Deuterated methyl protection glucose and preparation method thereof

Abstract

The preparation method comprises the following steps: firstly, preparing a double-chamber electrolytic tank which comprises a cathode chamber, an anode chamber and an electrolyte membrane which is arranged between the cathode chamber and the anode chamber and is used for isolating oxygen; preparing a gas diffusion electrode containing a ruthenium catalyst as a cathode and a metal electrode as an anode; then mixing methyl protected glucose, deuterium water and electrolyte to prepare a substrate reaction solution, respectively adding the substrate reaction solution into a cathode chamber and an anode chamber, inserting a cathode and an anode, connecting the cathode and the anode to an electrochemical workstation to carry out an electrolytic catalytic reaction for a period of time, and in the electrolytic process, generating adsorbed deuterium on the surface of the ruthenium-containing catalyst under the action of an electric field; carrying out exchange reaction on the adsorbed deuterium and hydrogen on the glucose to generate deuterated methyl protection glucose; and after the electrolytic catalytic reaction is finished, collecting a product in the cathode chamber to obtain a deuterated methyl protected glucose solution. The method is relatively mild in reaction condition, simple to operate and lower in cost, and the deuteration efficiency of the key sites reaches 99% or above.

Description

Technical Field

[0001] This invention relates to the field of tracer technology, and more particularly to a deuterated methyl protected glucose and its preparation method. Background Technology

[0002] Glucose (C6H) 12 Oxygen (O6) is the most essential energy substance and carbon source in living organisms. Its metabolic pathways, including glycolysis, the pentose phosphate pathway, and the tricarboxylic acid cycle, form the core network for cellular energy supply and biomolecule synthesis. Dynamic and quantitative studies of glucose metabolism are of vital importance for understanding the fundamental laws of life activities and revealing the mechanisms of disease development (such as cancer, diabetes, and neurodegenerative diseases).

[0003] Currently, the most commonly used technique for studying glucose metabolism is to use isotopically labeled glucose as a tracer. Among these, radioactive isotope labeling, such as carbon-14 (…), is employed. 14 C) or fluorine ( 18 F)-labeled glucose has been used for a long time due to its high sensitivity. However, radioactive isotopes have inherent drawbacks such as radioactive hazards, limited half-life, difficulties in waste disposal, and incompatibility with operators and the environment, which greatly limit their application in clinical diagnosis, especially in repeated human testing.

[0004] To overcome the shortcomings of radioactive tracers, stable isotope labeling technology has emerged. Especially carbon-13 (… 13 C) Labeled glucose, detected by nuclear magnetic resonance (NMR) or mass spectrometry (MS), has become an important tool in metabolomics research. However, 13 C-NMR technology has relatively low sensitivity, while utilizing 13 When C-labeling is combined with mass spectrometry analysis, the background interference caused by its natural abundance (approximately 1.1%) cannot be ignored, posing a challenge to trace detection and accurate quantification.

[0005] deuterium( 2 Hydrogen (H or D) is a stable isotope of hydrogen, non-radioactive, and naturally abundant in very low quantities (approximately 0.015%). Replacing specific hydrogen atoms in a glucose molecule with deuterium atoms yields deuterated glucose. 13 Compared to C-labeled glucose, deuterated glucose has the following potential advantages: Non-radioactive: High safety profile, suitable for repeated studies involving clinical patients, children, and pregnant women.

[0006] Mass spectrometry offers high detection sensitivity: There is a significant mass difference between deuterated and non-deuterated molecules, which can be easily distinguished and accurately quantified using mass spectrometry, with extremely low background interference.

[0007] NMRH test is fast and accurate: After deuteration, the hydrogen nuclear magnetic signal changes significantly, and by integrating the peak area, the accurate deuteration rate of different sites can be accurately obtained.

[0008] Multiple tracing pathways: By deuterating at different positions (such as C1, C2, C6, C7, etc.), different branch pathways of glucose metabolism can be specifically traced. For example, [1- 2 H]-glucose can be used to study glycolytic flux, while [6,6- 2 H2]-glucose is often used to evaluate the overall rate of glucose oxidation.

