Degradation impurity detection method of teprenone capsule
By using acetone solutions of n-hexane, ethyl acetate and benzoic acid as the developer in thin-layer chromatography, the problem that the prior art cannot detect degraded impurities such as oxidation and light in teprene capsules is solved, and the rapid and accurate detection of these impurities is achieved to ensure the quality of the drug.
Patent Information
- Application Number
- CN202510132523.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-06
- Publication Date
- 2025-04-15
AI Technical Summary
The prior art cannot effectively detect degraded impurities such as oxidation and light in teprene capsules.
The acetone solution of n-hexane, ethyl acetate and benzoic acid was used as the developer to detect impurities degraded by tepreneone by thin-layer chromatography.
It realizes rapid and accurate detection of degraded impurities in teprene capsules to ensure that the quality of the drug meets the limit requirements.
Smart Images
Figure CN119827695A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to a method for detecting drug degradation impurities, in particular to a method for detecting degradation impurities of teprenone capsules. Background Art
[0002] Teprenone is a protective drug for gastric mucosa. Its efficacy and function is to protect gastric mucosa. It is used to treat acute gastritis, chronic gastritis, gastric ulcer and other diseases. Teprenone promotes mucus synthesis and secretion in subcutaneous cells, which can promote cell protection. After taking the drug, it can maintain the stability of mucosal cell proliferation, inhibit lipid peroxidation, and alleviate mucosal damage.
[0003] During the synthesis process of teprenone API and the production and storage of capsules, various impurities are generated. Among them, mono-cis isomer, di-cis isomer and all-cis isomer are generated during the synthesis process of teprenone and are process impurities; teprenone degradation impurities (including oxidation, light and other impurities) are impurities generated during the production and storage process and are degradation impurities. In the known literature, the detection method in the national drug standard (standard number: YBH05822009) can detect isomeric impurities. At present, no detection method has been found that can detect oxidation, light and other degradation impurities. Summary of the invention
[0004] Purpose of the invention: The purpose of the present invention is to provide a method for detecting degradation impurities in teprenone capsules, so as to solve the problem of how to detect degradation impurities in teprenone.
[0005] Technical solution: The present invention discloses a method for detecting degradation impurities in teprenone capsules, wherein a mixed solvent consisting of n-hexane, ethyl acetate and additives is used as a developing agent, and thin layer chromatography is used to separate and detect teprenone degradation impurities;
[0006] The additive is an acetone solution of benzoic acid; the volume ratio of n-hexane, ethyl acetate and the additive is 80-90:9-15:1-5.
[0007] Preferably, the concentration of benzoic acid in the acetone solution of benzoic acid is 0.1-1.5 mol / L.
[0008] The invention solves the problem that the existing developing agent cannot effectively separate teprenone and its degradation products by adding an acetone solution of benzoic acid into a conventional developing agent, so that the thin layer chromatography method can be used for rapid separation and detection of teprenone degradation impurities.
[0009] Preferably, the specific detection method comprises the following steps:
[0010] (1) dissolving the contents of the teprenone capsule in an organic solvent, filtering the filtrate to obtain a test solution, and diluting the test solution with an organic solvent to obtain a control solution;
[0011] (2) spotting the test solution and the control solution on the thin layer plate respectively, and developing the plate with the developing agent;
[0012] (3) Take out the developed thin layer plate and let it dry. Spray the developer on the thin layer plate and inspect it under ultraviolet light after heating.
[0013] Preferably, in step (1), the organic solvent is n-hexane.
[0014] Preferably, in step (1), the concentration of teprenone in the test solution is 0.05-0.15 g / mL, and the control solution is obtained by diluting the test solution 500-2000 times.
[0015] Preferably, in step (1), the concentration of teprenone in the control solution is at least 50 μg / mL.
[0016] Preferably, in step (2), the thin layer plate is a silica gel thin layer plate, and the unfolded extension distance is 7-10 cm.
[0017] Preferably, in step (3), the developer is a 10% sulfuric acid ethanol solution.
[0018] Preferably, in step (3), the heating condition is 100-110° C. for 3-7 min.
[0019] Preferably, the wavelength of the ultraviolet light is 365 nm.
