Method for chiral resolution of cyflumetofen enantiomer
By using ultra-high liquid chromatography and polysaccharide derivative chiral chromatography columns in the separation process of fuflomide and using methanol and deionized water as mobile phases, the efficient separation of fuflomide enantiomers was achieved, and problems such as high equipment cost, complex mobile phase selection and control in the prior art were solved, and the effects of simple operation, high sensitivity and environmental protection were achieved.
Patent Information
- Application Number
- CN202411933291.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-26
- Publication Date
- 2025-04-08
AI Technical Summary
The prior art methods for chiral separation of fuflomide enantiomers have disadvantages such as high equipment cost, complex mobile phase selection and control, difficult method development and optimization, limited detection sensitivity, high sample pretreatment requirements, resulting in high experimental costs, narrow application scope, low resolution, and harmful flow to the environment and human body.
Chiral resolution was performed by ultra-high liquid chromatography, and a chiral chromatography column of polysaccharide derivatives was used. The mobile phase consisted of methanol and deionized water. The separation of the enantiomer of butylfluoride was achieved through gradient elution and isometric elution.
The operation conditions are gentle and simple, the sensitivity is high, the separation speed is fast, the separation monomer is high, and the mobile phase is green and environmentally friendly, and the shortcomings of the prior art are overcome.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of chemical synthesis, and particularly relates to a method for chiral separation of cyflumetofen enantiomers. Background Art
[0002] Enantiomers refer to isomers whose atomic structures are mirror images of each other but cannot be superimposed. Ecological environmental pollution caused by non-point source pollution of pesticides has been recognized as an extremely important and concerning environmental issue. The increase in pesticide use and inefficient application are important causes of pesticide pollution. Chiral pesticides account for 40% of total pesticides, and this proportion is gradually increasing with the continuous development of new compounds. However, most chiral pesticides are sold and used as racemates, and only 7% exist as single enantiomers. Stereoisomers of chiral compounds usually show significant differences in biological activity, toxicity and environmental behavior. In a sense, it is necessary to strengthen the evaluation of chiral pesticides at the enantiomeric level, screen for high-efficiency and low-risk forms, and promote the development of chiral pesticides.
[0003] Agricultural pest mites are one of the most difficult biological groups to control in the world. They have a strong destructive ability to economic crops such as fruit trees, vegetables, and flowers. Due to the frequent use of miticides and improper human use, mites have developed varying degrees of resistance to some existing miticides. It is urgent to develop new and efficient miticides with novel structures and unique mechanisms of action. Against this background, Otsuka Chemical Co., Ltd. of Japan has developed cyflumetofen. Cyflumetofen is a new acyl acetonitrile miticide with no cross-resistance to existing insecticides. It was first approved for registration and sale in Japan in 2007, and has now been registered and sold in Japan, Brazil, the United States, China, South Korea, the European Union and other countries and regions. It is used to control the main mites parasitic on plants in crops and flowers such as fruit trees, vegetables, and tea trees. This product is a 20% suspension. It is effective against eggs, larvae, nymphs, and adults of spider mites, especially nymphs and larvae. It can effectively control a variety of pest mites, such as citrus red spider, cotton red spider, tumor-skinned red spider, red spider, and pan-clawed spider. According to experimental comparison, the effects of cyfluthrin are superior to those of spirodiclofen and avermectin in all aspects.
[0004] The mechanism of action of cyflumetofen is to inhibit the respiration of mite mitochondria to exert its acaricidal effect. When entering the mite body, it will undergo a deesterification reaction in the body to form a metabolically active substance AB-1 with a hydroxyl structure. AB-1 specifically interferes with and inhibits protein complex II in mite mitochondria. Mitochondrial protein complex II is a key component of the electron transport chain involved in cellular respiration, and its normal function is essential for cellular energy metabolism. Due to the inhibitory effect of AB-1 on mitochondrial protein complex II, the transfer process of electrons (hydrogen) in the respiratory chain is blocked, thereby destroying the phosphorylation reaction related to it. Phosphorylation is an important process for cells to produce energy substances adenosine triphosphate (ATP). When it is destroyed, the mite body cannot obtain enough energy for normal life activities. When energy metabolism is blocked, the physiological function of the harmful mite gradually becomes disordered, eventually leading to paralysis and death. At present, there are few literature reports on the different efficacy and toxicological studies of the R and S configuration enantiomers of cyflumetofen, so it is particularly important to establish a chiral separation method for cyflumetofen enantiomers for further research.
