Synthesis method of stable isotope labeled thiamphenicol and thiamphenicol amine

Abstract

The invention discloses a method for synthesizing thiamphenicol and thiamphenicolamine labeled with a stable isotope. Thiamphenicol is used as a raw material, hydrolyzed to obtain thiamphenicolamine, hydrogen-deuterium exchanged under alkaline conditions to obtain stable isotope-labeled thiamphenicolamine, and further acylated to obtain stable isotope-labeled thiamphenicol. The invention has a short synthesis route, high atomic utilization, excellent yield, and the raw materials are cheap and easy to obtain. The chemical purity and deuterium isotope abundance of the prepared stable isotope-labeled thiamphenicol and thiamphenicolamine are both above 98.0%, and can be used as an isotope internal standard for the detection of thiamphenicol residues in food.

Description

Technical Field

[0001] The invention relates to the field of stable isotope labeled compound production, and in particular to a method for synthesizing stable isotope labeled thiamphenicol and thiamphenicol amine. Background Art

[0002] Thiamphenicol is a derivative of chloramphenicol. It is an amide alcohol broad-spectrum antibacterial drug synthesized by replacing the para-nitro group in the molecular structure of chloramphenicol with a methylsulfone group. It is widely used in clinical practice to prevent and treat infectious diseases in animals. However, due to its toxic side effects on the blood system, its residues in animal foods may endanger human health. In order to ensure the safety of animal-derived food, GB 31650.1-2022 stipulates that the maximum residue limit of thiamphenicol in animal foods is 10μg / kg. In recent years, the number of residual analysis and detection methods for chloramphenicol drugs in food has continued to increase, among which high-performance liquid chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry isotope internal standard method are the main detection methods for chloramphenicol drugs.

[0003] Chinese invention patent CN107827791 discloses a thiamphenicol-d 3 The preparation method comprises the following steps: using p-bromobenzaldehyde and isotope-labeled dimethyl sulfoxide as raw materials to synthesize isotope-labeled p-methylthiobenzaldehyde, further oxidizing to obtain isotope-labeled p-methylsulfonylbenzaldehyde, condensing with diphenylamine to form imine, and then constructing ethyl diazoacetate structure fragment with (R)-2,2'-diphenyl-3,3'-(4-biphenylphenol) and triphenyl borate, and finally opening the ethyl imine under dichloroacetic acid conditions and reducing the ester group to synthesize thiamphenicol-d 3 The experimental procedure requires six steps of reaction, which is cumbersome and has a low total yield of only 38%. 4 and intermediate thiamphenicolamine-d 3 No literature reports found.

[0004] From the above-mentioned prior art, it can be seen that the stable isotope labeling of thiamphenicol is very difficult and still far from commercialization, and the synthesis method needs to be further optimized and improved. Therefore, the art still needs to develop a low-cost, simple-step, and effective synthesis method of thiamphenicol with stable isotope labeling, requiring the purity of the target compound and the abundance of the labeled isotope to be above 98%, meeting the requirements for its use as an analytical internal standard. Summary of the invention

[0005] The purpose of the present invention is to provide a method for synthesizing thiamphenicol and thiamphenicol amine labeled with stable isotopes, which has low cost, simple steps, high atomic utilization rate and good effect, and the purity and isotopic abundance of the target compound are both above 98%, meeting the requirements for use as an analytical internal standard.

[0006] Therefore, one aspect of the present invention relates to a method for synthesizing stable isotope-labeled thiamphenicol amine, comprising:

[0007] Thiamphenicolamine, alkali and heavy water are subjected to hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0008] Another aspect of the present invention relates to a stable isotope labeled thiamphenicol-d 3 A synthesis method comprising:

[0009] Take thiamphenicol-d 3 , dissolved in a non-deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate to obtain thiamphenicol-d 3 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0010] Another aspect of the present invention relates to another stable isotope labeled thiamphenicol-d 4 A synthesis method comprising:

[0011] Take thiamphenicol-d 3 , dissolved in a deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate, and acylated to obtain thiamphenicol-d 4 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 2-4h. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 is a compound of the invention according to Example 1 of the present invention. 3 H NMR spectrum.

