Method for detecting cariprazine hydrochloride intermediate and related substances thereof

Abstract

According to the method for detecting the cariprazine hydrochloride intermediate and the related substances thereof, the sample solubility is high, the separation degree is good, the detection efficiency and the detection precision can be improved, and a method guarantee is provided for guaranteeing the medicine quality and the medication safety.

Description

Technical Field

[0001] This invention relates to the field of drug detection, specifically to a method for detecting cariprazine hydrochloride intermediates and related substances. Background Technology

[0002] Cariprazine is an atypical antipsychotic drug jointly developed by Forest Laboratories and Gedeon Richter Ltd. in the United States. It is a partial agonist of dopamine D2 and D3 receptors and is mainly used to treat schizophrenia and bipolar mania. It was approved for marketing by the U.S. Food and Drug Administration on September 17, 2015. Its chemical name is trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-ethyl}-N,N-dimethylformyl-cyclohexylamine.

[0003] trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine is a key intermediate in the synthesis of caliraline hydrochloride, and is described in CN113527227A with the following structural formula:

[0004]

[0005] Studies have found that this intermediate generates several related substances during its synthesis, such as 1-(2,3-dichlorophenyl)piperazine. Excessive introduction of these related substances into the finished drug product could lead to potential drug safety issues. Therefore, it is necessary to detect and strictly control the intermediate trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine and its related substances. Summary of the Invention

[0006] The purpose of this invention is to solve the above-mentioned technical problems and provide a method for detecting intermediates of cariprazine hydrochloride and related substances. The method has high sample solubility, good peak separation, good peak shape, and accurate and reliable detection results.

[0007] The applicant of this invention discovered that the intermediate trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine has poor solubility, with solubility below 0.5 mg / ml in solvents such as water, DMF, and DMSO. It exhibits good solubility only in solvents such as methanol and acetonitrile. However, when using methanol or acetonitrile as diluents to prepare intermediate solutions for high-performance liquid chromatography (HPLC) detection, related substances contained in the intermediate, such as 1-(2,3-dichlorophenyl)piperazine, are easily detected with two or more peaks, leading to inaccurate detection results. This invention addresses the problem that conventional solvent systems cannot simultaneously satisfy both good solubility of the intermediate and accurate detection of related substances, developing a more suitable method.

[0008] This invention provides a method for detecting calciporazine hydrochloride intermediates and related substances, comprising the following steps:

[0009] (1) Preparation of test solution: Dissolve the intermediate sample in a solution composed of methanol, dichloromethane and water to obtain the test solution;

[0010] (2) Detection: The test solution is injected into the liquid chromatograph and detected according to the high performance liquid chromatography (HPLC) conditions. The HPLC conditions are as follows: the column is an octadecylsilane bonded silica column, the mobile phase A is trifluoroacetic acid aqueous solution-acetonitrile, the mobile phase B is trifluoroacetic acid aqueous solution-acetonitrile, and gradient elution is used.

[0011] The present invention provides a method for detecting cariprazine hydrochloride intermediates and related substances, wherein the solution is composed of methanol, dichloromethane and water, wherein the volume ratio of methanol, dichloromethane and water is (50-75):(5-20):(20-40); preferably, the volume ratio of methanol, dichloromethane and water is (50-75):(5-10):(20-40), or (50-75):(5-10):(20-35), or (51-70):(10-15):(20-35), or (53-65):(15-20):(20-34); more preferably, the volume ratio of methanol, dichloromethane and water is 60:10:30.

[0012] The present invention provides a method for detecting cariprazine hydrochloride intermediates and related substances, wherein the mobile phase A and mobile phase B contain an aqueous solution of trifluoroacetic acid, wherein the concentration of trifluoroacetic acid is 0.03%-0.08%, preferably 0.05%.

[0013] The present invention provides a method for detecting caliracil hydrochloride intermediates and related substances, wherein the mobile phase A is a trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (85-90):(10-15), and the mobile phase B is a trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (10-20):(80-90). Preferably, the mobile phase A is trifluoroacetic acid aqueous solution-acetonitrile = 90:10, and the mobile phase B is trifluoroacetic acid aqueous solution-acetonitrile = 10:90.

