A method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultrahigh performance liquid chromatography

Abstract

The present invention discloses a method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultrahigh performance liquid chromatography. The method uses an Agilent Infinity II Prime 1260 ultrahigh performance liquid chromatograph and a VWD detector, a Shim-pack GISTC18, 250 mm × 4.6 mm, 5 μm chromatographic column, and 0.1% aqueous phosphoric acid solution and acetonitrile as mobile phases to detect 2-amino-3,4-difluorobenzaldehyde. The linear relationship of the 2-amino-3,4-difluorobenzaldehyde in the range of 0.500 mg / L to 10.0 mg / L is good, the linear equation is y=90.74x+1.16, and the linear correlation coefficient R is 0. 2 =0.9998; the LOD of 2-amino-3,4-difluorobenzaldehyde in test water is 0.0108 mg / L, and the LOQ is 0.0359 mg / L; the LOD in algae culture medium is 0.0100 mg / L, and the LOQ is 0.0334 mg / L, which can meet the requirements for detecting trace sample concentrations. The detection method of the present invention has the characteristics of strong specificity, high accuracy, short peak time, good reproducibility, and simple operation.

Description

Technical Field

[0001] The invention belongs to the technical field of organic compound detection, and particularly relates to a method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium by utilizing ultrahigh performance liquid chromatography. Background Art

[0002] 2-Amino-3,4-difluorobenzaldehyde (FDFA), with CAS number 1602097-79-9, molecular formula C7H5F2NO, molecular weight 157.1, and structural formula as follows:

[0003]

[0004] 2-Amino-3,4-difluorobenzaldehyde is an important chemical raw material, widely used as a pesticide intermediate. Currently, there are few reports on 2-amino-3,4-difluorobenzaldehyde, and no quality control methods for 2-amino-3,4-difluorobenzaldehyde have been published. Therefore, establishing a detection method for 2-amino-3,4-difluorobenzaldehyde is crucial for its industrial application and quality control during application. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium by using ultrahigh performance liquid chromatography, so as to achieve quality control of 2-amino-3,4-difluorobenzaldehyde.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] A method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultrahigh performance liquid chromatography (UHPLC) comprises the following steps: (1) drawing a standard curve: accurately weighing 0.05015 g of a 2-amino-3,4-difluorobenzaldehyde standard into a 50 mL volumetric flask, dissolving it with acetonitrile and diluting it to the mark, shaking it evenly to prepare a 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 1000 mg / L, taking an appropriate amount of the standard stock solution, diluting it with acetonitrile to prepare a series of standard working solutions with a concentration of 0.500 to 10.0 mg / L, using the solvent as a blank control, determining the peak area under UHPLC conditions, drawing a standard curve, and obtaining a linear regression equation; (2) preparing a test solution: accurately weighing 1.30371 g of each of the two solutions; and The 2-amino-3,4-difluorobenzaldehyde sample was dissolved in acetonitrile in a 25 mL volumetric flask and diluted to the mark, and then shaken to obtain a 2-amino-3,4-difluorobenzaldehyde sample stock solution with a concentration of 52003 mg / L; an appropriate amount of the sample stock solution was respectively drawn into two 100 mL volumetric flasks, and diluted with test water and algae culture medium, respectively, to obtain the test sample solution to be tested, and then subjected to ultra-high performance liquid chromatography determination; (3) Determination of the test sample: the instrument operating conditions of the ultra-high performance liquid chromatography were set, and after the instrument was stable, the test sample solution was subjected to ultra-high performance liquid chromatography determination, and the content of 2-amino-3,4-difluorobenzaldehyde in the test sample solution was calculated according to the linear regression equation.

[0008] In step (1), the concentrations of the series of standard working solutions are 0.500 mg / L, 1.00 mg / L, 2.00 mg / L, 4.00 mg / L, 8.00 mg / L, and 10.0 mg / L, respectively.

[0009] The ultrahigh performance liquid chromatography (UHPLC) method of the present invention adopts an Agilent Infinity II Prime 1260 chromatograph, and the instrument operating conditions of the UHPLC are as follows: chromatographic column: Shim-pack GIST C18, 250 mm×4.6 mm, 5 μm; injection volume: 2.0 to 8.0 μL; flow rate: 0.4 to 0.80 mL / min; detection wavelength: 218 to 223 nm; mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; column temperature: 32 to 38°C.

