CN113607836B - Analysis method for content of indoxacarb key intermediate - Google Patents
Analysis method for content of indoxacarb key intermediate Download PDFInfo
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- CN113607836B CN113607836B CN202110836227.XA CN202110836227A CN113607836B CN 113607836 B CN113607836 B CN 113607836B CN 202110836227 A CN202110836227 A CN 202110836227A CN 113607836 B CN113607836 B CN 113607836B
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- 239000005907 Indoxacarb Substances 0.000 title claims abstract description 21
- VBCVPMMZEGZULK-NRFANRHFSA-N indoxacarb Chemical compound C([C@@]1(OC2)C(=O)OC)C3=CC(Cl)=CC=C3C1=NN2C(=O)N(C(=O)OC)C1=CC=C(OC(F)(F)F)C=C1 VBCVPMMZEGZULK-NRFANRHFSA-N 0.000 title claims abstract description 21
- 238000004458 analytical method Methods 0.000 title claims abstract description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 96
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 57
- 239000000126 substance Substances 0.000 claims abstract description 39
- 229960000583 acetic acid Drugs 0.000 claims abstract description 26
- 238000001514 detection method Methods 0.000 claims abstract description 21
- 239000012362 glacial acetic acid Substances 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 19
- 239000007864 aqueous solution Substances 0.000 claims abstract description 17
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 12
- 238000004364 calculation method Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 62
- 239000012488 sample solution Substances 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 21
- 239000012086 standard solution Substances 0.000 claims description 19
- 238000002347 injection Methods 0.000 claims description 12
- 239000007924 injection Substances 0.000 claims description 12
- 238000010812 external standard method Methods 0.000 claims description 5
- 238000005070 sampling Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 239000000575 pesticide Substances 0.000 abstract description 6
- 239000012535 impurity Substances 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 3
- 230000014759 maintenance of location Effects 0.000 abstract description 3
- 238000003908 quality control method Methods 0.000 abstract description 3
- 238000000926 separation method Methods 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract description 2
- 239000013067 intermediate product Substances 0.000 abstract description 2
- 239000012071 phase Substances 0.000 description 30
- 238000012360 testing method Methods 0.000 description 16
- 238000005303 weighing Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- 238000011084 recovery Methods 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 6
- 238000007865 diluting Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000004090 dissolution Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- YFVYPTZIEXOAIT-UHFFFAOYSA-N 2-o-benzyl 4a-o-methyl 7-chloro-3,5-dihydroindeno[1,2-e][1,3,4]oxadiazine-2,4a-dicarboxylate Chemical compound C1OC2(C(=O)OC)CC3=CC(Cl)=CC=C3C2=NN1C(=O)OCC1=CC=CC=C1 YFVYPTZIEXOAIT-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- -1 7-chloroindeno [1 Chemical compound 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000004973 liquid crystal related substance Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- QMEQBOSUJUOXMX-UHFFFAOYSA-N 2h-oxadiazine Chemical compound N1OC=CC=N1 QMEQBOSUJUOXMX-UHFFFAOYSA-N 0.000 description 1
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 1
- 241000426497 Chilo suppressalis Species 0.000 description 1
- 241000008892 Cnaphalocrocis patnalis Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000256259 Noctuidae Species 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241000500437 Plutella xylostella Species 0.000 description 1
- VQXSOUPNOZTNAI-UHFFFAOYSA-N Pyrethrin I Natural products CC(=CC1CC1C(=O)OC2CC(=O)C(=C2C)CC=C/C=C)C VQXSOUPNOZTNAI-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- HYJYGLGUBUDSLJ-UHFFFAOYSA-N pyrethrin Natural products CCC(=O)OC1CC(=C)C2CC3OC3(C)C2C2OC(=O)C(=C)C12 HYJYGLGUBUDSLJ-UHFFFAOYSA-N 0.000 description 1
- VJFUPGQZSXIULQ-XIGJTORUSA-N pyrethrin II Chemical compound CC1(C)[C@H](/C=C(\C)C(=O)OC)[C@H]1C(=O)O[C@@H]1C(C)=C(C\C=C/C=C)C(=O)C1 VJFUPGQZSXIULQ-XIGJTORUSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
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Abstract
The invention relates to the technical field of chemical analysis, in particular to an analysis method for the content of a key intermediate of indoxacarb, which adopts high performance liquid chromatography to detect the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, wherein a chromatographic column is an ODS-C18 reversed phase chromatographic column, the temperature of the chromatographic column is 30-50 ℃, a mobile phase is a mixed system of methanol and glacial acetic acid aqueous solution, the detection wavelength is 230-300 nm, the separation of impurities and main peaks is complete, the shape of the chromatographic peak is good, the retention time is stable, the integral calculation result is accurate, the repeatability is good, and the reliability of the obtained result is high, and the method is particularly suitable for quality control of an intermediate product of a pesticide original drug.
