A high performance liquid phase detection and content determination method of chlorohydrin imine

Abstract

The invention discloses a high-performance liquid phase detection and content determination method of chlorohydrin-imine, which adopts a ZORBAXSB-C18 chromatographic column and a PAD detector, takes 0.1% phosphoric acid solution + acetonitrile as the mobile phase, and quantifies by an external standard method. The concentration of chlorohydrin-imine of the invention has a good linear relationship within the range of 0.05 to 1.00 mg / L, and the linear equation is Y=128852X+787.02, and the linear correlation coefficient R 2 =0.9998. The LOD of this method for chlorohydrin imine is 0.05 mg / L, and the LOQ is 0.10 mg / L. The target peak RSD R.T. 0.04%, RSD Area The method of the invention has the characteristics of simple and rapid operation, good separation effect, strong specificity, high precision and accuracy.

Description

Technical Field

[0001] The invention belongs to the technical field of organic compound detection, and in particular relates to a high performance liquid phase detection and content determination method of chlorohydrin imine. Background Art

[0002] The chemical name of chlorohydrin imine is (2S)-1-chloro-3-[[(4-chlorophenyl)methylene]amino]-2-propanol, and its molecular formula is C 10 H 11 Cl2NO, CAS registration number is 1345879-87-9, and its chemical structure is as follows:

[0003]

[0004] Chlorohydrin-imidazole is a chemical raw material, commonly used as a pharmaceutical intermediate, and has broad application prospects in the fields of pesticides, biopharmaceuticals, and chemicals. Chemical raw materials must be quality tested before production and application to ensure their application effect and safety. At present, there is no literature report on the qualitative and quantitative detection method of chlorohydrin-imidazole.

[0005] The invention utilizes high performance liquid phase technology to establish a quantitative and qualitative detection method for chlorohydrin imines which is simple to operate, efficient, has good separation, high accuracy and strong specificity. Summary of the invention

[0006] The invention aims to overcome the fact that the quality control technology of chlorohydrin-imines in the prior art is still blank, and to provide a high performance liquid phase detection and content determination method of chlorohydrin-imines.

[0007] The present invention provides a high-performance liquid phase detection and content determination method for chlorohydrin-imine, and the detection steps are as follows: (1) Preparation of standard working solution: weigh an appropriate amount of chlorohydrin-imine standard, add a solvent to dissolve and dilute to prepare a series of standard working solutions. (2) Preparation of test solution: weigh an appropriate amount of chlorohydrin-imine sample to be tested, add a solvent to dissolve and dilute to prepare a test solution. (3) Determination and calculation: inject a blank solvent, a chlorohydrin-imine test solution and a chlorohydrin-imine standard solution into a high-performance liquid chromatograph for analysis, record the chromatogram, and calculate the content of chlorohydrin-imine in the test solution by an external standard method.

[0008] The solvents were acetonitrile, ultrapure water or n-octanol.

[0009] The HPLC used a ZORBAXSB-C18 column and a PAD detector. The analysis conditions of the HPLC were as follows:

[0010] Injection volume: 25-35 μL;

[0011] Flow rate: 0.8~1.5mL / min;

[0012] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0013] Column temperature: 26-35°C;

[0014] Detection wavelength: 250~260nm.

[0015] Preferably, the injection volume is 28-32 μL, the flow rate is 1.0-1.2 mL / min, and the column temperature is 28-32° C.

[0016] Further preferably, the injection volume is 30 μL, the flow rate is 1.0 mL / min, and the column temperature is 30° C.

[0017] Preferably, the detection wavelength is 254 nm.

[0018] Preferably, the concentration of the sample solution is 0.10 to 10.0 mg / L.

[0019] Preferably, the concentration of the standard working solution is 0.05-10.00 mg / L.

[0020] A high performance liquid phase detection and content determination method of chlorohydrin imine, the content determination specific steps are as follows:

[0021] ① Preparation of standard working solution: Accurately weigh 0.10025g of chlorohydrin imine standard into a 100mL volumetric flask, dissolve it with acetonitrile to the mark, shake well to obtain a standard mother solution with a concentration of 1002.5mg / L, take 1.00mL of the standard mother solution into a 100mL volumetric flask, dissolve it with acetonitrile to the mark, shake well to obtain a standard reserve solution with a concentration of 10.00mg / L, take an appropriate amount of the standard reserve solution and dilute it with acetonitrile to prepare a series of standard working solutions with chlorohydrin imine concentrations of 0.05, 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00mg / L.