[0005]
[0006] So far, the separation methods of cyflumetofen are supercritical fluid chromatography and ultra-high performance liquid chromatography normal phase system. Supercritical fluid chromatography uses Trefoil AMY 1 column produced by Waters Corporation. However, supercritical fluid chromatography has disadvantages such as higher equipment cost, complex mobile phase selection and control, difficulty in method development and optimization, limited detection sensitivity, and high sample pretreatment requirements. The normal phase system of ultra-high performance liquid chromatography uses a coated chiral chromatographic column AD-H column produced by Daicel Corporation. However, the storage and use conditions of the normal phase chromatographic column are relatively harsh, and the service life is relatively short. The chromatographic column needs to be replaced more frequently, which increases the analysis cost. Secondly, the mobile phase of the normal phase system is usually a non-polar or weakly polar solvent, and the types of these solvents are relatively few, and the selection range is narrow. Moreover, the volatility of non-polar solvents is generally strong, which is easy to cause volatilization loss of the mobile phase, which not only affects the composition and proportion of the mobile phase, resulting in changes in retention time, but also may cause harm to the environment and human health. In addition, the cost of using these organic solvents is relatively high. The current separation and detection of cyfluthrin enantiomers has high experimental costs, a narrow scope of application, low separation, mobile phases that are harmful to the environment and human body, and no prospect of being scaled up to a production scale. Summary of the invention
[0007] In order to overcome the technical defects of the methods for chiral separation of cyflumetofen in the prior art, the object of the present invention is to provide a method for preparing cyflumetofen by chiral separation.
[0008] The technical solution adopted by the present invention is:
[0009] A method for chiral separation of cyfluthrin enantiomers comprises the following steps:
[0010] 1) Preparing a test solution: weighing cyflumetofen and dissolving it in an organic solvent to prepare a cyflumetofen test solution;
[0011] 2) using ultra high performance liquid chromatography to separate the enantiomers of cyflumetofen in the cyflumetofen test solution, the stationary phase is a polysaccharide derivative chiral chromatographic column, the mobile phase A is an organic solvent, the mobile phase B is deionized water, within 0-5 minutes, the mobile phase B is eluted by gradient at a volume percentage of 15-5%, within 5-10 minutes, the mobile phase B is eluted by isocratic at a volume percentage of 5%, the column temperature is 35° C., and the flow rate is 0.7 mL / min;
[0012] 3) The detector is used for measurement, and the chiral monomer fraction of cyfluthrin is collected in a fraction collector.
[0013] Furthermore, in the above-mentioned method for chiral separation of cyflumetofen enantiomers, in step 1), the organic solvent is one of methanol, ethanol, acetonitrile and isopropanol.
[0014] Preferably, in step 1), the organic solvent is acetonitrile.
[0015] Furthermore, in the above-mentioned method for chiral resolution of cyflumetofen enantiomers, in step 1), the concentration of the cyflumetofen test solution is 100 ng / mL.
[0016] Furthermore, in the above-mentioned method for chiral separation of cyfluthrin enantiomers, in step 2), the organic solvent is one of methanol, ethanol, acetonitrile and isopropanol.
[0017] Furthermore, in the above-mentioned method for chiral separation of cyflumetofen enantiomers, in step 2), the polysaccharide derivative chiral chromatographic column is a polysaccharide derivative chiral chromatographic column having amylose-tris(3-chloro-5-methylcarbamate) covalently bonded to the surface of silica gel.
[0018] Preferably, in step 2), the organic solvent is methanol.
[0019] Furthermore, in the above-mentioned method for chiral resolution of cyflumetofen enantiomers, in step 3), the detector is an ultraviolet detector or a mass spectrometer detector.
[0020] Compared with the prior art, the technical solution of the present application has the following beneficial effects: the present invention adopts ultra high performance liquid chromatography and uses a reverse phase chromatography column to directly chirally separate cyfluthrin, and its mobile phase consists of methanol and deionized water. This method overcomes the shortcomings of the prior art and has the characteristics of mild and simple operating conditions, high sensitivity, fast separation speed, high purity of separated monomers, and green and environmentally friendly mobile phase. 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. DETAILED DESCRIPTION
[0024] The advantages of the present invention are further described below in conjunction with the accompanying drawings and specific embodiments.
[0025] 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.
[0026] 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.
[0027] Example 1
[0028] A method for chiral separation of cyfluthrin enantiomers comprises the following steps:
[0029] 1) Weigh a certain amount of cyflumetofen, dissolve it in acetonitrile and make up to volume to prepare a cyflumetofen test solution with a concentration of 100 ng / mL.
[0030] 2) Chiral separation using ultra-high performance liquid chromatography
[0031] The ultra-high performance liquid chromatography (UHPLC) system used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IG chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with amylose-tris (3-chloro-5-methylcarbamate) covalently bonded to the surface of silica gel. The specification is 250mm×4.6mm and the particle size is 5μm.
[0032] Mobile phase A is methanol, mobile phase B is deionized water, within 0-5 minutes, mobile phase B is gradient eluted at a volume percentage of 20-15%, and within 5-15 minutes, mobile phase B is isocratically eluted at a volume percentage of 15%. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.7mL / min; and the injection volume is 5μL.
[0033] 3) HP-UV2040 was used for determination, and the chiral monomer fraction of cyfluthrin was collected in a fraction collector.