[0014] The present invention provides a method for detecting caliprazine hydrochloride intermediates and related substances, wherein the gradient elution procedure is as follows:

[0015] Time T / min 0 3 10 25 28 35 Mobile phase A% 70-80 70-80 15-25 15-25 70-80 70-80 Mobile phase B% 20-30 20-30 75-85 75-85 20-30 20-30

[0016] The gradient elution process is preferably as follows:

[0017] Time T / min 0 3 10 25 28 35 Mobile phase A% 75 75 20 20 75 75 Mobile phase B% 25 25 80 80 25 25

[0018] The method for detecting calirazine hydrochloride intermediates and related substances provided by the present invention comprises a solution of methanol, dichloromethane and water, which is prepared by first mixing methanol and dichloromethane to obtain a mixed solution, and then adding water to the mixed solution.

[0019] The present invention provides a method for detecting caliprazine hydrochloride intermediates and related substances, wherein the test solution is prepared by: taking an intermediate sample, accurately weighing it, dissolving and diluting it with a solution composed of methanol, dichloromethane and water to prepare a solution containing approximately 0.5 mg to 0.6 mg per 1 ml.

[0020] The present invention provides a method for detecting cariprazine hydrochloride intermediates and related substances, wherein the high performance liquid chromatography (HPLC) conditions are: wavelength 254 nm, column temperature 20-40 °C, injection volume 10-15 μL, and flow rate 0.8-1.2 mL / min.

[0021] This invention further provides a method for detecting calciporazine hydrochloride intermediates and related substances, comprising the following steps:

[0022] (1) Preparation of test solution: Take an appropriate amount of intermediate sample, weigh it accurately, dissolve and dilute it with a solution composed of methanol, dichloromethane and water to prepare a solution containing about 0.5 mg per ml; wherein the volume ratio of methanol, dichloromethane and water in the solution is (50-75):(5-10):(20-35);

[0023] (2) Detection: Inject the test solution into the liquid chromatograph and detect it according to the high-performance liquid chromatography (HPLC) conditions. The HPLC conditions are as follows: the column is an octadecylsilane-bonded silica column; mobile phase A is 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 90:10; mobile phase B is 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 10:90; gradient elution is used; the mobile phase flow rate is 0.8-1.2 ml / min; the column temperature is 20-40℃; the sample injection volume is 10-15 μl; and a photodiode array detector is used with a detection wavelength of 254 nm. The gradient elution program is as follows:

[0024] Time T / min 0 3 10 25 28 35 Mobile phase A% 75 75 20 20 75 75 Mobile phase B% 25 25 80 80 25 25

[0025] The present invention provides a method for detecting caliprazine hydrochloride intermediates and related substances, wherein the intermediate is trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine; and the related substances include 1-(2,3-dichlorophenyl)piperazine.

[0026] Through extensive experimentation and exploration, the inventors of this application have finally established a high-performance liquid chromatography (HPLC) method that can efficiently separate and detect cariprazine hydrochloride intermediates and related substances. This method has high sample solubility, good peak separation, and good peak shape, which can improve detection efficiency and accuracy, and provide a methodological guarantee for ensuring drug quality and medication safety. Attached Figure Description

[0027] Figure 1 These are high-performance liquid chromatograms of Test Examples 1-2 of this invention.

[0028] Figure 2 These are high-performance liquid chromatograms of Test Examples 1-5 of this invention.

[0029] Figure 3-1 The high-performance liquid chromatograms of Test 1, Test Examples 1-7 of this invention are shown below.

[0030] Figure 3-2 The high performance liquid chromatograms are for the blank solvents corresponding to Experiment 1, Examples 1-7 of this invention.

[0031] Figure 3-3 This is a superimposed image of the high performance liquid chromatograms of Test 1, Examples 1-7 of the present invention and the high performance liquid chromatograms of blank solvent.