Description
Technical Field
The invention relates to the technical field of chemical analysis, in particular to an analysis method for the content of a key intermediate of indoxacarb.
Background
Indoxacarb is a novel, efficient and low-toxicity oxadiazine pesticide developed by DuPont in the United states, has double effects of contact killing and stomach toxicity, and can effectively solve resistant pests. Has no cross resistance with other pesticides such as pyrethrin, organophosphorus and carbamate, and can well solve the problems of rice leaf rollers, chilo suppressalis and resistant plutella xylostella which are difficult to prevent in the current market. In addition, indoxacarb has extremely wide insecticidal spectrum, one drug is multi-proof, and has good inhibition effect on plant bug and the like while preventing and controlling noctuid pests, so that the indoxacarb is a good comprehensive treatment tool and can well solve the problems of residue and environmental pollution after the prior various pesticides are mixed. Indoxacarb has broad market prospect due to the unique action mechanism.
The analysis method for the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester is an intermediate of indoxacarb, and the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester is still in a blank state, and the content accuracy of the 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2-benzyl ester directly influences the accuracy of downstream products of the 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2-benzyl ester, and further influences the content accuracy of indoxacarb. The analysis method for stably and accurately detecting the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester is found to have important effect and practical significance for guaranteeing the quality of indoxacarb products.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides the analysis method for the content of the key intermediate of indoxacarb, which fills the technical blank, has the advantages of strong specificity, good precision, high recovery rate, high reliability and good repeatability, and is particularly suitable for quality control of pesticide raw medicine products.
The technical scheme of the invention is as follows:
the method for analyzing the content of the key intermediate of indoxacarb comprises the steps of adopting a high performance liquid chromatography analysis method to analyze and determine the content of the key intermediate of indoxacarb, namely 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, and specifically comprises the following steps:
(1) Respectively dissolving a standard substance and a sample to be tested by using methanol as a solvent to prepare a standard substance solution and a sample solution to be tested, wherein the concentration ranges of the standard substance solution and the sample solution to be tested are 0.5-1mg/ml;
(2) Setting the detection wavelength of high performance liquid chromatography to 230-300 nm, sequentially sampling according to the sequence of a standard substance, a sample to be detected and the standard substance after the instrument baseline is stable, and respectively calculating the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester of the standard substance solution and the sample to be detected;
(3) 7-chloroindeno [1,2-E in sample solution to be tested according to external standard method formula][1,3,4]Mass fraction X of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid 4A-methyl ester-2-benzyl ester 1 The calculation is performed according to the following specific formula:
in the method, in the process of the invention,
A 1 7-chloroindeno [1,2-E ] in a Standard solution][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
A 2 7-chloroindeno [1,2-E ] in the sample solution to be tested][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
m 1 the quality of the standard substance is that of the standard substance,
m 2 the mass of the sample to be measured,
P 1 7-chloroindeno [1,2-E ] in standard][1,3,4]Mass fraction of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl ester-2-benzyl ester;
wherein, the high performance liquid chromatography conditions include:
the chromatographic column is ODS-C18 reversed phase chromatographic column with temperature of 30deg.C-50deg.C, and mobile phase is mixed system of methanol and glacial acetic acid water solution. Through a large number of experiments, the inventor discovers that the choice of the mobile phase adopts the methanol and glacial acetic acid aqueous solution, so that the target substance and impurities can be effectively separated, and the economic applicability is superior to other solvents.
