A method for detecting the content of benzene sulfonic acid ester impurities in amlodipine besylate
By combining high-performance liquid chromatography (HPLC) and mass spectrometry (MS) detectors, the sensitivity problem of detecting benzene sulfonate impurities in amlodipine benzenesulfonate has been solved, achieving efficient detection of benzene sulfonate impurities and meeting the requirements of drug safety monitoring.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- 珠海润都制药股份有限公司
- Filing Date
- 2024-12-09
- Publication Date
- 2026-06-09
AI Technical Summary
Existing technologies are insufficient for efficiently and sensitively detecting the content of benzene sulfonate impurities in amlodipine besylate, especially methyl besylate, ethyl besylate, and isopropyl besylate, thus failing to meet the requirements for drug safety monitoring.
A high-performance liquid chromatography (HPLC) method combined with a mass spectrometry detector was used to detect benzenesulfonate impurities using specific solution preparation and gradient elution methods. This included setting the solution preparation, chromatographic conditions, and mass spectrometry detection conditions, and the detection was performed using the SIM method.
It achieves highly sensitive detection of benzenesulfonate impurities in amlodipine benzenesulfonate, meeting the limits specified by ICH M7 and satisfying the precision and specificity requirements for drug quality monitoring.
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Figure CN122171696A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical analysis, and specifically to a method for detecting the content of benzenesulfonate impurities in amlodipine benzenesulfonate. Background Technology
[0002] Amlodipine besylate is a classic antihypertensive drug. Studies have found that benzene ester impurities may be present in the amlodipine besylate raw material, with typical examples including methyl besylate, ethyl besylate, and isopropyl besylate. According to ICH guidelines, the risk of impurities is estimated based on the target molecular structure fragment to assess drug safety. Benzene ester molecules are typical genotoxic impurities, requiring content monitoring and control to meet product safety requirements. According to ICH M7, the TTC value for genotoxic impurities is 1.5 µg / day. The maximum daily dose of amlodipine in amlodipine besylate tablets is 10 mg / day, which translates to a maximum daily dose of 13.87 mg / day for amlodipine besylate, resulting in a limit of 108 ppm. Therefore, a method with sufficient precision and sensitivity is needed to detect the content of genotoxic impurities such as benzene esters in amlodipine besylate to meet the quality control requirements of pharmaceutical production. Summary of the Invention
[0003] Based on the above technical background, this invention provides a method for detecting the content of benzene sulfonates in amlodipine benzenesulfonate, wherein the benzene sulfonates are methyl benzenesulfonate, ethyl benzenesulfonate, and isopropyl benzenesulfonate, and the detection is performed by high performance liquid chromatography, including the following steps: 1. Solution preparation: Take methyl benzyl sulfonate, ethyl benzyl sulfonate, and isopropyl benzyl sulfonate reference standards respectively, and prepare reference standard solutions with a mixed solution of acetonitrile:formic acid = 100:1 (v / v); separately take amlodipine benzyl sulfonate sample to be tested, and prepare test solution with a mixed solution of acetonitrile:formic acid = 100:1 (v / v); 2. Inject the test solution and reference solution into a high-performance liquid chromatograph (HPLC). The chromatographic conditions are as follows: octadecylsilane-bonded silica gel column (Agilent Proshell 120 EC-C18 4.6 × 100 mm, 2.7 μm); injection volume: 3 µl; column temperature: 30℃; flow rate: 0.5 mL / min; mobile phase A: 10 mM ammonium formate solution; mobile phase B: methanol; gradient elution method. Elution gradient:
[0004] The detection was performed using a mass spectrometer detector, under the following conditions:
[0005] SIM method:
[0006] A more specific operating procedure is as follows: Diluent: Acetonitrile: Formic acid = 100:1 (v / v); Blank solution: diluent; Methyl benzenesulfonate stock solution: Weigh approximately 27 mg of methyl benzenesulfonate reference standard accurately, place it in a 25 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; Ethyl benzenesulfonate stock solution: Weigh approximately 27 mg of ethyl benzenesulfonate reference standard accurately, place it in a 25 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; Isopropyl benzenesulfonate stock solution: Weigh approximately 27 mg of isopropyl benzenesulfonate reference standard accurately, place it in a 25 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well. Reference standard mixed stock solutions: Accurately measure 100 μl of methyl benzenesulfonate stock solution, ethyl benzenesulfonate stock solution and isopropyl benzenesulfonate stock solution respectively, place them in 20 ml volumetric flasks, add diluent to the mark, and shake well; Reference solution: Accurately measure 500 μl of the mixed stock solution of reference standard, place it in a 10 ml volumetric flask, add diluent to the mark, and shake well; Sensitivity solution: Accurately measure 3.0 ml of the reference solution, place it in a 10 ml volumetric flask, add diluent to the mark, and shake well; Test solution: Weigh approximately 25 mg of the test sample accurately, place it in a 10 ml volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; After the system stabilizes, inject the sample and record the spectrum. The results are calculated using the following formula: (ppm) = (RU / RS) × (CS / CU) Wherein: RU: peak area of each target impurity in the chromatogram of the test solution; RS: average peak area of each target impurity in the chromatogram of the reference solution; CS: concentration of each target impurity in the reference solution (ng / ml); CU: concentration of the test solution (mg / ml).
[0007] The verification of the method is summarized as follows:
[0008] Beneficial effects: The present invention provides a method for detecting genotoxic impurities such as methyl benzyl sulfonate, ethyl benzyl sulfonate and isopropyl benzyl sulfonate in amlodipine benzyl sulfonate using liquid chromatography. The method has high detection sensitivity, and the verification results show that the system applicability and specificity of this method meet the requirements. Attached Figure Description
[0009] Figure 1 This is a chromatogram of a sensitivity solution; Figure 2 Sensitivity solution chromatogram - continued; Figure 3 This is the chromatogram of the reference solution; Figure 4 Chromatogram of reference solution - continued; Figure 5 This is a chromatogram of a selective solution; Figure 6 Selective solution chromatogram - continued; Figure 7 This is a chromatogram of a precision solution; Figure 8 Chromatogram of precision solution - continued. Detailed Implementation Example
[0011] System Applicability Solution preparation: Diluent: Acetonitrile: Formic acid = 100:1 (v / v); Blank solution: Diluent.
[0012] Reference stock solutions: Weigh approximately 27 mg each of methyl benzenesulfonate, ethyl benzenesulfonate, and isopropyl benzenesulfonate impurity reference standards, place them in separate 25 ml volumetric flasks, dissolve and dilute to the mark with diluent, and shake well (methyl benzenesulfonate concentration: 1.0452 mg / ml; ethyl benzenesulfonate concentration: 1.0993 mg / ml; isopropyl benzenesulfonate concentration: 1.0747 mg / ml).
[0013] Reference solutions: Accurately measure 100 µl of each reference stock solution, mix them in a 25 mL volumetric flask, dilute to the mark with diluent, and shake well. Take 500 µl of the above solution and dilute to the mark in a 10 mL volumetric flask to obtain the following concentrations: methyl benzenesulfonate: 261.2880 ng / mL; ethyl benzenesulfonate: 274.8190 ng / mL; isopropyl benzenesulfonate: 268.6752 ng / mL.
[0014] Sensitivity solution: Accurately measure 3 ml of the reference stock solution and place it in a 10 ml volumetric flask. Dilute to the mark with diluent and shake well (methyl benzenesulfonate concentration: 78.3864 ng / ml; ethyl benzenesulfonate concentration: 82.4457 ng / ml; isopropyl benzenesulfonate concentration: 80.6026 ng / ml).
[0015] The above solution was injected under the following chromatographic conditions: Octadecylsilane-bonded silica gel column (Agilent Proshell 120 EC-C18, 4.6 × 100 mm, 2.7 μm); injection volume: 3 µl; column temperature: 30℃; flow rate: 0.5 ml / min; mobile phase A: 10 mM ammonium formate solution; mobile phase B: methanol; gradient elution method was used. The elution gradient was:
[0016] The detection was performed using a mass spectrometer detector, under the following conditions:
[0017] SIM method:
[0018] The sensitivity solution chromatogram is shown below. Figure 1-2 As shown, the chromatogram of the reference solution is as follows: Figure 3-4 As shown, the system suitability test results are as follows.
[0019] Example
[0020] Specificity experiment Solution preparation: Diluent: Acetonitrile: Formic acid = 100:1 (v / v); Blank solution: Diluent.
[0021]
[0022] The above solution was injected under the same chromatographic conditions as in Example 1; the chromatogram of the reference solution is shown below. Figure 3-4 As shown, the selective solution chromatogram is as follows: Figure 5-6 As shown; the detection conditions are the same as in Example 1. The test results are shown in the table below.
[0023] Example
[0024] Precision test Solution preparation: The diluent, test solution, and reference solution are the same as in Example 1; Preparation of precision solutions:
[0025] The above solution was injected under the same detection conditions as in Example 1; the chromatogram of the precision solution is shown below. Figure 7-8 As shown in the figure; the experimental results are shown in the table below.
[0026]
[0027] This invention provides a method for detecting the content of benzenesulfonate esters in amlodipine benzenesulfonate using liquid chromatography. The impurities are methyl benzenesulfonate, ethyl benzenesulfonate, and isopropyl benzenesulfonate. These impurities are genotoxic and should be strictly controlled according to drug quality standards. As shown in the table above, the system suitability, specificity, precision, limit of quantitation, limit of detection, linearity, range, and accuracy of the detection method of this invention all meet the pharmacopoeia requirements.
Claims
1. A method for detecting the content of benzenesulfonate impurities in amlodipine benzenesulfonate, wherein the method employs liquid chromatography, characterized in that... The detected benzene sulfonate impurities were methyl benzenesulfonate, ethyl benzenesulfonate, and isopropyl benzenesulfonate. Includes the following steps: (1) Solution preparation: Take methyl benzenesulfonate, ethyl benzenesulfonate and isopropyl benzenesulfonate reference standards respectively, and prepare reference standard solutions with a mixed solution of acetonitrile and formic acid with a volume ratio of 100:1; take amlodipine benzenesulfonate sample to be tested, and prepare test solution with a mixed solution of acetonitrile and formic acid with a volume ratio of 100:
1. (2) Inject the test solution and the reference solution into the high performance liquid chromatograph. The chromatographic conditions are as follows: octadecylsilane bonded silica column Agilent Proshell 120 EC-C18 4.6×100mm, 2.7 μm, injection volume: 3µl, column temperature: 30℃, flow rate: 0.5ml / min, mobile phase A: 10mM ammonium formate solution, mobile phase B: methanol, and gradient elution method.
2. The method for detecting benzenesulfonate impurities in amlodipine benzenesulfonate as described in claim 1, wherein the elution gradient of the gradient elution is: 。