CN116068101A - Method for quantitatively analyzing dimethyl sulfate content by derivatization method - Google Patents
Method for quantitatively analyzing dimethyl sulfate content by derivatization method Download PDFInfo
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- CN116068101A CN116068101A CN202211670467.8A CN202211670467A CN116068101A CN 116068101 A CN116068101 A CN 116068101A CN 202211670467 A CN202211670467 A CN 202211670467A CN 116068101 A CN116068101 A CN 116068101A
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- VAYGXNSJCAHWJZ-UHFFFAOYSA-N dimethyl sulfate Chemical compound COS(=O)(=O)OC VAYGXNSJCAHWJZ-UHFFFAOYSA-N 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 43
- 238000001212 derivatisation Methods 0.000 title claims abstract description 24
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical compound COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 claims abstract description 56
- NESLWCLHZZISNB-UHFFFAOYSA-M sodium phenolate Chemical compound [Na+].[O-]C1=CC=CC=C1 NESLWCLHZZISNB-UHFFFAOYSA-M 0.000 claims abstract description 44
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 10
- 239000012488 sample solution Substances 0.000 claims abstract description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 42
- 238000007865 diluting Methods 0.000 claims description 25
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 24
- 238000002360 preparation method Methods 0.000 claims description 12
- 239000000523 sample Substances 0.000 claims description 11
- 239000006184 cosolvent Substances 0.000 claims description 8
- 238000012360 testing method Methods 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 39
- 238000006243 chemical reaction Methods 0.000 abstract description 13
- 239000000126 substance Substances 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 239000013076 target substance Substances 0.000 abstract 2
- 239000000243 solution Substances 0.000 description 66
- 239000011550 stock solution Substances 0.000 description 32
- 239000013558 reference substance Substances 0.000 description 27
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 238000007789 sealing Methods 0.000 description 13
- 238000005303 weighing Methods 0.000 description 11
- 239000000047 product Substances 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- FVAUCKIRQBBSSJ-UHFFFAOYSA-M sodium iodide Chemical compound [Na+].[I-] FVAUCKIRQBBSSJ-UHFFFAOYSA-M 0.000 description 6
- 238000004817 gas chromatography Methods 0.000 description 5
- 229960004048 pantoprazole sodium Drugs 0.000 description 5
- YNWDKZIIWCEDEE-UHFFFAOYSA-N pantoprazole sodium Chemical compound [Na+].COC1=CC=NC(CS(=O)C=2[N-]C3=CC=C(OC(F)F)C=C3N=2)=C1OC YNWDKZIIWCEDEE-UHFFFAOYSA-N 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 3
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 3
- JZMJDSHXVKJFKW-UHFFFAOYSA-M methyl sulfate(1-) Chemical compound COS([O-])(=O)=O JZMJDSHXVKJFKW-UHFFFAOYSA-M 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 238000010533 azeotropic distillation Methods 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 238000004255 ion exchange chromatography Methods 0.000 description 2
- XKHUUPWVDVIRHM-UHFFFAOYSA-N methylsulfinylmethane;propan-2-ol Chemical compound CC(C)O.CS(C)=O XKHUUPWVDVIRHM-UHFFFAOYSA-N 0.000 description 2
- 150000002894 organic compounds Chemical class 0.000 description 2
- 235000009518 sodium iodide Nutrition 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 101100400452 Caenorhabditis elegans map-2 gene Proteins 0.000 description 1
- 101150064138 MAP1 gene Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000001738 genotoxic effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 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
- 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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- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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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
- G01N2030/067—Preparation by reaction, e.g. derivatising the sample
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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Abstract
The invention belongs to the technical field of chemical detection, and particularly relates to a method for quantitatively analyzing dimethyl sulfate content by a derivatization method. Sodium phenolate is used as a derivative reagent to form derivative anisole with dimethyl sulfate, and the content of dimethyl sulfate is calculated by measuring the peak area of anisole in a sample solution through a gas chromatograph-mass spectrometer. Compared with the prior art, the analysis method has the advantages that (1) the derivative product prepared by the method is stable in 11h, the peak areas of the target substances are 32224, 33873, 31960 and 31560 respectively in 0, 2, 6 and 11h, the RSD is 3.14%, the peak shape is good, and the peak shape of the target substances is not greatly changed due to temperature programming; (2) The method has high sensitivity, and the detection shows that the quantitative limit of dimethyl sulfate is 0.1057ng and the detection limit is 0.0032ng, so that the detection requirement is completely met; (3) The derivatization reaction can be immediately carried out between the derivatization reagent and the dimethyl sulfate, so that the overall detection efficiency is greatly improved, and the operation is simple.
