Method for chiral resolution of fluoxazole amide enantiomer

Abstract

The invention provides a method for chiral resolution of a fluoxazole amide enantiomer. A high performance liquid chromatography-mass spectrometry technology is adopted to separate the fluoxazole amide enantiomer, a chromatographic column is a polysaccharide derivative chiral chromatographic column, a mobile phase A is an organic solvent, a mobile phase B is 0.05-0.1% formic acid water, the volume ratio of the mobile phases A to the mobile phase B is 60: 40-100: 0, gradient elution is performed, the column temperature is 30-35 DEG C, and the flow velocity is 0.3-0.6 mL / min. The invention provides a method for effectively splitting the chiral fluoxazole amide enantiomer, and the method has the advantages of high sensitivity, simplicity in operation, high separation speed and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical synthesis, and particularly relates to a method for chiral separation of fluorooxazole amide enantiomers. Background Art

[0002] With the emergence and overuse of pesticides, pesticide residues exist in vegetables, fruits, crops, soil and water bodies, which not only have adverse effects on non-target organisms, but also cause certain harm to human health and environmental safety. Among the pesticides currently used, chiral pesticides account for about 40% of the total pesticides, most of which are racemic mixtures of enantiomers. Enantiomers have the same physicochemical properties, but they usually show different pharmacokinetic and pharmacodynamic characteristics, and show stereoselective differences in biological activity, ecotoxicity and environmental behavior. Therefore, studying chiral pesticides at the enantiomeric level is of great significance for the development of environmentally friendly, efficient, low-toxic optically pure monomeric pesticides.

[0003] Fluxametamide is an isoxazole insecticide developed by Nissan Chemical Co., Ltd. in 2007. It has a novel mechanism of action and is a γ-aminobutyric acid (GABA)-gated chloride channel allosteric modulator, a neurotoxin. It inhibits the neurotransmission of target insects through stomach poison and contact killing, causing the insects to twitch and eventually die. Fluxametamide has no cross-resistance with conventional insecticides and is mainly used in crops such as vegetables, fruit trees, cotton, and tea trees to control pests and mites such as thrips, whiteflies, leaf miners, beetles, red spiders, and rust mites. At present, there are no literature reports on the toxicity and environmental risks of the R and S configuration enantiomers of fluxametamide, so it is particularly important to establish a chiral separation method for the enantiomers of fluxametamide for further research.

[0004] Summary of the invention

[0005] The invention provides a method for chiral separation of fluorooxazole amide enantiomers, and uses high performance liquid chromatography-mass spectrometry technology to achieve separation of R and S configurations of fluorooxazole amide.

[0006] The technical solution adopted by the present invention is:

[0007] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0008] 1) Prepare the test solution: weigh oxazolidinone and dissolve it in an organic solvent to prepare a oxazolidinone test solution with a concentration of 100-500 ng / mL;

[0009] 2) Separating the enantiomers of oxazolidinone in the oxazolidinone test solution by high performance liquid chromatography-mass spectrometry, wherein the stationary phase is a polysaccharide derivative chiral chromatographic column, the mobile phase A is an organic solvent, the mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-100:0, gradient elution, the column temperature is 30-35° C., and the flow rate is 0.3-0.6 mL / min;

[0010] 3) Determine using an ultraviolet detector or a mass spectrometer.

[0011] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 1), the organic solvent is one of methanol and acetonitrile.

[0012] Furthermore, in the above-mentioned method for chiral resolution of fluorooxazolamide enantiomers, in step 1), the concentration of the fluorooxazolamide test solution is 100 ng / mL.

[0013] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), the organic solvent is one of methanol and acetonitrile.

[0014] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), the polysaccharide derivative chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface.

[0015] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), the organic solvent is methanol.

[0016] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), mobile phase A is methanol, mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-85:15, and gradient elution is performed.

[0017] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), the column temperature is 35° C. and the flow rate is 0.3-0.6 mL / min.

[0018] Furthermore, in the above-mentioned method for chiral separation of fluorooxazole amide enantiomers, in step 2), mobile phase A is methanol, mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-85:15, gradient elution, column temperature is 35°C, and the flow rate is 0.6mL / min.

