Method for detecting related substances in dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone

Abstract

The invention provides a method for detecting related substances in dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone. According to the method, the quality condition of the dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone can be accurately and effectively represented by optimizing detection parameters, and the method is simple to operate, has the advantages of high sensitivity, high precision, accurate content determination result, good specificity and linear relationship and the like, is beneficial to quality control of the dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone, and has a wide application prospect in the field of detection of the dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone and application of the dibenzo [b, f] [1, 4] thiazepine-11-[10H] ketone. Therefore, the quality, safety and effectiveness of the raw material medicines and related preparation products are guaranteed.

Description

Technical Field

[0001] This invention relates to a dibenzo[ b , f [1,4]Thio-11-

[10] H Methods for detecting related substances in ketones, particularly involving a dibenzo[] b , f [1,4]Thio-11-

[10] H Methods for detecting the content of specific impurities in ketones. Background Technology

[0002] Quetiapine Fumarate (11-[4-[2-(2-hydroxyethoxy)ethyl]-1-piperazinyl]dibenzo[ b , f [1,4] Thionido-zirconium hemifumarate, C 21 H 25 N3O2S1 / 2C4H4O4 is an atypical antipsychotic drug primarily used to treat schizophrenia, bipolar disorder (manic and depressive episodes), and as adjunctive therapy for depression. Its mechanism of action involves antagonism of multiple neurotransmitter receptors (such as dopamine D2 and 5-HT2A), thereby regulating brain chemical homeostasis.

[0003]

[0004] dibenzo[ b , f [1,4]Thio-11-

[10] H [[Ketoconol] is a key starting material for the synthesis of quetiapine fumarate. Analysis of its synthetic route and a review of the impurity profile revealed seven main types of impurities, including process byproducts and degradation products. To ensure the safety and efficacy of quetiapine fumarate API, the quality of its starting materials must be studied and controlled. However, there is currently no specific method for controlling the quality of dibenzo[]] b , f [1,4]Thio-11-

[10] H Methods for detecting specific impurities in ketones.

[0005] Summary of the Invention

[0006] Purpose of the invention: This invention aims to provide an analytical method for the effective elution, separation, and quantification of dibenzo[a][b ... b , f [1,4]Thio-11-

[10] HMethods for detecting related substances in ketones.

[0007] Technical solution: The dibenzo[ b , f [1,4]Thio-11-

[10] H The detection method for related substances in ketones includes the following steps: (1) Prepare the sample solution; (2) Detection: The sample solution prepared in step (1) was subjected to gradient elution by high performance liquid chromatography using a phenylhexylsilane-bonded silica column, with 0.1% V trifluoroacetic acid solution as mobile phase A and acetonitrile as mobile phase B. The elution program, based on the volume fraction of mobile phase A, included 0~30 min: decreasing from 70%~80% to 10%~30%, and 30~35 min: maintaining at 10%~30%. (3) Analyze the content of each target component to be tested in step (2).

[0008] Preferably, the target components to be detected by the detection method are as follows: .

[0009] Preferably, the elution process in step (2) further includes 35 to 35.1 min, based on the volume fraction of mobile phase A, increasing from 10% to 30% to 70% to 80%.

[0010] Further preferably, the elution program in step (2) further includes 35.1 to 45 minutes, based on the volume fraction of mobile phase A, maintaining at 70% to 80%.

[0011] Preferably, in the elution process described in step (2), the initial volume fraction of mobile phase A is 73% to 77%.

[0012] Preferably, the elution procedure described in step (2) is as follows: .

[0013] Preferably, the chromatographic column in step (2) is a GL Sciences InertSustain Phenylhexyl column with a length of 150 mm, a diameter of 4.6 mm, and a packing particle size of 3 μm.

[0014] Preferably, in step (2), an ultraviolet absorption detector is used for detection, with a detection wavelength of 268~272nm and a column temperature of 38~42℃.

[0015] Further preferred, the detection wavelength is 270 nm and the column temperature is 40 °C.