Preferably, the detection wavelength of the high performance liquid chromatography is 280nm. The 280nm detection wavelength is the most stable ultraviolet absorption wavelength for 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester, and the measurement error is small at the wavelength.
Preferably, the column length of the chromatographic column is 150mm, the column inner diameter is 4.5-5mm, more preferably 4.6mm, and the column particle size is 3.5 μm.
Preferably, the temperature of the column is 40 ℃.
Preferably, the volume fraction of the aqueous glacial acetic acid solution in the mixed system is 20-50%, more preferably 25%.
Preferably, the high performance liquid chromatography conditions further include: the sample volume per sample injection was 5. Mu.L and the flow rate of the mobile phase was 1ml/min.
The invention has the advantages that,
(1) The invention adopts high performance liquid chromatography to detect the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, and comprises a chromatographic column which is an ODS-C18 reversed phase chromatographic column, the temperature of the chromatographic column is 30-50 ℃, the mobile phase is a mixed system of methanol and glacial acetic acid aqueous solution, the detection wavelength is 230-300 nm, the content can be accurately measured, and the technical blank in the corresponding field in the prior art is filled;
(2) The mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester is stable and accurate, the detection precision and stability are improved, the complete separation of impurities and main peaks can be realized, the chromatographic peak shape is good, the retention time is stable, the integral calculation result is accurate, the repeatability is good, the reliability of the obtained result is high, the method is particularly suitable for quality control of pesticide raw material intermediate products, and the method has important effect and practical significance in guaranteeing the quality of final products; meanwhile, the obtained result is more accurate and timely, and powerful data support is provided for the production of indoxacarb.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the description of the embodiments or the prior art will be briefly described below, and it will be obvious to those skilled in the art that other drawings can be obtained from these drawings without inventive effort.
FIG. 1 is a chromatogram of the standard solution in example 1;
FIG. 2 is a chromatogram of the sample solution to be tested in example 1;
FIG. 3 is a chromatogram of the standard solution in example 2;
FIG. 4 is a chromatogram of the sample solution to be tested in example 2;
FIG. 5 is a chromatogram of the standard solution in example 3;
FIG. 6 is a chromatogram of the sample solution to be tested in example 3;
FIG. 7 is a graph showing the linear relationship in test example 4;
FIG. 8 is a graph showing a linear relationship between the wavelength of 260nm in comparative example 1;
FIG. 9 is a graph showing a linear relationship between the wavelength of 300nm in comparative example 1;
FIG. 10 shows the ratio of the mobile phases in comparative example 2 as methanol: chromatogram at 0.8% aqueous acetic acid = 90:10;
FIG. 11 shows the ratio of the mobile phases in comparative example 2 as methanol: chromatogram at 0.8% aqueous acetic acid = 60:40;
wherein, the liquid crystal display device comprises a liquid crystal display device,
in the chromatograms of fig. 1 to 6, the abscissa represents time and the ordinate represents absorbance;
in the linear graphs of FIGS. 8-9, the abscissa represents the standard solution concentration (in g/L) and the ordinate represents the peak area;
in the chromatograms of fig. 10 to 11, the abscissa represents time and the ordinate represents absorbance.
Detailed Description
In order to make the technical solution of the present invention better understood by those skilled in the art, the technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
Example 1
210 batches in production gave 1960 kg of a solid sample of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl 2-benzyl ester, and the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl 2-benzyl ester in the product was analyzed, the method comprising the steps of:
(1) Accurately weighing 7-chloroindeno [1,2-E][1,3,4]The oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester standard is placed in a 100ml volumetric flask, 90ml of methanol is added, after shaking and dissolution, the standard solution is obtained by diluting the standard solution to the scale with the methanol for standby, and 7-chloroindeno [1,2-E in the standard solution][1,3,4]Mass fraction P of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl 2-benzyl ester 1 ＝98.6％；
Accurately weighing a sample to be measured, placing the sample into a 100ml volumetric flask, adding 90ml of methanol, oscillating for dissolution, and diluting the solution to a scale with the methanol to obtain a sample solution to be measured for later use;
(2) Adopting a high performance liquid chromatograph, wherein the chromatographic column is an ODS-C18 chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%; the sample volume of each sample injection is 5 mu L, and the flow rate of the mobile phase is 1ml/min; the detection wavelength is set to 280nm;
(3) After instrument baseline stabilization, sample injection is sequentially carried out according to the sequence of a standard substance, a sample to be detected and the standard substance, and the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester of the standard substance solution and the sample solution to be detected is calculated respectively, wherein the detection data are shown in the following table 1:
table 1 example 1 test results
(4) According to an external standard method formula, calculating the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in a sample to be detected, and calculating to obtain the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester in the sample to be detected, wherein the mass fraction is 98.6%.