[0020] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:
[0021] The present invention adopts thin layer chromatography to measure the degradation impurities in the teprenone capsule, and compares with the control solution. According to the ICH Q3B guideline, the content of each degradation impurity in the preparation shall not exceed 0.2%, and whether the degradation impurities in the test sample meet the limit requirements can be clearly compared. The developing agent used in the present invention can effectively separate teprenone and its degradation impurities or process impurities, and can be used in the teprenone thin layer chromatography detection in a targeted manner, so as to facilitate the rapid and accurate detection of various impurities in the contents of the teprenone capsule to judge the quality of the drug. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 The thin layer chromatogram of the contents of the teprenone capsules detected in Example 1;
[0023] Figure 2The thin layer chromatogram of detecting teprenone and its forced degradation impurities in Example 2;
[0024] Figure 3 The thin layer chromatogram of the light-damaged test sample in Example 3 is shown;
[0025] Figure 4 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 1;
[0026] Figure 5 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 2;
[0027] Figure 6 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 3;
[0028] Figure 7 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 4;
[0029] Figure 8 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 5;
[0030] Fig. 9 This is a thin layer chromatogram of the light-damaged test sample in Comparative Example 6;
[0031] Fig.10 This is a graph showing the detection limit determination results of the present invention. DETAILED DESCRIPTION
[0032] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.
[0033] Example 1: A method for detecting impurities in teprenone capsules is as follows:
[0034] (1) The preparation methods of each test solution and control solution are as follows:
[0035] Blank excipient test solution: Take about 0.7g of blank excipient of teprenone capsules, accurately add 5mL of n-hexane, dissolve by ultrasonic for 5min, filter, and take the filtrate. The blank excipient is mainly composed of corn starch, mannitol, racemic a-tocopherol, gel silica, polyethylene glycol 8000, glycine, and talc.
[0036] Capsule content test solution: Take an appropriate amount of teprenone capsule content (equivalent to 0.5 g of teprenone), accurately add 5 mL of n-hexane, sonicate for 5 min to dissolve teprenone, filter, and take the filtrate.
[0037] Capsule content control solution: Accurately measure 0.1 mL of the test solution of the capsule content, dilute it 500 times with n-hexane and shake well.
[0038] API test solution: Take 0.5 g of teprenone API, place in a 20 mL volumetric flask, accurately measure 5 mL of solvent, and sonicate to dissolve teprenone.
[0039] API control solution: Dilute the API solution 500 times with n-hexane and shake well.
[0040] (2) The three liquids were fully mixed in a volume ratio of n-hexane: ethyl acetate: benzoic acid acetone solution of 82:15:3, and the resulting mixed solvent was used as a developing solvent. In this example, the concentration of benzoic acid in the benzoic acid acetone solution was 1.0 mol / L.
[0041] (3) Take 5 μL of each of the above test solution and control solution, spot them on the same silica gel GF254 thin layer plate, develop them to about 8 cm with the developer prepared in step (2), take them out, dry them, spray them with 10% sulfuric acid ethanol solution, heat them at 105°C for 5 min, and examine them under ultraviolet light (365 nm). The results are as follows: Figure 1 As shown, from left to right: 1. n-hexane; 2. blank excipient test solution; 3. API control solution; 4. API test solution; 5. capsule content control solution; 6. capsule content test solution; Figure 1 The larger and clearer bright spot in the upper middle is teprenone, and the several discrete points below are process impurities or degradation impurities. Figure 1 The results showed that the developing solvent composed of n-hexane, ethyl acetate, benzoic acid and acetone could well separate teprenone and its impurities, and the blank solvent (n-hexane) and blank excipients had no interference with the sample detection.
[0042] Example 2: A method for detecting degradation impurities in teprenone capsules is as follows:
[0043] (1) The method for preparing the test solution sample is as follows:
[0044] Undegraded test solution: Take an appropriate amount of the contents of teprenone capsules (equivalent to 0.5 g of teprenone), place in a 20 mL volumetric flask, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0045] Control solution: dilute the undegraded test sample solution 500 times with n-hexane and shake well.
[0046] Acid destruction of the test solution: Take an appropriate amount of the contents of teprenone capsules (equivalent to 0.5 g of teprenone), place in a 20 mL volumetric flask, add 1 mL of 0.1 mol / L hydrochloric acid aqueous solution to destroy at room temperature for 1 day, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0047] Alkali destruction of the test solution: Take an appropriate amount of the contents of teprenone capsules (equivalent to 0.5 g of teprenone), place in a 20 mL volumetric flask, add 1 mL of 0.1 mol / L sodium hydroxide aqueous solution to destroy at room temperature for 1 day, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0048] Oxidative destruction of the test solution: Take an appropriate amount of the contents of teprenone capsules (equivalent to 0.5 g of teprenone), place in a 20 mL volumetric flask, add 1 mL of 1.0% hydrogen peroxide solution, destroy at room temperature for 1 day, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0049] High temperature destruction of the test solution: Take an appropriate amount of the contents of the teprenone capsule (equivalent to 0.5 g of teprenone), place it at 60°C for 1 day, cool it to room temperature, place it in a 20 mL volumetric flask, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0050] Light destruction of the test solution: Take an appropriate amount of the contents of the teprenone capsule (equivalent to 0.5 g of teprenone) and set the illumination to 5500 lx for visible light and 100 μw / cm for near-ultraviolet energy. 2 Destroy under the conditions for 4 days, place in a 20mL volumetric flask, accurately add 5mL of n-hexane, ultrasonicate for 5min to dissolve the solid, filter, and take the filtrate.