[0013] Figure 2 is a compound of the invention according to Example 1 of the present invention. 3 LC-MS spectrum of

[0014] Figure 3 is a compound of the invention according to Example 1 of the present invention. 3 H NMR spectrum.

[0015] Figure 4 is a compound of the invention according to Example 1 of the present invention. 3 HPLC spectrum of.

[0016] Figure 5 is a compound of the invention according to Example 1 of the present invention. 3 LC-MS spectrum of

[0017] Figure 6 is a compound of the invention according to Example 2 of the present invention. 4 H NMR spectrum.

[0018] Figure 7 is a compound of the invention according to Example 2 of the present invention. 4 HPLC spectrum of.

[0019] Figure 8 is a compound of the invention according to Example 2 of the present invention. 4 LC-MS spectrum of DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following general description and definition of the terms and uses mentioned in the specification and claims are provided. Unless otherwise specified, the technical and scientific terms used herein are the common meanings understood by those skilled in the art for the present invention. In case of conflict, the definitions in this specification shall prevail.

[0021] In this article, all features defined in the form of numerical ranges or percentage ranges, such as values, quantities, contents and concentrations, are only for brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be considered to have included and specifically disclosed all possible secondary ranges and individual values ​​within the range (including integers and fractions).

[0022] The above-mentioned features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this specification can be used in combination with any combination form. As long as there is no contradiction in the combination of these features, all possible combinations should be considered to be within the scope of this specification. Each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0023] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly described below in conjunction with the specific embodiments of the present invention and the corresponding drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0024] The inventors of the present invention have found through research that, in terms of isotope exchange, the exchange efficiency of thiamphenicol amine with heavy water and the abundance of the labeled isotope of the final product are significantly better than those of thiamphenicol, and when thiamphenicol amine is converted into thiamphenicol through an acylation reaction, its abundance of the labeled isotope is not affected. The present invention is completed on this basis.

[0025] 1. Stable isotope labeled thiamphenicol amine

[0026] The synthesis method of stable isotope-labeled thiamphenicol amine of the present invention comprises:

[0027] Thiamphenicolamine, alkali and heavy water are subjected to hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0028] The thiamphenicol amine is prepared by heating thiamphenicol under reflux under acidic conditions.

[0029] As a preferred embodiment of the present invention, the acidic condition is selected from 0.8 to 1.3 mol / L hydrochloric acid aqueous solution, preferably 1 mol / L hydrochloric acid aqueous solution.

[0030] As a preferred embodiment of the present invention, the reaction conditions of heating under reflux are heating under reflux at 80-100°C, preferably at 85-95°C, more preferably at 88-92°C for 4-8 hours, preferably 5-7 hours, more preferably 6 hours.

[0031] Suitable bases include barium oxide, sodium deuteride, potassium carbonate or cesium carbonate.

[0032] As a preferred embodiment of the present invention, the reaction temperature of the hydrogen-deuterium exchange reaction is 100-120°C, preferably the 100-120°C includes 105-120°C, 110-120°C, 115-120°C, 100-115°C, 105-115°C, 110-115°C, 100-110°C, 105-110°C, 100-105°C, etc.

[0033] As a preferred embodiment of the present invention, the reaction time of the hydrogen-deuterium exchange reaction is 46 to 50 hours, preferably the 46 to 50 hours includes 47 to 50 hours, 48 ​​to 50 hours, 49 to 50 hours, 46 to 49 hours, 47 to 49 hours, 48 ​​to 49 hours, 47 to 48 hours, etc.

