Method for detecting related substances of perampanel oral soluble film

Abstract

The invention provides a method for detecting related substances of a perampanel oral soluble film. According to the method for detecting the related substances of the perampanel oral soluble membrane, high performance liquid chromatography is adopted, and the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is taken as a filler of a chromatographic column, the column temperature is 38-42 DEG C, a mobile phase A and a mobile phase B are taken as mobile phases, and the mobile phase A is a phosphate buffer solution; the mobile phase B is acetonitrile; the volume ratio of the mobile phase A to the mobile phase B is set as 0 min, 75%: 25% in the elution procedure; 40 min, 50%: 50%; 50%: 50% when the time is 55min; 60 min, 75%: 25%; 80 min, 75%: 25%. According to the present invention, the HPLC method is adopted, the specific formula mobile phase is combined with the specific elution procedure and the specific chromatographic condition to determine the contents of the impurity B, the impurity C and the impurity D in the perampanel oral soluble film, the detection method has advantages of good separation effect, sensitivity, accuracy and the like, and has advantages of simpleness, strong specificity, high sensitivity, good durability, good repeatability, high detection sensitivity and the like; and the preparation quality of the perampanel oral soluble film is better guaranteed.

Description

Technical Field

[0001] The invention relates to the field of chemical drug quality detection, and in particular to a method for detecting related substances of a perampanel orodispersible film. Background Art

[0002] Perampanel is an anti-epileptic drug with a new mechanism of action. It is the first approved AMPA receptor (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) antagonist. It inhibits glutamate-induced excessive neurotransmission to treat epilepsy by non-competitively binding to AMPA receptors on the postsynaptic membrane. The original formulation of perampanel was developed by Eisai of Japan and was first approved for marketing in the European Union in July 2012 under the trade name Fycompa. In October 2012, it was approved for marketing in mainstream countries such as the United States and Switzerland. In September 2019, perampanel tablets were approved for marketing in China under the trade name Fycompa, with specifications of 2mg and 4mg. It is suitable for the treatment of partial seizures in adults and children aged 4 years and above (with or without secondary generalized seizures).

[0003] Perampanel orodispersible film is a film preparation with perampanel as the active ingredient and certain excipients. The current specifications include 4 mg and 2 mg. The preparation process and selected excipients of the preparation have an important impact on the content and stability of its related substances. At present, the pharmacopoeias of various countries have not yet included the detection methods of related substances of perampanel orodispersible film. There are only the registration standards for imported drugs for ordinary tablets and the quality standards of raw material suppliers. The standardization of the detection methods of related substances of perampanel orodispersible film is not conducive to the quality control of perampanel orodispersible film. Therefore, it is of great significance to carry out research on the detection methods of related substances of perampanel orodispersible film. Summary of the invention

[0004] In view of this, the present invention proposes a method for detecting related substances of perampanel orodispersible film to solve the above problems.

[0005] The technical solution of the present invention is achieved as follows: a method for detecting related substances of perampanel orodispersible film, using high performance liquid chromatography, the chromatographic conditions are: the filler of the chromatographic column is octadecylsilane bonded silica gel, the column temperature is 38-42°C, mobile phase A and mobile phase B are used as mobile phases, mobile phase A is phosphate buffer; mobile phase B is acetonitrile; the mobile phase elution program is:

[0006] Time (minutes) Mobile phase A (%, v / v) Mobile phase B (%, v / v) 0 75 25 40 50 50 55 50 50 60 75 25 80 75 25 .

[0007] Further, the phosphate buffer solution: potassium dihydrogen phosphate is taken, dissolved and diluted with water to prepare a concentration of 1-1.5 g / L, and the pH value is adjusted to 4.0-5.0.

[0008] Furthermore, the pH value is adjusted using phosphoric acid or 0.01-0.03 mol / L potassium hydroxide solution.

[0009] Furthermore, the detection solution includes a test solution, a control solution, and a system suitability solution, and the solvent for preparing the solution is acetonitrile-water with a volume ratio of 40-60:40-60.

[0010] Further, the test sample solution: take the test sample, add solvent, dissolve it by ultrasound, dilute it with solvent, and prepare a concentration of 0.45-0.55 mg / ml.

[0011] Further, control solution: accurately measure the test solution, dilute it 400-500 times with solvent to prepare a control solution.

