Method for detecting genotoxic impurities in brexpiprazole

Abstract

The invention relates to the technical field of drug analysis and detection, and particularly discloses a method for detecting genotoxic impurities in brexpiprazole. According to the method, high performance liquid chromatography is adopted for detection, and chromatographic conditions are as follows: a chromatographic column is a pentafluorophenyl chromatographic column; the detection wavelength is 258-262 nm, a mobile phase A is a phosphate buffer solution with the pH value of 2.0-2.4, a mobile phase B is methanol, and gradient elution is carried out. According to the method, effective separation of brexpiprazole from DDQ and DHQ is achieved, effective control over the quality of the brexpiprazole raw material is facilitated, therefore, the quality of brexpiprazole and the quality of a preparation of the brexpiprazole are guaranteed, monitoring of the synthesis process of the brexpiprazole is facilitated, and the method has very important significance on improvement of medication safety and has high practical value.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical analysis and detection technology, and in particular to a method for detecting genotoxic impurities in birepiperazole. Background Technology

[0002] Buripiperazole, also known as ipiperazole, has the chemical name 7-[4-(4-benzo[B]thiophene-4-yl-1-piperazine)butoxy]-2(1H)-quinolinone, with the chemical formula C. 25 H 27 N3O2S, molecular formula: C 11 H6ClN3O6 2C4H 11 NO3, with the chemical structure shown below. Biriperazole is a novel multi-target mechanism of action drug for the treatment of mental disorders. It exerts its pharmacological activity through partial agonistic effects on dopamine D2, D3, and serotonin 1A receptors, and antagonistic effects on serotonin 2A and serotonin 2B receptors. Clinically, it is mainly used to treat schizophrenia and as an adjunct therapy for major depressive disorder. It is also used to treat agitation in Alzheimer's patients, post-traumatic stress disorder, and borderline personality disorder. With the rising incidence of mental illnesses and the increasing demand for clinical medication, the market application of biriperazole continues to expand, and its drug quality and safety have received widespread attention from the global pharmaceutical industry.

[0003]

[0004] The preparation of birepiperazole may generate some impurity compounds, affecting drug quality and safety. Detection of these compounds is a crucial step in ensuring product quality. According to the birepiperazole production process, DDQ (4,5-dichloro-5,6-dicyanobenzoquinone) is used as the dehydrogenation reagent in the intermediate preparation, generating DHQ (4,5-dichloro-3,6-dihydroxyphthalonitrile) during the reaction. The structures of DDQ and DHQ reveal that they contain halogenated olefins and quinones. The EU JRC, in its article "Development of structural alerts for the in vivo micronucleus assay in rodents," explicitly states that halogenated olefins and quinones are warning structures. The European Medicines Agency (EMEA), the US Food and Drug Administration (FDA), and the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) have successively issued guidance documents on the control of genotoxic impurities, recommending the use of the Toxicological Concern Threshold (TTC) to control drug risks and keep mutagenic impurities at reasonable levels to reduce the risk of carcinogenicity in humans.

[0005] Based on the TTC calculation method, the acceptable daily intake (TC) for the most stringent lifetime dosing is 1.5 μg / day. The maximum daily dose of birepiperazole, as found in the product information, is 4 mg. Therefore, the limits for DDQ and DHQ must not exceed 1.5 μg / 4 mg = 0.0375% (i.e., 375 ppm). Due to the poor stability, low control limits, and difficulty in detecting DDQ, there is currently a lack of accurate methods for determining DDQ and DHQ in birepiperazole raw materials. To ensure patient safety, strict control and accurate detection of genotoxic impurities DDQ and DHQ in birepiperazole are necessary. Therefore, it is essential to develop a simple, highly sensitive, low-limit-of-detection, and reproducible method for detecting genotoxic impurities DDQ and DHQ in birepiperazole. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides a method for detecting genotoxic impurities in birepiperazole.