[0022] ② Preparation of test solution: Weigh 0.01-1.00 mg of chlorohydrin imine sample to be tested into a 100 mL volumetric flask, dilute to the mark with ultrapure water or n-octanol, mix well, and obtain the chlorohydrin imine test solution.

[0023] ③ Drawing of standard curve: Pipette 25-35 μL of the series of standard working solutions in step ① for HPLC determination, and draw a standard curve with the concentration of the series of standard working solutions as the abscissa and the peak area of ​​chlorohydrin imine as the ordinate, and calculate the regression equation and correlation coefficient.

[0024] ④ Sample determination and content calculation: Pipette 25-35 μL of the test solution in step ② for HPLC determination to obtain the peak area of ​​chlorohydrin imine in the test sample. Substitute the peak area into the linear regression equation in the above step ③ for calculation to obtain the concentration of chlorohydrin imine in the test sample.

[0025] The linear regression equation of the chlorohydrin imine in this method is Y=128852X+787.02, and the linear correlation coefficient R 2 =0.9998, linear range is 0.05~1.00mg / L.

[0026] The beneficial effects of the present invention are as follows:

[0027] 1. The present invention adopts LC2030CSDPlus high performance liquid chromatograph and uses ZORBAX SB-C 18 The chromatographic column and PAD detector carry out high performance liquid chromatography analysis on chlorohydrin imine, and the present invention can accurately detect the chlorohydrin imine in acetonitrile, ultrapure water and n-octanol, and the chlorohydrin imine in the sample is assayed by an external standard method, which provides a technical reference for the quality control of the chlorohydrin imine production and application process, and ensures that its quality is controllable. The chlorohydrin imine of the present invention peaks at 4.3min, and the blank sample has no interference peak at this time point. The detection method of the present invention is efficient, has good target peak separation, and is low in cost. The method is suitable for qualitative and quantitative detection of chlorohydrin imine in large-scale production, and is also suitable for detection and analysis in laboratories.

[0028] 2. The validation test of the analytical method of the present invention shows that the linear relationship is good in the range of 0.05-1.00 mg / L of chlorohydrin imine concentration, the linear equation is Y=128852X+787.02, and the linear correlation coefficient R 2 =0.9998. The detection limit LOD of this method for chlorohydrin imine is 0.05mg / L, and the LOQ is 0.10mg / L. Target peak RSD R.T. 0.04%, RSD Area The recovery rate of chlorohydrin imine in ultrapure water is 94.4% to 98.3%, and the recovery rate in n-octanol is 90.0% to 98.9%. The method of the invention has the advantages of simple operation, rapidity, good separation effect, strong specificity, high precision and accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is the linear relationship diagram of chlorohydrin imine;

[0030] Figure 2 This is a typical spectrum of chlorohydrin imine;

[0031] Figure 3 is the blank spectrum of acetonitrile;

[0032] Figure 4 is the water blank spectrum;

[0033] Figure 5 This is the blank spectrum of n-octanol;

[0034] Figure 6 Typical spectrum of water recovery spike solution (low concentration);

[0035] Figure 7 This is a typical spectrum of the spiked solution for water recovery (high concentration);

[0036] Figure 8 This is a typical spectrum of the spiked solution for n-octanol recovery (low concentration);

[0037] Fig. 9 This is a typical spectrum of the spiked solution for n-octanol recovery (high concentration). DETAILED DESCRIPTION

[0038] The present invention is further described below in conjunction with the accompanying drawings and embodiments, but is not intended to limit the protection scope of the present invention.

[0039] The main instruments, equipment, reagents and solutions used in the present invention are as follows:

[0040] 1. Main instruments and equipment

[0041] (1) High performance liquid chromatograph: LC2030CSDPlus;

[0042] (2) Electronic balance: Mettler-Toledo (China) Co., Ltd., model: XSE105DU;

[0043] (3) Gas bath constant temperature oscillator: Jiangsu Jintan Honghua Instrument Factory, model: ZD-85A, LHY190-1;

[0044] (4) Xiangyi desktop high-speed centrifuge: Hunan Xiangyi Laboratory Instrument Development Co., Ltd., model: H1850.