[0010] Preferably, the injection volume is 5.00 μL.

[0011] Preferably, the flow rate is 0.60 mL / min.

[0012] Preferably, the detection wavelength is 221 nm.

[0013] Preferably, the volume ratio of mobile phase A to mobile phase B is 30:70.

[0014] Preferably, the column temperature is 35°C.

[0015] The linear regression equation of the present invention is y=90.74x+1.16, and the linear correlation coefficient R 2 is 0.9998.

[0016] The LOD of the present invention in test water is 0.0108 mg / L, and the LOQ is 0.0359 mg / L; the LOD in algae culture medium is 0.0100 mg / L, and the LOQ is 0.0334 mg / L.

[0017] The beneficial effects of the present invention are:

[0018] 1. The present invention provides for the first time a method for detecting 2-amino-3,4-difluorobenzaldehyde using ultrahigh performance liquid chromatography, which provides a technical reference for quality control in the subsequent production and application process of 2-amino-3,4-difluorobenzaldehyde and related research. The detection method of the present invention has strong specificity, high accuracy, short peak time, good reproducibility and simple operation.

[0019] 2. The present invention has been investigated through linear tests and has a good linear relationship in the range of 0.500 mg / L to 10.0 mg / L. The linear equation is y=90.74x+1.16 and the linear correlation coefficient R 2 =0.9998.

[0020] 3. In the specificity test of the present invention, by comparing the spectra of blank samples of test water and algae culture medium and samples with added recovery rate, 2-amino-3,4-difluorobenzaldehyde peaked at 6.9 minutes, and the blank sample had no interference peak at this time point, indicating good specificity.

[0021] 4. According to the detection limit and quantification limit tests of the present invention, the LOD of 2-amino-3,4-difluorobenzaldehyde in the test water is 0.0108 mg / L, and the LOQ is 0.0359 mg / L; the LOD in the algae culture medium is 0.0100 mg / L, and the LOQ is 0.0334 mg / L, which can meet the detection requirements of trace sample concentrations.

[0022] 5. The recovery rate and repeatability test results of the present invention show that the average addition recoveries of 2-amino-3,4-difluorobenzaldehyde at two different concentrations in test water are 87.7% and 97.1%, respectively, and the relative standard deviations of the recoveries are 4.28% and 0.201%, respectively; the average addition recoveries of 2-amino-3,4-difluorobenzaldehyde at two different concentrations in algae culture medium are 92.2% and 102%, respectively, and the relative standard deviations of the recoveries are 3.89% and 0.164%, respectively. The present invention has high recovery rate and good repeatability. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1This is the linear fitting curve of 2-amino-3,4-difluorobenzaldehyde in acetonitrile;

[0024] Figure 2 This is a typical spectrum of 2-amino-3,4-difluorobenzaldehyde (4.00 mg / L);

[0025] Figure 3 This is the blank spectrum of test water;

[0026] Figure 4 is the blank atlas of algae culture medium;

[0027] Figure 5 This is a typical spectrum of the low-concentration recovery spiked solution of 2-amino-3,4-difluorobenzaldehyde in test water;

[0028] Figure 6 This is a typical spectrum of the high concentration recovery spiked solution of 2-amino-3,4-difluorobenzaldehyde in test water;

[0029] Figure 7 This is a typical spectrum of the spiked solution with low recovery rate of 2-amino-3,4-difluorobenzaldehyde in algae culture medium;

[0030] Figure 8 This is a typical spectrum of a high-concentration recovery spiked solution of 2-amino-3,4-difluorobenzaldehyde in algae culture medium. DETAILED DESCRIPTION

[0031] The present invention is further described below with reference to specific examples, which are not intended to limit the scope of protection of the present invention.

[0032] Unless otherwise specified, the methods, raw materials, reagents, and instruments involved in the present invention can be prepared by conventional methods or purchased from the market.

[0033] The reagents and instruments used in the following examples include but are not limited to:

[0034] 1. Reagents and solvents

[0035] (1) Acetonitrile: HPLC grade, Shanghai Anpu Laboratory Technology Co., Ltd.;

[0036] (2) Phosphoric acid: HPLC grade, Honeywell (China) Co., Ltd.