[0051] (2) The three liquids were fully mixed in a volume ratio of n-hexane: ethyl acetate: benzoic acid acetone solution of 80:15:5, and the resulting mixed solvent was used as a developing solvent. In this embodiment, the concentration of benzoic acid in the benzoic acid acetone solution was 0.7 mol / L.
[0052] (3) Take 5 μL of each of the above test solution and control solution, spot them on the same silica gel GF254 thin layer plate, develop them to about 8 cm with the developer prepared in step (2), take them out, dry them, spray them with 10% sulfuric acid ethanol solution, heat them at 105°C for 5 min, and examine them under ultraviolet light (365 nm). The results are as follows: Figure 2 As shown, from left to right, they are: 1. Control solution; 2. Undegraded test solution; 3. Light-damaged test solution; 4. Acid-damaged test solution; 5. Alkali-damaged test solution; 6. Oxidation-damaged test solution; 7. High-temperature-damaged test solution. Among them, only the test result of 3. Light-damaged test solution showed the phenomenon of increased brightness of discrete points in the impurity area below. Figure 2The results show that the contents of teprenone capsules showed no significant changes compared with the undamaged samples under acid, alkali, oxidation and high temperature conditions, and there was no increase in degradation impurities, indicating that teprenone is relatively stable under acid, alkali, oxidation and high temperature conditions; degradation impurities were produced under light damage conditions and were effectively detected, proving that the method of the present invention has good specificity.
[0053] Embodiment 3: The rest is the same as Embodiment 2, except that:
[0054] (1) The method for preparing the test solution sample is as follows:
[0055] Undegraded test solution: Take an appropriate amount of the contents of teprenone capsules (equivalent to 0.5 g of teprenone), place in a 20 mL volumetric flask, accurately add 5 mL of n-hexane, ultrasonicate for 5 min to dissolve the solid, filter, and take the filtrate.
[0056] Undegraded control solution: dilute the undegraded test sample solution 500 times with n-hexane and shake well.
[0057] Light destruction of the test solution: Take an appropriate amount of the contents of the teprenone capsule (equivalent to 0.5 g of teprenone) and set the illumination to 5500 lx for visible light and 100 μw / cm for near-ultraviolet energy. 2 Destroy under the conditions for 4 days, place in a 20mL volumetric flask, accurately add 5mL of n-hexane, ultrasonicate for 5min to dissolve the solid, filter, and take the filtrate.
[0058] Photodamage control solution: Dilute the photodamage test solution 500 times with n-hexane and shake well.
[0059] (2) The three liquids were fully mixed in a volume ratio of n-hexane: ethyl acetate: benzoic acid acetone solution of 90:9:1, and the resulting mixed solvent was used as a developing solvent. In this embodiment, the concentration of benzoic acid in the benzoic acid acetone solution was 1.0 mol / L.
[0060] (3) Take 5 μL of each of the above test solution and control solution, spot them on the same silica gel GF254 thin layer plate, develop them to about 8 cm with the developer prepared in step (2), take them out, dry them, spray them with 10% sulfuric acid ethanol solution, heat them at 105°C for 5 min, and examine them under ultraviolet light (365 nm). The results are as follows: Figure 3 As shown, from left to right: 1. Undegraded control solution; 2. Undegraded damaged test solution; 3. Light-damaged control solution; 4. Light-damaged test solution. Figure 3 The results show that the developing agent in this example can also separate the degradation impurities of teprenone, and after comparing with the undegraded test solution, it can be clearly found that the photo-degraded impurities of teprenone are significantly increased.
[0061] Example 4: Determination of the detection limit of TLC detection method for teprenone
[0062] Teprenone reference substance stock solution: Take 20 mg of teprenone reference substance into a 100 mL volumetric flask, add an appropriate amount of solvent, sonicate to dissolve teprenone, and dilute to the mark with n-hexane. Shake well to obtain a stock solution containing about 0.2 mg / mL of teprenone.
[0063] Solutions for detection limit determination: Take appropriate amounts of the teprenone reference stock solution and dilute with n-hexane to prepare solutions containing teprenone at concentrations of 25 μg / mL, 50 μg / mL, 100 μg / mL, 150 μg / mL, and 200 μg / mL.