[0016] Preferably, the flow rate of the mobile phase in step (2) is 0.9~1.1 ml / min.

[0017] Further preferably, the flow rate of the mobile phase is 1.0 ml / min.

[0018] Preferably, the detection linear range of each target component is as follows: .

[0019] Further preferably, the detection linear equations for each target component are as follows: .

[0020] Preferably, the detection limits for each target component are as follows: .

[0021] Preferably, the limits of quantitation for each target component are as follows: .

[0022] Preferably, in step (3), the area normalization method is used to analyze the content of each target component.

[0023] Preferably, the sample solution prepared in step (1) is as follows: Preparation method: Diluent: 1% acetonitrile phosphate solution; take 100 ml of acetonitrile, add 1 ml of phosphoric acid, mix well, and the solution is ready; Impurity stock solution: Take appropriate amounts of impurity 1 reference standard, impurity 2 reference standard, impurity 3 reference standard, impurity 4 reference standard, impurity 5 reference standard, impurity 6 reference standard and impurity 7 reference standard respectively, dissolve and dilute them with diluent to prepare a solution containing 100 μg of each impurity per 1 ml; Test solution: Take dibenzo[ b , f [1,4]Thio-11-

[10] H A suitable amount of ketone was prepared to contain dibenzo[] per 1 ml. b , f [1,4]Thio-11-

[10] H A solution of 0.5 mg of ketone; Impurity Mixed Solution: Take appropriate amounts of the above-mentioned impurity stock solutions and prepare a mixed solution containing 10 μg of each impurity per 1 ml; System suitability solution: Take dibenzo[ b , f [1,4]Thio-11-

[10] H Accurately weigh an appropriate amount of ketone reference standard, dissolve it in a diluent, and add an appropriate amount of the above-mentioned impurity mixture solution to prepare a solution containing 2.5 μg of each impurity and dibenzo[] per 1 ml. b , f [1,4]Thio-11-

[10] H A mixed solution of 0.5 mg of ketone.

[0024] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The dibenzo[ b , f [1,4]Thio-11-

[10] H The determination method for related substances in ketones can accurately and effectively characterize dibenzo[] b , f [1,4]Thio-11-

[10] H The method for determining the quality of ketones is simple to operate and has advantages such as high sensitivity, high precision, accurate content determination results, good specificity and linearity, which is beneficial for dibenzo[ b , f [1,4]Thio-11-

[10] H Quality control of ketones ensures the quality, safety, and efficacy of active pharmaceutical ingredients and related formulations. Attached Figure Description

[0025] Figure 1 This is the HPLC chromatogram of the system suitability solution from Example 1; Figure 2 The HPLC chromatogram of the test solution in Example 1 is shown below. Figure 3 The HPLC chromatogram of the impurity mixture solution in Comparative Example 1 is shown. Figure 4 The HPLC chromatogram of the system suitability solution for Comparative Example 2; Figure 5 The HPLC chromatogram is for the system suitability solution of Comparative Example 3. Detailed Implementation

[0026] The technical solution of the present invention will be further described below with reference to the embodiments. Example

[0027] (1) Experimental methods Column: GL Sciences InertSustain Phenylhexyl, 4.6 mm × 150 mm, 3 μm; Mobile phase A: 0.1% V trifluoroacetic acid solution; Mobile phase B: Acetonitrile; Flow rate: 1.0 ml / min; Column temperature: 40℃; Wavelength: 270nm; Table 1 Gradient elution procedure of Example 1 .

[0028] Solution preparation: Diluent: Take 100ml of acetonitrile, add 1ml of phosphoric acid, mix well, and you have the diluent.

[0029] Mobile phase A: Take 1000ml of water, add 1ml of trifluoroacetic acid, mix well, and degas by sonication to obtain the mobile phase A.

[0030] Mobile phase B: Acetonitrile.