[0032] Figure 4-1 These are high-performance liquid chromatograms of Test Examples 1-12 of this invention.

[0033] Figure 4-2 The high performance liquid chromatograms of the blank solvents corresponding to Experiment 1, Examples 1-12 of this invention are shown.

[0034] Figure 4-3 This is a superimposed image of the high performance liquid chromatograms of Test 1, Test Examples 1-12 of the present invention and the high performance liquid chromatograms of blank solvent.

[0035] Figure 5-1 This is a high-performance liquid chromatogram of the system suitability solution under condition 7 in Examples 1-3 of the present invention.

[0036] Figure 5-2 The high-performance liquid chromatograms are the blank solvents corresponding to the system suitability solutions in conditions 7 of Examples 1-3 of the present invention.

[0037] Figure 5-3 This is an overlay of the high-performance liquid chromatogram (HPLC) of the system suitability solution under condition 7 in Examples 1-3 of the present invention and the HPLC chromatogram of the blank solvent.

[0038] Figure 6-1 This is a high-performance liquid chromatogram of the test solution in Example 2 of the present invention.

[0039] Figure 6-2 This is a high-performance liquid chromatogram of the blank solvent corresponding to the test solution in Example 2 of the present invention.

[0040] Figure 6-3 This is an overlay of the high-performance liquid chromatogram of the test solution and the high-performance liquid chromatogram of the blank solvent in Example 2 of the present invention.

[0041] Figure 7-1 This is a high-performance liquid chromatogram of the system suitability solution in Example 2 of the present invention.

[0042] Figure 7-2 This is a high-performance liquid chromatogram of the blank solvent corresponding to the system suitability solution in Example 2 of the present invention. Figure 7-3 This is an overlay of the high-performance liquid chromatogram of the system suitability solution and the high-performance liquid chromatogram of the blank solvent in Example 2 of the present invention. Detailed Implementation

[0043] The present invention will be further described in detail through the following embodiments, but these embodiments should not be used to limit the scope of protection of the present invention.

[0044] I. Medicines and Reagents

[0045] The intermediate trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine (IM01) used in the experiments of this invention were all prepared in-house. The related substances 1-(2,3-dichlorophenyl)piperazine (IM01-1) and 1-hydroxybenzotriazole (IM01-3) reference standards were purchased externally. Specific information on the drugs and reagents is shown in Table 1.

[0046] Table 1 List of Medicines and Reagents

[0047]

[0048] II. Main Instruments

[0049] Table 2 List of Main Instruments

[0050] Instrument Name Instrument Model coding factory HPLC Agilent 1260 II JC271 Agilent HPLC Agilent 1260 II JC269 Agilent HPLC Agilent 1260 II JC287 Agilent Electronic balance BP211D JC047 Sartorius

[0051] Experiment 1: Effects of different diluents on the solubility of IM01 and on the liquid chromatography detection results of IM01-1

[0052] 1. Solubility

[0053] Accurately weigh 10 mg of intermediate sample IM01 and place it in 20 ml of different diluents (see Table 4). Observe whether the sample dissolves completely. If it dissolves completely, it means that the sample has a solubility of more than 0.5 mg / ml in the diluent. If it does not dissolve completely, it means that the sample has a solubility of less than 0.5 mg / ml in the diluent.

[0054] 2. High-performance liquid chromatography (HPLC) detection

[0055] Based on the solubility test results, select a diluent with a solubility greater than 0.5 mg / ml for IM01, and test the related substance IM01-01 reference solution according to the following method. Record the liquid chromatogram and the chromatographic detection results.

[0056] Detection method:

[0057] (1) Preparation of IM01-1 reference solution

[0058] Take an appropriate amount of impurity IM01-1 reference standard, accurately weigh it, dissolve and dilute it with diluent to prepare a solution containing about 0.5 mg per ml, as a stock solution. Take an appropriate amount of the stock solution, dilute it with diluent to prepare a solution containing about 0.01 mg per ml, as the impurity IM01-1 reference standard solution.

