阅读说明：本技术 一种邻氯苄基氯化镁含量的测定方法 (Method for measuring content of o-chlorobenzyl magnesium chloride ) 是由 张丽荣 赵娜 齐建敏 崔占卫 贾成国 于 2021-09-13 设计创作，主要内容包括：本发明涉及一种邻氯苄基氯化镁含量的测定方法,属于化学分析技术领域,反应液中的邻氯苄基氯化镁与衍生试剂反应得到邻氯苄基氯化镁衍生物,然后采用气相色谱法或液相色谱法对衍生物进行含量测定,根据衍生物的含量计算邻氯苄基氯化镁的含量。本发明将活泼的格氏试剂邻氯苄基氯化镁衍生后生成稳定的邻氯苄基氯化镁丙酮衍生物,然后再进行定量分析可以提高定量的准确性,避免了常规滴定过程中可能存在部分被破坏的邻氯苄基氯化镁也被滴定且滴定终点易存在有反色现象进而操作较难控制导致结果准确性较低的问题。(The invention relates to a method for measuring the content of o-chlorobenzyl magnesium chloride, which belongs to the technical field of chemical analysis. According to the method, the active Grignard reagent o-chlorobenzyl magnesium chloride is derived to generate the stable o-chlorobenzyl magnesium chloride acetone derivative, and then the quantitative analysis is carried out to improve the quantitative accuracy, so that the problems that the o-chlorobenzyl magnesium chloride which is possibly partially damaged in the conventional titration process is also titrated, the titration end point is easy to have a reverse color phenomenon, and the operation is difficult to control, so that the result accuracy is lower are solved.)

1. a method for measuring the content of o-chlorobenzyl magnesium chloride is characterized by comprising the following steps: reacting o-chlorobenzyl magnesium chloride in the reaction solution with a derivatization reagent to obtain an o-chlorobenzyl magnesium chloride derivative, then measuring the content of the derivative by adopting gas chromatography or liquid chromatography, and calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative.

2. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 1, which comprises the following steps: the derivatization reagent is acetone, and the o-chlorobenzyl magnesium chloride derivative is an o-chlorobenzyl magnesium chloride acetone derivative.

3. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 2, which comprises the following steps: in the reaction process, under the protection of nitrogen, firstly transferring a certain amount of acetone into a vacuum reaction bottle, then transferring a certain amount of reaction liquid into the vacuum reaction bottle, uniformly shaking, finally transferring a certain amount of acid, and uniformly shaking to obtain the o-chlorobenzyl magnesium chloride acetone derivative reaction liquid.

4. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 2, which comprises the following steps: the gas chromatography assay comprises the steps of:

(1) preparing an internal standard stock solution;

(2) preparing a standard solution;

(3) preparing a sample solution;

(4) performing gas chromatography detection, and calculating the content of the derivative by an internal standard method.

5. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 4, which comprises the following steps: the gas chromatography conditions of the step (4) are as follows:

a chromatographic column: DM-624, 30m × 0.53mm, 3.0 um;

nitrogen gas: 10mL/min, air: 300mL/min, hydrogen: 30 mL/min;

sample introduction amount: 1 mu L of the solution;

vaporization temperature: 250 ℃;

detecting the temperature: 250 ℃;

column temperature: the initial temperature is 65-75 ℃, the temperature is kept for 0-3min, the temperature is increased to 160 ℃ at the temperature increasing rate of 10 ℃/min, then the temperature is increased to 238 ℃ at the temperature increasing rate of 40 ℃/min, and the temperature is kept for 13 min;

a detector: FID.

6. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 2, which comprises the following steps: the liquid chromatography assay comprises the steps of:

(1) preparing a standard solution;

(2) preparing a sample solution;

(3) performing liquid chromatography detection, and calculating the content of the derivative by an external standard method.

7. The method for determining the content of o-chlorobenzyl magnesium chloride according to claim 6, which comprises the following steps: the liquid chromatography conditions of the step (3) are as follows:

a chromatographic column: the stationary phase is octadecylsilane chemically bonded silica;

mobile phase: 0.03-0.1% trifluoroacetic acid aqueous solution-methanol, wherein the proportion of 0.03-0.1% trifluoroacetic acid aqueous solution is 10-30%, and the proportion of methanol is 90-70%;

wavelength: 210 nm;

flow rate: 1.0 mL/min;

column temperature: 35 ℃;

sample introduction amount: 5 μ L.

