A method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography

Abstract

The invention discloses a method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography, and belongs to the technical field of pharmaceutical analysis. The invention adopts high performance liquid chromatography to determine the content of impurities in sertraline hydrochloride capsules, and sertraline can be well separated from various impurities; the auxiliary material peak and the solvent peak do not interfere with the detection of the main component and impurities; and the samples destroyed under various conditions are tested for peak purity, and the spectral purity is greater than 990, and the impurities generated by the destruction under various conditions can be well separated from the main peak, and the detection specificity of related substances is good; at the same time, when the chromatographic condition parameters are slightly changed, the separation and inspection results of related substances and impurities in Aprelet tablets are not affected, and the durability is good. The present application can quickly, effectively, accurately and reliably separate and detect related substances in sertraline hydrochloride capsules, which is beneficial to improving the product quality of sertraline hydrochloride capsules and improving the safety of patients' medication.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical analysis, and more specifically, relates to a method for determining related substances in sertraline hydrochloride capsules by high performance liquid phase method. Background Art

[0002] Sertraline is the main ingredient of sertraline hydrochloride capsules. Sertraline is used to treat symptoms related to depression, including depression with anxiety, with or without a history of mania. After satisfactory efficacy, continued use of sertraline can effectively prevent the recurrence and recurrence of depression. Sertraline is also used to treat obsessive-compulsive disorder. After the initial treatment responds, sertraline maintains its effectiveness, safety and tolerability in the treatment of obsessive-compulsive disorder for two years.

[0003] The method under the item of related substances of Sertraline Hydrochloride Capsules in the 2020 edition of the Chinese Pharmacopoeia uses isocratic elution of ammonium dihydrogen phosphate solution-methanol (35:65), which has a low detection ability for impurities and only controls total impurities, and the control requirements for each characteristic impurity are unclear. Therefore, it is necessary to improve the inspection method for related substances of Sertraline Hydrochloride Capsules and establish a rapid and effective detection method, which is conducive to improving the product quality of Sertraline Hydrochloride Capsules and improving the safety of patients' medication. Summary of the invention

[0004] In view of the above problems existing in the prior art, the technical problem to be solved by the present invention is to provide a method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0006] A method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography, characterized in that the content of impurities in sertraline hydrochloride capsules is determined by high performance liquid chromatography, specifically comprising the following steps:

[0007] (1) Prepare test solution, control solution and sensitivity solution;

[0008] (2) Setting HPLC detection conditions;

[0009] (3) Accurately pipette the test solution and control solution separately, inject them into the liquid chromatograph, record the chromatogram, and calculate the content of each impurity in the test sample.

[0010] Furthermore, the detection wavelength of the liquid chromatograph in the high performance liquid method is 220 nm.

[0011] Furthermore, a chromatographic column with octadecyl bonded silica as a filler is used, the column temperature is 33-37°C; ammonium acetate solution-methanol is used as a mobile phase, the flow rate of the mobile phase is 0.95-1.05 mL / min, the pH is 4.9-5.1, and the volume ratio of the ammonium acetate solution to methanol is 43:57-47:53.

[0012] Furthermore, the column temperature of the chromatographic column is 35°C.

[0013] Furthermore, the flow rate of the mobile phase is 1 mL / min, and the pH is 5.0.

[0014] Furthermore, the volume ratio of the ammonium acetate solution to methanol is 45:55.

[0015] Furthermore, the chromatographic column has a length of 150 mm, an inner diameter of 4.6 mm, and a filler particle size of 3 μm.

[0016] Further, 30 mg of the contents of Sertraline Hydrochloride Capsules were placed in a 20 mL volumetric flask, dissolved and diluted to the scale with mobile phase A, shaken, filtered, and the filtrate was taken as the test solution; 1 mL of the test solution was transferred to a 100 mL volumetric flask, diluted to the scale with mobile phase A, shaken, and used as the control solution; 5 mL of the test solution was transferred to a 100 mL volumetric flask, diluted to the scale with mobile phase A, shaken, and used as the sensitivity solution.