[0059] (2) Liquid chromatography detection conditions

[0060] The chromatographic column was an Inertsil ODS-3250 × 4.6 mm 5 μm column; mobile phase A was 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 90:10, and mobile phase B was 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 10:90, with gradient elution; the mobile phase flow rate was 1.0 ml / min, the column temperature was 30 ℃, the sample injection volume was 10 μl, and a photodiode array detector was used with a detection wavelength of 254 nm; the gradient elution program is shown in Table 3.

[0061] Table 3 Gradient elution program

[0062] Time T / min 0 3 10 25 28 35 Mobile phase A% 75 75 20 20 75 75 Mobile phase B% 25 25 80 80 25 25

[0063] (3) Determination: Accurately measure 10 μl each of the IM01-01 reference solution and its corresponding diluent (as blank solvent), inject them into the liquid chromatograph, and determine them by high performance liquid chromatography.

[0064] The solubility of IM01 in different diluents and the HPLC results of the IM01-1 reference solution are shown in Table 4. The HPLC chromatograms of the IM01-1 reference solutions in Examples 1-2 and 1-5 are shown below. Figure 1-2 The high performance liquid chromatograms of the IM01-1 reference solution and its corresponding blank solvent in test examples 1-7 and 1-12 are shown in Figure 3-4.

[0065] Table 4. Solubility of IM01 in different solvents and liquid chromatography analysis of IM01-1 reference solution.

[0066]

[0067]

[0068] The liquid chromatography chromatogram data of the reference solutions in Test Examples 1-7, 1-8, 1-9, and 1-12IM01-1 are shown in Table 5.

[0069] Table 5. Liquid chromatography data of reference solutions in test examples 1-7, 1-8, 1-9, and 1-12IM01-1

[0070] Test case IM01-1 Retention Time IM01-1 peak area IM01-1 peak height 1-7 5.265 78.969 11.05 1-8 5.239 95.963 13.41 1-9 5.262 92.751 13.07 1-12 5.591 115.876 15.80

[0071] As shown in Table 5, compared with Experimental Examples 1-12, the peak areas of the main peaks detected in Experimental Examples 1-7, 1-8 and 1-9 are smaller, and the detection results of Experimental Example 1-12 are more accurate.

[0072] Experiment 2: Effects of different methanol-dichloromethane-water ratios on the solubility of IM01 and its effect on the liquid phase detection results of IM01-1

[0073] Solutions of methanol-dichloromethane-water in different proportions were prepared as diluents (see Table 6). The preferred preparation method was to first mix methanol and dichloromethane to obtain a mixture, and then add water to the mixture. The solubility of intermediate sample IM01 was tested according to the method described in Experiment 1, and a reference solution of IM01-1 was prepared according to the method described in Experiment 1 and analyzed by high performance liquid chromatography. The results are shown in Table 6.

[0074] Table 6. Solubility of IM01 in methanol-dichloromethane-water at different ratios and liquid chromatography detection of IM01-1 reference solution.

[0075]

[0076] As a result, when the volume ratio of methanol, dichloromethane, and water was (50-75):(5-20):(20-40), the resulting solution exhibited a solubility of IM01 greater than 0.5 mg / ml. Simultaneously, the IM01-1 reference solution showed a single peak with good peak shape in liquid chromatography, indicating accurate detection results. Preferably, the volume ratio of methanol, dichloromethane, and water was (50-75):(5-10):(20-40), or (50-75):(5-10):(20-35), or (51-70):(10-15):(20-35), or (53-65):(15-20):(20-34); more preferably, 60:10:30.

[0077] Example 1-1

[0078] This embodiment examines the ratio of mobile phases A and B, and the solution preparation, chromatographic conditions, and determination methods are as follows:

[0079] (1) Solution preparation

[0080] Diluent: Measure 600ml of methanol and mix it with 100ml of dichloromethane to obtain a mixture. Then measure 300ml of water and mix it with the mixture.

[0081] IM01-1 stock solution: Weigh 10.20 mg of IM01-1 reference standard accurately, place it in a 20 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well.