Technical Field

The invention relates to a method for measuring the content of o-chlorobenzyl magnesium chloride, belonging to the technical field of chemical analysis.

Background

The o-chlorobenzyl magnesium chloride is a key intermediate for preparing the bactericide prothioconazole. Prothioconazole is a novel spectrum triazolethione bactericide developed by Bayer companies, is mainly used for preventing and treating diseases of cereal crops such as cereals, wheat, beans and the like, and particularly has excellent prevention and treatment effects on wheat stripe rust and powdery mildew. The mechanism of action is to inhibit the demethylation of lanosterol, the precursor of sterol, or the 14-position of 2, 4-methylenedihydrolanostane in fungi. Because of low toxicity and no toxicity to embryos, the compound is safe to people and environment and is generally concerned by people.

The synthesis of the high-content o-chlorobenzyl magnesium chloride Grignard reagent is the key of the prothioconazole synthesis process, and whether the content of the o-chlorobenzyl magnesium chloride is accurate or not determines the scientificity of reaction feeding, so that the accuracy of the o-chlorobenzyl magnesium chloride content determination has important significance. The conventional method for measuring the content of the o-chlorobenzyl magnesium chloride is a titration method, and because the Grignard reagent has high activity and cannot contact water, air and the like, the o-chlorobenzyl magnesium chloride needs to be carefully removed by an injector with good tightness in the measuring process and then titrated, the o-chlorobenzyl magnesium chloride which is possibly partially damaged in the titration process is also titrated, so that the result deviation is caused, the titration end point is easy to have a reverse color phenomenon, and the operation is difficult to control, so that the result accuracy is low.

Disclosure of Invention

The invention aims to provide a method for measuring the content of o-chlorobenzyl magnesium chloride, which is accurate and reliable.

In order to achieve the purpose, the invention adopts the technical scheme that:

a method for measuring the content of o-chlorobenzyl magnesium chloride comprises the steps of reacting o-chlorobenzyl magnesium chloride in reaction liquid with a derivative reagent to obtain an o-chlorobenzyl magnesium chloride derivative, measuring the content of the derivative by adopting a gas chromatography or a liquid chromatography, and calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative.

The technical scheme of the invention is further improved as follows: the derivatization reagent is acetone, and the o-chlorobenzyl magnesium chloride derivative is an o-chlorobenzyl magnesium chloride acetone derivative.

The technical scheme of the invention is further improved as follows: in the reaction process, under the protection of nitrogen, firstly transferring a certain amount of acetone into a vacuum reaction bottle, then transferring a certain amount of reaction liquid into the vacuum reaction bottle, uniformly shaking, finally transferring a certain amount of acid, and uniformly shaking to obtain the o-chlorobenzyl magnesium chloride acetone derivative reaction liquid.

The technical scheme of the invention is further improved as follows: the gas chromatography assay comprises the steps of:

(1) preparing an internal standard stock solution;

(2) preparing a standard solution;

(3) preparing a sample solution;

(4) performing gas chromatography detection, and calculating the content of the derivative by an internal standard method.

The technical scheme of the invention is further improved as follows: the gas chromatography conditions of the step (4) are as follows:

a chromatographic column: DM-624, 30m × 0.53mm, 3.0 um;

nitrogen gas: 10mL/min, air: 300mL/min, hydrogen: 30 mL/min;

sample introduction amount: 1 mu L of the solution;

vaporization temperature: 250 ℃;

detecting the temperature: 250 ℃;

column temperature: the initial temperature is 65-75 ℃, the temperature is kept for 0-3min, the temperature is increased to 160 ℃ at the temperature increasing rate of 10 ℃/min, then the temperature is increased to 238 ℃ at the temperature increasing rate of 40 ℃/min, and the temperature is kept for 13 min;

a detector: FID.

The technical scheme of the invention is further improved as follows: the liquid chromatography assay comprises the steps of:

(1) preparing a standard solution;

(2) preparing a sample solution;

(3) performing liquid chromatography detection, and calculating the content of the derivative by an external standard method.

