membrane@PTFE tube; the ZIF-8 membrane synthesis solution is prepared by dissolving 5.21g of 2-methylimidazole, 1.45g of sodium formate and 1.01g of ZnCl2 in 80ml of methanol.

[0026] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0027] The present invention provides a method for hyperpolarization 129 The device and method for Xe magnetic resonance molecular probe flow sampling include a hyperpolarized Xe gas supply device, an air inlet pipeline, an air return pipeline, a sample tube, a liquid flow device, a liquid inlet pipeline, and a liquid return pipeline; one end of the air inlet pipeline is connected to the hyperpolarized Xe gas supply device, and the other end extends into the sample tube; the sample tube is connected to the hyperpolarized Xe gas supply device through the air return pipeline; one end of the liquid inlet pipeline is connected to the liquid flow device, and the other end extends into the sample tube; the sample tube is connected to the liquid flow device through the liquid return pipeline. 129 Xe magnetic resonance molecular probe flow sampling device to obtain the probe's hyperpolarization under simulated blood physiological conditions 129 Xe NMR data. The simulated blood has a viscosity similar to that of human blood. The peristaltic pump allows the simulated blood to have a certain flow rate. The homemade liquid NMR tube matches the diameter and size of the blood vessel. Without the need for complex live animal experiments, the hyperpolarization of the probe in the physiological environment of blood can be obtained. 129 Xe magnetic resonance spectroscopy and other related data can simulate hyperpolarization 129 The state of Xe magnetic resonance molecular probe in blood vessels can obtain the hyperpolarization of the probe in the physiological environment of blood without complex living animal experiments 129 Xe magnetic resonance spectroscopy and other related data do not require multiple complex live animal experiments, saving researchers a lot of time, energy and money, allowing them to focus on more technical work and spend time planning and developing new projects. 129 The Xe magnetic resonance molecular probe flow sampling device has strong adaptability. In addition to simulating human blood, it can also use simulated or real blood from other animals. The flow rate of the peristaltic pump can be adjusted to match the blood flow rate of different experimental animals, meeting different experimental needs and achieving diversified simulation. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The present invention provides a method for hyperpolarization 129 Schematic diagram of the structure of the Xe magnetic resonance molecular probe flow sampling device;

[0029] Figure 2 Based on the hyperpolarization 129Hyperpolarization of simulated blood at rest or in flow obtained by Xe magnetic resonance molecular probe flow sampling method 129 Xe magnetic resonance molecular probe 129 Xe spectrum;

[0030] Figure 3 yes Figure 1 Schematic diagram of a local enlarged structure;

[0031] in:

[0032] 1-Hyperpolarized Xe gas line; 11-Xenon gas bottle; 12-Gas pressure reducing valve; 13-Gas impurity removal device; 14-Hyperpolarized 129 Xe generator; 15-gas pipeline; 151-inlet pipeline; 152-return pipeline; 2-sample tube; 21-hyperpolarization 129 Xe air inlet; 22-hyperpolarized 129 Xe air outlet; 23-liquid inlet; 24-liquid outlet; 3-liquid flow device; 31-peristaltic pump; 32-liquid pipeline; 321-liquid inlet pipeline; 322-liquid return pipeline. DETAILED DESCRIPTION

[0033] The present invention adopts hyperpolarization 129 The Xe magnetic resonance molecular probe ZIF-8 membrane@PTFE tube ("ZIF-8 membrane@PTFE tube" refers to the PTFE tube modified with ZIF-8 in the form of a membrane) was taken as the research object. The synthesis steps are as follows: a PTFE tube (Shenzhen Hongjiexin Technology Co., Ltd., HST-PTFE (1.7:1) type Teflon heat shrink tubing, inner diameter before heat shrinkage is 1.0-1.2mm) is placed in a Tris-HCl buffer solution with a concentration of 10mM and a pH of 8.5, dopamine hydrochloride DA with a final concentration of 2g / L and ZnCl2 with a final concentration of 4g / L are added, and the reaction is stirred at room temperature for 24h; the PTFE tube is taken out, cleaned and dried, and then placed in a 1.5M 2-methylimidazole aqueous solution for activation at room temperature for 5h; finally, the PTFE tube is placed in a reactor containing ZIF-8 membrane synthesis solution (5.21g of 2-methylimidazole, 1.45g of sodium formate and 1.01g of ZnCl2 dissolved in 80ml of methanol) and reacted at 120°C for 4h. After the reaction is completed, the tube is taken out and washed with deionized water and placed in an oven for drying to obtain a hyperpolarized membrane. 129 Xe magnetic resonance molecular probe ZIF-8 membrane@PTFE tube.