[0082] IM01-3 stock solution: Weigh 10.07 mg of IM01-3 reference standard accurately, place it in a 20 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well.

[0083] IM01 stock solution: Weigh 10.13 mg of IM01 reference standard accurately, place it in a 20 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well.

[0084] Positioning solutions: Take 0.1 ml of each stock solution and place them in 10 ml volumetric flasks respectively. Add diluent to dissolve and dilute to the mark to obtain the positioning solutions for each impurity.

[0085] System suitability solution: Weigh 10.35 mg of IM01 reference standard accurately and place it in a 20 ml volumetric flask. Add 0.2 ml each of IM01-3 and IM01-1 stock solutions, dissolve and dilute to the mark with diluent to obtain the system suitability solution.

[0086] (2) Chromatographic conditions

[0087] The HPLC detection conditions described in Experiment 1 are the same, except that the ratio of mobile phase A and mobile phase B is different (see Table 7 for mobile phase conditions).

[0088] (3) Determination: Accurately measure 10 μl each of diluent (i.e., as blank solvent), impurity positioning solution and system suitability solution, inject them into the liquid chromatograph, and determine them by high performance liquid chromatography.

[0089] The test results for the system suitability solution are shown in Table 7.

[0090] Table 7 Investigation of mobile phase ratio

[0091]

[0092] The results showed that when mobile phase A was trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (85-90):(10-15) and mobile phase B was trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (10-20):(80-90), the detected chromatographic peaks had good peak shapes, the resolution between the main peak and adjacent peaks was ≥1.5, and the resolution between impurities was ≥1.0, indicating good resolution. IM01-1 was detected as a single peak with good peak shape.

[0093] Examples 1-2

[0094] This embodiment examines the acid concentration of the mobile phase. The solution preparation and determination methods are the same as in Examples 1-1. The chromatographic conditions are the same as those described in Experiment 1 for high performance liquid chromatography detection. The only difference is the acid concentration of the mobile phase in the chromatographic conditions (acid conditions are shown in Table 8). The detection results of the system suitability solution are shown in Table 8.

[0095] Table 8. Investigation of mobile phase acid concentration

[0096]

[0097] The results showed that when the concentration of trifluoroacetic acid in mobile phases A and B was 0.03%-0.08%, the detected chromatographic peaks had good peak shapes, the resolution between the main peak and adjacent peaks was ≥1.5, and the resolution between impurities was ≥1.0, indicating good resolution. Compared with Examples 1-1, the number of impurities detected was consistent, the impurity content RSD was ≤10%, and IM01-1 was detected as a single peak with good peak shape.

[0098] Examples 1-3

[0099] This embodiment examines the gradient elution method. The solution preparation and determination methods are the same as in Examples 1-1, and the chromatographic conditions are the same as those described in Experiment 1 for high performance liquid chromatography (HPLC). The only difference is the gradient elution method used (gradient elution conditions are shown in Table 9). The detection results of the system suitability solution are shown in Table 9. The HPLC chromatograms of the system suitability solution and the blank solvent in Condition 7 are shown in Figure 5.

[0100] Table 9. Investigation of Gradient Elution Methods

[0101]

[0102] The results show that the gradient elution procedure is as follows:

[0103] Time T / min 0 3 10 25 28 35 Mobile phase A% 70-80 70-80 15-25 15-25 70-80 70-80 Mobile phase B% 20-30 20-30 75-85 75-85 20-30 20-30

[0104] At that time, the detected chromatographic peaks had good peak shapes, the resolution between the main peak and adjacent peaks was ≥1.5, and the resolution between impurities was ≥1.0, indicating good resolution. Compared with Example 1-1, the number of impurities detected was consistent, the impurity content RSD was ≤10%, and IM01-1 was detected as a single peak with good peak shape.

[0105] Examples 1-4

[0106] This embodiment examines the chromatographic column, solution preparation and determination methods are the same as in Examples 1-1, and the chromatographic conditions are the same as the high performance liquid chromatography detection conditions described in Experiment 1, the only difference being that the chromatographic column is HY-LC-0028 (250×4.6mm 5μm). The detection results of the system suitability solution are shown in Table 10.

