Method for determining content and impurity limit of imatinib mesylate based on HPLC-DAD (High performance liquid chromatography-diode array detection) method

Abstract

The invention discloses a method for determining the content and the impurity limit of imatinib mesylate based on a HPLC-DAD (High performance liquid chromatography-diode array detection) method. A C18 chromatographic column is used; a methanol-triethylamine water solution is used as a flowing phase; the volume percentage concentration of the triethylamine water solution is 0.5-1 percent; the pH value of the triethylamine water solution is 7.5-11; the flowing speed is 0.8-1.2 ml / min and the detection wavelength is 200-400 nm during a gradient elution process; the process of the gradient elution lasts for 0-55 minutes and the volume percentage concentration of the triethylamine water solution is 65-15 percent. Under the chromatographic condition, the separation degrees among all components are greater than 2.0; the linear range ranges from 9.09 to 90.92 microgram.ml<-1> (r=0.9998); the detection limit is 0.3ng. The method is simple, accurate, high in sensitivity and good in repeatability, and avoids the influence on agents and the service life of the chromatographic column due to octane sulfonic acid sodium ions.

Description

technical field

[0001] The invention relates to a method for determining the content of imatinib mesylate and its impurity limit based on the HPLC-DAD method. Background technique

[0002] Imatinib mesylate, the chemical name is 4-[(4-methyl-1-piperazine)methyl]-N-{4-methyl-3-[[4-(3- Pyridyl)–2-pyrimidinyl]amino]phenyl}benzamide methanesulfonate, whose trade name is Gleevec, has the following structural formula, and is a tyrosine kinase inhibitor drug developed by Novartis, Switzerland . It has obtained orphan drug status in the United States, the European Union, Japan and other countries, and is used for the treatment of alpha-interferon (interfereon–alfa) treatment failure blast crisis disease, accelerated disease or chronic myelogenous leukemia and gastrointestinal stroma cell tumor.

[0003] From the review of imatinib analysis methods by Zhang Xiaoshun et al., it can be seen that the current domestic and foreign analysis methods for imatinib mesylate (HPLC-MS, HPLC-M...

Examples

Embodiment 1

[0050] Preparation of reference substance and test solution for content determination: Take 25 mg of imatinib mesylate reference substance and sample respectively in a 50 mL volumetric flask, dissolve with methanol, dilute to the mark with solvent, and shake well. Take 1mL each and place in a 10mL volumetric flask, dilute to the mark with solvent, and shake well.

[0051] Preparation of test solution for related substance inspection: Take imatinib mesylate sample 25mg and place it in a 50mL volumetric flask, dissolve it with methanol, dilute the solvent to the mark, and shake up.

[0052] Preparation of reference substance solution for related substance inspection: Take imatinib mesylate related substance inspection test solution 1mL in a 100mL volumetric flask, dilute to the mark with a solvent, and shake well.

Embodiment 2

[0054] Get the imatinib mesylate reference substance and prepare the imatinib mesylate solution according to the method of Example 1, and get the imatinib mesylate reference substance and a single impurity and prepare the system suitability according to the method of embodiment 3 After testing the solution, accurately measure 10 μL and inject it into the liquid chromatograph, and perform DAD scanning in the range of 200nm to 400nm (see figure 1 ). The results show that imatinib mesylate and various impurities have maximum absorption at 230nm and 268nm, while triethylamine has a certain absorption at 230nm, which significantly increases the baseline noise and reduces the detection sensitivity, so 268nm is selected as the detection wavelength .

Embodiment 3

[0056] Take the appropriate amount of imatinib mesylate reference substance, impurity A, impurity B, impurity C, impurity D, impurity E, and impurity F, weigh them accurately, add methanol to ultrasonic treatment to dissolve, and quantitatively dilute with a solvent to make each 1mL A mixed solution containing 0.5 mg of imatinib mesylate and 50 μg of various impurities is used as a system suitability test solution. According to the above chromatographic conditions, 10 μL of sample was injected for analysis, the number of theoretical plates of imatinib mesylate was greater than 10,000, the number of theoretical plates of each impurity was greater than 5,000, the tailing factor was between 0.95 and 1.05, and the components were separated Degrees are greater than 2.0 (see figure 2 ).

technical field

[0001] The invention relates to a method for determining the content of imatinib mesylate and its impurity limit based on the HPLC-DAD method. Background technique

[0002] Imatinib mesylate, the chemical name is 4-[(4-methyl-1-piperazine)methyl]-N-{4-methyl-3-[[4-(3- Pyridyl)–2-pyrimidinyl]amino]phenyl}benzamide methanesulfonate, whose trade name is Gleevec, has the following structural formula, and is a tyrosine kinase inhibitor drug developed by Novartis, Switzerland . It has obtained orphan drug status in the United States, the European Union, Japan and other countries, and is used for the treatment of alpha-interferon (interfereon–alfa) treatment failure blast crisis disease, accelerated disease or chronic myelogenous leukemia and gastrointestinal stroma cell tumor.

[0003] From the review of imatinib analysis methods by Zhang Xiaoshun et al., it can be seen that the current domestic and foreign analysis methods for imatinib mesylate (HPLC-MS, HPLC-M...

Examples

Embodiment 1

[0050] Preparation of reference substance and test solution for content determination: Take 25 mg of imatinib mesylate reference substance and sample respectively in a 50 mL volumetric flask, dissolve with methanol, dilute to the mark with solvent, and shake well. Take 1mL each and place in a 10mL volumetric flask, dilute to the mark with solvent, and shake well.

[0051] Preparation of test solution for related substance inspection: Take imatinib mesylate sample 25mg and place it in a 50mL volumetric flask, dissolve it with methanol, dilute the solvent to the mark, and shake up.

[0052] Preparation of reference substance solution for related substance inspection: Take imatinib mesylate related substance inspection test solution 1mL in a 100mL volumetric flask, dilute to the mark with a solvent, and shake well.

Embodiment 2

[0054] Get the imatinib mesylate reference substance and prepare the imatinib mesylate solution according to the method of Example 1, and get the imatinib mesylate reference substance and a single impurity and prepare the system suitability according to the method of embodiment 3 After testing the solution, accurately measure 10 μL and inject it into the liquid chromatograph, and perform DAD scanning in the range of 200nm to 400nm (see figure 1 ). The results show that imatinib mesylate and various impurities have maximum absorption at 230nm and 268nm, while triethylamine has a certain absorption at 230nm, which significantly increases the baseline noise and reduces the detection sensitivity, so 268nm is selected as the detection wavelength .

Embodiment 3

[0056] Take the appropriate amount of imatinib mesylate reference substance, impurity A, impurity B, impurity C, impurity D, impurity E, and impurity F, weigh them accurately, add methanol to ultrasonic treatment to dissolve, and quantitatively dilute with a solvent to make each 1mL A mixed solution containing 0.5 mg of imatinib mesylate and 50 μg of various impurities is used as a system suitability test solution. According to the above chromatographic conditions, 10 μL of sample was injected for analysis, the number of theoretical plates of imatinib mesylate was greater than 10,000, the number of theoretical plates of each impurity was greater than 5,000, the tailing factor was between 0.95 and 1.05, and the components were separated Degrees are greater than 2.0 (see figure 2 ).

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