[0007] To solve the above-mentioned technical problems, the technical solution provided by the present invention is as follows: A method for detecting genotoxic impurities in birepiperazole, wherein the genotoxic impurities are 4,5-dichloro-5,6-dicyanobenzoquinone (DDQ) and 4,5-dichloro-3,6-dihydroxyphthalonitrile (DHQ), and the detection is performed by high-performance liquid chromatography (HPLC) under the following chromatographic conditions: Chromatographic column: Pentafluorophenyl column; UV detector, with a detection wavelength of 258nm~262nm; Mobile phase A: phosphate buffer solution with pH 2.0~2.4; mobile phase B: methanol; wherein the phosphate buffer solution is a mixed solution of ammonium dihydrogen phosphate, phosphate and triethylamine. The elution method is gradient elution, and the elution procedure is as follows: 0 min, 48%~52% mobile phase A, 52%~48% mobile phase B; 10 min, 48%~52% mobile phase A, 52%~48% mobile phase B; 15 min, 30% mobile phase A, 70% mobile phase B; 20 min, 30% mobile phase A, 70% mobile phase B; 20.1 min, 48%~52% mobile phase A, 52%~48% mobile phase B; 30 min, 48%~52% mobile phase A, 52%~48% mobile phase B.

[0008] It should be noted that the synthetic route of the birepiperazole detected in this invention is as follows.

[0009]

[0010] During the research on the synthesis process of birepiperazole active pharmaceutical ingredient, this invention discovered the potential presence of DDQ and DHQ genotoxic impurities. Information on these impurities is shown in Table 1. Table 1 Impurity Information Table

[0011] DDQ and DHQ, two genotoxic impurities, have similar physicochemical properties, making separation difficult during detection. This invention provides a method for detecting genotoxic impurities in birepiperazole. Using a pentafluorophenyl column and a phosphate buffer solution (pH 2.0-2.4) and methanol as the mobile phase, high-performance liquid chromatography (HPLC) with a specific gradient elution method achieves effective separation of birepiperazole from DDQ and DHQ. Furthermore, methodological studies and validations of specificity and sensitivity have shown that this method is sensitive, accurate, and reproducible. It provides a simple and rapid method for the accurate quantitative detection of genotoxic impurities DDQ and DHQ in birepiperazole, overcoming the limitations of existing methods for quantitative detection of genotoxic impurities in birepiperazole. This provides a reliable guarantee for improving the quality of birepiperazole products and is of great significance for enhancing medication safety.

[0012] Furthermore, the chromatographic column has dimensions of 250mm*4.6mm and a diameter of 5μm.

[0013] Furthermore, the chromatographic column is a Hypersil GOLD. TM PFP.

[0014] The optimal column specifications can provide excellent peak shape, resolution and detection sensitivity for each component, with minimal baseline interference. This facilitates the effective separation of the raw material from DDQ and DHQ in biriperazole, and results in accurate, reliable and reproducible results.

[0015] Furthermore, the degree of elution is: 0 min, 50% mobile phase A, 50% mobile phase B; 10 min, 50% mobile phase A, 50% mobile phase B; 15 min, 30% mobile phase A, 70% mobile phase B; 20 min, 30% mobile phase A, 70% mobile phase B; 20.1 min, 50% mobile phase A, 50% mobile phase B; 30 min, 50% mobile phase A, 50% mobile phase B.

[0016] Optimizing the gradient elution sequence can improve the separation and detection sensitivity of buriperazole with DDQ and DHQ, resulting in accurate quantitative results with high precision.

[0017] Furthermore, the detection wavelength is 260nm.

[0018] The selected detection wavelength can significantly enhance the chromatographic peak response signal of the target analyte, enabling the full detection of trace amounts of genotoxic impurities. It can also effectively avoid interference from solvents, excipients, and other matrix components, reduce baseline noise, and ensure the accuracy of the detection results.

[0019] Furthermore, the pH of the phosphate buffer solution is 2.2.

[0020] The optimal pH value of mobile phase A can reduce band tailing and improve peak shape, thereby improving the separation between components and resulting in higher accuracy and precision of the detection results.

[0021] Furthermore, the concentration of triethylamine is 0.5 mL / L to 1.5 mL / L, and the concentration of dihydrogen phosphate is 38 mmol / L to 42 mmol / L.

