Method for chiral resolution of metazosulfuron enantiomer

Abstract

The invention provides a method for chiral resolution of a metazosulfuron enantiomer. According to the method, metazosulfuron enantiomers are separated by adopting a high performance liquid chromatography-mass spectrometry technology, a chromatographic column is a polysaccharide derivative chiral chromatographic column, a mobile phase A is an organic solvent, a mobile phase B is pure water, the volume ratio of the mobile phase A to the mobile phase B is (40: 60)-(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 method for chiral resolution of metazosulfuron provided by the invention has the advantages of high separation degree, short analysis time and high separation speed.

Description

Technical Field

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

[0002] In recent decades, the use of chiral pesticides has become more and more common. The types of chiral pesticides registered for use in my country account for 40% of all pesticide types, and this proportion has been on an upward trend. After the 1990s, due to the impact of pesticides on the environment, it is not allowed to release ineffective forms into the environment. If the selectivity of the two optically active isomers of the racemate is different, that is, one of the isomers is inefficient or ineffective, or has different modes of action and sites of action, the pesticide administrative department tends to select only the required isomer to a large extent.

[0003] Azoxystrobin was developed by Nissan Chemical Co., Ltd. in 2004 and is a sulfonylurea herbicide. Azoxystrobin is an acetolactate synthase (ALS) inhibitor that inhibits the biosynthesis of key amino acids (valine, leucine, isoleucine), thereby blocking cell division, inhibiting plant growth, and ultimately killing weeds. It is mainly used to control weeds in rice and wheat fields, such as Amaranthus retroflexus, Echinochloa crus-galli, and Sagittaria sagittifolia.

[0004] Chiral pesticide enantiomers differ in biological activity, toxicity, environmental behavior, etc. Fully understanding the stereoselectivity of enantiomers is of great significance for the development of highly active pesticides and reduced use. At present, there are no reports on the separation method of the R and S configuration enantiomers of pyrazosulfuron-methyl, as well as the biological activity, toxicity and environmental risks of the R and S configuration enantiomers of pyrazosulfuron-methyl. Therefore, it is particularly important to establish a chiral separation method for pyrazosulfuron-methyl enantiomers for further research.

[0005] Summary of the invention

[0006] The invention provides a method for chiral separation of azine-pyrazosulfuron-methyl, and uses high performance liquid chromatography-mass spectrometry technology to achieve rapid separation of the R and S configurations of azine-pyrazosulfuron-methyl.

[0007] The technical solution adopted by the present invention is: a method for chiral separation of pyrazosulfuron-methyl enantiomers, comprising the following steps:

[0008] 1) Prepare a test solution: weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in an appropriate amount of organic solvent, and prepare a test solution of azine-pyrazosulfuron-methyl with a concentration of 100-500 ng / ml;

[0009] 2) Chiral separation using high performance liquid chromatography-mass spectrometry: The enantiomers of pyrazosulfuron-methyl in the pyrazosulfuron-methyl test solution were separated by high performance liquid chromatography-mass spectrometry, the stationary phase was a polysaccharide derivative chiral chromatographic column, the mobile phase A was an organic solvent, the mobile phase B was pure water, the mobile phase was calculated by volume ratio, A: B was 40:60-100:0, the column temperature was 20-35° C., and the flow rate was 0.3-0.6 ml / min;

[0010] Furthermore, in the above-mentioned method for chiral resolution of pyrazosulfuron-methyl enantiomers, in step 1), the organic solvent is selected from one of methanol and acetonitrile.

[0011] Furthermore, in the above-mentioned method for chiral resolution of azine-pyrazosulfuron-methyl enantiomers, in step 1), the concentration of the azine-pyrazosulfuron-methyl test solution is 100 ng / ml.

[0012] Furthermore, in the above-mentioned method for chiral separation of pyrazosulfuron-methyl 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.

[0013] Furthermore, in the above-mentioned method for chiral resolution of pyrazosulfuron-methyl enantiomers, in step 2), the organic solvent is selected from one of methanol and acetonitrile.

[0014] Furthermore, in the above-mentioned method for chiral separation of pyrazosulfuron-methyl enantiomers, mobile phase A is acetonitrile, mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-80:20, gradient elution, column temperature is 35°C, and flow rate is 0.3-0.6 ml / min.

[0015] Furthermore, in the above-mentioned method for chiral separation of pyrazosulfuron-methyl enantiomers, mobile phase A is acetonitrile, mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-80:20, gradient elution, column temperature is 35°C, and the flow rate is 0.6 ml / min.

[0016] Furthermore, in the above-mentioned method for chiral resolution of pyrazosulfuron-methyl enantiomers, in step 4), the detector is a mass spectrometer detector.

[0017] 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 pyrazosulfuron-methyl, and its mobile phase consists of acetonitrile and pure water. The method is simple to operate, has mild conditions, and takes a short separation time. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1It is the chromatographic separation pattern of Example 1.

[0019] Figure 2 It is the chromatographic separation pattern of Example 2.

[0020] Figure 3 It is the chromatographic separation pattern of Example 3.

[0021] Figure 4 It is the chromatographic separation pattern of Example 4.

[0022] Figure 5 It is the chromatographic separation pattern of Example 5.

[0023] Figure 6 It is the chromatographic separation pattern of Example 6. 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 resolution of pyrazosulfuron-methyl enantiomers comprises the following steps:

[0029] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0030] 2) Chiral separation using high performance liquid chromatography-mass spectrometry

[0031] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IG chiral chromatographic column is a chiral chromatographic column with covalent bonds on the silica surface and polysaccharide derivatives of amylose-tris (3-chloro-5-methylcarbamate), with specifications of 250mm×4.6mm and a particle size of 5μm.

[0032] The mobile phase A is acetonitrile and the mobile phase B is pure water. The mobile phase is calculated by volume ratio, A:B is 40:60-100:0, and gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6 mL / min; and the injection volume is 20 μL.

[0033] 3) Measured using a mass spectrometer.

[0034] The results are as follows Figure 1 .Depend on Figure 1 It can be seen that the enantiomers of pyrazosulfuron-methyl were not effectively separated.

[0035] Example 2

[0036] A method for chiral resolution of pyrazosulfuron-methyl enantiomers comprises the following steps:

[0037] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0038] 2) Chiral separation using HPLC-MS

[0039] The high performance liquid chromatography mass spectrometer 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] The mobile phase A is acetonitrile and the mobile phase B is pure water. The mobile phase is calculated by volume ratio, A:B is 60:40-100:0, and gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6 mL / min; and the injection volume is 20 μL.

[0041] 3) Measured using a mass spectrometer.

[0042] The results are as follows Figure 2 .according to Figure 2 It can be seen that the enantiomers of pyrazosulfuron-methyl were not effectively separated.

[0043] Example 3

[0044] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0045] 2) Chiral separation using HPLC-MS

[0046] The HPLC-MS / MS instrument used was an Agilent system, and the stationary phase was a polysaccharide derivative chiral chromatographic column. IA chiral chromatographic column is a polysaccharide derivative chiral chromatographic column with straight-chain starch-tris(3,5-dimethylphenylcarbamate) covalently bonded to the silica surface. The specification is 250mm×4.6mm and the particle size is 5μm.

[0047] The mobile phase A is acetonitrile, the mobile phase B is 5 mM ammonium acetate water, the mobile phase is calculated by volume ratio, A:B is 40:60-70:30, and the gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6 mL / min; and the injection volume is 20 μL.

[0048] 3) Measured using a mass spectrometer.

[0049] The results are as follows Figure 3 .according to Figure 3 It can be seen that the chromatographic peaks of the enantiomers of pyrazosulfuron-methyl cannot be effectively separated.

[0050] Example 4

[0051] A method for chiral resolution of pyrazosulfuron-methyl enantiomers comprises the following steps:

[0052] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0053] 2) Chiral separation using HPLC-MS

[0054] The HPLC-MS / MS instrument 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.

[0055] The mobile phase A is acetonitrile, the mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60, and the elution is isocratic. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6 mL / min; and the injection volume is 20 μL.

[0056] 3) Measured using a mass spectrometer.

[0057] The results are as follows Figure 4 .Depend on Figure 4 It can be seen that the enantiomers of pyrazosulfuron-methyl were not effectively separated.

[0058] Example 5

[0059] A method for chiral resolution of pyrazosulfuron-methyl enantiomers comprises the following steps:

[0060] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0061] 2) Chiral separation using HPLC-MS

[0062] The HPLC-MS / MS instrument 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.

[0063] The mobile phase A is acetonitrile, the mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 50:50, and the elution is isocratic. 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.

[0064] 3) Measured using a mass spectrometer.

[0065] The results are as follows Figure 5 .Depend on Figure 5 It can be seen that the enantiomers of pyrazosulfuron-methyl were not effectively separated.

[0066] Example 6

[0067] A method for chiral resolution of pyrazosulfuron-methyl enantiomers comprises the following steps:

[0068] 1) Weigh a certain amount of azine-pyrazosulfuron-methyl, dissolve it in acetonitrile and make up to volume to prepare a 100 ng / ml azine-pyrazosulfuron-methyl test solution.

[0069] 2) Chiral separation using HPLC-MS

[0070] The HPLC-MS / MS instrument 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.

[0071] The mobile phase A is acetonitrile and the mobile phase B is pure water. The mobile phase is calculated by volume ratio, A:B is 40:60-80:20, and gradient elution is performed. The column temperature of the polysaccharide derivative chiral chromatographic column is 35°C; the flow rate is 0.6 mL / min; and the injection volume is 20 μL.

[0072] 3) Measured using a mass spectrometer.

[0073] The results are as follows Figure 6 .according to Figure 6 It can be seen that the chromatographic peaks of the enantiomers of pyrazosulfuron-methyl are effectively separated, and the calculated separation degree is R=1.81, achieving effective baseline separation.

[0074] In summary, the present application adopts high performance liquid chromatography-mass spectrometry technology. Only under specific chromatographic conditions, that is, mobile phase A is acetonitrile, mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-80:20, gradient elution, column temperature is 35°C, flow rate is 0.6 ml / min, can effective chiral separation of azine pyrazosulfuron-methyl be achieved, the chromatographic peaks of azine pyrazosulfuron-methyl enantiomers are effectively separated, the calculated separation degree reaches more than 1.5, and effective baseline separation is achieved.

[0075] The separated pyrazosulfuron-methyl 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 enantiomers of pyrazosulfuron-methyl, It is characterized in that The following steps are involved: 1) Prepare a test solution: weigh a certain amount of oxazolidinone pyrazosulfuron-methyl and dissolve it in an organic solvent to prepare a oxazolidinone pyrazosulfuron-methyl test solution; 2) using high performance liquid chromatography-mass spectrometry to separate the enantiomers of pyrazosulfuron-methyl in the pyrazosulfuron-methyl 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 pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-100:0, 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 resolution of enantiomers of pyrazosulfuron-methyl as claimed in claim 1, It is characterized in that In step 1), the organic solvent is selected from one of methanol and acetonitrile.

3. A method for chiral resolution of enantiomers of pyrazosulfuron-methyl as claimed in claim 1, It is characterized in that In step 1), the concentration of the pyrazosulfuron-methyl test solution is 100 ng / ml.

4. A method for chiral resolution of enantiomers of pyrazosulfuron-methyl as claimed in claim 1, 2 or 3, It is 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.

5. A method for chiral resolution of enantiomers of pyrazosulfuron-methyl as claimed in claim 4, It is characterized in that In step 2), the organic solvent is selected from one of methanol and acetonitrile.

6. A method for chiral resolution of enantiomers of pyrazosulfuron-methyl as claimed in claim 5, It is characterized in that Mobile phase A is acetonitrile, mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-80:20, gradient elution, column temperature is 35°C, and flow rate is 0.3-0.6 ml / min.

7. A method for chiral resolution of pyrazosulfuron-methyl enantiomers as claimed in claim 6, It is characterized in that Mobile phase A is acetonitrile, mobile phase B is pure water, the mobile phase is calculated by volume ratio, A:B is 40:60-80:20, the column temperature is 35° C., and the flow rate is 0.6 ml / min.

8. A method for chiral resolution of pyrazosulfuron-methyl enantiomers according to claim 1, It is characterized in that In step 4), the detector is a mass spectrometer detector.

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