[0034] As a preferred embodiment of the present invention, the molar ratio of thiamphenicol amine, base and heavy water is 1:0.3-0.7:200-400, preferably 1:0.4-0.6:250-350, and more preferably 1:0.45-0.55:280-320.

[0035] 2. Thiamphenicol-d 3 Synthesis

[0036] The present invention also relates to a thiamphenicol-d 3 A synthesis method comprising:

[0037] Take thiamphenicol-d 3 , dissolved in a non-deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate to obtain thiamphenicol-d 3 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0038] The non-deuterated solvent suitable for the above method of the present invention can be selected from alcohol solvents and / or nitrile solvents. In a preferred embodiment of the present invention, the solvent is selected from methanol and / or acetonitrile.

[0039] The reaction temperature suitable for the above method of the present invention is 30-50°C, preferably 32-48°C, more preferably 35-45°C, and preferably 38-42°C.

[0040] The reaction time suitable for the above method of the present invention is 22 to 26 hours, preferably 23 to 25 hours.

[0041] 3. Thiamphenicol-d 4 Synthesis

[0042] The present invention also relates to a thiamphenicol-d 4 A synthesis method comprising:

[0043] Take thiamphenicol-d 3 , dissolved in a deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate, and acylated to obtain thiamphenicol-d 3 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0044] The deuterated solvent suitable for the above method of the present invention can be selected from deuterated alcohol solvents. In a preferred embodiment of the present invention, the solvent is selected from methanol-d 4 and / or methanol-d 1 .

[0045] The reaction temperature suitable for the above method of the present invention is 30-50°C, preferably 32-48°C, more preferably 35-45°C, and preferably 38-42°C.

[0046] The reaction time suitable for the above method of the present invention is 22 to 26 hours, preferably 23 to 25 hours.

[0047] In one example of the present invention, the synthesis method of the labeled thiamphenicol comprises:

[0048] (1) subjecting thiamphenicolamine, alkali and heavy water to a hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0049] (2) Take thiamphenicol amine-d 3 , dissolved in a non-deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate to obtain thiamphenicol-d 3 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0050] In one example of the present invention, the synthesis method of the labeled thiamphenicol comprises:

[0051] (1) subjecting thiamphenicolamine, alkali and heavy water to a hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0052] (2) Take thiamphenicol amine-d 3 , dissolved in a deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate, and acylated to obtain thiamphenicol-d 4 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0053] In one example of the present invention, the synthesis method of the labeled thiamphenicol comprises:

[0054] (1) heating thiamphenicol under acidic conditions at 80-110° C. for 6 h to obtain thiamphenicol amine;

[0055] (2) subjecting thiamphenicolamine, alkali and heavy water to a hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0056] (3) Take thiamphenicol amine-d 3 , dissolved in a non-deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate to obtain thiamphenicol-d 3 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0057] In one example of the present invention, the synthesis method of the labeled thiamphenicol comprises:

[0058] (1) heating thiamphenicol under acidic conditions at 80-110° C. for 6 h to obtain thiamphenicol amine;

[0059] (2) subjecting thiamphenicolamine, alkali and heavy water to a hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 100-120°C and the reaction time is 46-50h.

[0060] (3) Take thiamphenicol amine-d 3 , dissolved in a deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate, and acylated to obtain thiamphenicol-d 4 ; wherein the acylation reaction temperature is 30-50°C and the reaction time is 22-26h.

[0061] The synthesis method of the present invention has cheap and readily available raw materials, low synthesis cost, only three reaction steps, a short synthesis route, and good economic efficiency. 3 , which greatly shortens the technical route and has an excellent yield of more than 90%. The target compound thiamphenicol-d 3 , thiamphenicol-d 4 The yield is high, and the purity and isotope abundance are both above 98%, and can be used as an internal standard for analysis and detection.

[0062] Based on this, the present invention provides a low-cost, simple process, high purity and isotopic abundance thiamphenicol-d 3 Thiamphenicol-d 4 The synthesis method.