[0022] Preferably, the concentration of triethylamine is 1 mL / L, and the concentration of ammonium dihydrogen phosphate is 40 mmol / L.

[0023] The optimal buffer salt, combined with a specific chromatographic column, can produce good peak shapes for biriperazole and DDQ / DHQ, and improve the separation of biriperazole from DDQ / DHQ, thereby achieving the goal of effectively and accurately detecting the genotoxic content in biriperazole.

[0024] Furthermore, the flow rate is 0.9 mL / min to 1.1 mL / min.

[0025] Furthermore, the flow rate was 1.0 mL / min.

[0026] The optimized flow rate allows sufficient retention and interaction time between the main component of buriperazole and the genotoxic impurities DDQ and DHQ within the chromatographic column, effectively separating the structurally similar DDQ and DHQ. At the same time, it avoids excessively long analysis cycles caused by slow flow rates, balancing separation effect and detection efficiency, and adapting to the needs of industrial batch detection.

[0027] Furthermore, the column temperature is 37℃~43℃.

[0028] Furthermore, the column temperature is 40℃.

[0029] Optimal column temperature and column combination help ensure effective sample separation and improve detection accuracy and sensitivity.

[0030] Furthermore, the injection volume is 28 μL to 32 μL.

[0031] Furthermore, the injection volume was 30 μL.

[0032] Furthermore, the preparation process of the test solution is as follows: accurately weigh the biriperazole test sample, dissolve it in solvent, and dilute it to prepare a 2 mg / mL test solution.

[0033] Specifically, the solvent used to prepare the test solution is a mixed solution of acetonitrile, water and acetic acid in a volume ratio of 50:50:1.

[0034] In summary, the detection method provided by this invention can accurately quantify DDQ and DHQ in biriperazole, which is beneficial for effectively controlling the quality of biriperazole raw materials, thereby ensuring the quality of biriperazole and its preparations. It is also beneficial for monitoring the biriperazole synthesis process, which is of great significance for improving drug safety and has high practical value. Attached Figure Description

[0035] Figure 1 This is the liquid chromatogram of the test solution in Example 1 of the present invention; Figure 2 This is a liquid chromatogram of the impurity reference solution in Example 1 of the present invention; Figure 3 This is a liquid chromatogram of the system suitability solution in Example 1 of the present invention; Figure 4 This is the liquid chromatogram of the blank solvent in Example 2 of the present invention; Figure 5 This is the liquid chromatogram of the spiked test solution in Example 2 of the present invention; Figure 6 This is a liquid chromatogram of the system suitability solution in Comparative Example 1 of the present invention; Figure 7 This is a liquid chromatogram of the system suitability solution in Comparative Example 2 of the present invention; Figure 8 This is a liquid chromatogram of the system suitability solution in Comparative Example 3 of the present invention; Figure 9 This is a liquid chromatogram of the system suitability solution in Comparative Example 4 of the present invention; Peak 1 is impurity DDQ, peak 2 is impurity DHQ, and peak 3 is biriperazole. Detailed Implementation

[0036] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0037] The reference standards for genotoxic impurities DDQ and DHQ were both commercially available and purchased from Walderson and QCC. The batch number of the DDQ reference standard was E133603, and the purity was 97.46%; the batch number of the DHQ reference standard was 17-JAN-22-34, and the purity was 99.62%.

[0038] The buripiperazole sample selected was a self-made product of our company, with batch number BRE-20250701 and a purity of 99.5%.

[0039] The high performance liquid chromatograph used in this invention is a Shimadzu LC-2050C.

[0040] Example 1 1.1 Solution Preparation Blank solvent: a mixed solution of acetonitrile, water and acetic acid in a volume ratio of 50:50:1.

[0041] Preparation of the test solution: Take an appropriate amount of biriperazole test sample, accurately weigh it, dissolve it in blank solvent and quantitatively dilute it to prepare a solution containing about 2.0 mg per 1 mL, thus obtaining the test solution.

