A quality control method for linagliptin

Abstract

The invention discloses a quality control method for linagliptin, which adopts high performance liquid chromatography, uses 0.1% to 0.2% V / V acidic solution as mobile phase A, uses organic solvent as mobile phase B, and performs gradient elution; wherein, based on the volume ratio of mobile phase A, the gradient elution program is 0 to 5 minutes: maintained at 75% to 80%, 5.1 to 20 minutes: reduced from 75% to 80% to 20% to 25%, and 20.1 to 26 minutes: maintained at 75% to 80%. The linagliptin quality control method established by the method of the invention can quickly and accurately detect the content of each impurity therein, has good specificity and repeatability, and is not only suitable for the quality control of the whole process of linagliptin raw material preparation, but also suitable for the quality control of preparations.

Description

Technical Field

[0001] The present invention relates to a quality control method for linagliptin, and in particular to a quality control method capable of effectively monitoring the impurity content of linagliptin. Background Art

[0002] Dipeptidyl peptidase 4 (DPP-4) can degrade incretin-like polypeptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). As a DPP-4 inhibitor, linagliptin can increase the concentration of active incretin hormones, stimulate insulin release in a glucose-dependent manner, and reduce circulating glucagon levels.

[0003] Linagliptin preparation uses a lot of raw materials, which will introduce some impurities. At the same time, a variety of impurities will be produced during the preparation process, which will affect the quality, safety and effectiveness of the API and preparations. If the impurities in linagliptin can be effectively detected or controlled, it will be more conducive to controlling its preparation process and product quality. However, the current detection method is complicated and has low separation, resulting in poor accuracy of the test results. Summary of the invention

[0004] Purpose of the invention: In view of the deficiency that existing detection methods cannot efficiently and accurately monitor the quality of linagliptin, the present invention aims to provide a convenient, stable and reliable quality control method for linagliptin.

[0005] Technical solution: The quality control method of linagliptin of the present invention comprises the following steps:

[0006] (1) Prepare reference substance and test substance solutions;

[0007] (2) High performance liquid chromatography was used, with an acid solution having a volume fraction of 0.1% to 0.2% v / v as mobile phase A and an organic solvent as mobile phase B, and gradient elution was performed; wherein, based on the volume ratio of mobile phase A, the gradient elution program was: 0 to 5 minutes: maintained at 75% to 80%, 5.1 to 20 minutes: reduced from 75% to 80% to 25% to 20%, and 20.1 to 26 minutes: 75% to 80%;

[0008] (3) Calculate the content of each impurity using the area normalization method.

[0009] Among them, the impurities targeted by the above quality control method are as follows:

[0010] Impurity 37

[0011]

[0012] Impurity 7

[0013]

[0014] Impurity C

[0015]

[0016] Impurity S

[0017]

[0018] Impurity 33

[0019]

[0020] Impurity 30

[0021]

[0022] Impurity 35

[0023]

[0024] Impurity 55

[0025]

[0026] The preferred gradient elution procedure is as follows:

[0027]

[0028] Specifically, the acidic solution is at least one of trifluoroacetic acid aqueous solution, formic acid aqueous solution, phosphoric acid aqueous solution, and perchloric acid aqueous solution, and the organic solvent is acetonitrile or methanol; the preferred acidic solution is 0.1% trifluoroacetic acid aqueous solution, and the organic solvent is acetonitrile.

[0029] Preferably, an alkylsilane bonded silica gel phase chromatographic column is used, the detection wavelength is 220-230 nm, the mobile phase flow rate is 0.6-0.8 ml / min, and the column temperature is 50° C.-60° C.

[0030] Among them, for the chromatographic conditions, the more preferred detection wavelength is 225nm, the mobile phase flow rate is 0.7ml / min, and the column temperature is 55°C; for the chromatographic column, it is more preferred to use a C18 chromatographic column or a C8 chromatographic column with a column length of 150-250mm, a column diameter of 2.0-4.6mm, and a column packing particle size of 3.0-5.0μm; the most preferred is a C18 chromatographic column with a column length of 150mm, a column diameter of 3.0mm, and a column packing particle size of 3.5μm, and specifically a Waters XBridge C18 column can be used.