[0031] Impurity stock solution: Accurately weigh approximately 2 mg of each of the following reference standards: impurity 1, impurity 2, impurity 3, impurity 4, impurity 5, impurity 6, and impurity 7. Place them in separate 20 ml volumetric flasks, add an appropriate amount of diluent, sonicate to dissolve, cool, dilute to the mark with diluent, and shake well.

[0032] Test solution: Take dibenzo[ b , f [1,4]Thio-11-

[10] H Take about 10 mg of ketone, place it in a 20 ml volumetric flask, add an appropriate amount of diluent, sonicate to dissolve, cool, dilute to the mark with diluent, and shake well.

[0033] Impurity mixed solution: Take 5 ml of each of the above impurity reference standard stock solutions, place them in a 50 ml volumetric flask, dilute to the mark with diluent, and shake well.

[0034] System suitability solution: Take dibenzo[ b , f [1,4]Thio-11-

[10] H Take about 10 mg of ketone reference standard, place it in a 20 ml volumetric flask, add an appropriate amount of diluent, sonicate to dissolve, cool, add 5 ml of the above impurity mixture solution, dilute to the mark with diluent, and shake well.

[0035] Sample determination: Measure 10 μl each of blank solution, test solution and system suitability solution and inject them into the liquid chromatograph.

[0036] (2) Experimental results Chromatogram of solution based on system suitability ( Figure 1 It can be seen that RT=12.623min is dibenzo[ b , f [1,4]Thio-11-

[10] H The chromatographic peaks of the ketone were as follows: RT=5.720 min for impurity 1, RT=6.773 min for impurity 2, RT=9.080 min for impurity 3, RT=9.950 min for impurity 4, RT=11.483 min for impurity 5, RT=17.260 min for impurity 6, and RT=23.837 min for impurity 7. Under this method, baseline separation was achieved between the main component peak and each impurity peak, with a minimum resolution of 4.01 and a relatively stable baseline.

[0037] From the chromatogram of the test sample solution ( Figure 2 It can be seen that RT=12.633min is dibenzo[ b , f [1,4]Thio-11-

[10] H The chromatographic peaks of the ketone were as follows: RT=5.727 min for impurity 1, RT=6.787 min for impurity 2, RT=9.097 min for impurity 3, RT=9.967 min for impurity 4, RT=11.493 min for impurity 5, RT=17.251 min for impurity 6, and RT=23.823 min for impurity 7. Under this method, the separation between the main component chromatographic peak and each impurity chromatographic peak was good.

[0038] Example 2: Methodology Validation Experiment (1) Experimental methods (1.1) Specificity and System Applicability Blank solutions, system suitability solutions, and test solutions were prepared according to the method in Example 1 for determination to examine method specificity and system suitability. The results showed that, using the chromatographic conditions of this invention, the blank solution was free of interference, and the main component and impurities, as well as among the impurities themselves, could be effectively separated, indicating good method specificity.

[0039] (1.2) Linearity and Range The impurity reference standard stock solutions were prepared according to the method in Example 1. Appropriate amounts of each impurity reference standard stock solution were accurately measured and diluted with a diluent to prepare a series of concentrations, serving as linear solutions for each component. The results showed that, using the chromatographic conditions of this invention, the linear relationships of each analyte were good.

[0040] (1.3) Limit of Quantitation and Limit of Detection Stock solutions of each impurity reference standard were prepared according to the method in Example 1. Appropriate amounts of each stock solution were accurately measured and gradually diluted with diluent for determination. The limit of quantitation was set at a signal-to-noise ratio of approximately 10:1, and the limit of detection was set at a signal-to-noise ratio of approximately 3:1. The detection sensitivity of each impurity was then investigated. The results showed that the method has good sensitivity.

[0041] (1.4) Repeatability Accurately weigh an appropriate amount of this product, add a 100% impurity limit reference solution, dissolve and dilute to the mark with diluent to prepare the spiked test solution (prepare 6 parallel aliquots). Accurately measure 10 μl of each of the above test solutions and inject them into the liquid chromatograph, record the chromatogram, and examine the repeatability of the method. The results show that the recoveries of each impurity are all between 80% and 120%, and the RSD is less than 10%, indicating that the method has good repeatability.