[0034] The results are as follows Figure 1 .according to Figure 1 It can be seen that the enantiomers of cyflumetofen can be effectively separated, with retention times of 10.157 and 13.552, respectively. The response values are 11101 and 10773, and the calculated resolution is 5.71.
[0035] Example 2
[0036] A method for chiral separation of cyfluthrin enantiomers comprises the following steps:
[0037] 1) Weigh a certain amount of cyflumetofen, dissolve it in acetonitrile and make up to volume to prepare a cyflumetofen test solution with a concentration of 100 ng / mL.
[0038] 2) Chiral separation using ultra-high performance liquid chromatography
[0039] The ultra-high performance liquid chromatography (UHPLC) system used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IG chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with amylose-tris (3-chloro-5-methylcarbamate) covalently bonded to the surface of silica gel. The specification is 250mm×4.6mm and the particle size is 5μm.
[0040] Mobile phase A is methanol, mobile phase B is deionized water, within 0-5 minutes, mobile phase B is gradient eluted at a volume percentage of 15-5%, and within 5-10 minutes, mobile phase B is isocratically eluted at a volume percentage of 5%. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.7 mL / min; and the injection volume is 5 μL.
[0041] 3) HP-UV2040 was used for determination, and the chiral monomer fraction of cyfluthrin was collected in a fraction collector.
[0042] The results are as follows Figure 2 .according to Figure 2 It can be seen that the enantiomers of cyfluthrin can be effectively separated, with retention times of 7.326 and 8.552, respectively. The response values are 14052 and 9931, and the calculated resolution is 3.75.
[0043] Example 3
[0044] A method for chiral separation of cyfluthrin enantiomers comprises the following steps:
[0045] 1) Weigh a certain amount of cyflumetofen, dissolve it in acetonitrile and make up to volume to prepare a cyflumetofen test solution with a concentration of 100 ng / mL.
[0046] 2) Chiral separation using ultra-high performance liquid chromatography
[0047] The ultra-high performance liquid chromatography (UHPLC) system used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IG chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with amylose-tris (3-chloro-5-methylcarbamate) covalently bonded to the surface of silica gel. The specification is 250mm×4.6mm and the particle size is 5μm.
[0048] Mobile phase A is methanol, mobile phase B is deionized water, mobile phase B is gradient eluted at 10-0% by volume within 0-4 minutes, and mobile phase B is isocratically eluted at 0% by volume within 4-10 minutes. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.7 mL / min; and the injection volume is 5 μL.
[0049] 3) HP-UV2040 was used for determination, and the chiral monomer fraction of cyfluthrin was collected in a fraction collector.
[0050] The results are as follows Figure 3 .according to Figure 3 It can be seen that the enantiomers of cyfluthrin can be effectively separated, with retention times of 6.072 and 6.860, respectively. The response values are 9317 and 8497, and the calculated resolution is 2.63.
Claims
1. A method for chiral separation of cyfluthrin enantiomers, characterized in that: The following steps are involved: 1) Preparing a test solution: weighing cyflumetofen and dissolving it in an organic solvent to prepare a cyflumetofen test solution; 2) using ultra high performance liquid chromatography to separate the enantiomers of cyflumetofen in the cyflumetofen test solution, the stationary phase is a polysaccharide derivative chiral chromatographic column, the mobile phase A is an organic solvent, the mobile phase B is deionized water, within 0-5 minutes, the mobile phase B is eluted by gradient at a volume percentage of 15-5%, within 5-10 minutes, the mobile phase B is eluted by isocratic at a volume percentage of 5%, the column temperature is 35° C., and the flow rate is 0.7 mL / min; 3) The detector is used for measurement, and the chiral monomer fraction of cyfluthrin is collected in a fraction collector.
2. The method for chiral separation of cyfluthrin enantiomers according to claim 1, characterized in that: In step 1), the organic solvent is one of methanol, ethanol, acetonitrile and isopropanol.
3. A method for chiral separation of cyflumetofen enantiomers according to claim 2, characterized in that: In step 1), the organic solvent is acetonitrile.
4. The method for chiral separation of cyflumetofen enantiomers according to claim 1, characterized in that: In step 1), the concentration of the cyfluthrin test solution is 100 ng / mL.
5. The method for chiral separation of cyflumetofen enantiomers according to claim 1, characterized in that: In step 2), the organic solvent is one of methanol, ethanol, acetonitrile and isopropanol.
6. A method for chiral separation of cyflumetofen enantiomers according to claim 5, characterized in that: In step 2), the polysaccharide derivative chiral chromatographic column is a polysaccharide derivative chiral chromatographic column having amylose-tris(3-chloro-5-methylcarbamate) covalently bonded to the surface of silica gel.
7. A method for chiral separation of cyflumetofen enantiomers according to claim 6, characterized in that: In step 2), the organic solvent is methanol.
8. The method for chiral separation of cyflumetofen enantiomers according to claim 1, characterized in that: In step 3), the detector is an ultraviolet detector or a mass spectrometer detector.