[0009] Potential kinetic isotope effect (KIE): There is a difference in chemical bond strength between deuterium and hydrogen atoms, which can cause changes in metabolic reaction rates. Although this effect is a disturbance factor in some cases, it can also be cleverly used to regulate metabolic flow, thus serving as a therapeutic strategy, such as delaying the rapid glycolysis of cancer cells through selective deuteration.

[0010] However, the preparation of deuterated glucose still faces technical bottlenecks: Existing chemical synthesis routes for deuterated glucose often have complex steps, low overall yield, and high cost. In particular, achieving high regioselectivity and high chemical purity for specific position deuteration is challenging. Thermal chemical deuteration synthesis methods require extreme conditions such as high-pressure hydrogen and high temperature, making it difficult to scale up production. Deuterated glucose synthesized by traditional isotopic exchange methods has a low deuteration rate (deuteration rate 40%-60%), which is difficult to meet actual needs. While biological synthesis methods (such as using deuterated medium to culture algae or microorganisms) can obtain fully deuterated products, they are difficult to achieve site-specific deuteration, and the product separation and purification process is complex.

[0011] Therefore, there is an urgent need in the art to develop a new method for efficiently, cost-effectively, and selectively synthesizing specific position deuterated glucose to overcome the shortcomings of existing technologies.

[0012] The prior art CN116813673A discloses a preparation method of five deuterium-substituted glucose, D-methyl glucoside is added to heavy water, a catalyst is added, and the reaction is carried out under a hydrogen atmosphere, at a suitable pressure and at a suitable temperature, after the reaction is completed, [2,3,4,6,6'-d5]-D-methyl glucoside is obtained after treatment; the [2,3,4,6,6'-d5]-D-methyl glucoside prepared in the above step is added to a suitable solvent, a suitable demethylation reagent is added, and the reaction is carried out at a suitable temperature, after the reaction is completed, the crude product is obtained after treatment, and the target compound, [2,3,4,6,6'-d5]-D-glucose, is obtained by recrystallization. Although the five deuterium-substituted glucose is synthesized, it needs to be in a hydrogen atmosphere, and needs to be pressurized. Although the temperature range recorded in the specification book is very wide, in fact, the catalytic temperature recorded in the Chinese examples is 80℃, and the catalytic efficiency is not good at a low temperature. After the catalytic reaction, the product also needs to be added with an acidic reagent for demethylation treatment in a high-temperature environment, and the technical complexity is relatively high.

[0013] The prior art CN110885985A discloses a preparation method of deuterium-substituted chemicals, a halogenated hydrocarbon compound, a catalyst, a deuterium source and a solvent are added to a reaction bottle, an electrode is placed in the reaction bottle, and a dehalogenation and deuterium addition reaction is carried out under an inert gas atmosphere at a reaction temperature of room temperature to 80℃ by applying a voltage, to obtain deuterium-substituted chemicals; wherein the deuterium source is one or more of deuterium water, deuterium alcohol compounds and deuterium acid compounds. The main principle of the technology is electrocatalytic dehalogenation and deuterium addition reaction, and the point selectivity of the deuterium addition reaction is changed by changing the voltage, which has certain technical value. However, the substrate is only suitable for halogenated hydrocarbon compounds, such as 2-bromoacetophenone, 3-bromoacetophenone, 4-bromoacetophenone, bromocoumarin, bromphenamine and other bromine compounds. The reaction mechanism is dehalogenation and deuterium addition for carbon-halogen bond (C-X). Under the action of an electric field, the deuterium source (such as deuterium water) is electrolyzed on the surface of the electrode to generate a highly active deuterium intermediate (such as D• or D - ). At the same time, under the action of a catalyst (such as a palladium-based homogeneous or heterogeneous catalyst), the carbon-halogen bond (C-X) is activated and broken to form a carbon radical or carbon anion intermediate. The deuterium intermediate combines with these carbon active species to complete the deuterium addition reaction. The electric field not only drives the decomposition of the deuterium source, but also regulates the reaction path to achieve site selectivity (for example, the reaction order of different halogen functional groups can be controlled by adjusting the voltage); it can be known from the above principle that the technology is difficult to directly deuterate glucose.