The technical scheme of the invention is further improved as follows: the liquid chromatography conditions of the step (3) are as follows:

a chromatographic column: the stationary phase is octadecylsilane chemically bonded silica;

mobile phase: 0.03-0.1% trifluoroacetic acid aqueous solution-methanol, wherein the proportion of 0.03-0.1% trifluoroacetic acid aqueous solution is 10-30%, and the proportion of methanol is 90-70%;

wavelength: 210 nm;

flow rate: 1.0 mL/min;

column temperature: 35 ℃;

sample introduction amount: 5 μ L.

Due to the adoption of the technical scheme, the invention has the following technical effects:

according to the method, the active Grignard reagent o-chlorobenzyl magnesium chloride is derived to generate the stable o-chlorobenzyl magnesium chloride acetone derivative, and then the quantitative analysis is carried out by adopting gas chromatography or liquid chromatography, so that the quantitative accuracy can be improved, and the problems that the o-chlorobenzyl magnesium chloride which is possibly partially damaged in the conventional titration process is also titrated, the titration end point is easy to have a reverse color phenomenon, and the operation is difficult to control, so that the result accuracy is lower are solved.

The invention carries out quantitative analysis after the derivatization of the Grignard reagent by introducing the derivatization reagent, provides reliable data for synthesis, can effectively utilize the Grignard reagent, reduces side reactions and improves the conversion rate of the synthesis reaction.

Drawings

FIG. 1 is a gas chromatogram of an o-chlorobenzyl magnesium chloride acetone derivative according to the present invention;

FIG. 2 is a liquid chromatogram of an o-chlorobenzyl magnesium chloride acetone derivative of the present invention.

Detailed Description

The present invention will be described in further detail with reference to the drawings and specific embodiments, and the present invention is not limited to the following embodiments, and all simple modifications and changes made in the content of the present specification, or the direct and indirect application to other related fields, are within the scope of the present invention.

The instrument comprises the following steps: an Agilent 7820 gas chromatograph, an FID detector and an Agilent workstation; agilent 1260 series high performance liquid chromatograph equipped with ultraviolet detector, autosampler, column oven, and Agilent workstation.

Reagent: dichlorobenzene (analytically pure), trifluoroacetic acid (chromatographically pure), methanol (chromatographically pure), water (newly distilled water twice), and a reaction solution sample (Hebei Wignefar Biochemical Co., Ltd.).

Preparing an o-chlorobenzyl magnesium chloride acetone derivative working standard: 20g of acetone and 10g of Grignard reaction solution are introduced into a 100ml vacuum reaction tube, the mixture is shaken up, 10g of chloroacetic acid is introduced again, the mixture is shaken up, and the obtained reaction solution is rotary evaporated at 30-40 ℃ to obtain 5.6g of crude product. And (3) carrying out column chromatography on the crude product, selecting 200-mesh 300-mesh silica gel to fill the column, eluting and receiving by adopting petroleum ether and ethyl acetate in a ratio of 50:1-5:1 after sample loading, simultaneously carrying out chromatographic monitoring, combining and desolventizing high-purity eluent to obtain 2.1g of the o-chlorobenzyl magnesium chloride acetone derivative with the content of 98.7%, and obtaining the o-chlorobenzyl magnesium chloride acetone derivative working standard product.

Preparing an o-chlorobenzyl magnesium chloride acetone derivative reaction solution: 10g of acetone and 0.5g of Grignard reaction solution are introduced into a 100ml vacuum reaction tube, and the mixture is shaken up, 1g of chloroacetic acid is introduced again, and the mixture is shaken up to obtain the o-chlorobenzyl magnesium chloride acetone derivative reaction solution.

The content of the o-chlorobenzyl magnesium chloride is measured by adopting gas chromatography or liquid chromatography:

the content of o-chlorobenzyl magnesium chloride is determined by gas chromatography:

(1) preparing an internal standard stock solution: weighing 1.3g of dichlorobenzene in a 200mL volumetric flask, dissolving with methanol, fixing the volume, and shaking up for later use;

(2) preparation of a standard solution: weighing 0.03g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative working standard in a 50mL volumetric flask, transferring 5mL of internal standard stock solution in the volumetric flask, dissolving with methanol, fixing the volume and shaking up;