[0064] 5 μL of each of the above detection limit determination solutions were taken and spotted on the same silica gel GF254 thin layer plate, developed to about 8 cm with the developing agent in Example 3, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C for 5 minutes, and examined under ultraviolet light (365 nm). The results are as follows Fig.10 As shown, Fig.10 From left to right in the figure: 1. blank solvent (n-hexane); 2. 200 μg / mL solution; 3. 150 μg / mL solution; 4. 100 μg / mL solution; 5. 50 μg / mL solution; 6. 25 μg / mL. The detection limit concentration of the present invention is 50 μg / mL.
[0065] Comparative Example 1: The rest is the same as Example 3, except that:
[0066] Hexane:ethyl acetate (volume ratio 90:10) was used as the developing solvent.
[0067] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0068] Test results such as Figure 4 As shown, Figure 4 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0069] Comparative Example 2: The rest is the same as Example 3, except that:
[0070] The developing solvent was prepared by replacing benzoic acid with acetic acid.
[0071] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0072] Test results such as Figure 5 As shown, Figure 5 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0073] Comparative Example 3: The rest is the same as Example 3, except that:
[0074] The developing solvent was prepared by replacing benzoic acid with salicylic acid.
[0075] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0076] Test results such as Figure 6 As shown, Figure 6 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0077] Comparative Example 4: The rest is the same as Example 3, except that:
[0078] Acetone was replaced with cyclohexanone to prepare a developing solvent.
[0079] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0080] Test results such as Figure 7 As shown, Figure 7 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0081] Comparative Example 5: The rest is the same as Example 3, except that:
[0082] The developing solvent was prepared by replacing n-hexane with petroleum ether.
[0083] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0084] Test results such as Figure 8 As shown, Figure 8 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0085] Comparative Example 6: The rest is the same as Example 3, except that:
[0086] A benzoic acid acetone solution with a concentration of 1.0 mol / L of benzoic acid was directly used as the developing solvent.
[0087] The test samples were only the light-damaged test solution and the light-damaged control solution.
[0088] Test results such as Fig. 9 As shown, Fig. 9 1 is the light-damaged control solution, and 2 is the light-damaged test solution.
[0089] Depend on Figure 4-9 and Figure 3The inspection results show that in the absence of benzoic acid and / or acetone, the existing n-hexane / ethyl acetate developing agent or petroleum ether / ethyl acetate developing agent cannot effectively separate teprenone and its degradation impurities. Benzoic acid and acetone are indispensable for the improvement of the n-hexane / ethyl acetate developing agent system. At the same time, Comparative Example 5 shows that the improvement of benzoic acid and acetone is selective and does not have an optimization effect on non-n-hexane / ethyl acetate systems. Comparative Example 6 shows that the benzoic acid acetone solution itself does not have a development and separation effect on teprenone and its degradation impurities, and it mainly works by assisting in fine-tuning the n-hexane / ethyl acetate main system.
Claims
1. A method for detecting degradation impurities in teprenone capsules, characterized in that: Using a mixed solvent consisting of n-hexane, ethyl acetate and additives as a developing solvent, thin layer chromatography was used to separate and detect teprenone degradation impurities. The additive is an acetone solution of benzoic acid; the volume ratio of n-hexane, ethyl acetate and the additive is 80-90:9-15:1-5.
2. The method for detecting degradation impurities in teprenone capsules according to claim 1, characterized in that: The concentration of benzoic acid in the acetone solution of benzoic acid is 0.1-1.5 mol / L.
3. The method for detecting degradation impurities in teprenone capsules according to claim 1, characterized in that: The steps include: (1) dissolving the contents of the teprenone capsule in an organic solvent, filtering the filtrate to obtain a test solution, and diluting the test solution with an organic solvent to obtain a control solution; (2) spotting the test solution and the control solution on the thin layer plate respectively, and developing the plate with the developing agent; (3) Take out the developed thin layer plate and let it dry. Spray the developer on the thin layer plate and inspect it under ultraviolet light after heating.
4. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (1), the organic solvent is n-hexane.
5. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (1), the concentration of teprenone in the test solution is 0.05-0.15 g / mL, and the control solution is obtained by diluting the test solution 500-2000 times.
6. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (1), the concentration of teprenone in the control solution is at least 50 μg / mL.
7. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (2), the thin layer plate is a silica gel thin layer plate, and the unfolded extension distance is 7-10 cm.
8. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (3), the developer is a 10% sulfuric acid ethanol solution.
9. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: In step (3), the heating condition is 100-110° C. for 3-7 min.
10. The method for detecting degradation impurities in teprenone capsules according to claim 3, characterized in that: The wavelength of the ultraviolet light is 365nm.