[0037] (3) UP water: resistivity, 18.2 MΩ×cm;

[0038] (4) Test water: homemade in the laboratory;

[0039] (5) Algae culture medium: homemade in the laboratory;

[0040] (6) 2-Amino-3,4-difluorobenzaldehyde standard, purity 99.72194%:

[0041] 2. Main instruments and equipment

[0042] (1) Ultra-high performance liquid chromatography: Agilent Infinity II Prime 1260, VWD detector;

[0043] (2) Chromatographic column: Shim-pack GIST C18, 250 mm × 4.6 mm, 5 μm;

[0044] (3) Electronic balance: METTLER TOLEDO, XS105, HFC0044;

[0045] Example 1:

[0046] (1) Drawing of the standard curve: Accurately weigh 0.05015 g of 2-amino-3,4-difluorobenzaldehyde standard in a 50 mL volumetric flask, dissolve it in acetonitrile and dilute to the mark, shake well to prepare a 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 1000 mg / L, take an appropriate amount of the standard stock solution, dilute it with acetonitrile to prepare a series of standard working solutions with concentrations of 0.500 mg / L, 1.00 mg / L, 2.00 mg / L, 4.00 mg / L, 8.00 mg / L, and 10.0 mg / L, respectively. Use the solvent as a blank control, determine the peak area according to the ultra-high performance liquid chromatography conditions, draw a standard curve, and obtain the linear regression equation;

[0047] (2) Preparation of test solution: Accurately weigh 1.30371 g of 2-amino-3,4-difluorobenzaldehyde sample into a 25 mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, shake well, and prepare a 2-amino-3,4-difluorobenzaldehyde sample stock solution with a concentration of 52003 mg / L; respectively, draw appropriate amounts of the sample stock solution into two 100 mL volumetric flasks, dilute with test water and algae culture medium, respectively, to obtain the test solution to be tested, and then determine it by ultra-high performance liquid chromatography;

[0048] (3) Determination of the test sample: Set the operating conditions of the ultra-high performance liquid chromatography instrument. After the instrument is stable, perform ultra-high performance liquid chromatography on the test sample solution and calculate the content of 2-amino-3,4-difluorobenzaldehyde in the test sample solution according to the linear regression equation.

[0049] The instrument operating conditions for the ultrahigh performance liquid chromatography in this example are as follows: an Agilent Infinity II Prime 1260 chromatograph is used, a Shim-pack GIST C18 chromatographic column with specifications of 250 mm × 4.6 mm, 5 μm; an injection volume of 5.0 μL; a flow rate of 0.6 mL / min; a detection wavelength of 221 nm; mobile phases: A is a 0.1% aqueous phosphoric acid solution, and B is acetonitrile, with a volume ratio of mobile phases A to B of 30:70; and a column temperature of 35°C.

[0050] Example 2:

[0051] (1) Drawing of the standard curve: Accurately weigh 0.05015 g of 2-amino-3,4-difluorobenzaldehyde standard in a 50 mL volumetric flask, dissolve it in acetonitrile and dilute to the mark, shake well to prepare a 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 1000 mg / L, take an appropriate amount of the standard stock solution, dilute it with acetonitrile to prepare a series of standard working solutions with concentrations of 0.500 mg / L, 1.00 mg / L, 2.00 mg / L, 4.00 mg / L, 8.00 mg / L, and 10.0 mg / L, respectively. Use the solvent as a blank control, determine the peak area according to the ultra-high performance liquid chromatography conditions, draw a standard curve, and obtain the linear regression equation;

[0052] (2) Preparation of test solution: Accurately weigh 1.30371 g of 2-amino-3,4-difluorobenzaldehyde sample into a 25 mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, shake well, and prepare a 2-amino-3,4-difluorobenzaldehyde sample stock solution with a concentration of 52003 mg / L; respectively, draw appropriate amounts of the sample stock solution into two 100 mL volumetric flasks, dilute with test water and algae culture medium, respectively, to obtain the test solution to be tested, and then determine it by ultra-high performance liquid chromatography;

[0053] (3) Determination of the test sample: Set the operating conditions of the ultra-high performance liquid chromatography instrument. After the instrument is stable, perform ultra-high performance liquid chromatography on the test sample solution and calculate the content of 2-amino-3,4-difluorobenzaldehyde in the test sample solution according to the linear regression equation.

