Method for measuring in-vitro dissolution curve of linagliptin tablet
By using 0.1 mol/L hydrochloric acid solution and basket dissolution method to determine the dissolution curve of linagliptin tablets, the problem of difficulty in reacting the intrinsic quality of linagliptin tablets in the prior art was solved, and effective evaluation and distinction of the dissolutionability and quality of linagliptin tablets was achieved.
Patent Information
- Application Number
- CN202510164772.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-14
- Publication Date
- 2025-06-20
AI Technical Summary
The prior art is difficult to effectively reflect the intrinsic quality of linagliptin tablets, and lacks a dissolution curve measurement method that distinguishes linagliptin tablets from different sources.
The dissolution curve of the linagliptin tablet was measured by using 900 ml of 0.1 mol/L hydrochloric acid solution as the dissolution medium and the basket method was dissolved at 50 rpm. The method includes the preparation of the test sample solution and the reference sample solution, respectively injecting into the high performance liquid chromatography system, recording the chromatogram and calculating the dissolution amount.
The dissolution, specificity, stability and difference between the homemade preparation and reference preparation of linagliptin tablets is achieved, which is universal and practical, and is easy to operate.
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Figure CN120177642A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of pharmaceutical analysis and detection, and specifically to a method for determining the in vitro dissolution curve of linagliptin tablets. Background Art
[0002] Linagliptin is a dipeptidyl peptidase 4 (DPP-4) inhibitor. DPP-4 can degrade the incretin hormone-like polypeptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). Linagliptin can increase the concentration of active incretin hormones, stimulate insulin release in a glucose-dependent manner, and reduce the level of glucagon in circulation. Both of these incretin hormones are involved in the physiological regulation of glucose homeostasis. The incretin secretion maintains a relatively low basal level throughout the day and increases immediately after a meal. Under normal or elevated glucose levels, GLP-1 and GIP can increase the biosynthesis and secretion of insulin by pancreatic β-cells. In addition, GLP-1 can also reduce the glucagon secretion of pancreatic α-cells and decrease the hepatic glucose output.
[0003] The original research formulation is a tablet, which is a drug for treating type 2 diabetes. The specification is 5 mg [1], and the Chinese trade name is "Onglyza". The manufacturer is West-Ward Columbus Inc., and the marketing authorization holder is Boehringer Ingelheim International GmbH. It has been imported into China.
[0004] Dissolution Profile: The dissolution rate-time curve plotted by the dissolution rates obtained at different time points of the preparation. The dissolution curve reflects the in vitro dissolution and dissolution process of the preparation, and the quality of generic drugs can be evaluated by comparing the similarity of the in vitro dissolution curves of generic preparations and original research preparations.
[0005] Currently, there are few studies on the dissolution behavior of linagliptin tablets, and this variety is not included in the pharmacopoeias of various countries. There is no dissolution curve data of linagliptin tablets in the Japanese Orange Book, and only the IF file shows that the dissolution method uses the basket method. The FDA database shows that the dissolution method for this variety refers to the "FDA Dissolution Guide" published in 2018. It describes a dissolution method for linagliptin tablets. This method uses 500 ml of 0.1 mol / L hydrochloric acid solution as the dissolution medium, the dissolution mode is selected as the basket method at 100 revolutions per minute, and the dissolution rate exceeds 85% at 10 minutes. It cannot effectively reflect the intrinsic quality of the product. Therefore, a dissolution curve determination method with discrimination ability needs to be established to obtain the dissolution behavior of linagliptin tablets from different sources and distinguish the intrinsic quality of the products.
[0006] Establishing a discriminative dissolution curve is of great significance for guiding the research and development of generic linagliptin tablets and evaluating the quality and efficacy consistency between the generic preparation and the original preparation. Summary of the Invention
[0007] Aiming at the deficiencies of the prior art, the present invention provides a method for determining the in vitro dissolution curve of linagliptin tablets, which has good dissolution properties for linagliptin tablets, strong specificity, good stability, can effectively distinguish the differences between self-made preparations and reference preparations, is more universal and practical, and has the advantages of being more simple and efficient in operation, solving the problems raised in the background technology.
[0008] The present invention provides the following technical solution: A method for determining the in vitro dissolution curve of linagliptin tablets, which comprises the following steps:
[0009] Step 1: Preparation of test solution: Take linagliptin tablets, use 900 ml of 0.1 mol / L hydrochloric acid solution as the dissolution medium, and perform dissolution according to the basket method of the dissolution apparatus, with the rotation speed of the basket method being 50 revolutions per minute; At different sampling time points, take an appropriate amount of the dissolution solution, filter it, and take the subsequent filtrate as the test solution;
[0010] Step 2: Preparation of reference solution: Take an appropriate amount of linagliptin reference substance, weigh it accurately, dissolve and dilute it with the dissolution medium to prepare a solution containing about 5.6 μg per 1 ml;
[0011] Step 3: Determination: Inject the test solution and the reference solution into a high performance liquid chromatograph respectively, record the chromatogram, and calculate the dissolution amount.