[0017] According to the method provided by the present invention, impurity B, impurity D, impurity C, sertraline and impurity F can be effectively separated, and the blank auxiliary material and solvent peak do not interfere with the impurity detection. According to the European Pharmacopoeia EP10.0, the impurity limit is formulated as follows:

[0018] If there are impurity peaks in the chromatogram of the test solution, the peak area of ​​impurity B shall not be greater than 0.2 times (0.2%) the main peak area of ​​the control solution, the sum of the peak areas of impurity D and impurity C shall not be greater than 0.8 times (0.8%) the main peak area of ​​the control solution, the peak area of ​​impurity F shall not be greater than 0.2 times (0.2%) the main peak area of ​​the control solution, the peak area of ​​a single impurity shall not be greater than 0.2 times (0.2%) the main peak area of ​​the control solution, the sum of the peak areas of all impurities shall not be greater than 1.5 times (1.5%) the main peak area of ​​the control solution, and chromatographic peaks smaller than the main peak area of ​​the sensitivity solution shall be ignored.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] Compared with the method for related substances under the sertraline hydrochloride capsules of the 2020 edition of the "Chinese Pharmacopoeia", the method of the present application can well separate sertraline from various impurities; the excipient peaks and solvent peaks do not interfere with the detection of the main components and impurities; and the samples destroyed under various conditions are tested for peak purity, and the spectral purity is greater than 990, and the impurities produced by the destruction under various conditions can be well separated from the main peak, and the detection specificity of related substances is good; at the same time, when the chromatographic condition parameters undergo slight changes, the separation and inspection results of the impurities of the related substances of sertraline hydrochloride are not affected, and the durability is good; the use of this high performance liquid chromatography method can quickly, effectively, accurately and reliably separate and detect the related substances in sertraline hydrochloride capsules, which is beneficial to improving the product quality of sertraline hydrochloride capsules and improving the safety of patient use. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The chromatogram is tested under the chromatographic conditions of the present invention;

[0022] Figure 2 The chromatogram was not destroyed in the destructive test;

[0023] Figure 3 It is the acid destruction chromatogram in the destructive test;

[0024] Figure 4 It is the chromatogram of alkaline destruction in destructive test;

[0025] Figure 5 It is the chromatogram of oxidation destruction in destructive test;

[0026] Figure 6 It is the light destruction chromatogram in destructive test;

[0027] Figure 7 Chromatogram for high temperature destruction in destructive tests;

[0028] Figure 8 It is the chromatogram of thermal decomposition in destructive test;

[0029] Fig. 9 The chromatogram was obtained with reference to the HPLC conditions of related substances in Sertraline Hydrochloride Capsules of the Chinese Pharmacopoeia 2020 edition. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described below in conjunction with specific embodiments.

[0031] The impurity content in sertraline hydrochloride capsules was determined by high performance liquid chromatography. The experimental conditions were as follows:

[0032] Chromatographic column: C18 chromatography (Agilent ZORBAX Eclipse XDB-C8 4.6×250mm, 5μm);

[0033] Flow rate: 1.0 mL / min;

[0034] Column temperature: 35°C;

[0035] Injection volume: 20 μL;

[0036] Mobile phase: Mobile phase A (ammonium acetate solution-methanol = 45:55) + mobile phase B (methanol)

[0037] Detection wavelength: 220nm

[0038] Impurity B is (1S, 4S)-4-phenyl-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF:C 17 H 19 N.HClMW: 237.3536.46, structural formula is as follows:

[0039] and enantiomers

[0040] Impurity C is (1S, 4S)-4-(4-dichlorophenyl)-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF: C 17 H 18 ClN.HCl MW: 271.7936.46, structural formula is as follows:

[0041] and enantiomers

[0042] Impurity D is (1S, 4S)-4-(3-dichlorophenyl)-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF: C 17 H 18 ClN.HCl MW: 271.7936.46, structural formula is as follows:

[0043] and enantiomers

[0044] Impurity F is (R)-4-(3,4-dichlorophenyl)=3,4-dihydronaphthalene=1(2H)=ketone, MF:C 16 H 12 Cl2O MW: 291.17, structural formula is as follows:

[0045]

[0046] Example 1

[0047] The impurity content in sertraline hydrochloride capsules was determined by high performance liquid chromatography, which specifically includes the following steps:

[0048] (1) Take 30 mg of the contents of sertraline hydrochloride capsules and place them in a 20 mL volumetric flask, dissolve and dilute to the mark with mobile phase A, shake well, filter, and take the filtrate as the test solution. Take 1 mL of the test solution and place it in a 100 mL volumetric flask, dilute to the mark with mobile phase A, shake well, and use it as the control solution. Take 5 mL of the test solution and place it in a 100 mL volumetric flask, dilute to the mark with mobile phase A, shake well, and use it as the sensitivity solution. Take appropriate amounts of sertraline hydrochloride, impurity B, impurity C, impurity D, and impurity F and dissolve and dilute them to obtain a mixed solution of about 0.5 mg / mL sertraline and 25 μg / mL impurities B, impurity C, impurity D, and impurity F, and use it as the system suitability solution.