[0054] The instrument operating conditions for the ultrahigh performance liquid chromatography in this example are as follows: an Agilent Infinity II Prime 1260 chromatograph is used, a Shim-pack GIST C18 chromatographic column with specifications of 250 mm × 4.6 mm, 5 μm; an injection volume of 2.0 μL; a flow rate of 0.4 mL / min; a detection wavelength of 218 nm; mobile phases: A is a 0.1% aqueous phosphoric acid solution, and B is acetonitrile, with a volume ratio of mobile phases A to B of 30:70; and a column temperature of 32°C.

[0055] Example 3:

[0056] (1) Drawing of the standard curve: Accurately weigh 0.05015 g of 2-amino-3,4-difluorobenzaldehyde standard in a 50 mL volumetric flask, dissolve it in acetonitrile and dilute to the mark, shake well to prepare a 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 1000 mg / L, take an appropriate amount of the standard stock solution, dilute it with acetonitrile to prepare a series of standard working solutions with concentrations of 0.500 mg / L, 1.00 mg / L, 2.00 mg / L, 4.00 mg / L, 8.00 mg / L, and 10.0 mg / L, respectively. Use the solvent as a blank control, determine the peak area according to the ultra-high performance liquid chromatography conditions, draw a standard curve, and obtain the linear regression equation;

[0057] (2) Preparation of test solution: Accurately weigh 1.30371 g of 2-amino-3,4-difluorobenzaldehyde sample into a 25 mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, shake well, and prepare a 2-amino-3,4-difluorobenzaldehyde sample stock solution with a concentration of 52003 mg / L; respectively, draw appropriate amounts of the sample stock solution into two 100 mL volumetric flasks, dilute with test water and algae culture medium, respectively, to obtain the test solution to be tested, and then determine it by ultra-high performance liquid chromatography;

[0058] (3) Determination of the test sample: Set the operating conditions of the ultra-high performance liquid chromatography instrument. After the instrument is stable, perform ultra-high performance liquid chromatography on the test sample solution and calculate the content of 2-amino-3,4-difluorobenzaldehyde in the test sample solution according to the linear regression equation.

[0059] The instrument operating conditions for the ultrahigh performance liquid chromatography in this example are as follows: an Agilent Infinity II Prime 1260 chromatograph is used, a Shim-pack GIST C18 chromatographic column with specifications of 250 mm × 4.6 mm, 5 μm; an injection volume of 8.0 μL; a flow rate of 0.8 mL / min; a detection wavelength of 223 nm; mobile phases: A is a 0.1% aqueous phosphoric acid solution, and B is acetonitrile, with a volume ratio of mobile phases A to B of 30:70; and a column temperature of 38°C.

[0060] In order to further verify the feasibility of the present invention, the inventors conducted a series of methodological verification experiments, some of which are excerpted as follows:

[0061] 1. Linear investigation test

[0062] The series of standard working solutions in Example 1 were taken and measured according to the ultrahigh performance liquid chromatography conditions of the present invention. Each concentration was injected twice in parallel. The results are shown in Table 1. A standard curve was plotted with the average value of the target concentration as the abscissa and the average value of the target peak area as the ordinate.

[0063] Table 1: Analysis results of 2-amino-3,4-difluorobenzaldehyde (FDFA) standard working solution

[0064]

[0065] The results showed that when the concentration of 2-amino-3,4-difluorobenzaldehyde was in the range of 0.500 mg / L to 10.0 mg / L, there was a linear relationship between y and x. Linear regression of the curve gave the equation of the test substance in acetonitrile as y=90.74x+1.16, and the linear correlation coefficient R 2 is 0.9998, and the linear fitting curve is shown in Figure 1 .

[0066] 2. Specificity test

[0067] Using the ultrahigh performance liquid chromatography (UHPLC) conditions of the present invention, blank samples of test water, recovery samples of test water, blank samples of algae culture medium, and recovery samples of algae culture medium were measured, and the chromatograms of the blank and recovery samples were compared. The retention time of 2-amino-3,4-difluorobenzaldehyde was 6.9 minutes, and the blank sample had no interfering peaks at this time point. The results demonstrate that this method has good specificity for 2-amino-3,4-difluorobenzaldehyde (FDFA).