[0107] Table 10 Chromatographic Column Investigation

[0108]

[0109] The results showed that the chromatographic peaks were well-shaped, with a resolution of ≥1.5 between the main peak and adjacent peaks and ≥1.0 between impurities, indicating good resolution. The number of impurities detected was consistent with that in Example 1-1, and the impurity content RSD was ≤10%. IM01-1 was detected as a single peak with a good peak shape.

[0110] Examples 1-5

[0111] This embodiment examines the flow rate, solution preparation, and determination methods as in Examples 1-1. The chromatographic conditions are the same as those described in Experiment 1 for high performance liquid chromatography, except that the flow rates are 0.8 ml / min and 1.2 ml / min. The detection results of the system suitability solution are shown in Table 11.

[0112] Table 11 Flow Velocity Investigation

[0113]

[0114] The results showed that the chromatographic peaks were well-shaped, with a resolution of ≥1.5 between the main peak and adjacent peaks and ≥1.0 between impurities, indicating good resolution. The number of impurities detected was consistent with that in Example 1-1, and the impurity content RSD was ≤10%. IM01-1 was detected as a single peak with a good peak shape.

[0115] Examples 1-6

[0116] This embodiment examines column temperature. The solution preparation and determination methods are the same as in Examples 1-1. The chromatographic conditions are the same as those described in Experiment 1 for high performance liquid chromatography detection, except that the column temperature is 20℃ and 40℃. The detection results of the system suitability solution are shown in Table 12.

[0117] Table 12 Column Temperature Investigation

[0118]

[0119] The results showed that the chromatographic peaks were well-shaped, with a resolution of ≥1.5 between the main peak and adjacent peaks and ≥1.0 between impurities, indicating good resolution. The number of impurities detected was consistent with that in Example 1-1, and the impurity content RSD was ≤10%. IM01-1 was detected as a single peak with a good peak shape.

[0120] Example 2

[0121] This embodiment measures the intermediate sample of carilarazine.

[0122] (1) Solution preparation

[0123] Diluent and system suitability solution: Same as in Example 1-1.

[0124] Test solution: Weigh 10.22 mg of cariprazine intermediate sample accurately, place it in a 20 ml volumetric flask, add diluent to dissolve and dilute to the mark to obtain the test solution.

[0125] (2) Chromatographic conditions

[0126] The same high-performance liquid chromatography (HPLC) detection conditions as described in Experiment 1.

[0127] (3) Determination: Accurately measure 10 μl each of diluent (i.e., as blank solvent), system suitability solution and test solution, inject them into the liquid chromatograph, and determine by high performance liquid chromatography.

[0128] The test results are shown in Table 13. The high-performance liquid chromatograms of the test solution and its blank solvent, as well as the system suitability solution and its blank solvent, are shown in Figures 6-7.

[0129] Table 13 Test Results of Test Sample Solutions and System Suitability Solutions

[0130] name IM01-3 IM01-1 IM01 Minimum resolution blank ND ND ND / Test solution 3.735min 5.321min 20.876min 8.1 System suitability solution 3.734min 5.309min 20.875min 5.4

[0131] The results showed that the cariprazine intermediate sample had good solubility in the diluent and was completely dissolved. The minimum resolution of each peak in the system suitability solution was 5.4, which met the requirements. The blank solvent peak did not interfere with the determination of known impurities and principal components. This method has good specificity.

[0132] Example 3

[0133] This embodiment examines the concentration of the test sample. The solution preparation, chromatographic conditions, and determination methods are the same as in Example 2, except that the concentration of the test sample is prepared to be 0.6 mg / mL. The results show that the peak shapes in each chromatogram are good, the resolution between the main peak and adjacent peaks is ≥1.5, and the resolution between impurities is ≥1.0, indicating excellent resolution. Compared with Example 2, the number of impurities detected is consistent, and IM01-1 is detected as a single peak with a good peak shape.