The specific formula is as follows,
in the method, in the process of the invention,
A 1 7-chloroindeno [1,2-E ] in a Standard solution][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
A 2 7-chloroindeno [1,2-E ] in the sample solution to be tested][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
m 1 the quality of the standard substance is that of the standard substance,
m 2 the mass of the sample to be measured,
P 1 7-chloroindeno [1,2-E ] in standard][1,3,4]The mass fraction of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
X 1 7-chloroindeno [1,2-E ] in sample to be tested][1,3,4]Mass fraction of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl ester-2-benzyl ester;
FIG. 1 is a chromatogram of a standard solution of this example showing the peak at the 7.923min position as characteristic of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester; FIG. 2 is a chromatogram of a sample solution to be tested measured in this example, showing a peak at a 7.924min position corresponding to 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl 2-benzyl ester.
Example 2
A small sample of 20201208 was prepared to give 98.4g of solid and the product was analyzed for 2-benzyl 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester content, the method comprising the steps of:
(1) Accurately weighing 7-chloroindeno [1,2-E][1,3,4]Oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-Placing methyl ester 2-benzyl ester standard in 100ml volumetric flask, adding 90ml methanol, shaking for dissolving, diluting with methanol to scale to obtain standard solution, and collecting 7-chloroindeno [1,2-E in standard solution][1,3,4]Mass fraction P of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl 2-benzyl ester 1 ＝98.6％；
Accurately weighing a sample to be measured, placing the sample into a 100ml volumetric flask, adding 90ml of methanol, oscillating for dissolution, and diluting the solution to a scale with the methanol to obtain a sample solution to be measured for later use;
(2) Adopting a high performance liquid chromatograph, wherein the chromatographic column is an ODS-C18 reversed phase chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%; the sample volume of each sample injection is 5 mu L, and the flow rate of the mobile phase is 1ml/min; the detection wavelength is set to 280nm;
(3) After instrument baseline stabilization, sample injection is sequentially carried out according to the sequence of a standard substance, a sample to be detected and the standard substance, and the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester of the standard substance solution and the sample solution to be detected is calculated respectively, wherein the detection data are shown in the following table 2:
table 2 example 2 test results
(4) According to an external standard method formula, the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in a sample solution to be detected is calculated, and the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in the sample solution to be detected is calculated to be 98.8%.
FIG. 3 is a chromatogram of the standard solution of this example, in which the peak at 7.368min position is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, and FIG. 4 is a chromatogram of the sample solution to be tested of this example, in which the peak at 7.342min position is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester.
Example 3
In batch 203, a solid sample of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester was obtained as 1820 kg, and the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester was analyzed, which comprises the steps of:
(1) Accurately weighing 7-chloroindeno [1,2-E][1,3,4]The oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester standard is placed in a 100ml volumetric flask, 90ml of methanol is added, after shaking and dissolution, the standard solution is obtained by diluting the standard solution to the scale with the methanol for standby, and 7-chloroindeno [1,2-E ] in the standard solution][1,3,4]Mass fraction P of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl 2-benzyl ester 1 ＝98.6％；
Accurately weighing a sample to be measured, placing the sample into a 100ml volumetric flask, adding 100ml of methanol, oscillating for dissolution, and diluting the sample to a scale with the methanol to obtain a sample solution to be measured for later use;
(2) Adopting a high performance liquid chromatograph with a diode array detector, wherein the chromatographic column is an ODS-C18 chiral chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%; the sample volume of each sample injection is 5 mu L, and the flow rate of the mobile phase is 1ml/min; the detection wavelength is set to 280nm;
(3) After instrument baseline stabilization, sample injection is sequentially carried out according to the sequence of a standard substance, a sample to be detected, the standard substance and the sample to be detected, the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester of the standard substance solution and the sample to be detected is calculated respectively, and detection data are shown in the following table 3:
TABLE 3 example 3 detection results
(4) According to an external standard method formula, the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in a sample to be detected is calculated, and the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in the sample to be detected is calculated to be 98.4%.