Description
Technical Field
The invention belongs to the technical field of chemical detection, and particularly relates to a method for quantitatively analyzing dimethyl sulfate content by a derivatization method.
Background
Dimethyl sulfate has mutagenic/genotoxic properties both in vitro and in vivo, and a toxicological attention threshold (TTC) of 1.5 μg/day, according to the ICH M7 regulation. Currently, common analytical methods for detecting dimethyl sulfate include Gas Chromatography (GC) and direct detection by gas chromatography-mass spectrometry (GC-MS). Because the dimethyl sulfate has poor stability at room temperature, the direct analysis and detection can generate great error or the detection process needs to be used newly, so that the test operation is complicated; dimethyl sulfate is used as a common methylation reagent, and has active chemical properties, so that the derivatization method is combined with a gas chromatograph to measure the dimethyl sulfate.
Regarding the method for measuring dimethyl sulfate, the following documents are disclosed:
the first prior art is: directly using gas chromatography or gas chromatography to determine dimethyl sulfate content; the disadvantage is that: however, dimethyl sulfate has poor stability at room temperature, is difficult to accurately measure, and is usually required to be prepared newly, and the test process is time-consuming and labor-consuming; for samples with lower limit requirements, the detection sensitivity of gas chromatography may not meet the detection requirements; CN 113848279A discloses a method for detecting dimethyl sulfate as a residual reagent in a drug, and the content of monomethyl sulfate is detected by ion chromatography: precisely weighing 250mg of pantoprazole sodium as a drug to be measured, placing the pantoprazole sodium into a 50ml measuring flask, dissolving the pantoprazole sodium in 30mmol/L sodium hydroxide solution, fixing the volume to a scale, and shaking the pantoprazole sodium until the pantoprazole sodium is uniform; filtering the solution by a solid phase extraction small column, and directly measuring the content of monomethyl sulfate by ion chromatography.
And the second prior art is as follows: CN111505182a discloses a method for measuring dimethyl sulfate in a medicament by using a derivatization gas chromatography-mass spectrometry, which uses sodium iodide and sodium thiosulfate as derivatization solutions after being dissolved in water, acetonitrile and dimethyl sulfate are added for reaction to generate methyl iodide, and then the methyl sulfate content is obtained by calculating the methyl iodide peak area through gas chromatography-mass spectrometry combined instrument headspace sample injection. The method has short derivatization reaction time: the derivative product methyl iodide is volatile substance for 30min, so that the interference of non-volatile substances on detection can be reduced;
the second disadvantage is that: the derivatization reagent sodium iodide usually needs to use water as a solvent, and the service life of an ion source of the gas chromatograph-mass spectrometer can be shortened by multiple sample injections; the stability of the derivative product is poor, the derivative reaction is difficult to control, an internal standard is usually required to be added to ensure the detection accuracy, and the test operation is complicated.
The third prior art is: CN 10364545C discloses a process for preparing anisole, which comprises the following steps: (1) An excess of 10% -30% dimethyl sulfate is reacted with sodium phenolate solution (prepared from 20% by weight sodium hydroxide solution and phenol in equimolar form) at 10-30 c for 1 hour, 40 c for half an hour, and at reflux temperature for half an hour; (2) Distilling anisole by azeotropic distillation, and returning the distilled water part to the reactor in the distillation process; (3) When anisole obtained by azeotropic distillation is close to 90% of theoretical yield, adding sodium hydroxide solution with the agriculture degree of 20% (weight) into the reactor, continuously reacting, distilling for half an hour to 1 hour, completely distilling anisole generated by the reaction, and then drying and redistilling anisole product.