[0012] Further, in the above Step 1, the preparation of 0.1 mol / L hydrochloric acid solution: Accurately measure 8.5 ml of hydrochloric acid, add purified water and dilute it to 1000 ml, shake well, and obtain it.
[0013] Further, in the above Step 1, the sampling time points include 5 min, 10 min, 15 min, 30 min, 45 min and 60 min.
[0014] Further, in the above Step 1, after taking out the dissolution solution, filter more than 1 ml and then collect the subsequent filtrate as the test solution.
[0015] Further, in the above Step 3, the chromatographic conditions of the high performance liquid chromatography method are as follows:
[0016] Chromatographic column: Packed with octadecylsilane chemically bonded silica gel;
[0017] Detector: VWD detector, detection wavelength is 295 nm;
[0018] Mobile phase: Phosphate buffer - acetonitrile (78:22) as the mobile phase;
[0019] Column temperature: 50 °C;
[0020] Flow rate: 1.0 ml / min;
[0021] Running time: 15 min;
[0022] Sample injection volume: 10 μL.
[0023] Preferably, the chromatographic column filled with octadecylsilyl silica gel is a ZORBAX Eclipse XDB-C18 column, with a length of 150 mm, an inner diameter of 4.6 mm, and the diameter of the chromatographic column packing particles is 5 μm.
[0024] Preferably, the phosphate buffer solution is prepared as follows: Take 2.6 g of ammonium dihydrogen phosphate, add 1000 ml of water to dissolve it, and adjust the pH value to 3.0 ± 0.1 with phosphoric acid.
[0025] Compared with the prior art, the present invention has the following beneficial effects:
[0026] The present invention provides a method for determining the in vitro dissolution curve of linagliptin tablets. Through the dissolution results of the active pharmaceutical ingredient and in combination with the in vitro-in vivo correlation, the present invention selects 900 ml of 0.1 mol / L hydrochloric acid solution as the dissolution medium, and the dissolution method is the basket method with a rotation speed of 50 revolutions per minute, and the obtained curve has excellent resolution.
[0027] At the same time, when the dissolution medium selected by the present invention is used for the dissolution detection of the original research formulation and the self-made formulation of linagliptin tablets, the f2 value of the dissolution curve is greater than 50, indicating that the in vitro dissolution curve determination method of the present invention is applicable to evaluating the consistency of the original research formulation and the self-made formulation.
[0028] In addition, through methodological verification, the linagliptin tablet dissolution curve detection method provided by the present invention has the characteristics of strong specificity, high precision, and good solution stability. Description of the Drawings
[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for use in the embodiments. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as limiting the scope of the present invention.
[0030] Figure 1 It is the full wavelength scanning diagram of linagliptin;
[0031] Figure 2 It is the dissolution curve diagram of the reference preparation AC1555 in Example 1;
[0032] Figure 3 It is the dissolution curve diagram in Example 2;
[0033] Figure 4 It is the chromatogram of the reference solution in Example 3;
[0034] Figure 5 It is the chromatogram of the test solution in Example 3. Detailed implementation manners
[0035] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.
[0036] Example 1:
[0037] A method for determining the in vitro dissolution curve of linagliptin tablets, comprising the following steps:
[0038] 1. Chromatographic conditions
[0039] Chromatographic column: ZORBAX Eclipse XDB-C18 (4.6mm x 250mm, 5μm); using phosphate buffer - acetonitrile (78:22) as the mobile phase; detection wavelength is 295nm; column temperature is 50°C; flow rate is 1.0ml / min; injection volume is 10μl.
[0040] 2. Solution preparation
[0041] 0.1mol / L hydrochloric acid solution: Accurately measure 8.5ml of hydrochloric acid and dilute it to 1000ml with purified water, shake well, and obtain.
[0042] pH4.5 acetate buffer solution: Weigh about 1.5g of crystalline sodium acetate, dilute it to 500ml with water, adjust the pH value to 4.5 with glacial acetic acid, shake well, and obtain.
[0043] pH6.8 phosphate buffer solution: Weigh about 3.4g of potassium dihydrogen phosphate and about 0.5g of disodium hydrogen phosphate, dilute it to 500ml with water, adjust the pH value to 6.8 with sodium hydroxide solution, shake well, and obtain.
[0044] Aqueous solution: Take purified water and degas it to obtain.
[0045] Reference solution: Take an appropriate amount of linagliptin reference substance, accurately weigh it, dissolve and dilute it with the dissolution medium to prepare a solution containing about 5.6μg per 1ml.