[0042] (1.5) Accuracy Accurately weigh an appropriate amount of this product, add impurity reference solutions with limits of 50%, 100%, and 150%, respectively, dissolve and dilute to the mark with diluent to prepare the spiked test solutions. Accurately measure 10 μl of each of the above test solutions and inject them into the liquid chromatograph, record the chromatograms, and calculate the recovery rate of each impurity as [(measured amount - background amount) / added amount]. The results show that the recoveries of each impurity at different concentration levels are all between 80% and 120%, and the RSDs are all less than 10%, indicating that this method has good accuracy and is suitable for the detection of related substances in this product.

[0043] (1.6) Solution stability At different time points, samples of the system suitability solution and the 100% spiked test solution were injected into the liquid chromatograph, and the chromatograms were recorded. The results showed that after 75 hours at room temperature, the peak area ratios of all impurities and the main component to the 0h peak in the system suitability solution were all within the range of 0.70–1.30; after 70.5 hours at room temperature, the peak area ratios of all impurities and the main component to the 0h peak in the 100% spiked test solution were all within the range of 0.80–1.20, indicating good solution stability in this method.

[0044] (1.7) Durability Blank solutions, system suitability solutions, and test solutions were prepared according to the method in Example 1 for determination, and the robustness of the method was investigated within a small range of chromatographic parameters. The results showed that under all chromatographic conditions, the blank solution caused no interference, and the main component and impurities, as well as the impurities themselves, could be effectively separated, indicating that the method has good robustness.

[0045] (2) Experimental results Table 2 Verification Results and Conclusions .

[0046] As shown in Table 2, the detection method of Example 1 was used to determine dibenzo[ b , f [1,4]Thio-11-

[10] H When dealing with substances related to ketones, it exhibits high specificity, high sensitivity, good linearity, high accuracy, and good solution stability and robustness, enabling the detection of dibenzo[ b , f [1,4]Thio-11-

[10] H Accurate detection of ketone samples and related substances.

[0047] Comparative Example 1 (1) Experimental methods Column: Agilent Eclipse XDB-C18, 4.6mm × 150mm, 5.5μm; Mobile phase A: 0.1% V trifluoroacetic acid solution; Mobile phase B: Acetonitrile; Flow rate: 1.0 ml / min; Column temperature: 40℃; Wavelength: 270nm; Test sample concentration: 0.5 mg / ml; Table 3 Gradient elution procedure for Comparative Example 1 .

[0048] The solution preparation method and detection method (except for the chromatographic column) of Example 1 were used to determine dibenzo[ b , f [1,4]Thio-11-

[10] H Retention time of ketones and impurities.

[0049] (2) Experimental results Chromatogram of impurity mixture solution ( Figure 3 As can be seen, the chromatographic peaks are as follows: RT=8.063 min for impurity 1, RT=11.823 min for impurity 2, RT=16.533 min for impurity 3, RT=19.860 min for impurity 4, RT=22.370 min for impurity 5, and RT=32.477 min for impurity 6. Although the separation of other impurities is relatively good, impurity 7 was not eluted under this method. Therefore, this detection method does not meet the quality control requirements.

[0050] Comparative Example 2 (1) Experimental methods Column: MicroPulite PHS XP Phenyl-Hexyl, 4.6mm × 150mm, 3μm; Mobile phase A: 0.1% V trifluoroacetic acid solution; Mobile phase B: Acetonitrile; Flow rate: 1.0 ml / min; Column temperature: 40℃; Wavelength: 270nm; Test sample concentration: 0.5 mg / ml; Table 4 Gradient elution procedure for Comparative Example 2 .