[0019] Furthermore, in the above-mentioned method for chiral resolution of fluorooxazole amide enantiomers, in step 3), the detector is a mass spectrometer detector.

[0020] The technical solution of the present application has the following beneficial effects: the present invention adopts high performance liquid chromatography-mass spectrometry technology to directly chirally separate fluorooxazole amide, and its mobile phase is composed of methanol and formic acid water. This method has the advantages of simple operation, high sensitivity, mild conditions, and short separation time. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is the chromatographic separation pattern of Example 1.

[0022] Figure 2 It is the chromatographic separation pattern of Example 2.

[0023] Figure 3 It is the chromatographic separation pattern of Example 3.

[0024] Figure 4 It is the chromatographic separation pattern of Example 4.

[0025] Figure 5 It is the chromatographic separation pattern of Example 5. DETAILED DESCRIPTION

[0026] The advantages of the present invention are further described below in conjunction with the accompanying drawings and specific embodiments.

[0027] Exemplary embodiments will be described in detail herein, examples of which are shown in the accompanying drawings. When the following description refers to the drawings, the same numbers in different drawings represent the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. Instead, they are merely examples of devices and methods consistent with some aspects of the present disclosure as detailed in the appended claims.

[0028] The terms used in this disclosure are for the purpose of describing specific embodiments only and are not intended to limit the disclosure. The singular forms of "a", "said" and "the" used in this disclosure and the appended claims are also intended to include plural forms unless the context clearly indicates otherwise. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0029] Example 1

[0030] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0031] 1) Weigh a certain amount of oxazolidinone, dissolve it in methanol and make up to volume to prepare a 100 ng / mL oxazolidinone test solution.

[0032] 2) Chiral separation using HPLC-MS

[0033] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IB chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface, with specifications of 250mm×4.6mm and a particle size of 5μm.

[0034] The mobile phase A is methanol, the mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 40:60-90:10, and the gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6mL / min; and the injection volume is 20μL.

[0035] 3) Measured using a mass spectrometer.

[0036] The results are as follows Figure 1 .Depend on Figure 1 It can be seen that the enantiomers of fluorooxazole amide were not effectively separated.

[0037] Example 2

[0038] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0039] 1) Weigh a certain amount of oxazolidinone, dissolve it in methanol and make up to volume to prepare a 100 ng / mL oxazolidinone test solution.

[0040] 2) Chiral separation using HPLC-MS

[0041] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IB chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface, with specifications of 250mm×4.6mm and a particle size of 5μm.

[0042] The mobile phase A is methanol, the mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 40:60-100:0, and the gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6mL / min; and the injection volume is 20μL.

[0043] 3) Measured using a mass spectrometer.

[0044] The results are as follows Figure 2 .Depend on Figure 2 It can be seen that the enantiomers of fluorooxazole amide were not effectively separated.

[0045] Example 3

[0046] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0047] 1) Weigh a certain amount of oxazolidinone, dissolve it in methanol and make up to volume to prepare a 100 ng / mL oxazolidinone test solution.

[0048] 2) Chiral separation using HPLC-MS

[0049] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IB chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface, with specifications of 250mm×4.6mm and a particle size of 5μm.

[0050] The mobile phase A is methanol, the mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 50:50-80:20, gradient elution. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6mL / min; the injection volume is 20μL.

[0051] 3) Measured using a mass spectrometer.

[0052] The results are as follows Figure 3 .according to Figure 3 It can be seen that the chromatographic peaks of the fluorooxazole amide enantiomers cannot be effectively separated.

[0053] Example 4

[0054] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0055] 1) Weigh a certain amount of oxazolidinone, dissolve it in methanol and make up to volume to prepare a 100 ng / mL oxazolidinone test solution.

[0056] 2) Chiral separation using HPLC-MS

[0057] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IB chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface, with specifications of 250mm×4.6mm and a particle size of 5μm.

[0058] The mobile phase A is methanol, the mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 85:15, isocratic elution. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6mL / min; and the injection volume is 20μL.

[0059] 3) Measured using a mass spectrometer.

[0060] The results are as follows Figure 4 .Depend on Figure 4 It can be seen that the enantiomers of fluorooxazole amide were not effectively separated.