[0045] 2. Main reagents and solutions

[0046] (1) Phosphoric acid: high purity, Tianjin Komiou Chemical Reagent Co., Ltd.;

[0047] (2) Acetonitrile: HPLC, Shanghai Anpu Laboratory Technology Co., Ltd.;

[0048] (3) Ultrapure water: resistivity, 18.2 MΩ×cm.

[0049] Embodiment 1:

[0050] The high performance liquid phase detection method of chlorohydrin imine in water has the following steps:

[0051] (1) Preparation of standard working solution: accurately weigh 0.10025 g of chlorohydrin imine standard into a 100 mL volumetric flask, dissolve it with acetonitrile to the mark, and shake well to obtain a standard mother solution with a concentration of 1002.5 mg / L; take 1.00 mL of the standard mother solution into a 100 mL volumetric flask, dissolve it with acetonitrile to the mark, and shake well to obtain a standard stock solution with a concentration of 10.00 mg / L; take an appropriate amount of the standard stock solution and dilute it with acetonitrile to prepare a series of standard working solutions with chlorohydrin imine concentrations of 0.05, 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00 mg / L.

[0052] (2) Preparation of test solution: Pipette 1.00 mL of the 10.0 mg / L standard stock solution prepared in step (1) into a 100 mL volumetric flask, dilute to volume with ultrapure water, and mix well to obtain a test solution with a theoretical concentration of 0.10 mg / L of chlorohydrin imine.

[0053] (3) Drawing of standard curve: 30 μL of the series of standard working solutions in step (1) was injected into a high performance liquid chromatograph for analysis. The results are shown in Table 1. The concentration of the series of standard working solutions was used as the abscissa and the peak area of ​​chlorohydrin imine was used as the ordinate to draw a standard curve. The regression equation and correlation coefficient were calculated. The linear regression equation of chlorohydrin imine is:

[0054] Y=128852X+787.02, linear correlation coefficient R2=0.9998, standard curve see Figure 1 .

[0055] Table 1: Test results of chlorohydrin imine series standard working solutions

[0056]

[0057] (4) Determination and calculation of sample content: 30 μL of acetonitrile blank solvent, ultrapure water blank solvent and the test solution in step (2) were injected into a high performance liquid chromatograph for analysis to obtain the peak area of ​​chlorohydrin imine in the test solution. The peak area was substituted into the above linear regression equation for calculation to obtain the concentration of chlorohydrin imine in ultrapure water.

[0058] (5) High performance liquid chromatography analysis conditions are:

[0059] Flow rate: 1.0 mL / min;

[0060] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0061] Column temperature: 30°C;

[0062] Detection wavelength: 254nm.

[0063] Embodiment 2:

[0064] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0065] Except for the different test sample concentration in step (2), other steps, conditions, and reagent dosages are the same as those in Example 1.

[0066] The preparation steps of the test solution are as follows: 1.00 mL of the standard mother solution with a concentration of 1002.5 mg / L in step (1) is taken into a 100 mL volumetric flask, the volume is made up to the mark with ultrapure water, and mixed evenly to obtain a test solution with a theoretical concentration of 10.0 mg / L of chlorohydrin imine.

[0067] Embodiment 3:

[0068] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0069] Except that the aspirated amount of the standard working solution, acetonitrile blank solvent, ultrapure water blank solvent and the test solution in step (2) was changed to 25 μL, and the HPLC analysis conditions were changed to: flow rate of 0.8 mL / min, column temperature of 26° C., and detection wavelength of 250 nm, the other steps, conditions, and reagent amounts were the same as those in Example 1.

[0070] Embodiment 4:

[0071] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0072] Except that the aspirated amount of the standard working solution, acetonitrile blank solvent, ultrapure water blank solvent and the test solution in step (2) was changed to 28 μL, and the HPLC analysis conditions were changed to: flow rate of 0.9 mL / min, column temperature of 28° C., and detection wavelength of 252 nm, the other steps, conditions, and reagent amounts were the same as in Example 1.