[0051] The solution preparation method and detection method (except for the chromatographic column and elution procedure) of Example 1 were used to determine dibenzo[ b , f [1,4]Thio-11-

[10] H Retention time of ketones and impurities.

[0052] (2) Experimental results Chromatogram of solution based on system suitability ( Figure 4 It can be seen that RT=15.080min is dibenzo[ b , f [1,4]Thio-11-

[10] H The chromatographic peaks of the ketone were as follows: RT=6.780 min for impurity 1, RT=9.093 min for impurity 2, RT=11.580 min for impurity 3, RT=12.553 min for impurity 4, RT=15.537 min for impurity 5, RT=20.050 min for impurity 6, and RT=25.223 min for impurity 7. However, the baseline of impurity 5 was not separated from that of the main component, therefore this detection method does not meet the quality control requirements.

[0053] Comparative Example 3 (1) Experimental methods Chromatographic column: Artchrom Phenylhexyl WP, 4.6mm × 150mm, 3.5μm; Mobile phase A: 0.1% V trifluoroacetic acid solution; Mobile phase B: Acetonitrile; Flow rate: 1.0 ml / min; Column temperature: 40℃; Wavelength: 270nm; Table 5 Gradient elution procedure for Comparative Example 3 .

[0054] The solution preparation method and detection method (except for the chromatographic column) of Example 1 were used to determine dibenzo[ b , f [1,4]Thio-11-

[10] H Retention time of ketones and impurities.

[0055] (2) Experimental results Chromatogram of solution based on system suitability ( Figure 5 It can be seen that RT=15.070min is dibenzo[ b , f [1,4]Thio-11-

[10] H The chromatographic peaks of the ketone were as follows: RT=9.063 min for impurity 1, RT=11.560 min for impurity 2, RT=12.567 min for impurity 3, RT=14.810 min for impurity 5, RT=20.117 min for impurity 6, and RT=25.297 min for impurity 7. However, under this method, the chromatographic peak of impurity 4 could not achieve baseline separation from the chromatographic peak of the main component. Therefore, this detection method does not meet the quality control requirements.

Claims

1. A dibenzo[ b , f [1,4]Thio-11-[10] H A method for detecting related substances in ketones, characterized in that, Includes the following steps: (1) Prepare the sample solution; (2) Detection: The sample solution prepared in step (1) was subjected to gradient elution by high performance liquid chromatography using a phenylhexylsilane-bonded silica column, with 0.1% V trifluoroacetic acid solution as mobile phase A and acetonitrile as mobile phase B. The elution program, based on the volume fraction of mobile phase A, included 0~30 min: decreasing from 70%~80% to 10%~30%, and 30~35 min: maintaining at 10%~30%. (3) Analyze the content of each target component to be tested in step (2).

2. The detection method according to claim 1, characterized in that, The target components to be detected are as follows: 。 3. The detection method according to claim 1 or 2, characterized in that, In the elution process described in step (2), the initial volume fraction of mobile phase A is 73%~77%.

4. The detection method according to claim 1 or 2, characterized in that, The elution procedure described in step (2) is as follows: 。 5. The detection method according to claim 1 or 2, characterized in that, The chromatographic column mentioned in step (2) is a GLSciences InertSustain Phenylhexyl column with a length of 150 mm, a diameter of 4.6 mm, and a packing particle size of 3 μm.

6. The detection method according to claim 1 or 2, characterized in that, In step (2), an ultraviolet absorption detector is used for detection, with a detection wavelength of 268~272nm and a column temperature of 38~42℃.

7. The detection method according to claim 6, characterized in that, The detection wavelength is 270 nm, and the column temperature is 40 °C.

8. The detection method according to claim 1 or 2, characterized in that, The flow rate of the mobile phase in step (2) is 0.9~1.1 ml / min.

9. The detection method according to claim 1 or 2, characterized in that, The detection linear range of each target component is as follows: 。 10. The detection method according to claim 1 or 2, characterized in that, In step (3), the area normalization method is used to analyze the content of each target component.

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