[0042] Preparation of impurity reference stock solutions: Take appropriate amounts of impurity DDQ and DHQ reference standards respectively, dissolve and dilute them with methanol to prepare solutions containing approximately 7.5 μg per 1 mL of solution, shake well, and obtain DDQ impurity reference stock solutions and DHQ impurity reference stock solutions respectively.

[0043] Preparation of impurity reference solution: Accurately measure 1 mL each of DDQ and DHQ impurity reference stock solutions into a 10 mL volumetric flask, dilute to the mark with blank solvent, and prepare a solution containing approximately 0.75 μg of DDQ and DHQ per mL to obtain the impurity reference solution.

[0044] Preparation of spiked test solution: Accurately weigh an appropriate amount of biriperazole test sample, add 1 mL each of DDQ and DHQ impurity reference standard stock solution to a 10 mL volumetric flask, dissolve and quantitatively dilute with blank solvent to prepare a solution containing approximately 0.75 μg of DDQ and DHQ and 2.0 mg of biriperazole per 1 mL, thus obtaining the spiked test solution.

[0045] Preparation of system suitability solution: Accurately weigh an appropriate amount of biriperazole reference standard, add 1 mL each of DDQ and DHQ impurity reference standard stock solutions to a 10 mL volumetric flask, dissolve and quantitatively dilute with blank solvent to prepare a solution containing approximately 0.75 μg of DDQ and DHQ and 2.0 mg of biriperazole per 1 mL, thus obtaining the system suitability solution.

[0046] 1.2 Conditions for high performance liquid chromatography: Column: Hypersil GOLD TMPFP (250mm*4.6mm, 5μm); Mobile phase A: Phosphate buffer (dissolve 2.3004 g of ammonium dihydrogen phosphate in 1000 mL of water, add 1 mL of triethylamine, and adjust the pH to 2.2 with phosphate). Mobile phase B: methanol; Flow rate: 1.0 mL / min; Detection wavelength: 260nm; Column temperature: 40℃; Injection volume: 30 μL.

[0047] Elution was performed according to the following gradient procedure:

[0048] Accurately measure 30 μL each of the blank solution, system suitability solution, impurity reference solution, and test solution, inject them into the liquid chromatograph, perform the determination under the above chromatographic conditions, record the chromatogram, and analyze the impurity content.

[0049] The liquid chromatogram of the test solution is shown below. Figure 1 As shown, the liquid chromatogram of the impurity reference solution is as follows: Figure 2 As shown, the liquid chromatogram of the system suitability solution is as follows: Figure 3 As shown.

[0050] Figures 1-3 In the sample, peak 1 is impurity DDQ, peak 2 is impurity DHQ, and peak 3 is birepiperazole.

[0051] Example 2 Methodological validation: 2.1 Specificity Inject blank solvent and system suitability solution under the above-described high-performance liquid chromatography (HPLC) conditions and record the chromatograms. The chromatographic peaks of the blank solvent are shown below. Figure 4 As shown, the chromatographic peaks of the spiked test solution are as follows: Figure 5 As shown, the chromatographic peaks of the test solution are as follows: Figure 1 As shown in Table 2, the experimental results of the spiked test solution are as follows.

[0052] Table 2 Peak information of spiked test solution

[0053] The experimental results show that the baseline is stable and interference-free, and the blank solvent does not interfere with the detection of impurities and the main component. The resolution between the main component and adjacent impurities is greater than 1.5, and the resolution between each component and each impurity is greater than 1.2. There is no interference between impurities, and the resolution between each component meets the requirements, making it suitable for the detection of genotoxic impurities. Therefore, the detection method provided by this invention has good specificity.

[0054] 2.2 Limit of Detection and Limit of Quantification The ibuprofen, DDQ, and DHQ reference standards were dissolved in blank solvent and serially diluted. The limit of detection (LOD) was set at a signal-to-noise ratio (SNR) greater than 3:1, and the limit of quantitation (LOQ) was set at a SNR greater than 10:1. The results are shown in Table 3. The LOQ solution was injected six times consecutively, and the RSD of the peak area was calculated. The results are shown in Table 4.