[0073] Embodiment 5:

[0074] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0075] Except that the aspirated amount of the standard working solution, acetonitrile blank solvent, ultrapure water blank solvent and the test solution in step (2) was changed to 32 μL, and the HPLC analysis conditions were changed to: flow rate of 1.2 mL / min, column temperature of 32° C., and detection wavelength of 256 nm, the other steps, conditions, and reagent amounts were the same as in Example 1.

[0076] Embodiment 6:

[0077] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0078] Except that the aspirated amount of the standard working solution, acetonitrile blank solvent, ultrapure water blank solvent and the test solution in step (2) was changed to 35 μL, and the HPLC analysis conditions were changed to: flow rate of 1.5 mL / min, column temperature of 35° C., and detection wavelength of 268 nm, the other steps, conditions, and reagent amounts were the same as in Example 1.

[0079] Embodiment 7:

[0080] High performance liquid chromatography method for detection of chlorohydrin imines in water.

[0081] Except that the detection wavelength of the HPLC analysis conditions was changed to 260 nm, other steps, conditions and reagent amounts were the same as those in Example 1.

[0082] Embodiment 8-14:

[0083] The high performance liquid chromatography detection method of chlorohydrin imine in n-octanol is as follows:

[0084] Except for the different solvents used in step (2), the other steps, chromatographic conditions, and reagent amounts are the same as those in Examples 1-6.

[0085] In Examples 7-12, the preparation steps of the test solution in step (2) are as follows: 1.00 mL of the standard stock solution with a concentration of 10.0 mg / L in step (1) of Example 1 is taken into a 100 mL volumetric flask, the volume is made up to the mark with n-octanol, and the solution is mixed evenly to obtain the test solution.

[0086] Experimental example:

[0087] In order to illustrate the beneficial effects of the present invention, the present invention provides the following test examples:

[0088] 1. High performance liquid chromatography analysis condition screening test

[0089] 1.1 Screening test with different injection volumes

[0090] An injection volume of 25 to 35 μL was selected for the screening test, and other chromatographic conditions were as follows:

[0091] Chromatographic column: ZORBAXSB-C18;

[0092] Flow rate: 1.0 mL / min;

[0093] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0094] Column temperature: 30°C;

[0095] Detection wavelength: 254nm.

[0096] Test results: Under the above chromatographic conditions, 25 μL, 28 μL, 30 μL, 32 μL, and 35 μL of the test solution in Example 1 were taken for measurement. The injection volume was within the range of 25 to 35 μL and the peak separation could meet the detection requirements with good reproducibility. The peak shape was better when the injection volume was 30 μL, so the preferred injection volume was 30 μL.

[0097] 1.2 Screening test at different flow rates

[0098] A flow rate of 0.8 to 1.5 mL / min was selected for the screening test, and other chromatographic conditions were as follows:

[0099] Chromatographic column: ZORBAXSB-C18;

[0100] Injection volume: 30 μL;

[0101] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0102] Column temperature: 30°C;

[0103] Detection wavelength: 254nm.

[0104] Test results: Under the above chromatographic conditions, the test solution in Example 1 was measured at flow rates of 0.8 mL / min, 1.0 mL / min, 1.2 mL / min, and 1.5 mL / min, respectively. The flow rate in the range of 0.8 to 1.5 mL / min can achieve the effect of detecting chlorohydrin imine, and the peak separation can meet the detection requirements with good reproducibility. When the flow rate is 1.0 mL / min, the target peak has the shortest peak time, so the preferred flow rate is 1.0 mL / min.

[0105] 1.3 Screening test at different column temperatures

[0106] A column temperature of 26-35°C was selected for the screening test, and other chromatographic conditions were as follows:

[0107] Chromatographic column: ZORBAXSB-C18;

[0108] Injection volume: 30 μL;

[0109] Flow rate: 1.0 mL / min;

[0110] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0111] Detection wavelength: 254nm.

[0112] Test results: Under the above chromatographic conditions, the column temperature was set at 26°C, 28°C, 30°C, 32°C, and 35°C, respectively, and the test solution in Example 1 was measured. The column temperature in the range of 26 to 35°C can achieve the effect of detecting chlorohydrin imine, and the peak separation can meet the detection requirements. When the column temperature is 30°C, the peak elution time of the target peak is the shortest, so the preferred column temperature is 30°C.

