Kit for detecting ABL tyrosine kinase inhibitor based on HPLC-MS/MS method and detection method

A technology of tyrosine kinases and inhibitors, applied in the field of compositions, can solve problems such as ineffective treatment of patients, achieve the effects of improving detection quality, promoting standardization and standardization, and improving accuracy

Inactive Publication Date: 2021-04-27
北京陆道培生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CML patients are often accompanied by BCR / ABL mutations, and Imatinib can specifically target and inhibit the tyrosine kinase activity of the oncoprotein encoded by the BCR / ABL gene, which improves the survival rate of CML patients. However, it has been reported that It is pointed out that in the process of applying Imatinib, about 15% to 20% of patients are ineffective or relapse, which may be related to its drug resistance.

Method used

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  • Kit for detecting ABL tyrosine kinase inhibitor based on HPLC-MS/MS method and detection method
  • Kit for detecting ABL tyrosine kinase inhibitor based on HPLC-MS/MS method and detection method
  • Kit for detecting ABL tyrosine kinase inhibitor based on HPLC-MS/MS method and detection method

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Experimental program
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Effect test

Embodiment 1

[0045] This embodiment is a kit for detecting ABL tyrosine kinase inhibitors based on the HPLC-MS / MS method. ABL tyrosine kinase inhibitors include imatinib, dasatinib, nilotinib, pana tinib and flumatinib;

[0046] The kit includes the following reagents:

[0047] Sample processing solution, standard mother solution, quality control product, internal standard solution and mobile phase, the mobile phase includes mobile phase A and mobile phase B

[0048] Mobile phase A is 0.1% formic acid methanol solution;

[0049] Mobile phase B is 2mmol·L -1 Ammonium acetate-0.1% formic acid aqueous solution;

[0050] The sample processing solution is 2mmol L -1 Ammonium acetate-0.1% formic acid aqueous solution;

[0051] Standard stock solutions include 100 μg / mL Imatinib methanol solution, 100 μg / mL Nilotinib methanol solution, 10 μg / mL Dasatinib methanol solution, 10 μg / mL Ponatinib methanol solution and 10 μg / mL Flumatinib methanol solution;

[0052] Quality control products inclu...

Embodiment 2

[0058] The present embodiment is the preparation method of the kit in Example 1, and the preparation method comprises the following steps:

[0059] 1. Preparation of standard products:

[0060] Preparation of standard product mother solution: 100 μg / mL Imatinib methanol solution Preparation: 1 mg Imatinib standard product, dissolved in methanol, and fixed to a 10 mL volumetric flask with methanol, ultrasonically mixed, dispensed for use; the same operation as above to obtain 100 μg / mL Nilotinib methanol solution;

[0061] 10μg / mL Dasatinib methanol solution preparation: 1mg Dasatinib standard, dissolved in methanol, and dilute to a 10mL volumetric flask with methanol, ultrasonically mixed, dispensed for use, that is, 100μg / mL Dasatinib, take 100μL 100μg / mL Dasatinib stock solution, Add 900 μL of methanol, shake and mix to obtain a 10 μg / mL Dasatinib methanol solution; operate as above to obtain a 10 μg / mL Ponatinib methanol solution and 10 μg / mL Flumatinib methanol solution; ...

Embodiment 3

[0087] This embodiment is a method for detecting ABL tyrosine kinase inhibitors using the above kit, the method comprising:

[0088] Take 100 μL of the sample to be tested and add 2 mmol·L -1 Ammonium acetate-0.1% formic acid aqueous solution 100μL, then add 300μL of internal standard solution containing ABL tyrosine kinase inhibitor, vortex and mix for 30-60S, high-speed centrifugation (13200rpm) for 8min, pipette 170μL of supernatant, put it on the machine for testing, Perform HPLC-MS / MS quantitative analysis;

[0089] Wherein, the high performance liquid chromatography conditions are as follows:

[0090] Using Ultimate XB-C18 chromatographic column (4.6×50mm, 5um), column temperature 60°C; flow rate 0.8mL min -1 ; Mobile phase A is 0.1% formic acid methanol solution, mobile phase B is 2mmol L -1 Ammonium acetate-0.1% formic acid aqueous solution, injection volume 5uL;

[0091] The elution method is gradient elution, and the gradient elution procedure is as follows:

[...

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Abstract

The invention discloses a kit for detecting an ABL tyrosine kinase inhibitor based on an HPLC-MS / MS method and a detection method. The ABL tyrosine kinase inhibitor comprises imatinib, dasatinib, nilotinib, panatinib and flumatinib. The kit comprises the following reagents: a sample treatment solution, a standard substance, a quality control substance, an internal standard solution and a mobile phase, wherein the mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is a 0.1% formic acid methanol solution; the mobile phase B is a 2 mmol.L <-1 > ammonium acetate and -0.1% formic acid aqueous solution; according to the invention, the concentration of the ABL tyrosine kinase inhibitor in the blood plasma is quantitatively detected based on an HPLC-MS / MS technology; the detection result is not interfered by common drugs and combined drugs, and the accuracy of quantitative detection of the ABL tyrosine kinase inhibitor is greatly improved; the kit is helpful for improving the detection quality of the ABL tyrosine kinase inhibitor and promoting the standardization and normalization of a detection method, so that the blood concentration of the clinical ABL tyrosine kinase inhibitor can be monitored more quickly, accurately and reliably.

Description

technical field [0001] The invention relates to the technical field of compositions, in particular to an antiaging composition and its application. Background technique [0002] Chronic myelogenous leukemia (CML) is a malignant tumor formed by the clonal proliferation of bone marrow hematopoietic stem cells, accounting for 15% of adult leukemia, and the global annual incidence rate is 1.6-2.0 / 100,000. In recent years, BCR / ABL-targeted tyrosine kinase inhibitors are the first choice for clinical treatment of CML patients. CML patients are often accompanied by BCR / ABL mutations, and Imatinib can specifically target and inhibit the tyrosine kinase activity of the oncoprotein encoded by the BCR / ABL gene, which improves the survival rate of CML patients. However, it has been reported that It is pointed out that in the process of applying Imatinib, about 15% to 20% of patients are ineffective or relapse, which may be related to its drug resistance. The second-generation ABL tyro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/08
CPCG01N30/08G01N30/88G01N2030/027G01N2030/045G01N2030/8809
Inventor 王磊刘红星刘瑞孙文利
Owner 北京陆道培生物技术有限公司
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