The third technical disadvantage is that: (1) The reaction that can take place in the reaction system has sodium hydroxide + phenol, sodium phenolate + dimethyl sulfate to still have sodium hydroxide + dimethyl sulfate, comparatively complicated, if calculate dimethyl sulfate content through anisole that this produced, will cause the testing result to be lower than actual conditions even appear false negative situation like this, can't carry out accurate quality control (2) its main objective to the sample and constantly prepare, reactant amount is more, so the reaction condition is more complicated, including steps such as backward flow, distillation, reaction temperature need be according to progress adjustment, and time consuming is long.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for quantitatively analyzing the content of dimethyl sulfate by a derivatization method. Sodium phenolate is used as a derivative reagent, reacts with dimethyl sulfate to generate anisole, and the content of the dimethyl sulfate is calculated by detecting the content of anisole (with low melting point and easy volatilization); and has the following advantages: the reaction product is stable, is not easily affected by temperature programming, has high sensitivity and completely meets the measurement requirement.
The invention provides a method for quantitatively analyzing dimethyl sulfate content by a derivatization method, which mainly comprises the steps of derivatizing dimethyl sulfate into anisole by taking sodium phenolate solution as a derivatization reagent, and detecting the content of anisole to obtain the content of dimethyl sulfate;
preferably, in the preparation process of the sodium phenolate solution, isopropanol is used as a cosolvent of sodium phenolate; in the experiment, the solubility of sodium phenolate in dimethyl sulfoxide is low, and experiments show that the solubility of sodium phenolate in an organic phase is poor, the base material interference is large when methanol, ethanol and acetone are used as cosolvent, and isopropanol is selected as cosolvent through optimal final determination.
Preferably, the sodium phenolate solution is prepared by adding isopropanol into sodium phenolate, adding dimethyl sulfoxide, dissolving by ultrasonic, and diluting by using dimethyl sulfoxide, wherein the adding ratio of the isopropanol to the dimethyl sulfoxide before ultrasonic treatment is 1:10.8-13.2.
Preferably, the addition ratio of the isopropanol to the dimethyl sulfoxide is 1:12.
The detection system of the invention is dimethyl sulfoxide-isopropanol, can meet the trace measurement of dimethyl sulfate in most organic compounds, and has wide applicability.
Preferably, in the method provided by the invention, the derivative product anisole is diluted by dimethyl sulfoxide to obtain a sample solution, and the dimethyl sulfate content is calculated by measuring the peak area of anisole in the sample solution by using a gas chromatograph-mass spectrometer.
A method for quantitatively analyzing the content of dimethyl sulfate by a derivatization method, which comprises the following steps:
(1) Preparation of sodium phenolate solution: weighing 0.48-0.52 g of sodium phenolate, placing into a 500ml measuring flask, adding 25ml of isopropanol, then adding 270-330 ml of dimethyl sulfoxide, ultrasonically dissolving, diluting to a scale with the dimethyl sulfoxide, and shaking uniformly to obtain 0.96-1.04 mg/ml of sodium phenolate solution;
(2) Preparation of control stock solution: taking 18-22 mg of dimethyl sulfate, placing into a 10ml measuring flask, diluting to a scale with 0.96-1.04 mg/ml of sodium phenolate solution, shaking uniformly, precisely measuring 1ml, placing into a 200ml measuring flask, diluting to the scale with 0.96-1.04 mg/ml of sodium phenolate solution, shaking uniformly, and obtaining the reference substance stock solution.
(3) Preparation of a control solution: measuring 0.95-1.05 ml of reference substance stock solution, placing the stock solution into a 10ml measuring flask, diluting the stock solution to a scale with 0.96-1.04 mg/ml of sodium phenolate solution, shaking the stock solution uniformly, precisely measuring 5ml of stock solution, placing the stock solution into a 20ml headspace flask, and sealing the headspace flask to obtain a reference substance solution.
(4) Preparation of blank group: precisely measuring 5ml of 0.96-1.04 mg/ml sodium phenolate solution, placing into a 20ml headspace bottle, and sealing.
(5) Preparation of a linear solution: precisely measuring (2) 0.1ml, 0.3ml, 0.5ml, 1.0ml, 1.5ml and 2.0ml of control stock solution, respectively placing into 10ml measuring bottles, diluting to scale with 1mg/ml sodium phenolate solution, shaking, precisely measuring 5ml, respectively placing into 20ml headspace bottles, and sealing.