[0061] Example 5

[0062] A method for chiral separation of fluorooxazole amide enantiomers comprises the following steps:

[0063] 1) Weigh a certain amount of oxazolidinone, dissolve it in methanol and make up to volume to prepare a 100 ng / mL oxazolidinone test solution.

[0064] 2) Chiral separation using HPLC-MS

[0065] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IB chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with cellulose-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica gel surface, with specifications of 250mm×4.6mm and a particle size of 5μm.

[0066] The mobile phase A is methanol, the mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-85:15, gradient elution. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6mL / min; the injection volume is 20μL.

[0067] 3) Measured using a mass spectrometer.

[0068] The results are as follows Figure 5 .Depend on Figure 5 It can be seen that the enantiomers of fluorooxazole amide are effectively separated. The calculated resolution is R=1.58, and effective baseline separation is achieved.

[0069] In summary, the present application adopts high performance liquid chromatography-mass spectrometry technology. Only under specific chromatographic conditions, that is, mobile phase A is methanol, mobile phase B is 0.1% formic acid water, the mobile phase is calculated by volume ratio, A: B is 60:40-85:15, gradient elution, column temperature is 35°C, flow rate is 0.6mL / min, can effectively achieve chiral separation of fluorooxazole amide, and the chromatographic peaks of fluorooxazole amide enantiomers are effectively separated. The calculated resolution reaches more than 1.5, and effective baseline separation is achieved.

[0070] The separated fluorooxazolidinone can be used for subsequent practical industrial applications. It should be noted that the embodiments of the present invention have better practicability and are not intended to limit the present invention in any form. Any technician familiar with the field may use the above disclosed technical content to change or modify it into an equivalent effective embodiment. However, any modification or equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A method for chiral separation of fluorooxazole amide enantiomers, characterized in that: The following steps are involved: 1) Prepare the test solution: weigh oxazolidinone and dissolve it in an organic solvent to prepare a oxazolidinone test solution with a concentration of 100-500 ng / mL; 2) Separating the enantiomers of oxazolidinone in the oxazolidinone test solution by high performance liquid chromatography-mass spectrometry, wherein the stationary phase is a polysaccharide derivative chiral chromatographic column, the mobile phase A is an organic solvent, the mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-100:0, gradient elution, the column temperature is 30-35° C., and the flow rate is 0.3-0.6 mL / min; 3) Determine using an ultraviolet detector or a mass spectrometer.

2. A method for chiral separation of fluorooxazole amide enantiomers according to claim 1, characterized in that: In step 1), the organic solvent is one of methanol and acetonitrile.

3. A method for chiral separation of fluorooxazole amide enantiomers according to claim 1, characterized in that: In step 1), the concentration of the fluoxetine amide test solution is 100 ng / mL.

4. A method for chiral separation of fluorooxazole amide enantiomers according to claim 1, characterized in that: In step 2), the organic solvent is one of methanol and acetonitrile.

5. A method for chiral separation of fluorooxazole amide enantiomers according to claim 4, characterized in that: In step 2), the polysaccharide derivative chiral chromatographic column is a polysaccharide derivative chiral chromatographic column in which cellulose-tris(3,5-dimethylphenylcarbamate) is covalently bonded to the surface of silica gel.

6. A method for chiral separation of fluorooxazole amide enantiomers according to claim 5, characterized in that: In step 2), the organic solvent is methanol.

7. A method for chiral separation of fluorooxazole amide enantiomers according to claim 6, characterized in that: In step 2), mobile phase A is methanol, mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-85:15, and gradient elution is performed.

8. A method for chiral separation of fluorooxazole amide enantiomers according to claim 7, characterized in that: In step 2), the column temperature is 35°C and the flow rate is 0.3-0.6 mL / min.

9. A method for chiral separation of fluorooxazole amide enantiomers according to claim 8, characterized in that: In step 2), mobile phase A is methanol, mobile phase B is 0.05-0.1% formic acid water, the mobile phase is calculated by volume ratio, A:B is 60:40-85:15, gradient elution, column temperature is 35° C., and flow rate is 0.6 mL / min.

10. The method for chiral separation of fluorooxazole amide enantiomers according to claim 1, characterized in that: In step 3), the detector is a mass spectrometer detector.

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