[0034] like Figure 1 as well as Figure 3 As shown, the present invention provides a method for hyperpolarization 129The device for Xe magnetic resonance molecular probe flow sampling comprises: a hyperpolarized Xe gas path 1, a sample tube 2 and a liquid flow device 3, wherein the hyperpolarized Xe gas path 1 comprises a xenon gas bottle 11, a gas pressure reducing valve 12 connected to the xenon gas bottle 11, a gas impurity removal device 13 connected to the outlet of the gas pressure reducing valve 12, and a hyperpolarized Xe gas path 11 connected to the outlet of the gas impurity removal device 13. 129 Xe generating device 14 and connecting sample tube 2 and hyperpolarized 129 The two gas pipelines 15 (the inlet pipeline 151 and the return pipeline 152) of the Xe generator 14; the sample tube 2 is a homemade sample tube modified on the basis of the commercial standard 10mm liquid nuclear magnetic tube, and the upper end of the sample tube 2 has two interfaces corresponding to the hyperpolarized 129 Xe air inlet 21, hyperpolarization 129 Xe gas outlet 22, the middle section of the sample tube 2 has two interfaces: liquid inlet 23 and liquid outlet 24, and the upper end of the sample tube has a hyperpolarized 129 The plane where the Xe inlet 21 is located (or the hyperpolarized 129 The plane where the Xe gas outlet 22 is located) is perpendicular to the plane where the liquid inlet 23 in the middle section is located (or the plane where the liquid outlet 24 is located), which can make full use of the space and avoid the sample tube being too long; the liquid flow device 3 includes a peristaltic pump 31 and a liquid pipeline 32, the liquid pipeline 32 has two sections (a liquid inlet pipeline 321 and a liquid return pipeline 322), the liquid return pipeline 322 connects the liquid outlet 24 of the sample tube and the inlet end of the peristaltic pump 31 pipe joint, the liquid inlet pipeline 321 connects the liquid inlet 23 of the sample tube 2 and the outlet end of the peristaltic pump 31 pipe joint, and the liquid pipeline 32 (the liquid inlet pipeline 321 and the liquid return pipeline 322) are both plastic hoses with stop valves.

[0035] The gas in the xenon cylinder 1 is 2% Xe( 129 The gas is depressurized by the gas pressure reducing valve 12, and then undergoes primary purification (removing trace amounts of O2 and H2O in the gas) at the gas impurity removal device 13 before entering the hyperpolarization 129 The Xe generator 14 is where the spin exchange laser optical pumping process occurs and hyperpolarized Xe gas is generated. The gas flow rate is set by a flow controller to continuously deliver the hyperpolarized Xe to the sample tube 2 located in the NMR spectrometer.

[0036] Hyperpolarization 129 Xe generator 14 is independently developed 129 Xe gas hyperpolarizer (Wuhan Zhongke Polarized Medical Technology Co., Ltd.). 129 Xe NMR experiments were performed on a Bruker Avance 400 MHz wide-cavity NMR spectrometer using a 10 mm transmitter / receiver 129 Xe volume coil.

[0037] The four interfaces of the sample tube 2, namely, the hyperpolarization 129 Xe air inlet 21, hyperpolarization 129 The Xe gas outlet 22, liquid inlet 23 and liquid outlet 24 are all openings with external threads. The gas pipeline 15 (gas inlet pipeline 151 and gas return pipeline 152) and liquid pipeline 32 (liquid inlet pipeline 321 and liquid return pipeline 322) are connected to the hyperpolarized gas flowmeter through sealing caps and sealing rings respectively. 129 Xe air inlet 21, hyperpolarization 129 The Xe gas outlet 22, liquid inlet 23 or liquid outlet 24 are sealed and connected. The gas pipeline and liquid pipeline are both soft plastic hoses. Figure 3 , hyperpolarization 129 Xe gas is sent into sample tube 2 through the air inlet line 151, then reaches the bottom of sample tube 2 through four elastic fused silica capillaries, and finally flows out of sample tube 2 through the return air line 152; simulated blood is sent to the bottom of sample tube 2 through the liquid inlet line 321, and then flows out of the sample tube through the liquid return line 322.

[0038] Based on the above device, the hyperpolarized 129 The specific operation steps of Xe magnetic resonance molecular probe ZIF-8 membrane@PTFE tube flow sampling are as follows:

[0039] 1) Loading samples and simulated blood

[0040] For example, five hyperpolarized 129 The Xe magnetic resonance molecular probe ZIF-8 membrane @ PTFE tube is fixed at the bottom of the sample tube 2, and two sections of liquid pipelines 32 connect the sample tube 2 and the peristaltic pump 31, forming a closed liquid loop of the peristaltic pump 31 pipe joint outlet → liquid pipeline 32 → sample tube liquid inlet 23 → sample tube 2 bottom → sample tube liquid outlet 24 → another section of liquid pipeline 32 → peristaltic pump tube 31 joint inlet → peristaltic pump 31 pipe joint outlet, from the hyperpolarized 129 The Xe gas inlet 21 is used to fill the sample tube 2 with simulated blood. The amount of simulated blood filled is based on the formation of a continuous liquid circuit without bubbles. The two gas pipelines connect the sample tube 2 and the hyperpolarized 129 The Xe generating device 14 is connected to form a hyperpolarized 129 Xe generator 14 → gas pipeline 15 → sample tube hyperpolarization 129 Xe gas inlet 21 → sample tube 2 → sample tube hyperpolarization 129 Xe gas outlet 22 → another section of gas pipeline 15 → hyperpolarization 129The gas circuit of the Xe generator 14. After the device is connected, the sample tube 2 is placed vertically in the appropriate position of the liquid nuclear magnetic resonance spectrometer probe. The simulated blood is a mixture of glycerol and saline in a volume ratio of 4:6. After the mixture is obtained, a final concentration of 3×10 -3 The viscosity of the simulated blood at 37°C is 3 mPa·s.

[0041] 2) Hyperpolarization 129 Xe magnetic resonance molecular probe sampling in static simulated blood

[0042] Before starting sampling, ensure that the power supply of the peristaltic pump 31 and the valves of the two sections of the liquid pipeline 32 are all in the closed state, and the simulated blood in the liquid circuit is in a static state. First, open the xenon bottle 11, the gas is reduced in pressure by the pressure reducing valve 12, and then purified by the gas impurity removal device 13 before entering the hyperpolarized 129 Xe generator 14, generates hyperpolarized 129 Xe gas. Set hyperpolarization 129 The Xe flow rate was 0.1 SLPM, and the hyperpolarized 129 Hyperpolarization generated by the Xe generator 14 129 Xe gas is hyperpolarized from the sample tube through the gas line 15 at a set flow rate. 129 Xe gas inlet 21 enters sample tube 2. After 60 seconds of ventilation, the ventilation of sample tube 2 is stopped. After 3 seconds, the Xe spectrum of the molecular probe is immediately collected. The number of Xe spectrum accumulations is set to 8 times.

[0043] 3) Hyperpolarization 129 Xe magnetic resonance molecular probe sampling in flowing simulated blood

[0044] Hyperpolarization 129 After the Xe magnetic resonance molecular probe completes sampling in the static simulated blood, the power supply of the peristaltic pump 31 and the valves of the two sections of the liquid pipeline 32 are turned on. The simulated blood in the liquid circuit begins to circulate at a certain flow rate under the action of the peristaltic pump 31, and begins to hyperpolarize. 129 Flow sampling of Xe magnetic resonance molecular probe. Setting hyperpolarization 129 The Xe flow rate was 0.1 SLPM, and the hyperpolarized 129 Hyperpolarization generated by the Xe generator 14 129 Xe gas is hyperpolarized by the sample tube through the gas line 15 129 Xe gas inlet 21 enters sample tube 2 and ventilates for 60 seconds, then stops venting to sample tube 2. Xe spectrum of the molecular probe is collected immediately after 3 seconds. The number of Xe spectrum accumulations is set to 16 times.

[0045] For the above hyperpolarization 129The raw data obtained by the Xe magnetic resonance molecular probe in static simulated blood were Fourier transformed, baseline corrected and phase corrected to obtain Figure 2 (Bottom) Xe spectrum of the probe in static simulated blood. 129 The raw data obtained by the Xe magnetic resonance molecular probe in the flowing simulated blood were Fourier transformed, baseline corrected and phase corrected to obtain Figure 2 (Top) Xe spectrum of the probe in flowing simulated blood. Figure 2 (Next) 129 The chemical shift of 82.3 ppm in the Xe spectrum is inside the cage of the molecular probe ZIF-8 membrane@PTFE tube. 129 Xe signal (i.e. 129 Xe@Molecular Probes), chemical shift 207.8 ppm indicates dissolved state 129 Xe signal (i.e. 129 Xe@simulated blood), 129 The signal-to-noise ratio of Xe@Molecular Probe is 10.09. Figure 2 (Top) Probe in flowing simulated blood 129 The chemical shift of the Xe@molecular probe did not change compared to that in static simulated blood. Although the number of Xe spectrum accumulations was increased, the signal-to-noise ratio decreased from 10.09 to 3.39, indicating that the probe signal decreased in flowing simulated blood, which was in line with expectations. The experimental results prove that the device and method can obtain the hyperpolarization of the probe under physiological conditions of blood. 129 Xe magnetic resonance spectroscopy data is helpful in evaluating the stability of the probe in the blood and can be used to replace repeated synthesis-animal intravascular tests, saving researchers a lot of time, energy and costs.

Claims

1. A method for hyperpolarization 129 The device for Xe magnetic resonance molecular probe flow sampling is characterized by: The invention comprises a hyperpolarized Xe gas supply device, an air inlet pipeline (151), an air return pipeline (152), a sample tube (2), a liquid flow device (3), a liquid inlet pipeline (321) and a liquid return pipeline (322); one end of the air inlet pipeline (151) is connected to the hyperpolarized Xe gas supply device, and the other end thereof extends into the sample tube (2); the sample tube (2) is connected to the hyperpolarized Xe gas supply device through the air return pipeline (152); one end of the liquid inlet pipeline (321) is connected to the liquid flow device (3), and the other end thereof extends into the sample tube (2); the sample tube (2) is connected to the liquid flow device (3) through the liquid return pipeline (322); The sample tube (2) is provided with a hyperpolarized 129 Xe air inlet (21), hyperpolarization 129 Xe gas outlet (22), liquid inlet (23) and liquid outlet (24); the sample tube (2) is a homemade sample tube modified on the basis of a commercial standard 10mm liquid nuclear magnetic tube. The homemade sample tube matches the diameter and size of the blood vessel. The upper end of the sample tube (2) has two interfaces, corresponding to the hyperpolarized 129 Xe air inlet (21), hyperpolarization 129 Xe gas outlet (22), the middle section of the sample tube (2) has two interfaces: liquid inlet (23) and liquid outlet (24), and the hyperpolarized 129 The plane where the Xe inlet (21) is located or the hyperpolarized 129 The plane where the Xe gas outlet (22) is located is perpendicular to the plane where the liquid inlet (23) of the middle section is located or the plane where the liquid outlet (24) is located; the gas inlet pipeline (151) is connected to the hyperpolarized 129 The Xe gas inlet (21) extends into the sample tube (2); the sample tube (2) is subjected to hyperpolarization. 129 The Xe gas outlet (22) and the return gas line (152) are connected to the hyperpolarized 129 Xe gas supply device; the liquid inlet pipeline (321) extends into the sample tube (2) through the liquid inlet (23); the sample tube (2) is connected to the liquid flow device (3) through the liquid outlet (24) and the liquid return pipeline (322); The hyperpolarized Xe gas supply device comprises a xenon gas bottle (11) and a hyperpolarized 129 Xe generating device (14); said hyperpolarized 129 The Xe generating device (14) is connected to the sample tube (2) via an air inlet line (151); the sample tube (2) is connected to the hyperpolarized 129 The Xe generating device (14) is connected; 129 Xe gas is fed into the sample tube (2) through the gas inlet pipe (151) and then reaches the bottom of the sample tube (2) through four elastic fused silica capillaries; Hyperpolarization 129 The Xe magnetic resonance molecular probe ZIF-8 membrane@PTFE tube is fixed at the bottom of the sample tube (2); In the device, the liquid flows sequentially through the liquid flow device (3), the liquid inlet pipe (321), the liquid inlet (23), the bottom of the sample tube (2), the liquid outlet (24), the liquid return pipe (322), and then flows back to the liquid flow device (3), forming a closed liquid loop; hyperpolarization 129 Xe gas flows through the hyperpolarized 129 Xe generating device (14), air inlet pipeline (151), hyperpolarization 129 Xe gas inlet (21), sample tube (2), hyperpolarization 129 Xe gas outlet (22), return gas line (152), and then return to the hyperpolarized 129 A Xe generating device (14) forms a gas loop; After the device is connected, the sample tube (2) is placed vertically into the appropriate position of the liquid nuclear magnetic resonance spectrometer probe.

2. The method for hyperpolarization according to claim 1 129 The device for Xe magnetic resonance molecular probe flow sampling is characterized by: The air intake pipe (151) is connected to the hyperpolarized 129 A sealing ring is provided between the Xe air inlet (21); the return air line (152) and the hyperpolarized 129 A sealing ring is provided between the Xe gas outlets (22).

3. The method for hyperpolarization according to claim 1 129 The device for Xe magnetic resonance molecular probe flow sampling is characterized by: A sealing ring is provided between the liquid inlet pipe (321) and the liquid inlet (23); and a sealing ring is provided between the liquid return pipe (322) and the liquid outlet (24).

4. The method for hyperpolarization according to claim 1 129 The device for Xe magnetic resonance molecular probe flow sampling is characterized by: The liquid flow device (3) is a peristaltic pump (31); and both the liquid inlet pipeline (321) and the liquid return pipeline (322) are provided with stop valves.