[0046] 3. Establishment of a discriminative dissolution test method
[0047] 3.1 Selection of dissolution medium
[0048] Preparation of the test solution: Weigh accurately about 20 mg of linagliptin raw material, place it in a 100-ml volumetric flask, add the 0.1 mol / L hydrochloric acid solution, pH 4.5 acetate buffer solution, pH 6.8 phosphate buffer solution, and aqueous solution under "2 Solution Preparation" respectively, and make up to the mark. Shake vigorously and observe the dissolution situation.
[0049] The results showed that linagliptin was soluble in the 0.1 mol / L hydrochloric acid solution, pH 4.5 acetate buffer solution, and pH 6.8 phosphate buffer solution, and was still not completely dissolved after shaking in the aqueous solution for 1 h.
[0050] According to the requirements of the "General Guidelines for the Determination and Comparison of Dissolution Curves of Oral Solid Dosage Forms" and the "Q&A on the Study of Dissolution Curves in the 'Technical Guidelines for the Study of Pharmaceutical Changes of Listed Chemical Drugs (Trial)'", and combined with the results of the saturated solubility of linagliptin, select the 0.1 mol / L hydrochloric acid solution under "2 Solution Preparation" as the most discriminatory dissolution curve.
[0051] 3.2 Determination of the curve
[0052] Take the reference preparation (batch number: AC1555, linagliptin tablets with the license holder being Boehringer Ingelheim International GmbH, trade name: Trajenta, specification: 5 mg), 12 tablets. According to the dissolution and release determination method (Method 1 in General Chapter 0931 of the Fourth Part of the Chinese Pharmacopoeia 2020 Edition), conduct a dissolution experiment in a 0.1 mol / L hydrochloric acid medium (temperature: 37 °C), the volume of the medium is 900 ml, at a rotation speed of 50 revolutions per minute. After 5 min, 10 min, 15 min, 30 min, 45 min, and 60 min, take 10 ml of the sample respectively. After filtering and discarding 1 ml, take the subsequent filtrate as the test solution. Take an appropriate amount of the reference substance and prepare the reference solution according to "2 Solution Preparation". Take the above two solutions and conduct the determination according to "1 Chromatographic Conditions". Calculate the cumulative dissolution at each time point by the external standard method and draw the dissolution curve.
[0053] The results are shown in Figure 1 , and the specific data are shown in Table 1:
[0054] Table 1 Summary of the dissolution curve results of the reference preparation AC1555
[0055]
[0056]
[0057] The dissolution curve of the AC1555 reference preparation in 0.1 mol / L hydrochloric acid medium showed that the average dissolution at 5 min was 25.9%, at 10 min was 77.9%, at 15 min was 98.4%. The average dissolution at subsequent 30 min, 45 min and 60 min were 98.7%, 98.5% and 98.6% respectively. The difference in dissolution at the latter three points was less than 5%, and the dissolution plateau was reached at 15 min. And there was no burst release in the curve of this medium. Therefore, it is considered that the dissolution method of 900 ml of 0.1 mol / L hydrochloric acid medium with a basket method of 50 revolutions per minute is suitable for evaluating the consistency between the reference and the original research.
[0058] Example 2:
[0059] The test articles provided in this example were self-made preparations and reference preparations. Other chromatographic conditions, preparation of test solution, preparation of reference solution, etc. were the same as those under "3.2 Determination of Curve" in Example 1.
[0060] The test articles selected this time were self-made preparation - batch 24072302 and reference preparation - batch AC1555.
[0061] The cumulative dissolution of this product at different time points is shown in Table 2, and the dissolution curve atlas is shown in Figure 4 .
[0062] Table 2 Summary Table of Cumulative Dissolution Data
[0063]
[0064] Example 3: Specificity Test
[0065] Blank solvent: Take an appropriate amount of 0.1 mol / L hydrochloric acid solution as the dissolution medium as the blank solvent.
[0066] Blank excipient solution: Take about 7.5 mg of blank excipient, place it in a 500 ml volumetric flask, dilute it to the mark with the dissolution medium, and shake well to obtain.
[0067] Test solution: Take the sample solution of the first tablet of the reference preparation in Example 2 at 15 min of dissolution as the test solution.
[0068] Reference solution: Take an appropriate amount of linagliptin reference substance, weigh it accurately, dissolve and dilute it with the dissolution medium to prepare a solution containing about 5.6 μg per 1 ml.
[0069] Precision measure 10 μL of each of the above solutions respectively, inject them into the liquid chromatographic system, and record the chromatogram. The results are shown in Table 3.
[0070] Table 3 Dissolution Methodology Validation - Specificity Results
[0071]
[0072] The results showed that neither the blank solvent nor the blank excipients interfered with the dissolution test.