FIG. 5 is a chromatogram of the standard solution of this example, in which the peak at 7.263min position is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester, and FIG. 6 is a chromatogram of the sample solution to be tested of this example, in which the peak at 7.267min position is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester.
Test example 1 stability test
Taking the standard solution in the example 1 as an investigation object, analyzing at room temperature at intervals for 5 times, and recording peak areas at the same time, wherein the analysis conditions comprise: adopting a high performance liquid chromatograph with a diode array detector, wherein the chromatographic column is an ODS-C18 reversed phase chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%; the sample volume of each sample introduction was 5. Mu.L, the flow rate of the mobile phase was 1ml/min, and the detection wavelength was 280nm. The results are shown in Table 4 below, where retention time of chromatographic peaks is stable, and comparison of peak areas yields, RSD less than 1%, indicating good stability of the assay method of the present invention.
TABLE 4 stability test results
Test example 2 precision test
Taking 20201208 batches of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester as an investigation object, weighing a standard substance and five parallel samples to be detected, respectively sampling according to the sequence of a standard substance solution, a sample solution to be detected and a standard substance solution, and calculating the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester in the five parallel samples to be detected;
the analysis conditions were: adopting high performance liquid chromatograph and diode array detector island body fluid phase chromatograph, wherein the chromatographic column is ODS-C18 reversed phase chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 μm; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid water, and the volume fraction of the glacial acetic acid water in the mixed system is 25%; the sample volume of each sample injection is 5 mu L, and the flow rate of the mobile phase is 1ml/min; the detection wavelength was set to 280nm.
As a result, as shown in Table 5 below, the mass fraction of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester was compared, and RSD was less than 1%, indicating that the analytical method of the present invention was excellent in precision.
TABLE 5 precision test results
Test example 3 recovery test
Sample 20201208 batches of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester were taken as small sample of example 2, the standard samples were weighed, five samples of different masses were simultaneously weighed, the standard samples of different masses were added, and the recovery rate of the added standard samples was calculated by sample introduction.
The analysis conditions were: adopting a high performance liquid chromatograph with a diode array detector, wherein the chromatographic column is an ODS-C18 chiral chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%, the sample volume of each sample injection is 5 mu L, and the flow rate of the mobile phase is 1ml/min; the detection wavelength of the high performance liquid chromatography was set to 280nm. And after the instrument baseline is stabilized, sampling is sequentially carried out according to the sequence of a standard substance, a sample to be detected, the standard substance and the sample to be detected, and the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester of the standard substance solution and the sample to be detected is calculated respectively, wherein the result is shown in the following table 6, the recovery rate is between 98% and 101%, and the average recovery rate is 99.38%, so that the experimental recovery rate meets the requirement.
TABLE 6 recovery test results
Test example 4 Linear test
Weighing a series of standard substances with different qualities, placing the standard substances into a 100ml volumetric flask, dissolving the standard substances with methanol, fixing the volume, and examining the relation between the peak area and the solution concentration after sample injection;
the analysis conditions included: adopting a high performance liquid chromatograph with a diode array detector, wherein the chromatographic column is an ODS-C18 reversed phase chromatographic column, the column length of the chromatographic column is 150mm, the column inner diameter is 4.6mm, and the column granularity is 3.5 mu m; the temperature of the chromatographic column is 40 ℃; the mobile phase is a mixed system of methanol and 0.8% glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%; the sample volume of each sample introduction was 5. Mu.L, the flow rate of the mobile phase was 1ml/min, and the detection wavelength was 280nm.
The results are shown in the following table 7 and fig. 7, and the correlation coefficient is 1, which indicates that the analysis method provided by the invention meets the requirement of linearity.
