CN113607836A - Method for analyzing content of indoxacarb key intermediate - Google Patents
Method for analyzing content of indoxacarb key intermediate Download PDFInfo
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- CN113607836A CN113607836A CN202110836227.XA CN202110836227A CN113607836A CN 113607836 A CN113607836 A CN 113607836A CN 202110836227 A CN202110836227 A CN 202110836227A CN 113607836 A CN113607836 A CN 113607836A
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- 239000005907 Indoxacarb Substances 0.000 title claims abstract description 23
- 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 23
- 238000000034 method Methods 0.000 title claims description 19
- 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 59
- 239000000126 substance Substances 0.000 claims abstract description 40
- 229960000583 acetic acid Drugs 0.000 claims abstract description 27
- 239000012362 glacial acetic acid Substances 0.000 claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 21
- 238000004458 analytical method Methods 0.000 claims abstract description 20
- 239000007864 aqueous solution Substances 0.000 claims abstract description 17
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 13
- 238000004364 calculation method Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 52
- 239000000243 solution Substances 0.000 claims description 27
- 239000012488 sample solution Substances 0.000 claims description 22
- 239000012086 standard solution Substances 0.000 claims description 13
- 238000010812 external standard method Methods 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 2
- 239000000575 pesticide Substances 0.000 abstract description 5
- 239000012535 impurity Substances 0.000 abstract description 4
- 230000014759 maintenance of location Effects 0.000 abstract description 3
- 238000003908 quality control method Methods 0.000 abstract description 3
- 239000013067 intermediate product Substances 0.000 abstract description 2
- 239000012071 phase Substances 0.000 description 29
- 238000012360 testing method Methods 0.000 description 19
- 239000007788 liquid Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 238000011084 recovery Methods 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 6
- 238000007865 diluting Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 231100000572 poisoning Toxicity 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000012795 verification 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
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 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
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 238000003912 environmental pollution Methods 0.000 description 1
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- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000002265 prevention Effects 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
- 238000001228 spectrum Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
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- 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
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- 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
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- 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
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Abstract
The invention relates to the technical field of chemical analysis, in particular to an indoxacarb key intermediate content analysis method, which adopts high performance liquid chromatography to detect the content of 7-chloro-indeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, 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, impurities and main peaks can be completely separated, the chromatographic peak shape is good, the retention time is stable, the integral calculation result is accurate, the repeatability is good, and the obtained result is high in reliability and is particularly suitable for quality control of pesticide intermediate products.
Description
Technical Field
The invention relates to the technical field of chemical analysis, in particular to an analytical method for the content of an indoxacarb key intermediate.
Background
Indoxacarb is a novel, high-efficiency and low-toxicity oxadiazine pesticide developed by DuPont, has double effects of contact poisoning and stomach poisoning, and can effectively solve resistant pests. Has no cross resistance with other insecticides such as pyrethrin, organic phosphorus and carbamate, and can well solve the problems of the rice leaf roller, chilo suppressalis and resistant plutella xylostella which are difficult to prevent in the current market. In addition, indoxacarb has extremely wide insecticidal spectrum, has more prevention effect when being used for preventing and treating noctuid pests, and has good inhibition effect on plant bug and the like, thereby being a good comprehensive treatment tool and well solving the problems of residue and environmental pollution after various pesticides are mixed at present. The indoxacarb has a wide market prospect due to the unique action mechanism.
The 7-chlorin indeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester is an intermediate of indoxacarb, the existing analytical method for accurately detecting the content of the 7-chlorin indeno [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 directly influences the accuracy of downstream products, and further influences the accuracy of the content of the indoxacarb. An analytical 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, which has important function and practical significance for ensuring the quality of indoxacarb products.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an indoxacarb key intermediate content analysis method, which fills up the technical blank, has strong specificity, good precision, high recovery rate, high reliability and good repeatability, and is particularly suitable for quality control of pesticide raw drug products.