[0113] 1.4 Screening test with different detection wavelengths

[0114] A detection wavelength of 250-260 nm was selected for the screening test, and other chromatographic conditions were as follows:

[0115] Chromatographic column: ZORBAXSB-C18;

[0116] Injection volume: 30 μL;

[0117] Flow rate: 1.0 mL / min;

[0118] Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; A:B=30:70 (v / v);

[0119] Column temperature: 30℃.

[0120] Test results: Under the above chromatographic conditions, the detection wavelengths of 250nm, 252nm, 254nm, 256nm, 258nm, and 260nm were selected to measure the test solution in Example 1. The detection wavelength can achieve the effect of detecting chlorohydrin imines within the range of 250 to 260nm, and the peak separation can meet the detection requirements. When the detection wavelength is 254nm, the separation effect and peak shape are the best, so the preferred detection wavelength is 254nm.

[0121] 2. Methodology validation test

[0122] 2.1 Exclusivity

[0123] The HPLC analysis method established by the present invention was used to measure the blank samples of solvent acetonitrile, water and n-octanol, and the spectra of the blank samples of acetonitrile, water and n-octanol were compared with those of the tested samples. The results are shown in Figure 2-Figure 5 , the peak of chlorohydrin imine appeared at 4.3min, and the blank sample had no interfering peak at this time point, indicating that this method has good specificity for chlorohydrin imine.

[0124] 2.2 Limit of detection and limit of quantification

[0125] The spectrum of the standard working solution with a concentration of 0.05 mg / L was analyzed. At this concentration, the signal-to-noise ratio (S / N) of the target peak was 42.03. Therefore, the LOD of this method can be considered to be 0.05 mg / L; after analyzing the spectrum of the standard working solution with a concentration of 0.10 mg / L, at this concentration, the signal-to-noise ratio (S / N) of the target peak was 114.03. Therefore, 0.10 mg / L can be used as the LOQ of this analytical method.

[0126] 2.3 Precision

[0127] The same standard working solution with a concentration of 0.40 mg / L was injected 6 times continuously to determine the precision of the analytical method. The RSD R.T. 0.04%, RSD Area The results show that the analytical precision of this analysis method for the same sample meets the requirements, and the precision test results are shown in Table 2.

[0128] Table 2: Precision test results

[0129]

[0130] 2.4 Recovery test

[0131] 2.4.1 Recovery rate test in water

[0132] Pipette 1.00 mL of the standard stock solution with a concentration of 10.0 mg / L into a 100 mL volumetric flask, dilute to the mark with ultrapure water, and mix well to obtain a low-concentration recovery test solution with a concentration of 0.10 mg / L of chlorohydrin imine. Repeat the above operation to prepare 5 portions of a low-concentration recovery test solution with a concentration of 0.10 mg / L.

[0133] Pipette 1.00 mL of the standard mother solution with a concentration of 1002.5 mg / L into a 100 mL volumetric flask, dilute to the mark with ultrapure water, and mix well to obtain a high-concentration recovery test solution with a concentration of 10.0 mg / L of chlorohydrin imine. Repeat the above operation to prepare 5 portions of a high-concentration recovery test solution with a concentration of 10.0 mg / L, and dilute 20 times with acetonitrile before measurement.

[0134] At the same time, prepare an ultrapure water blank sample.

[0135] The above-mentioned low-concentration recovery test solution, high-concentration recovery test solution, and ultrapure water blank sample were sampled and tested according to the high-performance liquid chromatography analysis conditions of the present invention, and the recovery was calculated. The results are shown in Table 3.

[0136] Table 3: Results of water recovery test

[0137]

[0138] 2.4.2 Recovery test in n-octanol

[0139] Pipette 1.00 mL of the standard stock solution with a concentration of 10.0 mg / L into a 100 mL volumetric flask, dilute to the mark with n-octanol, mix well, and obtain a low-concentration recovery test solution with a concentration of 0.10 mg / L of chlorohydrin imine. Repeat the above operation to prepare 5 portions of a low-concentration recovery test solution with a concentration of 0.10 mg / L.

[0140] Pipette 1.00 mL of the standard mother solution with a concentration of 1002.5 mg / L into a 100 mL volumetric flask, dilute to the mark with n-octanol, and mix well to obtain a high-concentration recovery test solution with a concentration of 10.0 mg / L of chlorohydrin imine. Repeat the above operation to prepare 5 portions of a high-concentration recovery test solution with a concentration of 10.0 mg / L, and dilute 20 times with acetonitrile before measurement.