[0012] Further, system suitability solution: take perampanel and impurity B, impurity C, and impurity D reference substances, add solvent, sonicate to dissolve, and dilute with solvent to prepare a mixed solution containing 0.45-0.55 mg of perampanel and 0.45-0.55 μg of each impurity per 1 ml.

[0013] Furthermore, the specification of the chromatographic column is 4.6 mm×250 mm, 5 μm, or a chromatographic column with equivalent performance.

[0014] Furthermore, the chromatographic column is a Waters Xbridge C18 chromatographic column.

[0015] Furthermore, the flow rate is 0.8-1.2 mL / min; the detection wavelength is 200-220 nm; and the injection volume is 15-25 μl.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] (1) The present invention adopts HPLC (high performance liquid chromatography), adopts a specific formula flow combined with a specific elution program and specific chromatographic conditions to determine the content of impurity B, impurity C and impurity D in the perampanel orodispersible film. The detection method has the advantages of good separation effect, sensitivity and accuracy, and better controls the quality of the perampanel orodispersible film.

[0018] (2) The method for detecting related substances of the perampanel orodispersible film of the present invention is simple, has strong specificity, high sensitivity, good durability, and good repeatability, which can better ensure the preparation quality of the perampanel orodispersible film and has good practicality. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Typical spectra of system suitability solutions of related substances. DETAILED DESCRIPTION

[0020] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0021] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0022] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0023] Reference substance / standard substance (perampanel orodispersible film, film)

[0024] Table 1 Summary of Reference Material Information

[0025]

[0026] Example 1 - Detection of related substances (perampanel orodispersible film, film)

[0027] Methods: High performance liquid chromatography.

[0028] (I) Chromatographic conditions:

[0029] Chromatographic column: Octadecylsilane bonded silica gel as filler (Waters Xbridge C18, 4.6 mm × 250 mm, 5 μm or a chromatographic column with equivalent performance);

[0030] Detection wavelength: 210nm; flow rate: 1.0ml / min; column temperature: 40℃; injection volume: 20μl;

[0031] Mobile Phase:

[0032] Mobile phase A: phosphate buffer (take 1.36 g of potassium dihydrogen phosphate, dissolve it in water and dilute it to 1000 ml, and adjust the pH value to 4.5 with phosphoric acid or 0.02 mol / L potassium hydroxide solution);

[0033] Mobile phase B: acetonitrile;

[0034] Elution procedure:

[0035] Table 2 Elution program

[0036]

[0037] (II) Solution preparation:

[0038] Solvent: acetonitrile-water (50:50, v / v)

[0039] Test solution: Place 5 tablets of 2 mg test sample in a 20 ml volumetric flask, or 3 tablets of 4 mg test sample in a 25 ml volumetric flask, add an appropriate amount of solvent, sonicate for 5 minutes to dissolve the perampanel, dilute to the scale with solvent, shake well, filter, and take the filtrate.

[0040] Control solution: Accurately measure 1 ml of the test solution, place it in a 100 ml volumetric flask, dilute it to the mark with solvent, and shake well. Accurately measure 5 ml, place it in a 25 ml volumetric flask, dilute it to the mark with solvent, and shake well.

[0041] System suitability solution: Take about 1 mg of each impurity B, impurity C, and impurity D, place in a 100 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, dilute to scale with solvent, shake well, and use as a mixed solution. Take about 10 mg of perampanel reference substance, place in a 20 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, accurately add 1 ml of the mixed solution, dilute to scale with solvent, and shake well.

[0042] System suitability requirements: The chromatogram of the system suitability solution should be consistent with the typical spectrum of the system suitability solution of the relevant substance ( Figure 1 ) are basically consistent, the separation between adjacent chromatographic peaks should meet the requirements, and the number of theoretical plates calculated based on the perampanel peak should not be less than 5000.