[0049] (2) Setting the HPLC detection conditions: using octadecyl bonded silica gel as the filler, ammonium acetate solution (take 3.85 g of ammonium acetate, add 5 mL of triethylamine, dissolve in water and dilute to 1000 mL, adjust the pH to 5.0 with acetic acid)-methanol = 45:55 as the mobile phase A, methanol as the mobile phase B, gradient elution at a flow rate of 1.0 mL / min, column temperature of 35°C; injection volume of 20 μL; detection wavelength of 220 nm;

[0050] (3) Accurately pipette the test solution and the control solution respectively, inject them into the liquid chromatograph, and record the chromatogram. Figure 1 As shown. Figure 1 It can be seen that under this chromatographic condition, the main component can be effectively separated from sertraline and various impurities at a detection wavelength of 220nm, and the solvent peak and blank excipient peak do not interfere with the determination of known impurities and unknown impurities, which meets the standards.

[0051] Example 2

[0052] The detection method of Example 1 was validated from several aspects, including system applicability, destructive testing, limit of quantification, limit of detection, linear relationship, membrane adsorption, precision, accuracy, and solution stability, as described in detail below.

[0053] (1) System applicability

[0054] Take an appropriate amount of sertraline hydrochloride, impurity B, impurity C, impurity D, impurity E and impurity F, accurately weigh, add methanol to dissolve and dilute to make each impurity stock solution containing about 0.5 mg of each impurity per 1 mL. Take an appropriate amount of each impurity stock solution and make a single fixed solution of each impurity containing 25 μg of sertraline hydrochloride, impurity B, impurity C, impurity D, impurity E and impurity F per 1 mL. Measure an appropriate amount of each stock solution and add an appropriate amount of sertraline hydrochloride to the same volumetric bottle, dissolve with solvent and make up to a mixed solution of about 0.5 mg / mL of sertraline and 25 μg / mL of impurity B, impurity C, impurity D, impurity E and impurity F as the system suitability solution. Take an appropriate amount of blank excipient according to the prescription, dissolve with solvent and dilute to 1 mg / mL, shake well, and filter as a blank excipient solution.

[0055] Take 20 μL of each of the blank solution, blank excipient, sertraline hydrochloride positioning solution, each impurity positioning solution and system suitability solution, inject them into the HPLC, and record the chromatogram at a detection wavelength of 220 nm to examine the separation of each component. : Under the chromatographic conditions, sertraline, each known impurity and other impurities can be effectively separated, and the solvent peak and blank excipient peak do not interfere with the determination of known impurities and unknown impurities, which meets the standard.

[0056] Table 1 Impurity location and separation

[0057]

[0058] (2) Destructive testing

[0059] Blank excipient solution: Take 10 mg of blank excipient and place it in a 10 mL volumetric flask, add solvent to dissolve to volume, mix well. Filter and take the filtrate as the blank excipient solution.

[0060] Unbroken ( Figure 2 ): Take about 30 mg of the contents of Sertraline Hydrochloride Capsules and place them in a 20 mL volumetric flask, add solvent to dissolve to volume, and mix well. Filter and take the filtrate as the unbroken solution.

[0061] Acid damage ( Figure 3 ): Take about 30 mg of the contents of sertraline hydrochloride capsules and place them in a 20 mL volumetric flask, add 1 mL of 1 mol / L hydrochloric acid solution, destroy it for 2 hours, add 1 mL of 1 mol / L sodium hydroxide solution to neutralize it, add solvent to dissolve and make up the volume, mix well, filter, and take the filtrate as the acid destruction solution.

[0062] Alkali damage ( Figure 4 ): Take about 30 mg of the contents of sertraline hydrochloride capsules and place them in a 20 mL volumetric flask, add 1 mL of 1 mol / L sodium hydroxide solution, destroy for 2 hours, add 1 mL of 1 mol / L hydrochloric acid solution to neutralize, add solvent to dissolve to the fixed volume, mix well, filter, and take the filtrate as the alkali destruction solution.

