A method for separating conitinib and related substances by liquid chromatography
A technology for conitinib and related substances, which is applied in the field of drug analysis and can solve problems such as adverse drug reactions
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Embodiment 1
[0082] Embodiment 1: Determination of detection wavelength
[0083] 1) Experimental steps:
[0084] Using methanol as a solvent, a total of 8 solutions containing conitinib and its 7 impurities (impurity I-impurity VII) were prepared respectively, and the concentration of each solution was about 10 μg / ml.
[0085] Using methanol solvent as a blank, the solution of conitinib and various related substances was scanned by ultraviolet detection, and the scanning wavelength range was 200-400nm.
[0086] 2) Experimental results: Conitinib and related substances have strong absorption in the wavelength range of 220-270nm, and each sample has strong absorption at 250nm wavelength.
[0087] Therefore, the present invention selects 250nm as the detection wavelength.
[0088] In addition, the detection wavelength has also been verified by destroying experimental samples.
Embodiment 2
[0089] Embodiment 2: the screening of mobile phase
[0090] 1) Chromatographic conditions
[0091] Chromatographic column: the chromatographic column filler is octylsilane bonded silica gel with a particle size of 3.5 μm, and the chromatographic column specification is 150mm×4.6mm, purchased from Waters;
[0092] Mobile phase flow rate: 1.0ml / min;
[0093] Injection volume: 5μl;
[0094] Column temperature: 30°C;
[0095] Detection wavelength: 250nm.
[0096] 2) Preparation of the test solution: take 30 mg of conitinib and 3 mg of its related substances respectively, mix and place in a 100 ml volumetric flask, add solvent (0.05% trifluoroacetic acid aqueous solution and 0.05% trifluoroacetic acid acetonitrile solution) According to the mixed solution prepared with a volume ratio of 30:70), dissolve and dilute to the mark, shake well, and the test solution is ready.
[0097] 3) Screening of the aqueous phase
[0098] Mobile phase A is water, mobile phase B is acetonitrile...
Embodiment 3
[0117] Embodiment 3: determination of limit of detection and limit of quantitation
[0118] 1) Chromatographic conditions
[0119] Chromatographic column: the chromatographic column filler is octylsilane bonded silica gel with a particle size of 3.5 μm, and the chromatographic column specification is 150mm×4.6mm, purchased from Waters;
[0120] Mobile phase: Mobile phase A is a 0.05% trifluoroacetic acid aqueous solution, and mobile phase B is a 0.05% trifluoroacetic acetonitrile solution, and the elution gradient is eluted as shown in Table 1 in Example 1:
[0121] Mobile phase flow rate: 1.0ml / min;
[0122] Injection volume: 5μl;
[0123] Column temperature: 30°C;
[0124]The detection wavelength is 250nm.
[0125] 2) Preparation of sample solution: Weigh 30 mg of the standard product conitinib and related substances, put them in 100 ml measuring bottles respectively, mix them with solvent (mobile phase A and mobile phase B prepared according to the volume ratio of 70:30...
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