[0055] Table 3 Results of Limit of Detection and Limit of Quantitation tests

[0056] Table 4 Results of repeatability tests at the limit of quantitation

[0057] As shown in Tables 3-4, the limit of quantitation (LOQ) for DDQ is 0.073 μg / mL, and the limit of detection (LOD) is 6.60 μg / mL; the LQ for DHQ is 0.153 μg / mL, and the LOD is 12.74 μg / mL. The RSD of the peak area at the LQ is less than 10%, indicating that the sensitivity of both the main component and each impurity in the method for detecting impurities in birepiperazole provided by this invention meets the detection requirements.

[0058] 2.3 Linear Range Linear stock solution: Take appropriate amounts of DDQ and DHQ reference standards, dissolve and dilute them with methanol to prepare a solution containing approximately 75 μg of DDQ and DHQ per 1 mL, shake well, and obtain the DDQ and DHQ stock solutions. Accurately measure appropriate amounts of each impurity stock solution, add blank solvent to the same 10 mL volumetric flask, and dilute to prepare a linear stock solution containing approximately 7.5 μg of DDQ and DHQ per 1 mL.

[0059] The linear stock solution was serially diluted and detected under the chromatographic conditions of Example 1. 30 μL of each reference solution was accurately measured and injected into the liquid chromatograph. The chromatogram was recorded and the peak area was measured. Linear regression was performed with peak area A as the ordinate and concentration C as the abscissa. The results are shown in Tables 5 and 6.

[0060] Table 5 DDQ Linearity Results

[0061] Table 6. DHQ Linearity Results

[0062] The above results show that, within the linear concentration range of this invention, there is a good linear relationship between the concentration and peak area of ​​each impurity.

[0063] 2.4 Accuracy Preparation of the mixed stock solution of reference standards: Take appropriate amounts of DDQ and DHQ reference standards, dissolve and dilute them with methanol to prepare a solution containing approximately 7.5 μg of DDQ and DHQ per 1 mL, thus obtaining the mixed stock solution of reference standards.

[0064] Accurately weigh approximately 20 mg of iripepiperazole reference standard and place it in a 10 mL volumetric flask. Weigh 9 portions in parallel, dissolve each portion in an appropriate amount of blank solvent, and then add the above-mentioned reference standard mixed stock solution. Dilute each portion to the mark with blank solvent. Prepare 3 portions of each concentration in parallel as the recovery test solution.

[0065] Accurately measure 30 µL of each of the above solutions, inject them into the liquid chromatograph, record the chromatogram, calculate the recovery rate of each impurity and the RSD value of the recovery rate. The results are shown in Tables 7 and 8 below.

[0066] Table 7 DDQ Recovery Results

[0067] Table 8 DHQ Recovery Results

[0068] The results above show that the recovery rates of each impurity are in the range of 90% to 110% at all concentrations, and the RSD values ​​are all less than 5%, indicating that the method has good accuracy.

[0069] 2.5 Repeatability Repeatability: The test solution was prepared according to the method provided in Example 1, and the test was repeated 6 times by the same experimenter using the same instrument according to the above detection method. The results are shown in Table 9.

[0070] Intermediate precision: The test solution was prepared according to the method provided in Example 1, and the test was performed by another experimenter using a Waters ARC high-performance liquid chromatograph with a Hypersil GOLD sensor, following the detection method described above. TM A PFP (250mm*4.6mm, 5μm) chromatographic column was used, and the detection was repeated 6 times. The results are shown in Table 10.

[0071] Table 9 Results of Repeatability Tests

[0072] Table 10 Results of intermediate precision test

[0073] The results showed that the RSD of DDQ and DHQ contents was less than 1% in the repeatability and intermediate precision tests, indicating that the method had good repeatability and intermediate precision.

[0074] 2.6 Durability The spiked test solution was prepared according to the method described in Example 1, and the robustness test was conducted under the test conditions shown in Table 11. The measurement results under different pH values ​​of mobile phase A, different initial proportions of mobile phase, and different column temperatures, flow rates, and detection wavelengths are shown in Tables 11-16.