[0063] Oxidative damage ( Figure 5 ): Take about 30 mg of the contents of sertraline hydrochloride capsules and place them in a 20 mL volumetric flask, add 1 mL of 30% hydrogen peroxide solution, destroy for 2 hours, add solvent to dissolve to volume, mix well, filter, and take the filtrate as the oxidation destruction solution.

[0064] Light damage ( Figure 6 ): Take about 30 mg of the contents of sertraline hydrochloride capsules and place them in a 20 mL volumetric flask, add solvent to dissolve to a fixed volume, and mix well. After exposure to ultraviolet light at 254 nm and 365 nm for 5 min, filter and take the filtrate as the light exposure solution.

[0065] High temperature damage ( Figure 7 ): Take about 30 mg of the contents of sertraline hydrochloride capsules and put them into a 20 mL volumetric flask, place it at a high temperature of 100°C for 2 hours, take it out and let it cool at room temperature, add solvent to dissolve to the fixed volume, mix well, filter, and take the filtrate as the high temperature destruction solution.

[0066] Pyrolysis destruction Figure 8 ): Take about 30 mg of the content of sertraline hydrochloride capsules and put it into a 20 mL volumetric flask. After adding solvent to dissolve, place it at high temperature of 100°C for 2 hours. After taking it out, let it cool at room temperature, add solvent to dissolve it to the fixed volume, mix it evenly, filter it, and take the filtrate as the high temperature destruction solution.

[0067] For the destruction of excipients by acid, alkali, oxidation, high temperature and pyrolysis, take an appropriate amount of blank excipient according to the prescription, and the destruction conditions are the same as those of the test samples; the light destruction conditions of the blank excipients are the same as those of the test samples, and the destruction time is 2h.

[0068] Acid / base blank: Take hydrochloric acid solution and sodium hydroxide solution and prepare them in the same way as the acid-base destruction experiment of the test sample.

[0069] Oxidation blank: Take a hydrogen peroxide solution and prepare it in the same way as the test sample for oxidation damage experiment.

[0070] Take 20 μL of the system suitability solution and the above test solution and inject them into the chromatograph, record the chromatogram, and analyze the changes of each impurity. For the test solution under various destruction conditions, use diode array measurement to measure the peak purity or spectral similarity.

[0071] Table 2 Material balance investigation results

[0072]

[0073] (3) Limit of quantification and limit of detection test

[0074] Take appropriate amount of each impurity stock solution under the specificity item to prepare a mixed solution containing 10 μg of sertraline hydrochloride, impurity B, impurity C, impurity D and impurity F per 1 mL. Use the stepwise dilution method to measure the limit of quantification (S / N ≥ 10) and the limit of detection (S / N ≥ 3).

[0075] Table 3 Detection limit and quantification limit

[0076]

[0077]

[0078] (4) Filter membrane adsorption

[0079] Take about 30 mg of the content of sertraline hydrochloride capsules and put it into a 20 mL volumetric flask, add solvent to dissolve and make up the volume, mix well, filter through a 0.45 μm nylon filter membrane, discard 1 mL, 2 mL, 3 mL, and 4 mL respectively, take the filtrate as the filtration sample, and compare the main peak area and impurity changes with the centrifuged sample of the supernatant obtained by direct centrifugation of the solution to examine the membrane adsorption. The results are shown in the table below.

[0080] Table 4 Filter membrane adsorption

[0081]

[0082] (5) Injection precision

[0083] Under the present chromatographic conditions, the samples were injected six times in succession, and the RSD of the peak areas of each known impurity and main peak were all ≤2.0%, indicating good injection precision.

[0084] Table 5 Injection precision results

[0085]

[0086]

[0087] (6) Solution stability

[0088] Under the chromatographic conditions, the samples were injected at 0h, 2h, 4h, 6h, 8h, 12h, 18h and 24h to investigate its intra-day stability.

[0089] Table 6 Stability results of test solution

[0090]

[0091] (7) Linearity and range

[0092] Under the present chromatographic conditions, linear solutions were prepared at limits of 0.008%, 50%, 70%, 100%, and 200%. The results are shown in the table below.