[0068] 3. Recovery and repeatability test

[0069] (1) Preparation of test water recovery rate concentration

[0070] Prepare 0.800 ml of a 100 mg / L 2-amino-3,4-difluorobenzaldehyde standard stock solution and pipette it into a 100 mL volumetric flask. Dissolve it in test water and dilute to the mark. Shake well to obtain a 0.800 mg / L test water low-concentration recovery test solution. Repeat the above steps to prepare five replicates.

[0071] Prepare and pipette 0.230 mL of 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 52003 mg / L into a 100 mL volumetric flask, dissolve it with test water and dilute to the scale, shake well to obtain a high-concentration recovery test solution with a concentration of 120 mg / L test water. Repeat the above steps to prepare 5 parallel solutions.

[0072] Take the blank matrix and the high and low concentration recovery test solutions of the test water, dilute the high concentration recovery test solution 20 times with acetonitrile, directly sample the low concentration recovery test solution, pass through a 0.45 μm filter membrane, and test according to the operating conditions of the ultra-high performance liquid chromatography instrument of the present invention. Calculate the recovery according to formula (1), and calculate the relative standard deviation (RSD) of the recovery according to formula (2) and formula (3).

[0073] (2) Preparation of recovery rate and concentration of algae culture medium

[0074] Prepare 0.800 ml of a 100 mg / L 2-amino-3,4-difluorobenzaldehyde standard stock solution and pipette it into a 100 mL volumetric flask. Dissolve it in algae culture medium and dilute to the mark. Shake well to obtain a 0.800 mg / L algae culture medium high-concentration recovery test solution. Repeat this procedure for five replicates.

[0075] Prepare and pipette 1.000 mL of 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 52003 mg / L into a 100 mL volumetric flask, dissolve it with algae culture medium and dilute to the mark, shake well to obtain a low-concentration recovery rate test solution of algae culture medium with a concentration of 520 mg / L. Repeat the above operation to prepare 5 parallel solutions.

[0076] The blank matrix and the high and low concentration recovery test solutions of the algae culture medium were taken. The high concentration recovery test solution was diluted 100 times with acetonitrile, and the low concentration recovery test solution was directly sampled and passed through a 0.45 μm filter membrane. The test was performed according to the operating conditions of the ultrahigh performance liquid chromatography instrument of the present invention. The recovery was calculated according to formula (1), and the relative standard deviation (RSD) of the recovery was calculated according to formula (2) and formula (3).

[0077] The precision of the analytical method is reflected by repeatability and expressed as relative standard deviation (RSD).

[0078] Repeatability was analyzed using the recovery test solution, and the RSDs of five levels of high and low concentrations were calculated.

[0079] The analysis results are shown in Tables 2 and 3. The average recovery rates of 2-amino-3,4-difluorobenzaldehyde (FDFA) at two different concentrations in the test water were 87.7% and 97.1%, respectively, and the relative standard deviations of the recoveries were 4.28% and 0.201%, respectively; the average recovery rates of 2-amino-3,4-difluorobenzaldehyde (FDFA) at two different concentrations in the algae culture medium were 92.2% and 102%, respectively, and the relative standard deviations of the recoveries were 3.89% and 0.164%, respectively.

[0080]

[0081] Where:

[0082] R——recovery rate, %;

[0083] Cd——measured concentration of target substance, mg / L;

[0084] Ca——theoretical added concentration of target substance, mg / L.

[0085]

[0086] Where:

[0087] S——standard deviation;

[0088] Xi——recovery rate obtained from the i-th measurement, %;

[0089] ——Average value of recovery rate, %;

[0090] N——The number of recovery rates involved in the calculation.

[0091]

[0092] Where:

[0093] RSD——Relative standard deviation, %.