[0014] Therefore, it is necessary to develop a technology for deuterating glucose as a substrate with high deuterium substitution rate at key sites, to realize high-efficiency deuterium substitution of glucose, and to obtain high-quality isotopically labeled glucose as a tracer. SUMMARY

[0015] The application provides a kind of deuterated methyl protected glucose and its preparation method, to solve the key point of deuterated methyl protected glucose in prior art is not high, reaction condition is harsh, the defect of preparation process is complex, realize the low cost production key point high deuterium glucose.The application generates adsorbed deuterium by electrochemistry at the reaction interface with deuterium atom in deuterium water, hydrogen-deuterium exchange occurs between adsorbed deuterium and hydrogen atom on glucose, and finally deuterated methyl protected glucose is obtained.

[0016] To solve the above technical problems, the technical scheme adopted by the application is as follows: In a first aspect, the application provides a method for preparing deuterated methyl protected glucose, comprising the following steps: Step 1, prepare a double-chamber electrolytic cell, which includes a cathode chamber, an anode chamber, and an electrolyte membrane arranged between the cathode and anode chambers for isolating oxygen; Step 2, prepare a gas diffusion electrode containing a ruthenium catalyst as the cathode, and prepare a metal electrode as the anode; Step 3, mix and configure the substrate reaction solution with methyl protected glucose, deuterium water, and electrolyte; Step 4, add the substrate reaction solution to the cathode chamber and the anode chamber respectively, then insert the cathode and the anode respectively (connect the cathode and the anode to the electrochemical workstation), and carry out electrolytic catalytic reaction for a period of time. During the electrolysis process, adsorbed deuterium is generated on the surface of the ruthenium-containing catalyst under the action of the electric field, and hydrogen-deuterium exchange reaction occurs between the adsorbed deuterium and the hydrogen on the glucose to produce deuterated methyl protected glucose; Step 5, after the electrolytic catalytic reaction is completed, collect the product in the cathode chamber to obtain a deuterated methyl protected glucose solution.

[0017] The above steps 1-3 do not have strict order requirements, and the numbering is only for ease of description.

[0018] In step 1 of the above method for preparing deuterated methyl protected glucose, the electrolyte membrane includes a cationic electrolyte membrane and an anionic electrolyte membrane.

[0019] The cationic electrolyte membrane is a perfluorosulfonic acid polymer (Nafion), and the anionic electrolyte membrane is a quaternized p-terphenyl-piperidine copolymer (QAPPT).

[0020] In step 2 of the above method for preparing deuterated methyl protected glucose, the catalyst type in the gas diffusion electrode containing a ruthenium catalyst includes a ruthenium catalyst or a palladium-ruthenium alloy catalyst.

[0021] For example, the ruthenium catalyst is a Ru / C catalyst with carbon as the carrier, and the metal loading of the Ru / C catalyst is 5%-50%; the ruthenium alloy catalyst is a PdRu / C catalyst with carbon as the carrier, and the total metal loading of the PdRu / C catalyst is 15%-40% (the atomic ratio of Pd and Ru is 1:1).

[0022] In step 3 of the preparation method of the deuterated methyl-protected glucose, the concentration of the substrate reaction solution has no obvious effect on the deuteration rate, and the common glucose concentration range can be 20 mmol / L-100 mmol / L, 20 mmol / L, 40 mmol / L, or 80 mmol / L.

[0023] The deuterated methyl-protected glucose in the application is glucose in which at least one hydroxyl group is protected by a methyl group. For example, the deuterated methyl-protected glucose is methyl β-D-glucopyranoside, as shown in the formula. Figure 2 As shown in the formula, the hydroxyl group at position 1 of the methyl β-D-glucopyranoside is protected by a methyl group.

[0024] In step 3 of the preparation method of the deuterated methyl-protected glucose, the electrolyte concentration in the substrate reaction solution has little effect on the deuteration rate, and the common concentration range is 20 mmol / L-500 mmol / L.

[0025] In step 3 of the preparation method of the deuterated methyl-protected glucose, the electrolyte in the substrate reaction solution is any one or several of sodium sulfate, potassium sulfate, potassium hydroxide, and sodium hydroxide.

[0026] In step 4 of the preparation method of the deuterated methyl-protected glucose, the catalyst loading in the gas diffusion electrode is 0.3-2.0 mg / cm 2 .

[0027] More preferably, the catalyst loading is 0.9-1.8 mg / cm 2 , and the range can be more preferably 1.2-1.5 mg / cm 2 .

[0028] In step 4 of the preparation method of the deuterated methyl-protected glucose, the temperature of the electrolytic catalytic reaction is 70-100℃.

[0029] More preferably, the temperature of the electrolytic catalytic reaction is 70-90℃.

[0030] In step 4 of the preparation method of the deuterated methyl-protected glucose, the reaction time of the electrolytic catalytic reaction is 12-72 hours.

[0031] More preferably, the reaction time is 24-72 hours.

[0032] More preferably, the reaction time is 24-60 hours.

[0033] In step 4 of the above method for preparing deuterated methyl-protected glucose, the process of the electrolytic catalytic reaction is alternating constant voltage electrolysis and static exchange until the total reaction time is reached. The present application quickly obtains a large amount of adsorbed deuterium on the cathode surface by constant voltage electrolysis, and improves the exchange degree of adsorbed deuterium and hydrogen in glucose by static reaction.

[0034] The time of static exchange is 1.5-2.5 times the time of constant voltage electrolysis.

[0035] For example, constant voltage electrolysis is performed for 10 minutes, and static exchange is performed for 20 minutes.

[0036] It should be noted that the constant voltage electrolysis can use common parameters in the prior art, for example, the constant voltage electrolysis can be performed at a cell voltage of 3.5 V.

[0037] It should be noted that the anode is a metal electrode, such as a Pt metal sheet, an Al sheet (the deuterium substitution effect is different), an iridium metal electrode, or a ruthenium electrode; the applicant found in the research process that if other anodes are used, such as some non-metal anodes, such as carbon paper, carbon felt, carbon paper loaded with Pt / C, etc., the deuterium substitution reaction cannot be performed.

[0038] In a second aspect, the present application provides a deuterated methyl-protected glucose prepared by the above preparation method.

[0039] In the present application, the cathode and anode are respectively placed in two electrolytic cells (H-shaped two-chamber electrolytic cells), and are connected to an electrochemical workstation. The electrolytic cells (both the cathode chamber and the anode chamber) contain a certain concentration of electrolyte solution (such as a sodium sulfate deuterium water solution) and a glucose deuterium water solution. The sodium sulfate is a supporting electrolyte (providing conductive migration ions), the deuterium water is the deuterium source for the deuterium substitution reaction, and the glucose is the reaction substrate. The two chambers are connected through a high-molecular polymer diaphragm. A potential is applied to the two electrodes, so that the cathode undergoes a hydrogen evolution reaction (HER), and the anode undergoes an oxygen evolution reaction (OER). After reacting for a period of time at a certain temperature, a deuterated methyl-protected glucose deuterium water solution is obtained in the cathode chamber.

[0040] The cathode reaction principle is as follows: First step: electrochemically generating active deuterium atoms (adsorbed deuterium) Electrolysis of deuterium water: in an electrolytic cell with a ruthenium (Ru) cathode, a solution containing deuterium water (D2O) is electrolyzed. A deuterium evolution reaction (HER) occurs on the cathode.

[0041] Deuterium evolution reaction: the reaction on the cathode surface is: Here, Active deuterium atom adsorbed on the surface of the ruthenium electrode.

[0042] Second step: adsorbed deuterium reacts with glucose to generate deuterium-substituted methyl-protected glucose Surface reaction: the generated adsorbed deuterium ( ) has high chemical reactivity, is attached to the electrode surface, and is close to the hydroxyl oxygen of the methyl-protected glucose in the solution near the electrode surface, coordinates with the Ru catalyst, and undergoes configuration inversion.

[0043] Deuterium-hydrogen exchange: the hydrogen atoms in some positions of the glucose molecule, especially the hydrogen on the hydroxyl group (-OH) connected to the oxygen atom, and the hydrogen (C-H) on the alpha-carbon activated by adjacent hydroxyl groups and the like have high acidity or activity; these active hydrogens can undergo deuterium-hydrogen replacement reaction with the active deuterium atom ( ) on the electrode surface to obtain deuterium-substituted methyl-protected glucose.