[0075] Table 11 Durability Test Conditions

[0076] Table 12 Experimental results of mobile phase A at different pH values

[0077] Table 13 Experimental results with different initial proportions of mobile phase

[0078] Table 14 Results of tests at different column temperatures

[0079] Table 15 Test results at different flow rates

[0080] Table 16 Test results at different detection wavelengths

[0081] The results showed that the separation degree of each component did not change significantly with changes in the initial mobile phase ratio, mobile phase pH, flow rate, column temperature, and wavelength.

[0082] Example 3 The method for detecting genotoxic impurities in birepiperazole provided in Example 1 was used to test actual samples. The specific details are as follows: Take appropriate amounts of birepiperazole raw materials from three batches (BRE-20250701, BRE-20250702, and BRE-20250703), dissolve them in blank solvent, and quantitatively dilute them to prepare a solution containing approximately 2.0 mg per mL, thus obtaining the test solution. The test solution was determined according to the method for detecting genotoxic impurities in birepiperazole provided in Example 1. The experimental results are shown in Table 17 below.

[0083] Table 17 Results of DDQ and DHQ Impurity Content Detection in Three Batches of Raw Materials

[0084] The experimental results show that the detection method of the present invention can accurately detect the genotoxic DDQ and DHQ in the buripiperazole raw material, providing a more effective basis for the safety study of buripiperazole preparations and the monitoring of the synthesis process.

[0085] Comparative Example 1 This comparative example provides a method for detecting bripiprazole genotoxicity. The only difference between this method and Example 1 is that the phosphoric acid in mobile phase A of Example 1 is replaced with acetic acid to adjust the pH to 2.2, while all other detection conditions are the same.

[0086] The system suitability solution provided in Example 1 was tested, and the chromatogram was recorded. The results are shown in [Figure Number]. Figure 6 And Table 18.

[0087] Table 18 System Adaptability Solution Chromatography Information

[0088] Depend on Figure 6 As can be seen from Table 18, when the system suitability solution was tested using the chromatographic conditions provided in this comparative example, the impurities DDQ and DHQ were poorly separated, failing to reach baseline separation, and their peak times were too early, making it impossible to guarantee the accurate determination of each impurity.

[0089] Comparative Example 2 This comparative example provides a method for detecting genotoxicity in buriperazole. The only difference between this method and Example 1 is that the mobile phase A in Example 1 is replaced with 0.1% phosphoric acid solution, while all other detection conditions remain the same.

[0090] The system suitability solution provided in Example 1 was tested, and the chromatogram was recorded. The results are shown in [Figure Number]. Figure 7 And Table 19.

[0091] Table 19 System Adaptability Solution Chromatography Information

[0092] Depend on Figure 7 As can be seen from Table 19, when the system suitability solution was tested using the chromatographic conditions provided in this comparative example, the DDQ peak showed tailing, and the peak shape of the test sample was poor, making it impossible to guarantee the accurate determination of each impurity.

[0093] Comparative Example 3 This comparative example provides a method for detecting genotoxicity in buriperazole. The only difference between this method and Example 1 is that the chromatographic column in Example 1 is replaced with a Waters Xselect® CSH column. TM C18 (250mm×4.6mm, 5μm) column, all other detection conditions were the same.

[0094] The system suitability solution provided in Example 1 was tested, and the chromatogram was recorded. The results are shown in [Figure Number]. Figure 8 And Table 20.

[0095] Table 20 System Adaptability Solution Chromatography Information

[0096] Depend on Figure 8 As can be seen from Table 20, when the system suitability solution was tested using the chromatographic conditions provided in this comparative example, DDQ was not detected, and DHQ did not achieve baseline separation from the main peak, which could not guarantee the accurate determination of each impurity.

[0097] Comparative Example 4 This comparative example provides a method for detecting genotoxicity in buriperazole. The only difference between this method and Example 1 is that the chromatographic column in Example 1 is replaced with an XBridge column. ® C18 (4.6×250mm, 5μm) column, all other parameters are the same.