(6) Preparation of detection limit solution: precisely measuring 0.1ml of the reference substance stock solution, placing the reference substance stock solution into a 10ml measuring flask, diluting the reference substance stock solution to a scale with 1mg/ml sodium phenolate solution, shaking the solution uniformly, precisely measuring 3ml of the solution, placing the solution into the 10ml measuring flask, diluting the solution to the scale with 1mg/ml sodium phenolate solution, shaking the solution uniformly, precisely measuring 5ml of the solution, placing the solution into a 20ml headspace bottle, and sealing the headspace bottle to obtain the detection limiting solution.
(7) Preparation of the stability solution: precisely measuring 5ml of the reference substance stock solution in (2), placing the reference substance stock solution in a 50ml measuring flask, diluting to a scale with 1mg/ml sodium phenolate solution, shaking uniformly, precisely measuring 5ml, placing the reference substance stock solution in a 20ml headspace bottle, sealing, taking 4 parts, and respectively taking samples from the headspace of 0h, 2h, 6h and 11h to examine the solution stability.
(8) Determination of the derivative products: weighing 19.5-20.5 mg of anisole, placing into a 10ml measuring flask, diluting to a scale with dimethyl sulfoxide, shaking uniformly, precisely measuring 0.1ml, placing into a 100ml measuring flask, diluting to a scale with dimethyl sulfoxide, shaking uniformly, precisely measuring 5ml, placing into a 20ml headspace bottle as a sample to be measured (sample solution), and sealing.
(9) And (3) detection: and (3) measuring the sample solution in the headspace bottle by using a gas chromatograph-mass spectrometer, obtaining the peak area, and calculating to obtain the content of the dimethyl sulfate.
The chemical reaction formula of the derivative is as follows:
namely 2C 6 H 5 NaO+C 2 H 6 O 4 S→2C 7 H 8 O；
Preferably, in the detection process (9), the program takes 38-42 ℃ as an initial temperature, keeps for 7-8.5 min, then heats up to 58-62 ℃ at the speed of 4.5-5.5 ℃/min and keeps for 1-1.5 min, and then heats up to 230-250 ℃ at the speed of 38-42 ℃/min and keeps for 1-2 min.
Preferably, (9) during the detection, the program is maintained at a starting temperature of 40℃for 8 minutes, then at a rate of 5℃per minute to 60℃for 1 minute, and then at a rate of 40℃per minute to 240℃for 1 minute.
Preferably, during the detection, the chromatographic conditions: HP-5ms UI chromatographic column, column length 30m, inner diameter 0.25mm, liquid film thickness 0.25 μm; the flow rate is 1.0ml/min; split ratio 20:1, a step of; the temperature of the sample inlet is 220 ℃.
Preferably, in the detection process, when the solution to be detected is placed in a headspace sample injector of a gas chromatograph-mass spectrometer for balancing, the balancing temperature is 100 ℃, and the balancing time is 30min.
The invention has the beneficial effects that:
(1) The derivative product prepared by the invention is stable in 11h, and the peak areas of the sample solution are 32224, 33873, 31960 and 31560 and the RSD is 3.14% respectively in 0, 2, 6 and 11 h; the peak shape of the target is good, and the peak shape of the target is not greatly changed due to temperature programming;
(2) The method has high sensitivity, and the detection shows that the quantitative limit of dimethyl sulfate is 0.1057ng and the detection limit is 0.0032ng, so that the detection requirement is completely met;
(3) The defect of multiple sample injection in the prior art is overcome, the quality of the instrument can be better maintained, and the service life is prolonged; meanwhile, the stability of the derivative product is good, and an internal standard is not needed to be added to ensure the detection accuracy, so that the test operation is simpler and more convenient;
(4) The derivatization reaction can be immediately carried out between the derivatization reagent and the dimethyl sulfate, so that the overall detection efficiency is greatly improved, and the operation is simple;
(5) The detection system of the invention is dimethyl sulfoxide-isopropanol, can meet the trace measurement of dimethyl sulfate in most organic compounds, and has wide applicability.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the description of the embodiments will be briefly described below.