[0043] (III) Determination method

[0044] Calculation method: Self-control method

[0045] Result calculation: Accurately measure 20 μl of the test solution and the control solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0046] Calculation formula:

[0047] Single impurity (%) = (A 单个杂质 ×0.2%)÷A 对

[0048] Total impurities (%) = (A 总峰－ A 主峰 )×0.2%÷A 对

[0049] Where:

[0050] A 单个杂质 : The peak area of ​​each impurity in the test solution;

[0051] A 对 : Peak area of ​​the main peak in the control solution;

[0052] A 总峰 : Peak area of ​​the total peak in the test solution; A 主峰 : Peak area of ​​the main peak in the test solution;

[0053] Specifications: 2mg, 4mg;

[0054] Judgment criteria: If there are impurity peaks in the chromatogram of the test solution, the area of ​​a single impurity peak shall not be greater than the main peak area of ​​the control solution (0.2%), and the sum of the areas of all impurity peaks shall not be greater than 5 times the main peak area of ​​the control solution (1.0%). Chromatographic peaks that are less than 0.1 times (0.02%) the main peak area of ​​the control solution are negligible.

[0055] A systematic methodological validation was carried out on the above-mentioned methods for related substances and the validation results are detailed in the table below.

[0056] 1 System suitability

[0057] 1.1 Acceptable Standards:

[0058] (1) The blank membrane should not interfere with the detection of the product and impurities;

[0059] (2) The chromatogram of the system suitability solution should be consistent with the typical spectrum of the system suitability solution of the relevant substance ( Figure 1 ) are basically consistent, the separation between adjacent chromatographic peaks should meet the requirements, and the number of theoretical plates calculated based on the perampanel peak should not be less than 5000.

[0060] 1.2 Verification Results

[0061] Table 3

[0062]

[0063] Conclusion: (1) The blank membrane does not interfere with the detection of this product and its impurities; (2) The chromatogram of the system suitability solution is similar to the typical spectrum of the system suitability solution of related substances ( Figure 1 ) are basically consistent, the separation between adjacent chromatographic peaks is greater than 13 (required to be greater than 1.5), with good separation, and the theoretical plate number calculated based on the perampanel peak is greater than 5000.

[0064] In summary, this method has good system applicability.

[0065] 2 Exclusiveness

[0066] 2.1 Acceptable Standards:

[0067] (1) Blank excipients should not interfere with the product and the detection of impurities;

[0068] (2) The separation between the main component and adjacent impurities and between known impurities should meet the requirements;

[0069] (3) The main impurities produced by degradation can be well separated, the purity of the main peak meets the requirements, and the material is conserved before and after degradation.

[0070] (4) The purity angle of the principal component peak should be less than the purity threshold.

[0071] 2.2 Verification Results

[0072] Table 4

[0073]

[0074]

[0075] in conclusion:

[0076] (1) Under all degradation conditions, the blank does not interfere with the determination of the main component and its impurities;

[0077] (2) The separation between perampanel and adjacent impurities is greater than 4 (required to be greater than 1.5), maintaining good separation;

[0078] (3) This product was destroyed under severe conditions such as high temperature oxidation and light degradation. The impurities produced under various degradation conditions were well separated from the main peak. The peak purity of the main peak of perampanel met the requirements, and the material was balanced before and after degradation.

[0079] 3. Durability

[0080] 3.1 Acceptable Standards:

[0081] When there are slight changes in the mobile phase pH and instrument conditions of chromatography:

[0082] (1) The chromatogram of the system suitability solution should be consistent with the typical spectrum of the system suitability solution of the relevant substance ( Figure 1 ) are basically consistent, the separation between adjacent chromatographic peaks should meet the requirements, and the number of theoretical plates calculated based on the perampanel peak should not be less than 5000.

[0083] (2) The number of impurities detected in the test solution should be consistent; when the impurity detection amount is less than 0.1%, the RSD should not exceed 40%.

[0084] 3.2 Verification Results

[0085] When the mobile phase pH of the chromatographic conditions is in the range of pH 4.0 to pH 5.0 and when changing to a different instrument (Shimadzu, Agilent):

[0086] (1) System suitability solution chromatogram, system suitability solution chromatogram and typical spectrum of system suitability solution of related substances ( Figure 1 ) were basically consistent, the separation between adjacent chromatographic peaks was greater than 10 (greater than 1.5), the separation was good, and the theoretical plate number calculated based on the perampanel peak was greater than 5000.

[0087] (2) The number of impurities detected in the test solution was consistent; the amount of impurities detected was less than 0.1%, and the RSD was less than 40%, indicating good durability.

[0088] 4 Sensitivity

[0089] 4.1 Acceptable Criteria: (1) The signal-to-noise ratio of the quantification limit solution should be between 10 and 30; the quantification limit concentrations of impurities B, C and perampanel should be less than 0.5 times the reporting limit (0.025%), and the quantification limit solution concentration of impurity D should be less than 30% of the limit concentration;

[0090] (2) The signal-to-noise ratio of the detection limit solution should be between 2 and 10; the concentration should be less than 0.5 times the quantification limit.

[0091] 4.2 Verification Results

[0092] Table 5

[0093]

[0094] in conclusion:

[0095] (1) In the quantification limit solution, the signal-to-noise ratios of perampanel and each known impurity were between 13 and 25 (all between 10 and 30), the quantification limits of impurities B, C and perampanel were all less than 0.5 times the reporting limit (0.025%), and the concentration of impurity D in the quantification limit solution was less than 30% of the limit concentration;

[0096] (2) In the detection limit solution, the signal-to-noise ratio of each component was between 4.8 and 9.8 (all between 2 and 10), and the concentration was less than 0.5 times the quantification limit.

[0097] In summary, this method has good sensitivity.

[0098] 5Injection precision

[0099] 5.1 Acceptable standard: The quantitative limit solution is injected 6 times continuously, and the RSD value of the retention time of each component should not exceed 1.0%, and the RSD value of the peak area should not exceed 5.0%.

[0100] 5.2 Verification Results

[0101] Table 6

[0102] name Retention time RSD% Peak area RSD% Impurity D 0.05 0.98 Impurity C 0.06 2.06 Impurity B 0.04 2.64 Perampanel 0.04 4.78

[0103] Conclusion: The quantitative limit solution was injected 6 times in succession, and the RSD values ​​of retention time of each component were less than 1.0%, and the RSD values ​​of peak area were less than 5.0%. This method has good injection precision.

[0104] 6 Linearity and Correction Factors

[0105] 6.1 Acceptable criteria: The concentration is within the range of the quantitative limit to the total impurity limit, the linear relationship between the concentration and the peak area is good, and the correlation coefficient R between the main component and each known impurity is 2 The values ​​should all be no less than 0.999, the Y-intercept should be within 25% of the 100% response value, and the RSD value of the response factor should be within 10%.

[0106] 6.2 Verification Results

[0107] Table 7

[0108]

[0109] Conclusion: (1) Impurities B and C are within the range of the limit of quantification to the limit of total impurities (1.0%), and impurity D is within the range of the limit of quantification to the measured concentration (0.05%). The correlation coefficient R2 values ​​of the main component and each known impurity are all greater than 0.999, and the Y-axis intercept is within 25% of the 100% response value; the RSD value of the response factor is within 10%, and the linearity is good. (2) The correction factors of impurities B and C are not within the range of 0.9 to 1.1, and the self-control method with correction factors is used for calculation. The correction factors of each impurity are basically consistent with the standards of the raw material supplier (impurity B: 0.5, impurity C: 1.3), so the correction factors are calculated according to the correction factors specified in the standards of the raw material supplier; the correction factor of impurity D is within the range of 0.9 to 1.1, and the self-control method without correction factors is used for calculation.

[0110] 7. Repeatability

[0111] 7.1 Acceptability criteria: Prepare six test sample solutions in parallel. The test results of the relevant substances should have no significant differences and meet the requirements of the following table.

[0112] Table 8

[0113] Impurity detection RSD% ＜0.1％ ≤40 0.1％≤y≤0.2％ ≤30 0.2％<y≤0.5％ ≤20 0.5％<y<2.0％ ≤10

[0114] 7.2 Verification results (test sample size is 4 mg)

[0115] Table 9

[0116]

[0117] Conclusion: There was no significant difference in the detection results of related substances when 6 test solutions were prepared in parallel. The RSD value of impurities with a detection amount of <0.1% was 0.86% (less than 40%), and the RSD value of impurities with a detection amount of 0.2% to 0.5% was 1.26% (less than 20%). This method has good repeatability.

[0118] 8. Accuracy

[0119] 8.1 Acceptable standard: Within the reporting limit (impurity B and impurity C are reporting limit concentrations, and impurity D is a limit concentration of 50%), the limit concentration of 100% and 150%, the mean value of the recovery rate determination results of impurities B and C should be within the range of 90.0% to 108.0%, and the mean value of the recovery rate determination results of impurity D should be within the range of 85.0% to 110.0%, and the RSD should not exceed 5.0%.

[0120] 8.2 Verification Results

[0121] Table 10

[0122] name Recovery rate mean % (n=9) Impurity D 94.1 Impurity C 102.3 Impurity B 99.5

[0123] Conclusion: Within the reporting limit (impurity B and impurity C are reporting limit concentrations, and impurity D is a limit concentration of 50%), limit concentrations of 100% and 150%, the mean recovery results of impurities B and impurity C are 99.5% to 102.3% (within the range of 90.0% to 108.0%), and the mean recovery result of impurity D is 94.1% (within the range of 85.0% to 110.0%), with RSDs less than 5.0%, and good accuracy.

[0124] Comparative Example 1

[0125] The difference from Example 1 is that the mobile phase program is adjusted to:

[0126] Table 11 Elution program

[0127]

[0128]

[0129] The results showed that the separation between the main peak and the impurity B peak was less than 1.5%, and the impurity peaks had poor symmetry.

[0130] Comparative Example 2

[0131] The difference from Example 1 is that the mobile phase A is replaced with 0.1% ammonium acetate solution (pH 4.0).

[0132] The results showed that the separation between the main peak and the impurity B peak was significantly reduced (less than 1.5%), and each impurity peak showed tailing.

[0133] Comparative Example 3

[0134] The difference from Example 1 is that the column temperature is adjusted to 30°C.

[0135] The results showed that the impurity B peak interfered with the perampanel peak.

[0136] Comparative Example 4

[0137] The difference from Example 1 is that the solvent is adjusted to: solvent: acetonitrile-water (70:30, v / v).

[0138] The results showed that the impurity peaks overlapped.

[0139] 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, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting related substances of perampanel orodispersible film, characterized in that: High performance liquid chromatography was used, and the chromatographic conditions were as follows: the filler of the chromatographic column was octadecylsilane bonded silica gel, the column temperature was 38-42°C, mobile phase A and mobile phase B were used as mobile phases, mobile phase A was phosphate buffer; mobile phase B was acetonitrile; the mobile phase elution procedure was: 。 2. The method for detecting related substances of the perampanel orodispersible film according to claim 1, characterized in that: The detection solution includes a test solution, a control solution and a system suitability solution. The solvent for preparing the solution is acetonitrile-water with a volume ratio of 40-60:40-60.

3. The method for detecting related substances of the perampanel orodispersible film according to claim 1, characterized in that: The phosphate buffer solution: potassium dihydrogen phosphate is taken, dissolved and diluted with water to prepare a concentration of 1-1.5 g / L, and the pH value is adjusted to 4.0-5.

0.

4. The method for detecting related substances of the perampanel orodispersible film according to claim 3, characterized in that: The pH value is adjusted using phosphoric acid or 0.01-0.03 mol / L potassium hydroxide solution.

5. The method for detecting related substances of the perampanel orodispersible film according to claim 1, characterized in that: Test sample solution: Take the test sample, add solvent, dissolve it by ultrasound, dilute it with solvent, and prepare a concentration of 0.45-0.55 mg / ml.

6. The method for detecting related substances of the perampanel orodispersible film according to claim 5, characterized in that: Control solution: Accurately measure the test sample solution and dilute it 400-500 times with solvent to prepare a control solution.

7. The method for detecting related substances of the perampanel orodispersible film according to claim 1, characterized in that: System suitability solution: Take perampanel and impurity B, impurity C, and impurity D reference substances, add solvent, sonicate to dissolve, and dilute with solvent to prepare a mixed solution containing 0.45-0.55 mg of perampanel and 0.45-0.55 μg of each impurity per 1 ml.

8. The method for detecting related substances of the perampanel orodispersible film according to claim 1, characterized in that: The specifications of the chromatographic column are 4.6 mm×250 mm, 5 μm.

9. The method for detecting related substances of the perampanel orodispersible film according to claim 8, characterized in that: The chromatographic column is a Waters Xbridge C18 chromatographic column.

10. The method for detecting related substances of the perampanel orodispersible film according to any one of claims 1 to 9, characterized in that: The flow rate is 0.8-1.2 mL / min; the detection wavelength is 200-220 nm; and the injection volume is 15-25 μl.

Images