[0098] The system suitability solution provided in Example 1 was tested, and the chromatogram was recorded. The results are shown in [Figure Number]. Figure 9 And Table 21.

[0099] Table 21 System Adaptability Solution Chromatography Information

[0100] Depend on Figure 9 As can be seen from Table 21, when the system suitability solution was tested using the chromatographic conditions provided in this comparative example, DDQ showed tailing and poor peak shape, which could not guarantee the accurate determination of each impurity.

[0101] As can be seen from the results of the above examples and comparative examples, the present invention uses a pentafluorophenyl column and adds triethylamine to the phosphate buffer to inhibit the interaction between silanol groups and basic compounds, thereby enhancing the retention time of the compounds and solving the problems of retention and separation of impurity peaks and main peaks.

[0102] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for detecting genotoxic impurities in brexpiprazole, the genotoxic impurities being 4,5-dichloro-5,6-dicyano-benzoquinone and 4,5-dichloro-3,6-dihydroxyphthalonitrile, characterized in that, The high performance liquid chromatography is used for detection, and the chromatographic conditions are as follows: A chromatographic column is used, and the chromatographic column is a pentafluorophenyl chromatographic column; An UV detector is used, and the detection wavelength is 258nm-262nm; A mobile phase A is a phosphate buffer solution with a pH of 2.0-2.4, and a mobile phase B is methanol; wherein the phosphate buffer solution is a mixed solution of ammonium dihydrogen phosphate, phosphoric acid and triethylamine; Gradient elution is used, and the elution program is as follows: 0min, 48%-52% of the mobile phase A and 52%-48% of the mobile phase B; 10min, 48%-52% of the mobile phase A and 52%-48% of the mobile phase B; 15min, 30% of the mobile phase A and 70% of the mobile phase B; 20min, 30% of the mobile phase A and 70% of the mobile phase B; 20.1min, 48%-52% of the mobile phase A and 52%-48% of the mobile phase B; 30min, 48%-52% of the mobile phase A and 52%-48% of the mobile phase B.

2. The method for detecting genotoxic impurities in brexpiprazole according to claim 1, wherein, The specification of the chromatographic column is 250mm*4.6mm, 5μm.

3. The method for detecting genotoxic impurities in brexpiprazole according to claim 2, wherein The chromatographic column is a Hypersil GOLD TM PFP.

4. The method for detecting genotoxic impurities in brexpiprazole according to claim 1, wherein The elution program is as follows: 0min, 50% of the mobile phase A and 50% of the mobile phase B; 10min, 50% of the mobile phase A and 50% of the mobile phase B; 15min, 30% of the mobile phase A and 70% of the mobile phase B; 20min, 30% of the mobile phase A and 70% of the mobile phase B; 20.1min, 50% of the mobile phase A and 50% of the mobile phase B; 30min, 50% of the mobile phase A and 50% of the mobile phase B.

5. The method for detecting genotoxic impurities in brexpiprazole according to claim 1, wherein The pH of the phosphate buffer solution is 2.

2.

6. The method for detecting genotoxic impurities in brexpiprazole according to claim 5, wherein, The concentration of the triethylamine is 0.5mL / L-1.5mL / L, and the concentration of the ammonium dihydrogen phosphate is 38mmol / L-42mmol / L.

7. The method for detecting genotoxic impurities in brexpiprazole according to claim 6, wherein The concentration of the triethylamine is 1mL / L, and the concentration of the ammonium dihydrogen phosphate is 40mmol / L.

8. The method for detecting genotoxic impurities in birepiperazole as described in claim 1, characterized in that, The flow rate is 0.9mL / min-1.1mL / min, the column temperature is 37℃-43℃, and the injection volume is 28μL-32μL.

9. The method for detecting genotoxic impurities in brexpiprazole according to claim 8, wherein, The flow rate is 1.0mL / min, the column temperature is 40℃, and the injection volume is 30μL.

10. The method for detecting genotoxic impurities in birepiperazole as described in claim 1, characterized in that, The concentration of the test sample solution is 2mg / mL.

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