Method for separating and determining palbociclib and impurities thereof

Abstract

A method for separating and determining palbociclib and impurities thereof comprises the following steps: 1) taking palbociclib or a preparation containing palbociclib, and adding a diluent to dissolve palbociclib or the preparation containing palbociclib to obtain a sample solution with a concentration of 0.1-10 mg / ml; 2) taking the sample solution obtained in the step 1), and adding a diluent todilute the sample solution by 50-1000 times to obtain a control solution; 3) adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filler, setting the flow velocity of a mobile phase to be 0.8 to 1.2 ml / min; wherein the mobile phase is composed of a mobile phase A and a mobile phase B, the mobile phase A being a buffer solution with the concentration of 0.0001-1.0mol / L, adjusting the pH value of the buffer solution to 1-5 with phosphoric acid, the mobile phase B being methanol, and the mobile phase entering a chromatographic column in a gradient elution mode;and 4) respectively injecting isopyknic sample solution in the step 1) and contrast solution in the step 2) into a high performance liquid chromatograph, detecting by utilizing the wavelength of 200nm to 280nm, and recording a chromatogram to finish the separation and determination of the impurities in the sample solution, wherein the sample injection amount is 5[mu]l to 100[mu]l.

Description

technical field

[0001] The invention relates to the field of analytical chemistry, in particular to a method for separating and measuring palbociclib and its impurities. Background technique

[0002] Palbociclib is a highly selective and reversible antagonist of the cyclin-dependent kinases CDK4 and CDK6. Blocks cell cycle progression from G1 phase to S phase by inhibiting CDK4 / 6, thereby inhibiting DNA synthesis. Preclinical research data show that palbociclib may have the effect of reducing tumor cells and inhibiting cell growth, and is a new breast cancer drug. The molecular formula of palbociclib is C 24 h 29 N 7 o 2 , chemical name: 6-acetyl-8-cyclopentyl-5-methyl-2-[[5-(piperazin-1-yl)pyridin-2-yl]amino]-8H-pyrido[2 ,3-D] pyrimidin-7-one, the structural formula is as shown in formula (a)

[0003]

[0004] In the process of synthesizing this compound, there are several important intermediates and unknown impurities that may affect the purity and quality of the...

Examples

Embodiment 1

[0040] Take impurity SM1, impurity SM2, impurity A1, impurity Z7, impurity Z8, impurity Z12, impurity Z16, impurity Z21, impurity Z23, impurity Z25, impurity Z26, impurity Z29 (the purity of each impurity is above 95%), each 25mg , accurately weighed, placed in a 25ml measuring bottle, dissolved in methanol-buffer solution (volume ratio 40:60) and diluted to the mark, shaken well, as impurity stock solution; take about 20mg of palbociclib, accurately weighed , placed in a 25ml measuring bottle, accurately add 0.25ml of impurity stock solution, add methanol-buffer solution (volume ratio 40:60) to dissolve and dilute to the mark, shake well, as a mixed control solution.

[0041] Take diluent methanol-buffer solution (volume ratio is 40:60) and mixed control solution respectively, carry out liquid chromatography analysis according to the above-mentioned chromatographic conditions, record the chromatogram, the result is as follows: figure 1 , figure 2 shown.

[0042] figure 1 ...

Embodiment 2

[0044] The determination of embodiment two palbociclib bulk drug (provided by Chongqing Sansheng Industrial Co., Ltd.)

[0045] Take palbociclib 20mg, weigh it accurately, place it in a 25ml measuring bottle, add methanol-buffer solution (volume ratio 40:60) for ultrasonic treatment to dissolve and dilute to the mark, shake well, and use it as the sample solution; Sample solution 1.0ml is placed in 100ml measuring bottle, diluted to scale with methanol-buffer solution (volume ratio is 40:60), shakes up, as contrast solution; Carry out liquid chromatography analysis by the chromatographic condition of embodiment one, record Chromatogram. If there are impurity peaks (except solvent peaks) in the chromatogram of the sample solution, calculate the impurity content according to the self-control method with a correction factor. The result is as image 3 , Figure 4 shown. The test results are shown in Table 1:

[0046] Table 1 palbociclib related substance detection results

...

Embodiment 3

[0049] The determination of embodiment three palbocicini capsules (provided by Chongqing Sansheng Industrial Co., Ltd.)