[0094] Table 2: Test results of recovery rate in test water

[0095]

[0096] Table 3: Recovery rate in algae culture medium

[0097]

[0098]

[0099] 4. Detection limit and quantification limit

[0100] The chromatograms obtained from the low-concentration recovery samples of the present invention were analyzed, and the results are shown in Tables 4 and 5. At this concentration, the average signal-to-noise ratio (S / N) of the target peak in the test water was 223, and the average signal-to-noise ratio (S / N) of the target peak in the algae culture medium was 239. Based on the formulas LOD = 3 × assay concentration / (S / N) and LOQ = 10 × assay concentration / (S / N), the LOD and LOQ of this method in the test water were 0.0108 mg / L and 0.0359 mg / L, respectively. The LOD and LOQ in the algae culture medium were 0.0100 mg / L and 0.0334 mg / L, respectively.

[0101] Table 4: LOD, LOQ test results - test water

[0102]

[0103] Table 5: LOD, LOQ test results - algae culture medium

[0104]

[0105]

[0106] Although the present invention has been described in detail above using general explanations, specific embodiments, and experiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein.

Claims

1. A method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultrahigh performance liquid chromatography, characterized in that: The steps include: (1) Drawing of the standard curve: Accurately weigh 0.05015 g of 2-amino-3,4-difluorobenzaldehyde standard into a 50 mL volumetric flask, dissolve it in acetonitrile and dilute to the mark, shake well to prepare a 2-amino-3,4-difluorobenzaldehyde standard stock solution with a concentration of 1000 mg / L, take an appropriate amount of the standard stock solution, dilute it with acetonitrile to a series of standard working solutions with concentrations of 0.500 to 10.0 mg / L, use the solvent as a blank control, measure the peak area according to ultra-high performance liquid chromatography conditions, draw a standard curve, and obtain the linear regression equation; (2) Preparation of test solution: Accurately weigh 1.30371 g of 2-amino-3,4-difluorobenzaldehyde sample into a 25 mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, shake well, and prepare a 2-amino-3,4-difluorobenzaldehyde sample stock solution with a concentration of 52003 mg / L; respectively, draw appropriate amounts of the sample stock solution into two 100 mL volumetric flasks, dilute with test water and algae culture medium, respectively, to obtain the test solution to be tested, and then determine it by ultra-high performance liquid chromatography; (3) Determination of the test sample: Set the operating conditions of the ultra-high performance liquid chromatography instrument. After the instrument is stable, perform ultra-high performance liquid chromatography on the test sample solution and calculate the content of 2-amino-3,4-difluorobenzaldehyde in the test sample solution according to the linear regression equation. In step (1), the concentrations of the series of standard working solutions are 0.500 mg / L, 1.00 mg / L, 2.00 mg / L, 4.00 mg / L, 8.00 mg / L, and 10.0 mg / L, respectively; The ultra-high performance liquid chromatography (UHPLC) was performed using an Agilent Infinity II Prime 1260 chromatograph. The instrument operating conditions for the UHPLC were as follows: chromatographic column: Shim-pack GIST C18, 250 mm × 4.6 mm, 5 μm; injection volume: 2.0 to 8.0 μL; flow rate: 0.4 to 0.80 mL / min; detection wavelength: 218 to 223 nm. Mobile phase: A is 0.1% phosphoric acid aqueous solution, B is acetonitrile; column temperature: 32-38°C; The volume ratio of the mobile phases A and B was 30:

70.

2. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultra high performance liquid chromatography according to claim 1, wherein: The injection volume was 5.0 μL.

3. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultra high performance liquid chromatography according to claim 1, characterized in that: The flow rate was 0.60 mL / min.

4. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultrahigh performance liquid chromatography according to claim 1, wherein: The detection wavelength is 221 nm.

5. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultra high performance liquid chromatography according to claim 1, characterized in that: The column temperature was 35°C.

6. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultra high performance liquid chromatography according to claim 1, characterized in that: Step (1), the linear regression equation is y=90.74x+1.16, and the linear correlation coefficient R 2 is 0.9998.

7. The method for detecting 2-amino-3,4-difluorobenzaldehyde in water and algae culture medium using ultra high performance liquid chromatography according to claim 1, characterized in that: The LOD of this method in test water is 0.0108 mg / L and LOQ is 0.0359 mg / L. The LOD in algae culture medium is 0.0100 mg / L and LOQ is 0.0334 mg / L.

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