[0044] Since oxygen is generated in the anode chamber, and oxygen will seriously interfere with the adsorbed deuterium, the present application needs to separate the cathode chamber and the anode chamber by an electrolyte membrane, only allowing cations or anions to pass through, and not allowing oxygen molecules to pass through, in order to avoid the influence of oxygen on the deuterium substitution reaction in the cathode chamber.

[0045] It should be noted that although there is a separation effect of the electrolyte membrane, part of the deuterium-substituted methyl-protected glucose in the cathode chamber still migrates to the anode chamber, and the deuterium-substituted methyl-protected glucose product obtained by the present application is obtained by sampling from the cathode chamber.

[0046] According to the above reaction principle, in order to efficiently perform the hydrogen-deuterium exchange reaction, the cathode catalyst must have high hydrogen evolution reaction activity and appropriate adsorption capacity. If the adsorption capacity of the cathode catalyst for adsorbed deuterium is too weak, not only the catalytic efficiency of the hydrogen evolution reaction is reduced, but also the generated adsorbed deuterium quickly escapes into the solution, and the local concentration required for high-efficiency hydrogen-deuterium exchange cannot be reached; if the adsorption capacity for adsorbed deuterium is too strong, a large amount of adsorbed deuterium will undergo molecular combination reaction to generate molecules, which reduces the efficiency of the hydrogen-deuterium exchange reaction or even does not occur. The applicant found through a large number of experimental studies that metal ruthenium as the active component of the cathode catalyst has just the appropriate adsorption capacity, has high hydrogen-deuterium exchange efficiency, and the metal Pt catalyst almost does not have hydrogen-deuterium exchange reaction.

[0047] Compared with the prior art, the present application has the following beneficial effects: 1. The present application is different from the method for synthesizing deuterium-substituted, which uses electrochemical means to obtain deuterium-substituted methyl-protected glucose in one step, has fewer reaction steps, and low cost. 2. The present application is different from the thermal chemical deuteration method, which generates adsorbed hydrogen on the electrode surface in situ by an electrochemical method, and can be carried out at a lower temperature (the thermal chemical method needs to pass high-pressure hydrogen to form adsorbed hydrogen on the surface of the catalyst); 3. Compared with the traditional isotope replacement method (deuteration rate 40%-60%), the deuteration rate of the key site of the present application can reach more than 99%, and the product is closer to the actual use requirement; 4. The working condition of the electrolytic cell of the present application is a relatively mild environment of 75℃ (20℃ up and down), which reduces the influence of high temperature on the polymer and the catalyst; 5. The preparation process of the present application is simple, compared with the existing deuteration glucose technology, only simple device and mild conditions are needed to realize the scale production, the overall equipment investment cost is less, and it has very high practical value; 6. The present application selects suitable catalyst and catalyst loading through a large number of studies, studies the influence of reaction temperature and time on the deuteration rate, and obtains the process parameters with high comprehensive deuteration rate and key point deuteration rate, which provides technical support for industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0048] In order to more clearly illustrate the technical solutions in the present application or prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0049] Figure 1 is a schematic diagram of the double-chamber electrolytic cell structure provided by the present application; Figure 2 is the magnetic spectrum and peak integration result diagram of the reaction product deuteration methyl-protected glucose in example 1 of the present application; Figure 3 is the variation diagram of the deuteration rate at different deuteration reaction times in example 1 of the present application, wherein Figure 3 (a) in the figure is the variation diagram of the total deuteration rate with reaction time, Figure 3 (b) in the figure is the variation diagram of the key site deuteration rate with reaction time; Figure 4 is the variation diagram of the deuteration rate of different cathode GDE catalyst loadings in example 2 of the present application, wherein Figure 4 (a) in the figure is the variation diagram of the total deuteration rate of different cathode GDE catalyst loadings, Figure 4 (b) in the figure is the variation diagram of the key site deuteration rate of different cathode GDE catalyst loadings.

[0050] Figure 5is a total deuterium substitution rate change graph of different cathode GDE catalysts in Example 3 of the present application.

[0051] Figure 6 is a total deuterium substitution rate change graph of different electrolyte membranes in Example 4 of the present application.

[0052] Figure 7 is a deuterium substitution rate change graph of different reaction temperatures in Example 5 of the present application, wherein Figure 7 in (a) is a total deuterium substitution rate change graph of different reaction temperatures, Figure 7 in (b) is a key site deuterium substitution rate change graph of different reaction temperatures.

[0053] 1-cathode, 2-cathode chamber, 3-gas inlet, 4-gas outlet, 5-first connecting chamber, 6-second connecting chamber, 7-anode chamber, 8-anode, 9-porous rubber plug, 10-electrolyte membrane. DETAILED DESCRIPTION

[0054] The embodiments of the present application will be further described below in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present application, but cannot be used to limit the scope of the present application.

[0055] In the description of the present application, it should be noted that the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the present application. In addition, the terms "first", "second" are only for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0056] In the description of the present application, it should be noted that unless otherwise explicitly specified and limited, the terms "connected", "connected" should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or integrally connected; it can be mechanically connected, or it can be electrically connected; it can be directly connected, or it can be indirectly connected through an intermediate medium. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0057] Example 1: Cathode GDE preparation: Take 2.25 mg of 40% commercial Ru / C catalyst, add 400 microliters of Nafion solution (Nafion is purchased from Dupont, the dilution solvent is ethanol, the concentration is about 5 mg / mL), and uniformly disperse it in an ice water bath for 1 hour. The above precursor ink is dropped on the gas diffusion layer (GDS 3250) under the condition of infrared lamp irradiation to obtain the catalytic layer, and the whole is the gas diffusion electrode (GDE) as the cathode. The size of the catalytic layer in the GDE is 1 cm×1 cm (i.e. the catalyst loading of the cathode is 0.9 mg / cm 2 ), and the surface of the cathode GDE occurs hydrogen evolution reaction (HER) when the electrolytic cell is working.

[0058] Anode electrode preparation: The anode electrode is a commercial Pt metal sheet with a size of 1 cm×1 cm. The surface of the anode electrode occurs oxygen evolution reaction (OER) when the electrolytic cell is working.

[0059] Preparation of double-chamber electrolytic cell: First, the structure of the double-chamber electrolytic cell used in the present application is introduced and described, as shown in Figure 1 , the double-chamber electrolytic cell is a H-shaped two-chamber electrolytic cell, which can be made of glass and other materials, which includes: Cathode chamber 2, the top is provided with a porous rubber plug 9 for fixing the cathode 1, and the middle and lower parts are provided with a first connecting chamber 5; Anode chamber 7, the top is provided with a porous rubber plug 9 for fixing the anode 8, and the middle and lower parts are provided with a second connecting chamber 6; The opposite faces of the first connecting chamber 5 and the second connecting chamber 6 are provided with flange sealing surfaces, and the electrolyte membrane 10 is arranged between the two flange sealing surfaces. The two electrolytic chambers are fastened by fasteners or clamped by clamping members, so as to fix the electrolyte membrane between the two flange sealing surfaces and realize the assembly of the cathode chamber and the anode chamber.

[0060] In addition, the cathode chamber 2 is also provided with an air inlet 3 and an air outlet 4, which is used for air replacement of the cathode chamber, so that the cathode chamber works in an inert gas atmosphere (such as nitrogen atmosphere), avoiding the influence of oxygen in the air on the adsorption of deuterium.

[0061] Preparation of substrate reaction solution: Mixing and configuring the substrate reaction solution by mixing glucose, deuterium water and electrolyte.

[0062] Preparation of double-chamber electrolytic cell: place the cathode GDE in the cathode chamber, and use perfluorosulfonic acid polymer (Nafion) as the electrolyte membrane. The anode electrode is a commercial Pt metal sheet, which is fixed in the anode chamber through a porous rubber plug; the double-chamber electrolytic cell is placed in a constant temperature environment or a water bath environment to maintain the required reaction temperature.

[0063] Electrolytic catalytic reaction: Discharge operation of electrolytic cell: Both cathode chamber and anode chamber are filled with a certain volume of substrate reaction solution (such as 100 mmol / L of glucose concentration), then nitrogen is introduced into the cathode chamber to replace and remove oxygen in the system; then the cathode and anode are connected to the negative electrode and positive electrode of the electrochemical workstation respectively, the double-chamber electrolytic cell is heated to 90 degrees Celsius, and constant voltage electrolysis is carried out at 3.5 V cell voltage for 10 minutes, and then static replacement reaction is carried out for 20 minutes, and the constant voltage electrolysis and static replacement reaction are cycled for 24-72 hours, and deuterated methyl-protected glucose solution is obtained in the cathode chamber.

[0064] NMR quantitative characterization of deuterium enrichment rate of deuterated methyl-protected glucose: After the deuterium enrichment reaction, 200 microliters of deuterated methyl-protected glucose solution in the cathode chamber is taken and placed in an NMR tube, and 200 microliters of 4.8 mmol / L dimethyl sulfoxide deuterium water solution is added as an internal standard. After the deuterium enrichment reaction, the NMR signal and peak integration results of the deuterated methyl-protected glucose in the product are shown in Figure 2 , wherein the deuterium enrichment at the 6 and 7 sites is the key deuterium enrichment site. The deuterium enrichment rate at the corresponding site is obtained by NMR integration. The examples focus on the total deuterium enrichment rate and the deuterium enrichment rate of the key site.

[0065] As the deuterium enrichment reaction time is prolonged, both the total deuterium enrichment rate and the deuterium enrichment rate of the key site are improved, as shown in Figure 3 , it can be seen that more than half of the deuterium enrichment reaction has occurred when the reaction time is 12 hours, and the deuterium enrichment reaction is basically completed when the reaction time is 24 hours, and the best state is reached when the reaction time is 48 hours. After more than 48 hours, the deuterium enrichment rate decreases, indicating that prolonging the time will not only not increase the deuterium enrichment rate, but will actually decrease it.

[0066] Example 2: The difference between it and Example 1 is that the catalyst loading of the cathode GDE is changed, which is achieved by changing the amount of ink added to change the catalyst loading. Other experimental conditions: deuterium enrichment reaction time is 24 hours, catalyst type is Ru / C, electrolyte membrane type is Nafion (Nafion is purchased from DuPont Company, dilution solvent is ethanol, concentration is about 5 mg / mL), reaction temperature is 90 degrees Celsius. The specific loading and deuterium enrichment rate change are shown in Figure 4 , the catalyst loading is 0.3 mg / cm 2 , with the increase of the loading, the total deuterium enrichment rate and the deuterium enrichment rate of the key site are greatly improved, indicating that the catalyst loading size has a great influence on the deuterium enrichment rate at this loading, and when the catalyst loading is 0.3 mg / cm 2 , the deuterium enrichment rate has exceeded 50% and is close to 60%, reaching the level of the prior art; when the catalyst loading is 0.9 mg / cm 2At 1.5 mg / cm 2 When the catalyst loading is further increased, although the total deuterium enrichment rate increases slightly, the key site deuterium enrichment rate decreases instead.

[0067] Example 3: The difference between it and Example 2 is that the catalyst type of the cathode GDE is changed, which is achieved by changing the catalyst type in the ink. Other experimental conditions: deuterium enrichment reaction time is 24 hours, catalyst loading is 0.3 mg / cm 2 , electrolyte membrane type is Nafion (Nafion is purchased from DuPont Company, the dilution solvent is ethanol, and the concentration is about 5 mg / mL), and the reaction temperature is 90 degrees Celsius. The specific catalyst types include 60% PtRu / C (the total amount of Pt and Ru is 60% of the total mass of the catalyst, the atomic ratio is 1:1, carbon is the carrier, and other catalysts are similar), 10% Pd / C, 20% PdRu / C, and 40% Ru / C. The four catalysts are common commercial catalyst types. The deuterium enrichment rates of different cathode catalysts are shown in Figure 5 It can be seen from the third and fourth catalysts that the higher the content of ruthenium in the catalyst, the higher the deuterium enrichment rate. From the second catalyst (60% PtRu / C), it can be seen that although the content of ruthenium is high, the metal platinum has a greater negative effect on the deuterium enrichment reaction. According to the reaction principle of the present application, the main reason is that the adsorption capacity of metal platinum is too strong, which makes most of the adsorbed deuterium convert into deuterium water. From the first catalyst, it can be seen that the total deuterium enrichment rate of the catalyst without ruthenium is 0, which means that the metal palladium Pd alone does not have the catalytic ability of deuterium enrichment reaction, fully demonstrating that among the currently known common catalysts, only metal ruthenium has good electrochemical catalytic performance for deuterium enrichment reaction.