[0031] The linear range of the above method for each quality control component is as follows:

[0032]

[0033] Furthermore, step (1) preparing the sample solution includes preparing an impurity mixed solution, an impurity localization solution and a test sample solution, and the preferred preparation solvent is an acetonitrile-water (30:70) solution;

[0034] Wherein, the preparation method of impurity mixed solution is as follows:

[0035] A. Preparation of impurity stock solution

[0036] Weigh an appropriate amount of each impurity reference substance, dissolve it in acetonitrile-water (30:70) and quantitatively dilute it to make a solution containing about 0.2 mg per 1 ml as the stock solution of each impurity;

[0037] B. Preparation of impurity mixed solution

[0038] Weigh an appropriate amount of linagliptin reference substance, dissolve it with an appropriate amount of acetonitrile-water (30:70), add an appropriate amount of the above-mentioned impurity stock solution, and quantitatively dilute it with acetonitrile-water (30:70) to prepare a solution containing approximately 0.1 mg of linagliptin and 2 μg of each impurity per 1 ml, as the impurity mixed solution.

[0039] The preparation method of impurity localization solution is as follows:

[0040] Take an appropriate amount of the above impurity stock solution and quantitatively dilute it with acetonitrile-water (30:70) to prepare a solution containing about 2 μg of each impurity per 1 ml, which serves as the impurity location solution.

[0041] The preparation method of the test solution is as follows:

[0042] Weigh an appropriate amount of linagliptin, dissolve it in an appropriate amount of acetonitrile-water (30:70), and then quantitatively dilute it with acetonitrile-water (30:70) to make a solution containing about 0.1 mg of linagliptin per 1 ml as the test solution. The test sample can be linagliptin raw material, intermediate product of preparation containing linagliptin, or finished product.

[0043] The injection volume of the test sample in step (2) is preferably 10 μl, and the preparation method of 0.1% trifluoroacetic acid solution is to take 1 ml of trifluoroacetic acid and dissolve it in 1000 ml of water.

[0044] The content of the relevant substance in step (3) is preferably calculated according to the area normalization method.

[0045] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0046] 1. A detection method for linagliptin-related substances has been established, which can quickly and accurately reflect the quality status of linagliptin, thereby effectively controlling the quality of the raw materials and their preparations;

[0047] 2. The method is easy to operate, with good specificity (the separation between each impurity and the main component is not less than 1.5, and the purity of the main peak is greater than 990), repeatability and accuracy (the linear correlation coefficient is greater than 0.999);

[0048] 3. The method has wide applicability and can be used for quality control of the entire preparation process of linagliptin API, as well as its preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 This is the chromatogram of the blank interference test;

[0050] Figure 2 is the chromatogram of the impurity mixed solution;

[0051] Figure 3 Chromatogram of the solution for locating impurity 37;

[0052] Figure 4 Chromatogram of the solution for locating impurity 7;

[0053] Figure 5 Chromatogram of the solution for locating impurity C;

[0054] Figure 6 Chromatogram of the solution for locating impurity S;

[0055] Figure 7 Chromatogram of the solution for locating impurity 33;

[0056] Figure 8 Chromatogram of the solution for locating impurity 30;

[0057] Fig. 9 Chromatogram of the solution for locating impurity 35;

[0058] Fig.10 Chromatogram of the solution for locating impurity 55;

[0059] Fig.11 It is the superimposed chromatogram of each damage condition of the forced degradation test;

[0060] Fig.12 Overlay chromatograms of repeatability tests. DETAILED DESCRIPTION

[0061] The technical solution of the present invention is further described below in conjunction with embodiments.

[0062] Example 1: System suitability test

[0063] 1. Prepare mixed impurity solution and each impurity location solution

[0064] Weigh an appropriate amount of each impurity reference substance, dissolve it in acetonitrile-water (30:70) and quantitatively dilute it to make a solution containing about 0.2 mg per 1 ml, which serves as the stock solution of each impurity.

[0065] Weigh an appropriate amount of linagliptin reference substance, dissolve it with an appropriate amount of acetonitrile-water (30:70), add an appropriate amount of the above-mentioned impurity stock solutions, and quantitatively dilute with acetonitrile-water (30:70) to prepare a mixed solution containing approximately 0.1 mg of linagliptin and 2 μg of each impurity per 1 ml, as the impurity mixed solution.

[0066] Take an appropriate amount of the above impurity stock solution and quantitatively dilute it with acetonitrile-water (30:70) to prepare a solution containing 2 μg of each impurity per 1 ml, which is used as the impurity location solution.

[0067] 2. Detection

[0068] According to the high performance liquid chromatography method, a Waters XBridge C18 (150×3.0 mm, 3.5 μm) column was used, 0.1% trifluoroacetic acid solution was used as mobile phase A, acetonitrile was used as mobile phase B, gradient elution was performed according to the table below, the flow rate was 0.7 ml per minute, the detection wavelength was 225 nm, and the column temperature was 55° C.

[0069]

[0070] Take the above impurity mixed solution and 10 μl of each impurity location solution, inject them into the liquid chromatograph, and record the chromatogram ( Figures 1 to 10 ).

[0071] 3. Conclusion

[0072] Depend on Figure 1 It can be seen that the blank solvent has no interference and the impurities in the system applicability are highly separated.

[0073] Example 2: Forced degradation test

[0074] 1. Preparation of test solution

[0075] Undamaged: Take an appropriate amount of linagliptin, dissolve it with an appropriate amount of acetonitrile-water (30:70), and then quantitatively dilute it with acetonitrile-water (30:70) to prepare a solution containing approximately 0.1 mg of linagliptin per 1 ml.

[0076] Acid destruction: Take an appropriate amount of linagliptin, add 1 ml of 1 mol / L HCL solution, leave at room temperature for 2 hours, neutralize with 1 mol / L NaOH, dissolve with an appropriate amount of acetonitrile-water (30:70), and quantitatively dilute with acetonitrile-water (30:70) to make a solution containing approximately 0.1 mg of linagliptin per 1 ml.

[0077] Alkaline destruction: Take an appropriate amount of linagliptin, add 1 ml of 1 mol / L NaOH solution, leave it at room temperature for 2 hours, then neutralize it with 1 mol / L HCL, dissolve it with an appropriate amount of acetonitrile-water (30:70), and quantitatively dilute it with acetonitrile-water (30:70) to make a solution containing approximately 0.1 mg of linagliptin per 1 ml.

[0078] Oxidative destruction: Take an appropriate amount of linagliptin, add 1 ml of 30% H2O2, and leave it at room temperature for 2 hours. Dissolve it with an appropriate amount of acetonitrile-water (30:70), and then quantitatively dilute it with acetonitrile-water (30:70) to make a solution containing approximately 0.1 mg of linagliptin per 1 ml.

[0079] 2. Detection

[0080] The above-mentioned test sample was tested by high performance liquid chromatography according to Example 1, and the chromatogram was recorded ( Fig.11 ).

[0081] 3. Conclusion

[0082] Under each destructive test condition, the peak purity of the main peak was greater than 990, and the separation between the main component, known impurity peak and adjacent impurity peak met the requirements. The specificity of this method met the detection requirements of the relevant substances.

[0083] Example 3: Repeatability test

[0084] 1. Preparation of test solution

[0085] Take an appropriate amount of linagliptin, dissolve it in an appropriate amount of acetonitrile-water (30:70), and then quantitatively dilute it with acetonitrile-water (30:70) to make a solution containing about 0.1 mg of linagliptin per 1 ml. Prepare 6 portions in the same way.