TABLE 7 Linear test results
As is clear from the above test examples 1 to 4, the method for analyzing the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester provided by the invention has high accuracy and good operability, and can be widely applied to the analysis and detection of the content of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester.
Comparative example 1 comparison of Linear detection results at different detection wavelengths
During the test, wavelengths of 260nm and 300nm were selected for linear verification, respectively, and the results are shown in tables 8-9. It was found that under the conditions of the wavelengths of 260nm and 300nm, although the linear correlation coefficient R 2 The value of (2) is also greater than 0.99, but the intercept is large, see fig. 8 and 9, so that the error is large if the sample analysis is performed by the single standard comparison method in the present invention.
TABLE 8 results of Linear experiments at wavelengths of 260nm
TABLE 9 results of Linear experiments at wavelengths of 300nm
Comparative example 2 case of chromatograms at different mobile phase ratios
Taking a small test sample, and taking methanol: 0.8% aqueous acetic acid = 90:10 The mixed system (or 60:40) is a mobile phase, the flow rate is 1mL/min, the sample volume of each sample injection is 5 mu L, and the peak condition of the chromatogram is inspected according to different mobile phase proportions.
In methanol: 0.8% aqueous acetic acid = 90: at 10, some small impurities have poor separation from the target, which affects the calculation of peak area and thus content, as shown in fig. 10.
In methanol: 0.8% aqueous acetic acid = 60:40, the time to peak is prolonged and the peak pattern is widened after the ratio of the organic phase is reduced, see fig. 11.
Although the present invention has been described in detail by way of preferred embodiments with reference to the accompanying drawings, the present invention is not limited thereto. Various equivalent modifications and substitutions may be made in the embodiments of the present invention by those skilled in the art without departing from the spirit and substance of the present invention, and it is intended that any person skilled in the art who is within the scope of the present invention shall fall within the technical scope of the present invention, and it is easy to think that the modifications and substitutions shall fall within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (5)
1. The analysis method for the content of the key intermediate of indoxacarb is characterized in that the key intermediate of indoxacarb is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, and the method adopts a high performance liquid chromatography analysis method and comprises the following specific steps:
(1) Respectively dissolving a standard substance and a sample to be tested by using methanol as a solvent to prepare a standard substance solution and a sample solution to be tested;
(2) Setting the detection wavelength of high performance liquid chromatography to 280nm, sequentially sampling according to the sequence of a standard substance, a sample to be detected and the standard substance after the baseline of an instrument is stable, and respectively calculating the peak area average value of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester in the standard substance solution and the sample to be detected;
(3) 7-chloroindeno [1,2-E in sample to be tested according to external standard method formula][1,3,4]Mass fraction X of oxadiazine-2, 4A (3 h,5 h) -dicarboxylic acid-4A-methyl ester-2-benzyl ester 1 The calculation is performed according to the following specific formula:
in the method, in the process of the invention,
A 1 7-chloroindeno [1,2-E ] in a Standard solution][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
A 2 7-chloroindeno [1,2-E ] in the sample solution to be tested][1,3,4]Average value of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester peak area,
m 1 the quality of the standard substance is that of the standard substance,
m 2 the mass of the sample to be measured,
P 1 7-chloroindeno [1,2-E ] in standard][1,3,4]The mass fraction of oxadiazine-2, 4A (3H, 5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
wherein, the high performance liquid chromatography conditions include:
the chromatographic column is an ODS-C18 reversed phase chromatographic column, the temperature of the chromatographic column is 30-50 ℃, the mobile phase is a mixed system of methanol and glacial acetic acid aqueous solution, and the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%.
2. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, wherein the concentration range of the standard substance solution and the solution to be measured in the step (1) is 0.5-1mg/ml.
3. The method for analyzing the content of the key intermediate of indoxacarb according to claim 1, wherein the column length of the chromatographic column is 150mm, the inner diameter of the column is 4.5-5mm, and the granularity of the column is 3.5 μm.
4. The method for analyzing the content of the key intermediate of indoxacarb according to claim 1, wherein the temperature of the chromatographic column is 40 ℃.
5. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, wherein the high performance liquid chromatography conditions further comprise: the sample volume per sample injection was 5. Mu.L and the flow rate of the mobile phase was 1ml/min.
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