[0093] Table 7 Linear relationship test results

[0094]

[0095]

[0096] (8) Repeatability

[0097] Under the present chromatographic conditions, the determination was repeated 6 times and the results are shown in the table below.

[0098] Table 8 Repeatability test results

[0099]

[0100] (9) Intermediate precision

[0101] The same batch of samples were checked for related substances by the same operator at different times using the repeatability test method; the same batch of samples were checked for related substances by different operators using the repeatability test method; the same batch of samples were checked for related substances by the same operator using different instruments. The results are shown in the table below.

[0102] Table 9 Intermediate precision test results

[0103]

[0104]

[0105] (10) Accuracy

[0106] Under the present chromatographic conditions, recovery solutions were prepared at 50%, 100%, and 200% limits. The results are shown in the table below.

[0107] Table 10 Sample recovery

[0108]

[0109]

[0110] (11) Durability

[0111] Under the chromatographic conditions, the durability of the method for the relevant substances was verified mainly from four aspects: different proportions of mobile phase, different flow rates, different column temperatures and different chromatographic columns. The results are shown in the table below.

[0112] Table 11 Durability test results

[0113]

[0114]

[0115] Comparative Example 1

[0116] Referring to the HPLC conditions of the relevant substances of Sertraline Hydrochloride Capsules in the 2020 edition of the Chinese Pharmacopoeia, the chromatogram was measured as follows Fig. 9 As shown, it can be seen that impurity C cannot be separated from impurity D, and impurity B cannot be separated from the solvent peak.

[0117] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography, characterized in that: The content of impurities in sertraline hydrochloride capsules was determined by high performance liquid chromatography, which specifically includes the following steps: (1) Prepare test solution, control solution and sensitivity solution; (2) Setting high performance liquid chromatography detection conditions; the detection wavelength of the liquid chromatograph in the high performance liquid chromatography method is 220 nm, a chromatographic column with octadecyl bonded silica gel as a filler is used, and the column temperature is 33 to 37° C.; ammonium acetate solution-methanol is used as a mobile phase, the flow rate of the mobile phase is 0.95 to 1.05 mL / min, the pH is 4.9 to 5.1, and the volume ratio of the ammonium acetate solution to methanol is 43:57 to 47:53; (3) Accurately pipette the test solution and the control solution respectively, inject them into the liquid chromatograph, record the chromatogram, and calculate the content of each impurity in the test sample; Impurity B is (1S, 4S)-4-phenyl-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF: C17H19N .HClMW: 237.3536.46, and the structural formula is as follows: and enantiomers; Impurity C is (1S, 4S)-4-(4-dichlorophenyl)-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF: C17H18ClN.HCl MW: 271.7936.46, and the structural formula is as follows: and enantiomers; Impurity D is (1S, 4S)-4-(3-dichlorophenyl)-1,2,3,4-tetrahydro-N-methyl-1-naphthylamine hydrochloride, MF: C17H18ClN.HCl MW: 271.7936.46, and the structural formula is as follows: and enantiomers; Impurity F is (R)-4-(3,4-dichlorophenyl)=3,4-dihydronaphthalene=1(2H)=ketone, MF: C16H12Cl2OMW: 291.17, and the structural formula is as follows: 。 2. The method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography according to claim 1, characterized in that: The column temperature of the chromatographic column is 35°C.

3. The method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography according to claim 1, characterized in that: The flow rate of the mobile phase is 1 mL / min, and the pH value is 5.

0.

4. The method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography according to claim 1, characterized in that: The volume ratio of the ammonium acetate solution to methanol is 45:

55.

5. The method for determining related substances in sertraline hydrochloride capsules by high performance liquid chromatography according to claim 1, characterized in that: The chromatographic column has a length of 250 mm, an inner diameter of 4.6 mm, and a filler particle size of 5 μm.

6. The method for determining the impurity content in sertraline hydrochloride capsules according to claim 1, characterized in that: Take 30 mg of the contents of Sertraline Hydrochloride Capsules and place it in a 20 mL volumetric flask, dissolve and dilute to the scale with mobile phase A, shake well, filter, and take the filtrate as the test solution; transfer 1 mL of the test solution to a 100 mL volumetric flask, dilute to the scale with mobile phase A, shake well, and use it as the control solution; transfer 5 mL of the test solution to a 100 mL volumetric flask, dilute to the scale with mobile phase A, shake well, and use it as the sensitivity solution.

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