[0086] 2. Detection

[0087] The above-mentioned test sample was tested by high performance liquid chromatography according to Example 1, and the chromatogram was recorded ( Fig.12 ).

[0088] 3. Conclusion

[0089] This method is easy to operate, has good reproducibility, and meets the detection requirements of relevant substances.

[0090] Example 4: Linearity and range test

[0091] 1. Sample preparation

[0092] Take appropriate amounts of impurity 37, linagliptin, impurity 7, impurity C, impurity S, impurity 33, impurity 30, impurity 35 and impurity 55 reference substances, add acetonitrile-water (30:70) to dissolve and quantitatively dilute to prepare a series of linear mixed solutions.

[0093] 2. Detection

[0094] The above samples were tested by high performance liquid chromatography according to Example 1. The results are shown in Tables 1 to 9.

[0095] Table 1. Linearity and range investigation results of linagliptin

[0096]

[0097]

[0098] Table 2. Linearity and range investigation results of impurity 37

[0099]

[0100] Table 3. Linearity and range investigation results of impurity 7

[0101]

[0102] Table 4. Linearity and range investigation results of impurity C

[0103]

[0104] Table 5. Linearity and range investigation results of impurity S

[0105]

[0106] Table 6. Linearity and range investigation results of impurity 33

[0107]

[0108]

[0109] Table 7. Linearity and range investigation results of impurity 30

[0110]

[0111] Table 8. Linearity and range investigation results of impurity 35

[0112]

[0113] Table 9. Linearity and range investigation results of impurity 55

[0114]

[0115] 3. Conclusion

[0116] The linear correlation coefficients of linagliptin and each impurity are all greater than 0.999, and the percentage of the Y-axis intercept and the 100% concentration level response value is within 25%, indicating that the linearity and range of the detection method of the present invention meet the detection requirements of the relevant substances.

Claims

1. A quality control method for linagliptin, characterized in that: The following steps are involved: (1) preparing sample solution; (2) High performance liquid chromatography was performed using a C18 column with a length of 150 mm, a diameter of 3.0 mm, and a particle size of 3.5 μm as the chromatographic column, a 0.1% V / V trifluoroacetic acid aqueous solution as the mobile phase A, and acetonitrile as the mobile phase B, and gradient elution was performed; wherein, based on the volume ratio of the mobile phase A, the gradient elution program was 0 to 5 minutes: maintained at 75% to 80%, 5.1 to 20 minutes: reduced from 75% to 80% to 20% to 25%, and 20.1 to 26 minutes: 75% to 80%; (3) Calculate the content of quality control impurities by area normalization method, and the quality control impurities are as follows: Impurity 37 Impurity 7 Impurity C Impurity S Impurity 33 Impurity 30 Impurity 35 Impurity 55 2. The quality control method according to claim 1, characterized in that: The procedure of the gradient elution is as follows:

3. The quality control method according to claim 1, characterized in that: The detection wavelength of the high performance liquid chromatography is 220-230 nm, the flow rate of the mobile phase is 0.6-0.8 ml / min, and the column temperature is 50° C.-60° C.

4. The quality control method according to claim 3, characterized in that: The detection wavelength was 225 nm, the mobile phase flow rate was 0.7 ml / min, and the column temperature was 55° C.

5. The quality control method according to claim 1, characterized in that: The chromatographic column is a Waters XBridge C18 column with a column length of 150 mm, a column diameter of 3.0 mm, and a column filler particle size of 3.5 μm.

6. The quality control method according to claim 1, characterized in that: The detection linear range of each quality control component is as follows:

7. The quality control method according to claim 6, characterized in that: The detection linear equations of each quality control component are as follows:

8. The quality control method according to claim 1, characterized in that: The sample solution is prepared using a 30:70 V / V acetonitrile-water solution.

9. The quality control method according to claim 1, characterized in that: The injection volume of the gradient elution was 10 μl.

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