[0073] Example 4: Solution Stability
[0074] Test solution: Take the sample solution of the first reference preparation in Example 2 at 60 min of dissolution as the test solution.
[0075] Precisely measure 10 μL of each of the above test solutions, and inject them into the liquid chromatography system at 0 h, 8 h, 20 h, 1 d up to 9 d, and record the chromatograms. The results are shown in Table 4.
[0076] Table 4 Results of dissolution methodology verification - stability of test solution
[0077]
[0078] The results showed that when the reference solution and the test solution were placed at room temperature (10 - 30 °C) for 9 d, the RSD value of the change in peak area was 0.4%, less than 5%. Therefore, the test solution was stable when placed at room temperature for 9 d.
[0079] Example 5: Filter membrane adsorption
[0080] Examine the filter membrane adsorption of the white filter head installed in the instrument during automatic sampling and the filter membrane used during manual filtration, respectively.
[0081] Test solution 1: Take the test solution at 5 min of the first dissolution point of the dissolution curve as the sample for investigating low - concentration filter membrane adsorption. Centrifuge this sample as test solution 1 - 1, filter and discard 1 ml, take the subsequent filtrate as test solution 1 - 2, filter and discard 3 ml, take the subsequent filtrate as test solution 1 - 3, filter and discard 5 ml, take the subsequent filtrate as test solution 1 - 4, sample this sample through the dissolution apparatus with a filter head as test solution 1 - 5, and after filtering through the filter head and sampling again, discard 5 ml and take the subsequent filtrate as test solution 1 - 6; a total of 6 samples.
[0082] Test solution 2: Take the test solution at 60 min of the last dissolution point of the dissolution curve as the sample for investigating high - concentration filter membrane adsorption. Centrifuge this sample directly as test solution 2 - 1; filter and discard 5 ml, take the subsequent filtrate as test solution 2 - 2, sample this sample through the dissolution apparatus with a filter head as test solution 2 - 3, and after filtering through the filter head and sampling again, discard 5 ml and take the subsequent filtrate as test solution 2 - 4; a total of 4 samples.
[0083] Precisely measure 10 μL of each of the above test solutions, inject them into the liquid chromatography system, and record the chromatograms. The results are shown in Table 5.
[0084] Table 5 Dissolution Methodology Validation - Filter Membrane Adsorption
[0085]
[0086] The results showed that the white filter head had no adsorption of linagliptin in the high - concentration test solution and the low - concentration test solution during automatic sampling by the instrument. When manually sampling and filtering off more than 1 ml, there was also no adsorption of linagliptin. Therefore, in subsequent dissolution experiments, the filtered white head or the subsequent filtrate after manually filtering off more than 1 ml can be used as the test solution during sampling.
[0087] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non - exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or elements inherent to such process, method, article or device.
[0088] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A method for determining the in vitro dissolution curve of linagliptin tablets, characterized in that: Step 1: Preparation of test solution: Take linagliptin tablets, use 900 ml of 0.1 mol / L hydrochloric acid solution as the dissolution medium, and dissolve according to the dissolution apparatus basket method, the basket speed is 50 rpm; at different sampling time points, take out an appropriate amount of dissolution solution, filter and take the filtrate as the test solution; Step 2: Preparation of reference solution: Take an appropriate amount of linagliptin reference substance, weigh accurately, dissolve and dilute with dissolution medium to make a solution containing about 5.6 μg per 1 ml; Step 3: Determination: Inject the test solution and reference solution into HPLC respectively, record the chromatogram and calculate the dissolution amount.
2. The method for determining the in vitro dissolution curve of linagliptin tablets according to claim 1, characterized in that: The dissolution medium is 0.1 mol / L hydrochloric acid solution.
3. The method for determining the in vitro dissolution curve of linagliptin tablets according to claim 1, characterized in that: The sampling time points include 5min, 10min, 15min, 30min, 45min and 60min.
4. The method for determining the in vitro dissolution curve of linagliptin tablets according to claim 1, characterized in that: After taking out the solution, filter and discard more than 1 ml.
5. The method for determining the in vitro dissolution curve of linagliptin tablets according to claim 1, characterized in that: The chromatographic conditions of the high performance liquid chromatography are: Chromatographic column: Octadecylsilane bonded silica gel is used as filler; Detector: VWD detector, detection wavelength is 295nm; Mobile phase: phosphate buffer-acetonitrile (78:22) was used as the mobile phase; Column temperature: 50°C; Flow rate: 1.0ml / min; Running time: 15min; Injection volume: 10 μL.
6. The method for determining the in vitro dissolution curve of linagliptin tablets according to claim 5, characterized in that: The chromatographic column using octadecylsilane bonded silica as filler is a ZORBAX Eclipse XDB-C18 column with a length of 250 mm, an inner diameter of 4.6 mm, and a diameter of the chromatographic column filler particles of 5 μm.