(3) preparation of sample solution: weighing about 0.5g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative reaction liquid into a 50mL volumetric flask, transferring 5mL of internal standard stock solution into the volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(4) performing gas chromatography detection under the following conditions:

a chromatographic column: DM-624, 30m × 0.53mm, 3.0 um;

nitrogen gas: 10mL/min, air: 300mL/min, hydrogen: 30 mL/min;

sample introduction amount: 1 mu L of the solution;

vaporization temperature: 250 ℃;

detecting the temperature: 250 ℃;

column temperature: the initial temperature is 65-75 deg.C, the temperature is maintained for 0-3min, the temperature is increased to 160 deg.C at a rate of 10 deg.C/min, then to 238 deg.C at a rate of 40 deg.C, and the temperature is maintained for 13min

A detector: FID;

calculating the content x of the derivative by an internal standard method₁The formula is as follows:

content of derivative

In the formula:

r₂-average value of peak area ratio of o-chlorobenzyl magnesium chloride acetone derivative to internal standard in sample solution;

r₁-average value of peak area ratio of o-chlorobenzyl magnesium chloride acetone derivative to internal standard in standard sample solution;

m₁-mass in g of o-chlorobenzyl magnesium chloride acetone derivative working standard;

m₂-mass in g of sample of o-chlorobenzyl magnesium chloride acetone derivative;

the content of the P-o-chlorobenzyl magnesium chloride acetone derivative as a working standard substance in unit percent;

(5) calculating the content x of the o-chlorobenzyl magnesium chloride according to the content of the derivative, wherein the calculation formula is as follows:

in the formula:

185.33-molecular weight of o-chlorobenzyl magnesium chloride;

184.66-molecular weight of O-chlorobenzyl magnesium chloride acetone derivative.

(II) determining the content of the o-chlorobenzyl magnesium chloride by liquid chromatography:

(1) preparation of a standard solution: accurately weighing 0.025g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative working standard in a 50ml volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(2) preparation of sample solution: accurately weighing about 1.0g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative-containing reaction liquid in a 50ml volumetric flask, dissolving with methanol, fixing the volume and shaking up;

(3) performing liquid chromatography detection under the following conditions:

a chromatographic column: c18(250 mm. times.4.6 mm, 5 um);

mobile phase: 0.03-0.1% trifluoroacetic acid aqueous solution-methanol (10-30: 90-70);

wavelength: 210 nm;

flow rate: 1.0 ml/min;

column temperature: 35 ℃;

sample introduction: 5 ul;

calculating the content x of the derivative by external standard method₁The formula is as follows:

content of derivative:

in the formula:

A₂-average value of the peak area of the o-chlorobenzyl magnesium chloride acetone derivative in the sample solution;

A₁-average value of peak area of o-chlorobenzyl magnesium chloride acetone derivative in standard sample solution;

m₁-mass in g of o-chlorobenzyl magnesium chloride acetone derivative working standard;

m₂-mass in g of sample of o-chlorobenzyl magnesium chloride acetone derivative;

the content of the P-o-chlorobenzyl magnesium chloride acetone derivative as a working standard substance in unit percent;

(4) calculating the content x of the o-chlorobenzyl magnesium chloride according to the content of the derivative, wherein the calculation formula is as follows:

content of o-chlorobenzyl magnesium chloride:

in the formula:

185.33-molecular weight of o-chlorobenzyl magnesium chloride

184.66 molecular weight of O-chlorobenzyl magnesium chloride acetone derivative

Example 1

Determining the content of o-chlorobenzyl magnesium chloride by gas chromatography:

(1) preparing an internal standard stock solution: weighing 1.3g of dichlorobenzene in a 200mL volumetric flask, dissolving with methanol, fixing the volume, and shaking up for later use;

(2) preparation of a standard solution: weighing 0.0310g of o-chlorobenzyl magnesium chloride acetone derivative working standard in a 50mL volumetric flask, transferring 5mL of internal standard stock solution in the volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(3) preparation of sample solution: weighing 0.5003g of o-chlorobenzyl magnesium chloride acetone derivative reaction liquid (batch number: 20210311) in a 50mL volumetric flask, transferring 5mL of internal standard stock solution in the volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(4) performing gas chromatography detection, calculating the content of the derivative by an internal standard method,

the gas chromatography conditions were as follows:

a chromatographic column: DM-624, 30m × 0.53mm, 3.0 um;

nitrogen gas: 10mL/min, air: 300mL/min, hydrogen: 30 mL/min;

sample introduction amount: 1 mu L of the solution;

vaporization temperature: 250 ℃;

detecting the temperature: 250 ℃;

column temperature: the initial temperature is 70 deg.C, the temperature is maintained for 1min, the temperature is increased to 160 deg.C at a rate of 10 deg.C/min, the temperature is increased to 238 deg.C at a rate of 40 deg.C, and the temperature is maintained for 13min