The technical scheme of the invention is as follows:
an analytical method for the content of an indoxacarb key intermediate, namely, the content of 7-chloro-indeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester of the indoxacarb key intermediate is analyzed and determined by adopting a high performance liquid chromatography analytical method, which comprises the following specific steps:
(1) respectively dissolving a standard substance and a sample to be detected by using methanol as a solvent to prepare a standard substance solution and a sample solution to be detected, wherein the concentration ranges of the standard substance solution and the sample solution to be detected are both 0.5-1 mg/ml;
(2) setting the detection wavelength of the high performance liquid chromatography to be within 230-300 nm, sequentially injecting samples according to the sequence of a standard substance, a sample to be detected and the standard substance after the baseline of the instrument is stable, and respectively calculating the peak area average values 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;
(3) according to an external standard method formula, 7-chloro-indeno [1,2-E ] in a sample solution to be detected][1,3,4]Weight fraction X of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid 4A-methyl ester-2-benzyl ester1The calculation is carried out according to the following specific formula:
in the formula (I), the compound is shown in the specification,
A1-7-Chloroindeno [1,2-E in Standard solution][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
A2-7-Chloroindeno [1,2-E in the sample solution to be tested][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
m1-the quality of the standard substance,
m2-the mass of the sample to be tested,
P1-7-Chloroindeno [1,2-E in Standard][1,3,4]Mass fraction of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester;
wherein, the high performance liquid chromatography conditions comprise:
the chromatographic column is an ODS-C18 reversed phase chromatographic column, the temperature of the chromatographic column is 30-50 ℃, and the mobile phase is a mixed system of methanol and glacial acetic acid aqueous solution. Through a large number of experiments, the inventor finds that in terms of selection of a mobile phase, a target substance can be effectively separated from impurities by selecting methanol and a glacial acetic acid aqueous solution, and the method is superior to other solvents in economic applicability.
Preferably, the detection wavelength of the high performance liquid chromatography is 280 nm. The detection wavelength at 280nm 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 at this wavelength is small.
Preferably, the column length of the chromatography column is 150mm, the column internal diameter is 4.5-5mm, more preferably 4.6mm, and the column particle size is 3.5. mu.m.
Preferably, the temperature of the column is 40 ℃.
Preferably, the volume fraction of the aqueous glacial acetic acid solution in the mixing system is 20-50%, more preferably 25%.
Preferably, the high performance liquid chromatography conditions further comprise: the sample volume per injection was 5. mu.L, and the flow rate of the mobile phase was 1 ml/min.
The beneficial effect of the invention is that,
(1) the invention adopts high performance liquid chromatography to detect the content of 7-chloro-indeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, 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, the detection wavelength is 230-300 nm, the content can be accurately determined, and the technical blank in the corresponding field in the prior art is filled;
(2) the method for detecting the mass fraction of the 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, 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 and good in repeatability, the obtained result has high reliability, the method is particularly suitable for quality control of intermediate products of pesticide raw medicines, and has important effect and practical significance on ensuring 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 or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a chromatogram of the standard solution in example 1;
FIG. 2 is a chromatogram of a 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 a 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 a 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 the linear relationship of the wavelength of 260nm in comparative example 1;
FIG. 9 is a graph showing the linear relationship of the wavelength of 300nm in comparative example 1;
fig. 10 shows the ratio of mobile phases in comparative example 2 as methanol: chromatogram for 0.8% aqueous acetic acid at 90: 10;
fig. 11 shows the ratio of mobile phases in comparative example 2 as methanol: chromatogram at 60:40 in 0.8% aqueous acetic acid;
wherein the content of the first and second substances,
in the chromatograms of fig. 1-6, the abscissa represents time and the ordinate represents absorbance;
in the linear relationship graphs of FIGS. 8-9, the abscissa represents the concentration of the standard solution (in g/L) and the ordinate represents the peak area;
in the chromatograms of fig. 10-11, the abscissa represents time and the ordinate represents absorbance.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the technical solution in the embodiment of the present invention will be clearly and completely described below with reference to the drawings in the embodiment of the present invention, and it is obvious that the described embodiment is only a part of the embodiment of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Production run 210 batches, giving 1960 kg of a solid sample of the product 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester, and analyzing the product for 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester content, the method comprising the steps of:
(1) accurately weighing 7-chloro-indeno [1,2-E][1,3,4]Putting the 2-benzyl ester standard substance of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester into a 100ml volumetric flask, adding 90ml of methanol, shaking to dissolve, diluting with methanol to a scale to obtain a standard substance solution for later use, wherein 7-chloroindeno [1,2-E ] in the standard substance][1,3,4]The mass fraction P of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester1＝98.6％；
Accurately weighing a sample to be detected, placing the sample to be detected in a 100ml volumetric flask, adding 90ml of methanol, after oscillation and dissolution, diluting the sample to a scale with the methanol to obtain a sample solution to be detected for later use;
(2) adopting a high performance liquid chromatograph, wherein a chromatographic column is an ODS-C18 chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 volume of the sample injected each time is 5 mu L, and the flow rate of the mobile phase is 1 ml/min; the detection wavelength is set to 280 nm;
(3) after the baseline of the instrument is stabilized, samples are sequentially injected according to the sequence of a standard substance, a sample to be detected, the sample to be detected and the standard substance, the peak area average values of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester of the solution of the standard substance and the solution of the sample to be detected are respectively calculated, and the detection data are shown in the following table 1:
table 1 example 1 test results
(4) And (3) calculating the mass fraction of the 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 according to an external standard method formula, and calculating to obtain the mass fraction of the 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 concrete formula is as follows,
in the formula (I), the compound is shown in the specification,
A1-7-Chloroindeno [1,2-E in Standard solution][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
A2-7-Chloroindeno [1,2-E in the sample solution to be tested][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
m1-the quality of the standard substance,
m2-the mass of the sample to be tested,
P1-7-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,
X1-7-Chloroindeno [1,2-E in the sample to be tested][1,3,4]Mass fraction of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester;
FIG. 1 is a chromatogram of a standard solution of this example, in which the peak at the 7.923min position is the characteristic peak 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 the chromatogram of the sample solution to be tested measured in this example, and the peak at the 7.924min position is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid 4A-methyl ester 2-benzyl ester.
Example 2
A small sample of 20201208 lots gave 98.4g of a solid which was analyzed for 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid 4A-methyl ester 2-benzyl ester content by the method comprising the steps of:
(1) accurately weighing 7-chloro-indeno [1,2-E][1,3,4]Putting the 2-benzyl ester standard substance of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester into a 100ml volumetric flask, adding 90ml of methanol, shaking to dissolve, diluting with methanol to a scale to obtain a standard substance solution for later use, wherein 7-chloroindeno [1,2-E ] in the standard substance][1,3,4]The mass fraction P of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester1＝98.6％；
Accurately weighing a sample to be detected, placing the sample to be detected in a 100ml volumetric flask, adding 90ml of methanol, after oscillation and dissolution, diluting the sample to a scale with the methanol to obtain a sample solution to be detected for later use;
(2) adopting a high performance liquid chromatograph, wherein a chromatographic column is an ODS-C18 reversed phase chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 volume of the sample injected each time is 5 mu L, and the flow rate of the mobile phase is 1 ml/min; the detection wavelength is set to 280 nm;
(3) after the baseline of the instrument is stabilized, samples are sequentially injected according to the sequence of a standard substance, a sample to be detected, the sample to be detected and the standard substance, the peak area average values of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester of the solution of the standard substance and the solution of the sample to be detected are respectively calculated, and the detection data are shown in the following table 2:
table 2 example 2 test results
(4) 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 tested is calculated according to an external standard method formula, the specific formula is the same as that in example 1, 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 tested is calculated to be 98.8%.
FIG. 3 is a chromatogram of the standard solution in this example, in which the peak at the position of 7.368min is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, FIG. 4 is a chromatogram of the sample solution to be tested in this example, and the peak at the position of 7.342min is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester.
Example 3
Batch 203 of the run, giving 1820 kg of 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, the 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid 4A-methyl ester-2-benzyl ester content of which was analyzed, the method comprising the following steps:
(1) accurately weighing 7-chloro-indeno [1,2-E][1,3,4]Putting the oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester standard substance into a 100ml volumetric flask, adding 90ml of methanol, shaking to dissolve, diluting with methanol to a scale to obtain a standard substance solution for later use, wherein 7-chloroindeno [1,2-E ] in the standard substance][1,3,4]The mass fraction P of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester1＝98.6％；
Accurately weighing a sample to be detected, placing the sample to be detected in a 100ml volumetric flask, adding 100ml of methanol, after oscillation and dissolution, diluting the sample to a scale with the methanol to obtain a sample solution to be detected for later use;
(2) adopting a high performance liquid chromatograph with a diode array detector, wherein a chromatographic column is an ODS-C18 chiral chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 volume of the sample injected each time is 5 mu L, and the flow rate of the mobile phase is 1 ml/min; the detection wavelength is set to 280 nm;
(3) after the baseline of the instrument is stabilized, samples are sequentially injected according to the sequence of the standard substance, the sample to be detected, the 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester 2-benzyl ester peak area average value of the standard substance solution and the sample solution to be detected is respectively calculated, and the detection data are shown in the following table 3:
table 3 example 3 test results
(4) 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 tested is calculated according to an external standard method formula, the specific formula is the same as that in example 1, 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 tested is calculated to be 98.4%.
FIG. 5 is a chromatogram of a standard solution of this example, in which the peak at the position of 7.263min is 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, FIG. 6 is a chromatogram of a sample solution to be tested of this example, in which the peak at the position of 7.267min 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 example 1 as a subject, analyzing the standard solution at room temperature once every certain time interval for 5 times, and simultaneously recording peak areas, 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 length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 water solution, and the volume fraction of the glacial acetic acid water solution in the mixed system is 25%; the volume of the sample injected each time is 5 mu L, the flow rate of the mobile phase is 1ml/min, and the detection wavelength is 280 nm. The results are shown in the following table 4, the retention time of the chromatographic peak is stable, the RSD is less than 1% when comparing the peak areas, which indicates that the analysis method of the present invention has good stability.
TABLE 4 stability test results
Test example 2 precision test
Taking 20201208 batches of the small samples in example 2 as subjects to be investigated, weighing a standard substance and five parallel samples to be tested, respectively injecting samples according to the sequence of the standard substance solution, the sample solution to be tested and the 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 tested;
the analysis conditions were: adopting a high performance liquid chromatograph and an Shimadzu liquid chromatograph of a diode array detector, wherein a chromatographic column is an ODS-C18 reversed phase chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 water, and the volume fraction of the glacial acetic acid water in the mixed system is 25%; the volume of the sample injected each time is 5 mu L, and the flow rate of the mobile phase is 1 ml/min; the detection wavelength was set at 280 nm.
As shown in Table 5 below, the results obtained by comparing the mass fractions of 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester and the RSD of less than 1% indicate that the analytical method of the present invention is excellent in precision.
TABLE 5 results of precision test
Test example 3 recovery test
A small sample of 20201208 from example 2, batch 7-chloroindeno [1,2-E ] [1,3,4] oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester, was weighed and a standard sample was weighed, five samples of different mass were weighed simultaneously and separately, and samples of different mass were added to calculate the recovery of the added standard sample.
The analysis conditions were: adopting a high performance liquid chromatograph with a diode array detector, wherein a chromatographic column is an ODS-C18 chiral chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 volume of a sample injected each time is 5 mu L, and the flow rate of the mobile phase is 1 ml/min; the detection wavelength of the high performance liquid chromatography was set to 280 nm. After the baseline of the instrument is stable, samples are sequentially injected according to the sequence of the standard substance, the sample to be tested, the standard substance and the sample to be tested, the peak areas of the 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 tested are respectively calculated, the results are shown in the following table 6, the recovery rates are respectively between 98% and 101%, and the average recovery rate is 99.38%, which indicates that the experimental recovery rate meets the requirements.
TABLE 6 recovery test results
Test example 4 Linear test
Weighing a series of standard products with different qualities, placing the standard products in a 100ml volumetric flask, dissolving the standard products with methanol, fixing the volume, and observing the relation between the peak area and the solution concentration after sample injection;
the analysis conditions include: adopting a high performance liquid chromatograph with a diode array detector, wherein the chromatographic column is an ODS-C18 reversed phase chromatographic column, the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.6mm, and the granularity of the chromatographic column 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 volume of the sample injected each time is 5 mu L, the flow rate of the mobile phase is 1ml/min, and the detection wavelength is 280 nm.
The results are shown in table 7 below and fig. 7, and the correlation coefficient is 1, which indicates that the analysis method provided by the present invention is linear and meets the requirements.
TABLE 7 results of the Linear test
As can be seen 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 present 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
In the test process, wavelengths of 260nm and 300nm are selected for linear verification respectively, and the results are shown in tables 8-9. The verification shows that under the conditions of the wavelengths of 260nm and 300nm, the linear correlation coefficient R is in spite of2Also greater than 0.99, but with a large intercept, see fig. 8 and 9, the error is large if the single standard comparison method of the present invention is used for sample analysis.
TABLE 8 wavelength at 260nm for linear test results
TABLE 9 Linear test results at 300nm wavelength
Comparative example 2 chromatogram for different mobile phase ratios
Taking a small test sample, adding methanol: 0.8% aqueous acetic acid solution ═ 90:10 (or 60: 40) mixed system is a mobile phase, the flow rate is 1mL/min, the sample volume of each sample injection is 5 mu L, and the peak appearance condition of the chromatogram map by different mobile phase ratio examples is examined.
In the presence of methanol: 0.8% aqueous acetic acid solution ═ 90: at 10, some small impurities have poor separation degree from the target, which affects the calculation of peak area and further the content, as shown in FIG. 10.
In the presence of methanol: 0.8% aqueous acetic acid 60: at 40, when the organic phase composition decreased, the time to peak was prolonged and the peak profile broadened, as shown in FIG. 11.
Although the present invention has been described in detail by referring to the drawings in connection with the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and the spirit of the present invention, and any changes or substitutions which are within the technical scope of the present invention and are easily conceived by those skilled in the art are within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (8)
1. An analytical method for content of a 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 a high performance liquid chromatography analytical method is adopted, and the specific method is as follows:
(1) respectively dissolving a standard substance and a sample to be detected by using methanol as a solvent, and preparing a standard substance solution and a sample solution to be detected;
(2) setting the detection wavelength of the high performance liquid chromatography to be within 230-300 nm, sequentially injecting samples according to the sequence of a standard substance, a sample to be detected and the standard substance after the baseline of the 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 solution of the standard substance and the solution of the sample to be detected;
(3) according to an external standard method formula, 7-chloro-indeno [1,2-E ] in a sample to be detected][1,3,4]Weight fraction X of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester1The calculation is carried out according to the following specific formula:
in the formula (I), the compound is shown in the specification,
A1-7-Chloroindeno [1,2-E in Standard solution][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
A2-7-Chloroindeno [1,2-E in the sample solution to be tested][1,3,4]Average value of peak area of oxadiazine-2, 4A (3H,5H) -dicarboxylic acid-4A-methyl ester-2-benzyl ester,
m1-the quality of the standard substance,
m2-the mass of the sample to be tested,
P1-7-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 comprise:
the chromatographic column is an ODS-C18 reversed phase chromatographic column, the temperature of the chromatographic column is 30-50 ℃, and the mobile phase is a mixed system of methanol and glacial acetic acid aqueous solution.
2. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the concentration range of the standard solution and the concentration range of the solution to be tested in step (1) are 0.5-1 mg/ml.
3. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the detection wavelength of the high performance liquid chromatography is 280 nm.
4. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the length of the chromatographic column is 150mm, the inner diameter of the chromatographic column is 4.5-5mm, and the particle size of the chromatographic column is 3.5 μm.
5. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the temperature of the chromatographic column is 40 ℃.
6. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 20-50%.
7. The method for analyzing the content of the indoxacarb key intermediate according to claim 6, characterized in that the volume fraction of the glacial acetic acid aqueous solution in the mixed system is 25%.
8. The method for analyzing the content of the indoxacarb key intermediate according to claim 1, characterized in that the high performance liquid chromatography conditions further comprise: the sample volume per injection was 5. mu.L, and the flow rate of the mobile phase was 1 ml/min.
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