[0050] Take an appropriate amount of Palbociclib capsules (approximately equivalent to 20 mg Palbociclib), put them in a 25ml measuring bottle, add methanol-buffer solution (40:60 by volume) to dissolve by ultrasonic treatment and dilute to the mark, shake well, filter After that, take the filtrate as the sample solution; accurately measure 1.0ml of the sample solution, place it in a 100ml measuring bottle, dilute it to the mark with methanol-buffer solution (40:60 in volume ratio), shake well, and use it as a control solution; press For the prescription ratio of Palbociclib capsules, take an appropriate amount of blank excipients, and prepare blank excipients for test solution in the same way as the sample solution; carry out liquid chromatography analysis according to the chromatographic conditions of Example 1. Record the chromatogram, the result is as f...

technical field

[0001] The invention relates to the field of analytical chemistry, in particular to a method for separating and measuring palbociclib and its impurities. Background technique

[0002] Palbociclib is a highly selective and reversible antagonist of the cyclin-dependent kinases CDK4 and CDK6. Blocks cell cycle progression from G1 phase to S phase by inhibiting CDK4 / 6, thereby inhibiting DNA synthesis. Preclinical research data show that palbociclib may have the effect of reducing tumor cells and inhibiting cell growth, and is a new breast cancer drug. The molecular formula of palbociclib is C 24 h 29 N 7 o 2 , chemical name: 6-acetyl-8-cyclopentyl-5-methyl-2-[[5-(piperazin-1-yl)pyridin-2-yl]amino]-8H-pyrido[2 ,3-D] pyrimidin-7-one, the structural formula is as shown in formula (a)

[0003]

[0004] In the process of synthesizing this compound, there are several important intermediates and unknown impurities that may affect the purity and quality of the...

Examples

Embodiment 1

[0040] Take impurity SM1, impurity SM2, impurity A1, impurity Z7, impurity Z8, impurity Z12, impurity Z16, impurity Z21, impurity Z23, impurity Z25, impurity Z26, impurity Z29 (the purity of each impurity is above 95%), each 25mg , accurately weighed, placed in a 25ml measuring bottle, dissolved in methanol-buffer solution (volume ratio 40:60) and diluted to the mark, shaken well, as impurity stock solution; take about 20mg of palbociclib, accurately weighed , placed in a 25ml measuring bottle, accurately add 0.25ml of impurity stock solution, add methanol-buffer solution (volume ratio 40:60) to dissolve and dilute to the mark, shake well, as a mixed control solution.

[0041] Take diluent methanol-buffer solution (volume ratio is 40:60) and mixed control solution respectively, carry out liquid chromatography analysis according to the above-mentioned chromatographic conditions, record the chromatogram, the result is as follows: figure 1 , figure 2 shown.

[0042] figure 1 ...

Embodiment 2

[0044] The determination of embodiment two palbociclib bulk drug (provided by Chongqing Sansheng Industrial Co., Ltd.)

[0045] Take palbociclib 20mg, weigh it accurately, place it in a 25ml measuring bottle, add methanol-buffer solution (volume ratio 40:60) for ultrasonic treatment to dissolve and dilute to the mark, shake well, and use it as the sample solution; Sample solution 1.0ml is placed in 100ml measuring bottle, diluted to scale with methanol-buffer solution (volume ratio is 40:60), shakes up, as contrast solution; Carry out liquid chromatography analysis by the chromatographic condition of embodiment one, record Chromatogram. If there are impurity peaks (except solvent peaks) in the chromatogram of the sample solution, calculate the impurity content according to the self-control method with a correction factor. The result is as image 3 , Figure 4 shown. The test results are shown in Table 1:

[0046] Table 1 palbociclib related substance detection results

...

Embodiment 3

[0049] The determination of embodiment three palbocicini capsules (provided by Chongqing Sansheng Industrial Co., Ltd.)

[0050] Take an appropriate amount of Palbociclib capsules (approximately equivalent to 20 mg Palbociclib), put them in a 25ml measuring bottle, add methanol-buffer solution (40:60 by volume) to dissolve by ultrasonic treatment and dilute to the mark, shake well, filter After that, take the filtrate as the sample solution; accurately measure 1.0ml of the sample solution, place it in a 100ml measuring bottle, dilute it to the mark with methanol-buffer solution (40:60 in volume ratio), shake well, and use it as a control solution; press For the prescription ratio of Palbociclib capsules, take an appropriate amount of blank excipients, and prepare blank excipients for test solution in the same way as the sample solution; carry out liquid chromatography analysis according to the chromatographic conditions of Example 1. Record the chromatogram, the result is as f...

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