A detector: FID;

content of derivative:

(5) calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative:

example 2

Determining the content of o-chlorobenzyl magnesium chloride by liquid chromatography:

(1) preparation of a standard solution: accurately weighing 0.0274g of o-chlorobenzyl magnesium chloride acetone derivative working standard substance in a 50ml volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(2) preparation of sample solution: 0.9071g of o-chlorobenzyl magnesium chloride acetone derivative reaction liquid (batch number: 20210311) is accurately weighed in a 50ml volumetric flask, dissolved by methanol and subjected to constant volume, and shaken up;

(3) performing liquid chromatography detection, calculating the content of the derivative according to an external standard method,

the liquid chromatography conditions were as follows:

a chromatographic column: c18(250 mm. times.4.6 mm, 5 um);

mobile phase: methanol-0.03% aqueous trifluoroacetic acid (80: 20);

wavelength: 210 nm;

flow rate: 1.0 ml/min;

column temperature: 35 ℃;

sample introduction: 5 ul;

content of derivative:

(4) calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative:

example 3

Determining the content of o-chlorobenzyl magnesium chloride by gas chromatography:

(1) preparing an internal standard stock solution: weighing 1.3g of dichlorobenzene in a 200mL volumetric flask, dissolving with methanol, fixing the volume, and shaking up for later use;

(2) preparation of a standard solution: weighing 0.0348g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative working standard in a 50mL volumetric flask, transferring 5mL of internal standard stock solution in the volumetric flask, dissolving with methanol, fixing the volume and shaking up;

(3) preparation of sample solution: weighing about 0.5205g (accurate to 0.0002g) of o-chlorobenzyl magnesium chloride acetone derivative reaction solution (batch number: 20210412) in a 50mL volumetric flask, transferring 5mL of internal standard stock solution in the volumetric flask, dissolving with methanol, fixing the volume and shaking up;

(4) performing gas chromatography detection, calculating the content of the derivative by an internal standard method,

the gas chromatography conditions were as follows:

a chromatographic column: DM-624, 30m × 0.53mm, 3.0 um;

nitrogen gas: 10mL/min, air: 300mL/min, hydrogen: 30 mL/min;

sample introduction amount: 1 mu L of the solution;

vaporization temperature: 250 ℃;

detecting the temperature: 250 ℃;

column temperature: the initial temperature is 75 ℃, the temperature is kept for 3min, the temperature is increased to 160 ℃ at the temperature increasing rate of 10 ℃/min, the temperature is increased to 238 ℃ at the temperature increasing rate of 40 ℃, and the temperature is kept for 13min

A detector: FID;

content of derivative:

(5) calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative:

example 4

Determining the content of o-chlorobenzyl magnesium chloride by liquid chromatography:

(1) preparation of a standard solution: accurately weighing 0.0254g of o-chlorobenzyl magnesium chloride acetone derivative working standard substance in a 50ml volumetric flask, dissolving with methanol, fixing the volume, and shaking up;

(2) preparation of sample solution: 1.0212g of o-chlorobenzyl magnesium chloride acetone derivative reaction liquid (batch number: 20210412) is accurately weighed in a 50ml volumetric flask, dissolved by methanol and subjected to constant volume, and shaken up;

(3) performing liquid chromatography detection, calculating the content of the derivative according to an external standard method,

the liquid chromatography conditions were as follows:

a chromatographic column: c18(250 mm. times.4.6 mm, 5 um);

mobile phase: methanol-0.1% trifluoroacetic acid in water (70:30)

Wavelength: 210 nm;

flow rate: 1.0 ml/min;

column temperature: 35 ℃;

sample introduction: 5 ul;

content of derivative:(4) calculating the content of the o-chlorobenzyl magnesium chloride according to the content of the derivative: