Method for detecting impurities of cibopadol
The detection of impurities in Cibopado by high performance liquid chromatography solves the problem of lack of effective detection methods in the prior art, achieves detection effects of high accuracy and good resolution, and ensures the safety of drug preparations.
Patent Information
- Application Number
- CN202311057500.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-22
- Publication Date
- 2025-02-25
AI Technical Summary
The prior art lacks effective detection methods to detect impurities in cibopado, affecting the safety of pharmaceutical preparations.
High performance liquid chromatography was used to achieve efficient detection of cibopado impurities through reverse phase chromatography columns, using phosphate buffer, acetonitrile and tetrahydrofuran as mobile phases, combined with gradient elution procedures.
This method has high accuracy, good resolution and stability, is easy to operate, has good repeatability and strong targeting, and can effectively detect impurities in cibopado and ensure the safety of drug preparations.
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Figure CN119510589A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting impurities of sibopadol. Background Art
[0002] Sibopadol (trans-6-fluoro-4,9-dihydro-N,N-dimethyl-4-phenyl-spiro[cyclohexane-1,1(3H)-pyrano[3,4-b]indol]-4-amine) is an analgesic nociceptin / orphanin FQ peptide (NOP) and an opioid receptor agonist. It is currently being developed internationally by Grünenthal GmbH, Germany, and its partner, the US pharmaceutical company DepoMed, for the treatment of a number of different acute and chronic pain states. Sibopadol has demonstrated highly potent analgesic and antihypertensive effects in a variety of different animal models of pain. Notably, it has also been found to be more potent in chronic neuropathic pain models than in acute nociceptive pain, compared to selective μ-opioid receptor agonists. Relative to morphine, tolerance to the analgesic effects of sibopadol has been found to be delayed (26 days vs. 11 days of full tolerance). Furthermore, unlike morphine, sibopadol has not been found to affect motor coordination or reduce breathing in animals at or above the analgesic dose range. Therefore, it may have improved and prolonged effectiveness and greater tolerability compared with currently available opioid analgesics.
[0003] The inventors found that there are impurities such as impurity 1, impurity 2, impurity 3 in Xibopadol, and there is currently no good detection method. In order to ensure the safety of subsequent drug preparations, impurities such as impurity 1, impurity 2, impurity 3 should be detected, and the impurity residues should be controlled within a reasonable limit. Summary of the invention
[0004] In order to solve the above technical problems, the purpose of the present invention is to provide a method for detecting impurities in sibopadol, and the detection method of the present invention has high accuracy, good separation, good stability, simple operation, good repeatability, strong specificity, and good application value.
[0005] The invention provides a method for detecting impurities of Sibopadol, which adopts a high performance liquid chromatography method, and the chromatographic conditions include: adopting a reverse phase chromatographic column, and taking phosphate buffer, acetonitrile and tetrahydrofuran as mobile phases.
[0006] In one embodiment, the chromatography column is a C18 chromatography column.
[0007] In one embodiment, the C18 chromatographic column has a specification of 4.6*250 mm and 5.0 μm.
[0008] In one embodiment, the column temperature is 25°C to 45°C, preferably, the column temperature is 30°C to 40°C.
[0009] In one embodiment, the mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is 0.01-0.03 mol / L phosphate buffer (pH 7.0-8.0)-acetonitrile (85-95:5-15), and mobile phase B is acetonitrile-tetrahydrofuran (92-98:2-8).
[0010] In one embodiment, mobile phase A is 0.02 mol / L phosphate buffer (pH 7.5)-acetonitrile (90:10), and mobile phase B is acetonitrile-tetrahydrofuran (95:5).
[0011] In one embodiment, the flow rate is 0.8-1.2 mL / min.
[0012] In one embodiment, detection is performed using an ultraviolet detector with a detection wavelength of 215-235 nm.
[0013] In one embodiment, the injection volume is 20-40 μL.
[0014] In one embodiment, high performance liquid chromatography is used, and the chromatographic conditions include: using a C18 chromatographic column; detecting with an ultraviolet detector, the detection wavelength is 215-235nm; mobile phase A is 0.02mol / L phosphate buffer (pH7.5)-acetonitrile (90:10), and mobile phase B is acetonitrile-tetrahydrofuran (95:5); column temperature is 30°C to 40°C; flow rate is 0.8 to 1.2mL / min; gradient elution. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 HPLC chromatogram of the test product. DETAILED DESCRIPTION
[0016] Example 1 Specificity Experiment
[0017] 1. Preparation of reference and test solutions
[0018] Specificity testing requires verification that the blank solution has no interference at the retention time of the main peak in the test and reference solutions, as well as the separation between impurities and the main component.
[0019] The following are the preparation methods of each impurity and main component:
[0020] Solvent 1: tetrahydrofuran;
[0021] Solvent 2: acetonitrile-water (50:50);
[0022] Blank: solvent 1 - solvent 2 (50:50).
[0023] Starting material - positioning solution: Take an appropriate amount of starting material - reference substance, weigh accurately, dissolve in solvent 2 and dilute to make a solution containing about 0.1 mg per 1 ml.
[0024] Starting material 2 positioning solution: Take an appropriate amount of starting material 2 reference substance, weigh accurately, dissolve in solvent 2 and dilute to make a solution containing about 0.1 mg per 1 ml.
[0025] Intermediate 1 positioning solution: Take an appropriate amount of intermediate 1 reference substance, accurately weigh it, add methanol to dissolve it and dilute it to make a solution containing about 0.1 mg per 1 ml.
[0026] Test solution: Take an appropriate amount of cebopadol citrate, weigh it accurately, add an appropriate amount of solvent 1 to dissolve it, and quantitatively dilute it with solvent 2 to make a solution containing about 0.1 mg per 1 ml.
[0027] Control solution: Accurately pipette 1 ml of the test solution into a 100 ml volumetric flask, dilute to the mark with 50% acetonitrile, and shake well.
[0028] Mixed solution: Take about 10 mg of the test sample, weigh it accurately, put it in a 100 ml volumetric flask, add 50 ml of solvent 1 to dissolve it, transfer 1 ml of each impurity location solution, add solvent 2 to the fixed volume and shake well to obtain.
[0029] 2. Detection chromatographic conditions
[0030] Chromatographic column: C18 (250 mm × 4.6 mm, 5 μm);
[0031] Detector: UV detector
[0032] Detection wavelength: 225nm;
[0033] Mobile phase: mobile phase A and mobile phase B; wherein mobile phase A is 0.02 mol / L phosphate buffer (pH 7.5)-acetonitrile (90:10), and mobile phase B is acetonitrile-tetrahydrofuran (95:5);
[0034] Use mobile phase A and mobile phase B to perform gradient elution according to Gradient Table 1;
[0035] Flow rate: 1.0ml / min;
[0036] Column temperature: 35°C;
[0037] Diluent: acetonitrile-water (50:50);
[0038] Injection volume: 30 μL.
[0039] Table 1 Gradient elution program table (volume ratio)
[0040] Time (min) Mobile phase A% Mobile phase B% 0 85 15 5 85 15 10 50 50 20 40 60 40 40 60 40.1 85 15 50 85 15
[0041] Referring to the above chromatographic detection conditions, 30 μL of each of the above solutions were accurately measured and injected into the liquid chromatograph, and the chromatogram was recorded. The results are shown in Table 2.
[0042] Table 2 Results of specificity investigation
[0043]
[0044]
[0045] As can be seen from Table 2, the blank solution has no interference at the retention time of the main peak in the test and reference solutions, and the separation between each impurity is greater than 1.5, indicating that the specificity of the detection method of the present application meets the requirements.
[0046] Example 2 Sensitivity test
[0047] Sibopadol reference solution: Take an appropriate amount of Sibopadol reference, weigh it accurately, add an appropriate amount of solvent 1 to dissolve it, and dilute it step by step with solvent 2 to the appropriate multiple. The solution with a signal-to-noise ratio ≥10:1 is used as the quantitative limit solution; the solution with a signal-to-noise ratio ≥3:1 is used as the detection limit solution.
[0048] Referring to the chromatographic conditions of Example 1, 30 μL of each of the above solutions were accurately measured and injected into a liquid chromatograph to record the chromatogram. The results are shown in Table 3.
[0049] Table 3 Results of detection limit and quantification limit
[0050]
[0051] As can be seen from Table 3, the detection limit sensitivity of the detection method of the present application is 0.01%, and the quantification limit sensitivity is 0.03%, which indicates that the sensitivity of the detection method of the present application meets the requirements.
[0052] Example 3 Solution stability test
[0053] Take the test solution and control solution prepared in Example 1, refer to the detection chromatographic conditions of Example 1, accurately measure 30 μL of each of the above solutions, inject them into the liquid chromatograph, record the chromatogram, and the results are shown in Table 4.
[0054] Table 4 Solution stability test results
[0055] Test solution 0h 3h 6h 10h 24h Maximum change Impurity 1 0.013％ 0.013％ 0.012％ 0.019％ 0.014％ 0.006％ Impurity 2 0.028％ 0.032％ 0.033％ 0.037％ 0.044％ 0.016％ Impurity 3 0.019％ 0.020％ 0.020％ 0.025％ 0.021％ 0.006％ Total Miscellaneous 0.060％ 0.066％ 0.065％ 0.081％ 0.079％ 0.021％ Control solution 0h 3h 6h 10h 24h RSD% Peak area 130861 131438 131724 133696 130812 0.89
[0056] As can be seen from Table 4, under room temperature conditions, within 24 hours, the changes in the impurity content of the test solution at different time points were all less than 0.05% compared with 0 hour, and the peak area RSD of the control solution was less than 3%, indicating that the stability of the detection method of the present application meets the requirements.
Claims
1. A method for detecting impurities in Xibopadol, characterized in that: The high performance liquid chromatography method is adopted, and the chromatographic conditions include: using a reverse phase chromatographic column and using phosphate buffer, acetonitrile and tetrahydrofuran as mobile phases.
2. The impurity detection method according to claim 1, characterized in that: The chromatographic column is a C18 chromatographic column.
3. The impurity detection method according to claim 2, characterized in that: The specifications of the C18 chromatographic column are 4.6*250 mm and 5.0 μm.
4. The impurity detection method according to claim 1, characterized in that: The column temperature is 25°C to 45°C, preferably, the column temperature is 30°C to 40°C.
5. The impurity detection method according to claim 1, characterized in that: The mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is 0.01-0.03 mol / L phosphate buffer (pH 7.5)-acetonitrile (85-95:5-15), and the mobile phase B is acetonitrile-tetrahydrofuran (92-98:2-8).
6. The impurity detection method according to claim 5, characterized in that: Mobile phase A was 0.02 mol / L phosphate buffer (pH 7.5)-acetonitrile (90:10), and mobile phase B was acetonitrile-tetrahydrofuran (95:5).
7. The impurity detection method according to claim 1, characterized in that: The flow rate is 0.8-1.2 mL / min.
8. The impurity detection method according to claim 1, characterized in that: Use an ultraviolet detector for detection, with a detection wavelength of 215-235nm.
9. The impurity detection method according to claim 1, characterized in that: The injection volume is 20-40 μL.
10. The impurity detection method according to any one of claims 1 to 9, characterized in that: High performance liquid chromatography was used, and the chromatographic conditions included: a C18 chromatographic column; detection by an ultraviolet detector with a detection wavelength of 215-235 nm; mobile phase A was 0.02 mol / L phosphate buffer (pH 7.5)-acetonitrile (90:10), and mobile phase B was acetonitrile-tetrahydrofuran (95:5); column temperature was 30°C to 40°C; flow rate was 0.8 to 1.2 mL / min; and gradient elution.