[0134] In summary, the analytical method described in this invention can efficiently separate and detect cariprazine hydrochloride intermediates and related substances. The method has high sample solubility, good peak separation, good peak shape, good robustness, and high sensitivity.

Claims

1. A method for detecting a cariprazine hydrochloride intermediate and its related substances, characterized by, Includes the following steps: (1) Preparation of test solution: Dissolve the intermediate sample in a solution composed of methanol, dichloromethane and water to obtain the test solution; (2) Detection: The test solution is injected into the liquid chromatograph and detected according to the high performance liquid chromatography (HPLC) conditions. The HPLC conditions are as follows: the column is an octadecylsilane bonded silica column, the mobile phase A is trifluoroacetic acid aqueous solution-acetonitrile, the mobile phase B is trifluoroacetic acid aqueous solution-acetonitrile, and gradient elution is used.

2. The detection method according to claim 1, characterized in that, The solution composed of methanol, dichloromethane, and water has a volume ratio of methanol, dichloromethane, and water of (50-75):(5-20):(20-40); preferably, the volume ratio of methanol, dichloromethane, and water is (50-75):(5-10):(20-40), or (50-75):(5-10):(20-35), or (51-70):(10-15):(20-35), or (53-65):(15-20):(20-34); more preferably, the volume ratio of methanol, dichloromethane, and water is 60:10:

30.

3. The method of claim 1, wherein, The trifluoroacetic acid aqueous solution in mobile phase A and mobile phase B, wherein the concentration of trifluoroacetic acid is 0.03%-0.08%, preferably 0.05%.

4. The method of claim 1, wherein, The mobile phase A is a trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (85-90):(10-15), and the mobile phase B is a trifluoroacetic acid aqueous solution-acetonitrile with a volume ratio of (10-20):(80-90). Preferably, the mobile phase A is trifluoroacetic acid aqueous solution-acetonitrile = 90:10, and the mobile phase B is trifluoroacetic acid aqueous solution-acetonitrile = 10:

90.

5. The method of claim 1, wherein, The gradient elution procedure is as follows: The gradient elution process is preferably as follows:

6. The method of claim 1, wherein, The solution composed of methanol, dichloromethane and water is prepared by first mixing methanol and dichloromethane to obtain a mixed solution, and then adding water to the mixed solution.

7. The detection method according to claim 6, characterized in that, The method for preparing the test solution is as follows: take an intermediate sample, weigh it accurately, dissolve and dilute it with a solution composed of methanol, dichloromethane and water to prepare a solution containing approximately 0.5 mg to 0.6 mg per 1 ml.

8. The method of claim 1, wherein, The high-performance liquid chromatography (HPLC) conditions are as follows: wavelength 254 nm, column temperature 20-40 °C, injection volume 10-15 μL, and flow rate 0.8-1.2 mL / min.

9. The assay of any one of claims 1-8, wherein, Includes the following steps: (1) Preparation of test solution: Take an appropriate amount of intermediate sample, weigh it accurately, dissolve and dilute it with a solution composed of methanol, dichloromethane and water to prepare a solution containing about 0.5 mg per ml; wherein the volume ratio of methanol, dichloromethane and water in the solution is (50-75):(5-10):(20-35); (2) Detection: Inject the test solution into the liquid chromatograph and detect it according to the high-performance liquid chromatography (HPLC) conditions. The HPLC conditions are as follows: the column is an octadecylsilane-bonded silica column; mobile phase A is 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 90:10; mobile phase B is 0.05% trifluoroacetic acid aqueous solution-acetonitrile = 10:90; gradient elution is used; the mobile phase flow rate is 0.8-1.2 ml / min; the column temperature is 20-40℃; the sample injection volume is 10-15 μl; and a photodiode array detector is used with a detection wavelength of 254 nm. The gradient elution program is as follows:

10. The detection method according to claim 1, characterized in that, The intermediate is trans-4-{2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-tert-butoxycarbonyl-cyclohexylamine; the related substances include 1-(2,3-dichlorophenyl)piperazine.

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