5. A method for hyperpolarization according to any one of claims 1 to 4 129 Xe magnetic resonance molecular probe flow sampling device for hyperpolarization 129 The method for flow sampling of Xe magnetic resonance molecular probe is characterized by: The method comprises the following steps: 1) Constructing a device for hyperpolarization according to any one of claims 1 to 4 129 Device for flow sampling of Xe magnetic resonance molecular probe; 2) preparing simulated blood for the device in step 1), and perfusing the simulated blood into the device constructed in step 1); 3) Preparation of hyperpolarization 129 Xe magnetic resonance molecular probe, the prepared hyperpolarized 129 The Xe magnetic resonance molecular probe is fixedly arranged at the bottom of the sample tube (2) of the device constructed in step 1); 4) Simulate blood under static conditions or simulate blood under flowing conditions to hyperpolarize the sample tube 129 Xe magnetic resonance molecular probe for sampling; The simulated blood is hyperpolarized in the sample tube under static conditions. 129 The specific implementation method of Xe magnetic resonance molecular probe sampling is: Simulated blood is perfused into the sample tube (2) through the liquid inlet line (321) and the liquid inlet (23). When the simulated blood overflows from the liquid outlet (24) to the liquid return line (322), the power supply of the peristaltic pump (31) is turned off, and the stop valves on the liquid inlet line (321) and the liquid return line (322) are closed; the xenon bottle (11) is opened, and the gas in the xenon bottle (11) is depressurized by the gas pressure reducing valve (12), and then purified by the gas impurity removal device (13) and enters the hyperpolarized gas chamber. 129 Xe generating device (14), through hyperpolarization 129 Hyperpolarization generated by the Xe generating device (14) 129 Xe gas passes through the inlet pipe (151) and the hyperpolarized 129 Xe enters the sample tube (2) through the air inlet (21), and after a period of ventilation, the ventilation of the sample tube (2) is stopped, and then the hyperpolarized Xe in the sample tube is immediately 129 Xe magnetic resonance molecular probe is used to sample and collect 129 Xe spectrum of Xe magnetic resonance molecular probe, completing the hyperpolarization of blood in the sample tube under static conditions 129 Xe magnetic resonance molecular probe for sampling; The simulated blood is hyperpolarized in the sample tube under flow conditions 129 The specific implementation method of Xe magnetic resonance molecular probe sampling is: The peristaltic pump (31) perfuses the sample tube (2) with simulated blood through the liquid inlet line (321) and the liquid inlet (23), and waits for the simulated blood to overflow from the liquid outlet (24) to the liquid return line (322) and flow to the peristaltic pump (31) to form a closed liquid circuit; the power supply of the peristaltic pump (31) is turned on, and the stop valves on the liquid inlet line (321) and the liquid return line (322) are opened; the simulated blood in the liquid circuit begins to circulate at a certain flow rate under the action of the peristaltic pump (31); the xenon bottle (11) is turned on, and the gas in the xenon bottle (11) is depressurized by the gas pressure reducing valve (12), and then purified by the gas impurity removal device (13) and enters the hyperpolarized gas chamber. 129 Xe generating device (14), through hyperpolarization 129 Hyperpolarization generated by the Xe generating device (14) 129 Xe gas passes through the inlet pipe (151) and the hyperpolarized 129 Xe enters the sample tube (2) through the air inlet (21), and after a period of ventilation, the ventilation of the sample tube (2) is stopped, and then the hyperpolarized Xe in the sample tube is immediately 129 Xe magnetic resonance molecular probe is used to sample and collect 129 Xe spectrum of Xe magnetic resonance molecular probe, completing the hyperpolarization in the sample tube 129 Xe magnetic resonance molecular probe sampling.

6. The method according to claim 5, characterized in that: In step 2), the simulated blood is prepared by mixing glycerol and physiological saline in a volume ratio of 4:6, and the mixture is added with a final concentration of 3×10 -3 The viscosity of the simulated blood at 37° C. is 3 mPa·s.

7. The method according to claim 5, characterized in that: In step 3), hyperpolarization 129 The preparation method of Xe magnetic resonance molecular probe is as follows: placing a PTFE tube in a Tris-HCl buffer solution with a concentration of 10mM and a pH of 8.5, adding dopamine hydrochloride with a final concentration of 2g / L and ZnCl2 with a final concentration of 4g / L, stirring at room temperature at a stirring speed of 550rpm-650rpm for 24h; after removing the PTFE tube, washing it with deionized water, drying it in a 60℃ oven for 5-8 hours, and then placing it in a 1.5M 2-methylimidazole aqueous solution for activation at room temperature for 5h; finally, placing the PTFE tube in a reactor containing ZIF-8 membrane synthesis solution and reacting it at 120℃ for 4h, taking it out after the reaction is completed, washing it with deionized water, and drying it in a 60℃ oven for 5-8 hours to obtain hyperpolarized 129 Xe magnetic resonance molecular probe, the hyperpolarized 129 The Xe magnetic resonance molecular probe is a ZIF-8 membrane@PTFE tube; the ZIF-8 membrane synthesis solution is prepared by dissolving 5.21g of 2-methylimidazole, 1.45g of sodium formate and 1.01g of ZnCl2 in 80ml of methanol.

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