[0068] Example 4: The difference between it and Example 2 is that the electrolyte membrane type of the H-type electrolytic cell is changed, and the electrolyte membrane is changed to quaternary ammoniumized p-phenylene-piperidine copolymer (QAPPT) (purchased from Billion Lithium Energy). Other experimental conditions: catalyst type is Ru / C, deuterium enrichment reaction time is 24 hours, catalyst loading is 0.3 mg / cm 2 , and the reaction temperature is 90 degrees Celsius. The specific electrolyte membrane types and deuterium enrichment rate changes are shown in Figure 6 It can be seen that the electrolyte membrane has little effect on the efficiency of the deuterium enrichment reaction. The reason is that the function of the electrolyte membrane in the present application is only to transfer conductive ions and isolate oxygen generated by the anode. Whether it is a cationic electrolyte membrane or an anionic electrolyte membrane can meet the requirements.

[0069] Example 5: The difference between it and example 2 is that the deuterium exchange reaction temperature is changed, which is realized by the heating temperature of the H-type electrolytic cell. Other experimental conditions: the catalyst type is Ru / C, the deuterium exchange reaction time is 24 hours, the catalyst loading is 0.3 mg / cm 2 , and the electrolyte membrane type is Nafion. The specific deuterium exchange reaction temperature and deuterium exchange rate change are shown in Figure 7 , from Figure 7 It can be seen that when the temperature is below 50 degrees Celsius, the electrolytic catalytic reaction of the present application cannot occur, and when the temperature is 70 degrees Celsius, the total deuterium exchange rate is almost at the latest level. When the temperature is further increased, the total deuterium exchange rate is slightly increased, but the key point deuterium exchange rate is greatly increased, about 20 percentage points, which shows that the higher the temperature, the higher the key point deuterium exchange rate. However, there is an upper limit to the temperature, and after the deuterium water solution boils, the electrolytic catalytic deuterium exchange reaction cannot proceed smoothly.

[0070] Finally, it should be pointed out that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit it; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for preparing deuterated methyl protected glucose, characterized in that, Includes the following steps: Prepare a dual-chamber electrolytic cell, which includes a cathode chamber, an anode chamber, and an electrolyte membrane disposed between the cathode and anode chambers to isolate oxygen; A gas diffusion electrode containing a ruthenium catalyst is prepared as the cathode, and a metal electrode is prepared as the anode; A substrate reaction solution was prepared by mixing methyl-protected glucose, deuterium water, and electrolytes. The substrate reaction solution was added to the cathode chamber and the anode chamber respectively, and then inserted into the cathode and anode respectively to carry out the electrolytic catalytic reaction for a period of time. During the electrolysis, an exchange reaction occurred to produce deuterated methyl protected glucose. After the electrolytic catalytic reaction is completed, the product from the cathode chamber is collected to obtain a deuterated methyl protected glucose solution.

2. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The electrolyte membrane includes a cationic electrolyte membrane and an anionic electrolyte membrane.

3. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The catalyst type in the gas diffusion electrode containing the ruthenium catalyst includes a ruthenium catalyst or a palladium-ruthenium alloy catalyst.

4. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The catalyst loading in the diffusion electrode in the gas is 0.3-2.0 mg / cm³. 2 .

5. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The electrolytic catalytic reaction is carried out at a temperature of 70-100℃ for 12-72 hours.

6. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The electrolytic catalytic reaction process involves alternating constant-voltage electrolysis and static exchange until the total reaction time is reached.

7. The method for preparing deuterated methyl protected glucose according to claim 6, characterized in that, The settling time is 1.5-2.5 times the constant voltage electrolysis time.

8. The method for preparing deuterated methyl protected glucose according to claim 1, characterized in that, The electrolyte in the substrate reaction solution is any one or more of sodium sulfate, potassium hydroxide, and sodium hydroxide.

9. The method for preparing deuterated methyl protected glucose according to claim 8, characterized in that, The electrolyte in the substrate reaction solution is sodium sulfate.

10. A deuterated methyl protected glucose, characterized in that, It is prepared by the preparation method described in any one of claims 1-9.

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