FIG. 1 is a derivative blank chromatogram;
FIG. 2 is a color chart of a control solution;
FIG. 3 is a color chart of anisole solution;
FIG. 4 is a linear graph;
FIG. 5 is a chromatogram of anisole with methanol as a co-solvent;
FIG. 6 is a chromatogram of anisole with ethanol as a co-solvent;
FIG. 7 is a chromatogram of anisole with acetone as a co-solvent;
FIG. 8 is a chromatogram of anisole with isopropanol as a co-solvent;
FIG. 9 is a chromatogram of anisole with dimethyl sulfoxide alone as solvent.
Detailed Description
The present invention will now be further described in connection with specific embodiments in order to enable those skilled in the art to better understand the invention.
Example 1
A method for quantitatively analyzing the content of dimethyl sulfate by a derivatization method, which comprises the following steps:
(1) Preparation of a derivatizing agent (sodium phenolate solution);
(2) Taking dimethyl sulfate as a raw material, and preparing anisole with a sodium phenolate solution;
(3) And diluting anisole with dimethyl sulfoxide to obtain solution to be detected, injecting sample into the headspace, and recording the map.
The chromatographic conditions and mass spectrum conditions of the detection process of the gas chromatograph-mass spectrometer are as follows:
method verification of chromatographic method:
1. specialization of
(1) Preparation of derivatizing agent (sodium phenolate solution): weighing 0.5g of sodium phenolate, placing into a 500ml measuring flask, adding 25ml of isopropanol, adding 300ml of dimethyl sulfoxide, performing ultrasonic dissolution, diluting to a scale with dimethyl sulfoxide, and shaking uniformly to obtain 1mg/ml sodium phenolate solution;
(2) Preparing a reference stock solution: taking 20mg of dimethyl sulfate reference substance, precisely weighing, placing into a 10ml measuring flask, diluting to a scale with 1mg/ml sodium phenolate solution, shaking uniformly, precisely weighing 1ml, placing into a 200ml measuring flask, diluting to a scale with 1mg/ml sodium phenolate solution, shaking uniformly, and obtaining a reference substance stock solution;
(3) Blank group: precisely measuring 5ml of 1mg/ml sodium phenolate solution, placing in a 20ml headspace bottle, sealing, and performing headspace sample injection to obtain a map 1;
(4) Determination of a reference substance solution: precisely measuring 1ml of reference substance stock solution, placing in a 10ml measuring flask, diluting to scale with 1mg/ml sodium phenolate solution, shaking, precisely measuring 5ml, placing in a 20ml headspace bottle, sealing, and performing headspace sampling to obtain a map 2;
at this time, the concentration of the control solution was 1ug/ml, and the desired sodium phenolate concentration was 1/126×2× 116.02 =1.84 ug/ml;
the linear peak concentration is about 2ug/ml, with a desired sodium phenolate concentration of 2/126 x 2 x 116.02 = 3.68ug/ml;
the concentration of sodium phenolate in the sodium phenolate solution actually used is 1mg/ml, which is far greater than the theoretical concentration, so that the complete derivatization is ensured;
(5) Determination of the derivative products: weighing anisole 20mg precisely, placing into a 10ml measuring flask, diluting to scale with dimethyl sulfoxide, shaking uniformly, precisely weighing 0.1ml, placing into a 100ml measuring flask, diluting to scale with dimethyl sulfoxide, shaking uniformly, precisely weighing 5ml, placing into a 20ml headspace bottle, sealing, and performing headspace sampling to obtain a map 3;
detection conclusion: the spectrum shows that the blank does not interfere with the measurement, and the specificity of the detection method is good.
2. Linear relationship and range investigation
Precisely weighing 0.1ml, 0.3ml, 0.5ml, 1.0ml, 1.5ml and 2.0ml of the reference substance stock solution prepared in the step (1), respectively placing the reference substance stock solution and the reference substance stock solution in a 10ml measuring flask, diluting to scale with 1mg/ml sodium phenolate solution, shaking uniformly, precisely weighing 5ml respectively, placing the reference substance stock solution in a 20ml headspace flask, sealing, and performing headspace sampling to obtain a map 4, wherein the linear and range results are shown in the following table.