[0141] At the same time, prepare an octanol blank sample.

[0142] The above-mentioned low-concentration recovery test solution, high-concentration recovery test solution, and n-octanol blank sample were sampled and tested according to the high-performance liquid chromatography analysis conditions of the present invention, and the recovery was calculated. The results are shown in Table 4.

[0143] Table 4: Recovery in n-octanol

[0144]

[0145]

[0146] Although the present invention has been described in detail above by combining preferred embodiments with reference to the accompanying drawings, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person skilled in the art who is familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which shall be within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be subject to the scope of protection of the claims.

Claims

1. A high performance liquid phase detection and content determination method of chlorohydrin-imines, the chemical structural formula of the chlorohydrin-imines is: It is characterized in that The steps of the high performance liquid phase detection and content determination method are as follows: (1) Preparation of standard working solution: weigh an appropriate amount of chlorohydrin imine standard, add solvent to dissolve and dilute to prepare a series of standard working solutions; (2) Preparation of test solution: Weigh an appropriate amount of the sample of chlorohydrin imine to be tested, add a solvent to dissolve and dilute it to prepare a test solution; (3) Determination and calculation: inject the blank solvent, the test solution of chlorohydrin-imine and the standard solution of chlorohydrin-imine into a high performance liquid chromatograph for analysis, record the chromatogram, and calculate the content of chlorohydrin-imine in the test solution by the external standard method; The solvent is acetonitrile, ultrapure water or n-octanol; The HPLC uses a ZORBAX SB-C18 chromatographic column and a PAD detector, and the analysis conditions of the HPLC are: Injection volume: 25-35 μL; Flow rate: 0.8~1.5mL / min; Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; the volume ratio of A to B is 30:70; Column temperature: 26-35°C; Detection wavelength: 250~260nm.

2. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 1, characterized in that, The injection volume is 28-32 μL, the flow rate is 1.0-1.2 mL / min, and the column temperature is 28-32° C.

3. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 2, characterized in that, The injection volume was 30 μL, the flow rate was 1.0 mL / min, and the column temperature was 30° C.

4. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 1, characterized in that, The detection wavelength is 254 nm.

5. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 1, characterized in that, The concentration of the sample solution is 0.10-10.0 mg / L.

6. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 1, characterized in that, The concentration of the standard working solution is 0.05~10.00mg / L.

7. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 1, characterized in that, The specific steps of content determination are as follows: ① Preparation of standard working solution: accurately weigh 0.10025g of chlorohydrin imine standard in a 100mL volumetric flask, dissolve it in acetonitrile to the mark, shake well to obtain a standard mother solution with a concentration of 1002.5mg / L, take 1.00mL of the standard mother solution in a 100mL volumetric flask, dissolve it in acetonitrile to the mark, shake well to obtain a standard reserve solution with a concentration of 10.00mg / L, draw an appropriate amount of the standard reserve solution and dilute it with acetonitrile to prepare a series of standard working solutions with chlorohydrin imine concentrations of 0.05, 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00mg / L; ② Preparation of test solution: weigh 0.01-1.00 mg of chlorohydrin imine sample to be tested into a 100 mL volumetric flask, dilute to the mark with ultrapure water or n-octanol, mix well, and obtain chlorohydrin imine test solution; ③ Drawing of standard curve: Pipette 25-35 μL of the series of standard working solutions in step ① for HPLC determination, use the concentration of the series of standard working solutions as the abscissa and the peak area of ​​chlorohydrin imine as the ordinate to draw a standard curve, and calculate the regression equation and correlation coefficient; ④ Sample determination and content calculation: Pipette 25-35 μL of the test solution in step ② for HPLC determination to obtain the peak area of ​​chlorohydrin imine in the test sample. Substitute the peak area into the linear regression equation in the above step ③ for calculation to obtain the concentration of chlorohydrin imine in the test sample.

8. A high performance liquid phase detection and content determination method of chlorohydrin imine according to claim 7, characterized in that, The linear regression equation of chlorohydrin imine is Y=128852X+787.02, and the linear correlation coefficient R2=0.9998.

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