[0063] The main compounds involved in the present invention are shown in the following table:

[0064]

[0065]

[0066] As used in the present invention, "a compound having a structure as shown in Formula I" and "a compound of Formula I" both refer to the compound numbered I in the above table. The same applies to compounds of other structures.

[0067] In the present invention, the thiamphenicol is a compound with a structure as shown in Formula I, and the thiamphenicol amine is a compound with a structure as shown in Formula IV; the target compound thiamphenicol-d 3 The compound shown in formula II, thiamphenicol-d 4 It is a compound with a structure as shown in formula III.

[0068] Preferably, the synthetic route of the present invention is as follows:

[0069]

[0070] In one embodiment of the present invention, thiamphenicol is dissolved in a 1 mol / L hydrochloric acid aqueous solution; the reaction temperature is 100° C. and heated under reflux for 6 h to obtain thiamphenicol amine; thiamphenicol amine, barium oxide, and heavy water are sequentially added to a sealed tube, stirred and dissolved, and the reaction temperature is 110° C. and heated for 48 h to obtain thiamphenicol amine-d 3 ; Weigh thiamphenicol amine-d 3 , dissolved in 40 mL of acetonitrile, added triethylamine and methyl dichloroacetate at 1-2 drops / second, heated at 40°C for 3 h to obtain thiamphenicol-d 3 .

[0071] In one embodiment of the present invention, thiamphenicol is dissolved in a 1 mol / L hydrochloric acid aqueous solution; the reaction temperature is 100° C. and heated under reflux for 6 h to obtain thiamphenicol amine; thiamphenicol amine, barium oxide, and heavy water are sequentially added to a sealed tube, stirred and dissolved, and the reaction temperature is 110° C. and heated for 48 h to obtain thiamphenicol amine-d 3 ; Weigh thiamphenicol amine-d 3 , dissolved in 40 mL methanol-d 1 , add triethylamine and methyl dichloroacetate at 1-2 drops / second, heat at 40℃ for 3h to obtain thiamphenicol-d 4 .

[0072] Purity and isotope abundance detection methods used in the examples:

[0073] Thiamphenicol-d 3 Purity testing

[0074] (1) Thiamphenicol-d 3 HPLC test conditions

[0075] Chromatographic column: Athena UHPLC C18 (100mm*2.1mm, 1.8μm)

[0076] Column temperature: 30°C

[0077] Mobile phase: A: acetonitrile; B: 0.1% phosphoric acid water

[0078] Gradient: 0min 5%A; 2min 5%A; 8min 100%A; 12min 100%A; 12.1

[0079] min 5% A; 15min 5% A

[0080] Flow rate: 0.3mL / min

[0081] Detector: DAD 226nm

[0082] Sample: 0.5 μL, 1.02 mg / mL in methanol

[0083] (2) The chromatographic purity was calculated using the HPLC area normalization method.

[0084] Thiamphenicol-d 3 Isotope abundance detection

[0085] Determination of thiamphenicol-d by the “Simplified Mass Cluster” method 3 The peak areas of the quantitative ion pairs 357-188, 356-187, 355-186, and 354-185 were measured by mass spectrometry, and were approximately equal to the peak areas of thiamphenicol-D3, thiamphenicol-D2, thiamphenicol-D1, and thiamphenicol-D0, respectively. Based on the detection basis that "the mass spectrometry peak intensity is proportional to the number of molecules", the isotopic abundance percentages of D3-D0 were normalized. Finally, the formula:

[0086]

[0087] Wherein, D%: represents isotope abundance; i: represents the number of D labels, i=0-3; Ri: represents the percentage of the peak area of ​​i D labels, taking 3 D labels as an example, the calculation formula is: R3=A(D3) / [A(D3)+A(D2)+A(D1)+A(D0)]; n: represents the total number of D labels, n=3.

[0088] Mass spectrometry conditions:

[0089] Detection mode: multiple ion scan

[0090] Ionization mode: Negative ion mode

[0091] Ion source voltage: 3500

[0092] Ion transfer tube temperature: 300°C

[0093] Steam temperature: 200℃

[0094] Scan range: 50-400 (m / z)

[0095] Flow rate: 20 μL / min

[0096] Qualifying ion collision energy: 15

[0097] Quantitative ion pairs: 354-185, 355-186, 356-187, 357-188

[0098] Thiamphenicol-d 4 Purity testing

[0099] (1) Thiamphenicol-d 4 HPLC test conditions

[0100] Chromatographic column: Athena UHPLC C18 (100mm*2.1mm, 1.8μm)

[0101] Column temperature: 30°C

[0102] Mobile phase: A: acetonitrile; B: 0.1% phosphoric acid water

[0103] Gradient: 0min 5%A; 2min 5%A; 8min 100%A; 12min 100%A; 12.1

[0104] min 5% A; 15min 5% A

[0105] Flow rate: 0.3mL / min

[0106] Detector: DAD 226nm

[0107] Sample: 0.5 μL, 1.02 mg / mL in methanol

[0108] (2) The chromatographic purity was calculated using the HPLC area normalization method.

[0109] Thiamphenicol-d 4 Isotope abundance detection

[0110] Determination of thiamphenicol-d by the “Simplified Mass Cluster” method 4 The isotopic abundance of 358-188, 357-187, 356-186, 355-185, and 354-184 was measured by mass spectrometry, and the peak areas of thiamphenicol-D4, thiamphenicol-D3, thiamphenicol-D2, thiamphenicol-D1, and thiamphenicol-D0 were approximated. Based on the detection basis that "the mass spectrometry peak intensity is proportional to the number of molecules", the isotopic abundance percentages of D4-D0 were normalized. Finally, the formula:

[0111]

[0112] Wherein, D%: represents isotope abundance; i: represents the number of D labels, i=0-4; Ri: represents the percentage of the peak area of ​​i D labels, taking 4 D labels as an example, the calculation formula is: R4=A(D4) / [A(D4)+A(D3)+A(D2)+A(D1)+A(D0)]; n: represents the total number of D labels, n=4.

[0113] Mass spectrometry detection conditions:

[0114] Detection mode: multiple ion scan

[0115] Ionization mode: Negative ion mode

[0116] Ion source voltage: 3500

[0117] Ion transfer tube temperature: 300°C

[0118] Steam temperature: 200℃

[0119] Scan range: 50-400 (m / z)

[0120] Flow rate: 20 μL / min

[0121] Qualifying ion collision energy: 15

[0122] Quantitative ion pairs: 358-188, 357-187, 356-186, 355-185, 354-184

[0123] Example 1 Thiamphenicol-d 3 Thiamphenicol-d 3 Synthesis method

[0124] A method for synthesizing thiamphenicol and thiamphenicol amine labeled with a stable isotope comprises the following steps:

[0125] (1) Thiamphenicol hydrolysis

[0126] Take a 250mL three-necked flask, dissolve thiamphenicol (3.56g, 10mmol) in 100mL dilute hydrochloric acid (1mol / L), reflux at 100℃ overnight, monitor the reaction completion by TLC, cool the reaction solution to room temperature, extract twice with ethyl acetate to remove impurities, and spin dry the aqueous phase to obtain thiamphenicol amine, 2.40g, yield 97.84%.

[0127] (2) Thiamphenicol-d 3 synthesis

[0128] Take 250mL sealed tube, stir and dissolve thiamphenicol amine (2.40g, 9.78mmol), barium oxide (750.09mg, 4.89mmol), and heavy water (78.38g, 3.91mol), heat and stir at 110℃ for 48h, cool to room temperature, filter to remove insoluble impurities, and spin dry the filtrate to obtain thiamphenicol amine-d 3 . Thiamphenicol-d 3 The isotopic abundance is 98.82%, D3: 96.64%, D2: 3.22%, D1: 0.09%, and D0: 0.05%.

[0129] (3) Thiamphenicol-d 3 synthesis

[0130] Thiamphenicol-d 3(1g, 4.03mmol) was dissolved in 40mL acetonitrile, and 1mL triethylamine and methyl dichloroacetate (1.73g, 12.08mmol) were added at 1-2 drops / second, and the reaction was heated at 40℃ for 3h. The reaction was complete after TLC monitoring. The reaction liquid was spin-dried, recrystallized from methanol / dichloromethane, and filtered after standing at low temperature. The filter cake was vacuum dried to obtain the target product thiamphenicol-d 3 , 1.43 g, yield 98.62%. The target product thiamphenicol-d 3 The chromatographic purity is 99.63% and the isotopic abundance is 98.83%.

[0131] Thiamphenicol-d 3 The mass spectrum of the negative ion mode is shown in the attached figure. Figure 5 As shown. Measured molecular ion peak [MH] - =357.08, which is consistent with the theoretical calculation result of 357.02 (C 12 H 11 D 3 Cl 2 NO 5 S - ) is consistent. The calculated thiamphenicol-d 3 The isotopic abundance value is 98.83atom%D, and there is no obvious dilution of isotopic abundance.

[0132] Example 2 Thiamphenicol-d 4 Thiamphenicol-d 3 Synthesis method

[0133] A method for synthesizing thiamphenicol and thiamphenicol amine labeled with a stable isotope comprises the following steps:

[0134] (1) Thiamphenicol hydrolysis

[0135] Take a 250mL three-necked flask, dissolve thiamphenicol (3.56g, 10mmol) in 100mL dilute hydrochloric acid (1mol / L), reflux at 100℃ overnight, monitor the reaction completion by TLC, cool the reaction solution to room temperature, extract twice with ethyl acetate to remove impurities, and spin dry the aqueous phase to obtain thiamphenicol amine, 2.40g, yield 97.84%.

[0136] (2) Thiamphenicol-d 3 synthesis

[0137] Take 250mL sealed tube, stir and dissolve thiamphenicol amine (2.40g, 9.78mmol), barium oxide (750.09mg, 4.89mmol), and heavy water (78.38g, 3.91mol), heat and stir at 110℃ for 48h, cool to room temperature, filter to remove insoluble impurities, and spin dry the filtrate to obtain thiamphenicol amine-d 3 .

[0138] (3) Thiamphenicol-d 4 synthesis

[0139] Thiamphenicol-d 3 (1 g, 4.03 mmol) was dissolved in 40 mL methanol-d 1 1 mL of triethylamine and methyl dichloroacetate (1.73 g, 12.08 mmol) were added at 1-2 drops / second, and the mixture was heated at 40°C for 3 h. The reaction was complete after monitoring by TLC. The reaction solution was dried by spin drying, and recrystallized from methanol / dichloromethane. After standing at low temperature, the mixture was filtered and the filter cake was dried in vacuo to obtain the target product thiamphenicol-d 4 , 1.19 g, yield 82.26%. The target product thiamphenicol-d 4 The chromatographic purity is 99.64% and the isotopic abundance is 99.12%.

[0140] Thiamphenicol-d 4 The mass spectrum of the negative ion mode is shown in the attached figure. Figure 8 As shown. Measured molecular ion peak [MH] - =358.04, which is consistent with the theoretical calculation result of 358.02 (C 10 H 10 D 4 NO 5 S - ) is consistent. The calculated thiamphenicol-d 4 The isotopic abundance value is 99.12atom%D, and there is no obvious dilution of isotopic abundance.

[0141] Comparative Example 1 Intermediate product thiamphenicol amine-d 3 Synthesis

[0142] Take 50mL sealed tube, stir and dissolve thiamphenicol amine (0.3g, 1.22mmol), barium oxide (93.76mg, 0.61mmol), and heavy water (9.80g, 0.49mol), heat and stir at 80℃ for 12h, cool to room temperature, filter to remove insoluble impurities, and spin dry the filtrate to obtain thiamphenicol amine-d 3 , the stable isotope abundance is 51.86%.

[0143] Comparative Example 2 Thiamphenicol-d 3 synthesis

[0144] Take 250mL sealed tube, stir and dissolve thiamphenicol (3.56g, 10.00mmol), barium oxide (1.07g, 7.00mmol), and heavy water (80.11g, 4.00mol), heat and stir at 110℃ for 48h, cool to room temperature, filter to remove insoluble impurities, and spin dry the filtrate to obtain thiamphenicol-d 3 The isotope abundance was detected to be 94.02%, D3: 83.26%, D2: 15.59%, D1: 1.09%, and D0: 0.05%.

Claims

1. A method for synthesizing stable isotope-labeled thiamphenicol amine, It is characterized in that Includes steps: Thiamphenicolamine, alkali and heavy water are subjected to hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 110°C and the reaction time is 48h; The base is selected from barium oxide; The thiamphenicolamine-d 3 The structural formula is:

2. A method for synthesizing stable isotope-labeled thiamphenicol, It is characterized in that Includes steps: Thiamphenicolamine, alkali and heavy water are subjected to hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 110°C and the reaction time is 48h; The base is selected from barium oxide; Take thiamphenicol-d 3 , dissolved in a non-deuterated solvent, and then mixed with triethylamine and methyl dichloroacetate, and subjected to an acylation reaction to obtain stable isotope-labeled thiamphenicol; wherein the reaction temperature of the acylation reaction is 30 to 50° C., and the reaction time is 2 to 4 hours; The stable isotope labeled thiamphenicol is thiamphenicol-d 3 , whose structural formula is:

3. The synthesis method according to claim 2, It is characterized in that The non-deuterated solvent is selected from an alcohol solvent, a nitrile solvent or a mixture thereof.

4. The synthesis method according to claim 3, It is characterized in that The solvent is selected from methanol and / or acetonitrile.

5. A method for synthesizing stable isotope-labeled thiamphenicol, It is characterized in that Includes steps: Thiamphenicolamine, alkali and heavy water are subjected to hydrogen-deuterium exchange reaction to obtain thiamphenicolamine-d 3 ; Wherein, the hydrogen-deuterium exchange reaction temperature is 110°C and the reaction time is 48h; The base is selected from barium oxide; Take thiamphenicol-d 3 , dissolved in a solvent, and then mixed with triethylamine and methyl dichloroacetate, and subjected to an acylation reaction to obtain stable isotope-labeled thiamphenicol; wherein the reaction temperature of the acylation reaction is 30 to 50° C., and the reaction time is 2 to 4 hours; Wherein, the solvent is selected from deuterated alcohol solvents; The stable isotope labeled thiamphenicol is thiamphenicol-d 4 , whose structural formula is:

6. The synthesis method according to claim 5, It is characterized in that The solvent is selected from methanol-d4 and / or methanol-d1.

7. The synthesis method according to any one of claims 1, 2 and 5, It is characterized in that The molar ratio of thiamphenicol amine, alkali and heavy water is 1:0.3-0.7:200-400.

8. The synthesis method according to claim 7, It is characterized in that The thiamphenicol amine is prepared by heating thiamphenicol under reflux at 80-110° C. for 4-8 hours under acidic conditions.

9. The synthesis method according to claim 8, It is characterized in that The thiamphenicol amine is prepared by heating thiamphenicol under reflux at 80-110° C. for 6 hours under acidic conditions.

10. The synthesis method according to claim 8, It is characterized in that The acidic condition is 0.8-1.3 mol / L hydrochloric acid aqueous solution.

11. The synthesis method according to claim 10, It is characterized in that The acidic condition is 1 mol / L hydrochloric acid aqueous solution.

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