Table 1 linear and range results:
3. quantitative limit and detection limit
(1) Precisely measuring 0.1ml of the reference substance stock solution prepared in the step (1), placing in a 10ml measuring flask, diluting to scale with 1mg/ml sodium phenolate solution, shaking uniformly, preparing 6 parts in parallel, precisely measuring 5ml respectively, placing in a 20ml headspace flask, and sealing to obtain quantitative limiting solution;
precisely measuring 0.1ml of the reference substance stock solution prepared in the step (1), placing the reference substance stock solution into a 10ml measuring flask, diluting the reference substance stock solution to a scale with 1mg/ml of sodium phenolate solution, shaking the solution uniformly, precisely measuring 3ml of the solution, placing the reference substance stock solution into the 10ml measuring flask, diluting the reference substance stock solution to the scale with 1mg/ml of sodium phenolate solution, shaking the solution uniformly, precisely measuring 5ml of the solution, placing the solution into a 20ml headspace bottle, and sealing the headspace bottle to obtain the detection limiting solution.
The quantitative limit and the detection limit result are shown in the following table:
LOQ-1 to LOQ-6 limit of quantification, LOD is limit of detection;
TABLE 2 quantitative limit and detection limit detection results
4. Investigation of solution stability
Preparation and detection of a stable solution: precisely measuring 5ml of the reference substance stock solution prepared in the step (1), placing the reference substance stock solution in a 50ml measuring flask, diluting to a scale with 1mg/ml sodium phenolate solution, shaking uniformly, precisely measuring 5ml, placing the reference substance stock solution in a 20ml headspace flask, sealing, taking 4 parts, respectively placing the reference substance stock solution in the headspace for 0h, 2h, 6h and 11h at normal temperature, and observing the stability of the solution.
The solution stability results are shown in the following table:
TABLE 3 solution stability results
From the stability of the peak area, the derivatization reaction is fully reacted, and the stability of the peak area is not affected after the reaction is placed at normal temperature; meanwhile, the stability of the derivative product is good, and an internal standard is not needed to be added to ensure the detection accuracy, so that the test operation is simpler and more convenient and efficient;
the derivatization reaction can be immediately carried out between the derivatization reagent and the dimethyl sulfate, so that the overall detection efficiency is greatly improved, and the operation is simple.
Claims (10)
1. A method for quantitatively analyzing the content of dimethyl sulfate by a derivatization method is characterized in that dimethyl sulfate is formed into derivative anisole by a derivatization reagent, and the content of dimethyl sulfate is obtained by detecting the content of anisole;
wherein isopropanol is used as a cosolvent in the preparation of the derivatizing agent.
2. The method of claim 1, wherein the derivatizing agent is a sodium phenolate solution.
3. The method of claim 1, wherein the derivatizing agent is a sodium phenolate solution of 0.96-1.04 mg/ml.
4. A method according to any one of claims 2 to 3, wherein the sodium phenolate solution is prepared by: firstly adding isopropanol into sodium phenolate, then adding dimethyl sulfoxide, and diluting with dimethyl sulfoxide after ultrasonic dissolution.
5. The method according to claim 4, wherein the addition ratio of the isopropanol to the dimethyl sulfoxide before ultrasonic treatment is 1: 10.8-13.2.
6. The method according to claim 5, wherein the addition ratio of the isopropanol to the dimethyl sulfoxide before ultrasonic treatment is 1:12.
7. the method of claim 1, wherein the dimethyl sulfate content is calculated by diluting the derivative anisole with dimethyl sulfoxide to obtain a sample solution and measuring the peak area of anisole in the sample solution by a gas chromatograph-mass spectrometer.
8. The method of claim 7, wherein during the detecting, the program is started at 38-42 ℃ for 7-8.5 min, then is heated to 58-62 ℃ at a speed of 4.5-5.5 ℃/min and is kept for 1-1.5 min, and then is heated to 230-250 ℃ at a speed of 38-42 ℃/min and is kept for 1-2 min.
9. The method of claim 1, wherein during the detecting, chromatographic conditions: HP-5ms UI chromatographic column, column length 30m, inner diameter 0.25mm, liquid film thickness 0.25 μm; the flow rate is 1.0ml/min; split ratio 20:1, a step of; the temperature of the sample inlet is 220 ℃.
10. The method of claim 7, wherein the solution to be measured is equilibrated in a headspace sample injector of a gas chromatograph at 100 ℃ for 30min during the testing process.
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