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A method for simultaneous determination of ticagrelor and its active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry

A technology of ticagrelor and liquid chromatography-mass spectrometry, applied in the field of medical testing, to achieve the effect of less blood sample consumption, good repeatability, and simple operation

Active Publication Date: 2022-07-22
SHANGHAI CHANGHAI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no relevant report about the method of the present invention for the simultaneous determination of ticagrelor, its active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry

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  • A method for simultaneous determination of ticagrelor and its active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry
  • A method for simultaneous determination of ticagrelor and its active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry
  • A method for simultaneous determination of ticagrelor and its active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Pre-experiment

[0048] 1. Experimental materials

[0049] 30% perchloric acid, 30% acetonitrile, 50% acetonitrile, 80% acetonitrile, 100% acetonitrile, ticagrelor, AR-C124910XX

[0050] 2. Experimental method

[0051] Adenosine in the sample to be tested is unstable, so the present invention selects 30% perchloric acid to precipitate protein, effectively removes enzymes in biological samples, and ensures that the concentration of adenosine is stable after pretreatment.

[0052] 3. Experimental results

[0053] In the present invention, the analyte of adenosine is a very polar compound, while ticagrelor and its active metabolite are very weak. Therefore, the present invention examines the ratio of organic solvents in different proportions. After dissolving perchloric acid, its extraction and recovery rate difference, see Table 1.

[0054] Table 1, different proportions of acetonitrile-aqueous solution dissolved perchloric acid to investigate the extraction...

Embodiment 2

[0058] Example 2 Selectivity and Linearity Test

[0059] 1. Experimental method

[0060] Biological blank matrix treatment: Put the plasma sample for a period of time until the adenosine disappears completely (the adenosine can disappear completely in 2 hours), add 50 μl of stabilizer to 1 ml of plasma, and mix the plasma for use.

[0061] Linear test:

[0062] Configure a series of standard curve plasma samples, including adenosine concentrations of 110, 55, 11, 5.5, 3.3, 2.2ng / mL; ticagrelor 1600, 800, 160, 80, 48, 32ng / mL; ticagrelor activity Metabolites 1108, 554, 110.8, 55.4, 33.2, 22.2 ng / mL. According to the plasma sample pretreatment operation. Take the concentration of the analyte (C, ng / mL) as the abscissa, and the ratio (As / Ai) of the peak area of ​​the analyte (As) to the peak area of ​​the internal standard (Ai) as the ordinate, using the weighting (weight coefficient: 1 / C 2 ) least squares method was used for linear regression, and the correlation coefficie...

Embodiment 3

[0069] Example 3 Extraction recovery and matrix effect

[0070] 1. Experimental method

[0071] The blank plasma treated with blank matrix was taken, except that the standard series solution and internal standard solution were not added, and the supernatant was prepared according to the "plasma sample pretreatment method" in Example 2. The corresponding concentration standard series solution and internal standard were added to the supernatant, and 5 samples were analyzed for each concentration, and the corresponding peak area value A was obtained. Take 200 μL of blank plasma treated with blank matrix, prepare standard series solutions of corresponding concentrations and internal standard solutions, operate according to the "plasma sample pretreatment method" in Example 2, and analyze 5 samples for each concentration to obtain the corresponding peak area value B. In addition to preparing the corresponding concentration standard series solution and internal standard solution, 5...

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Abstract

The invention relates to the technical field of medical testing, in particular to a method for simultaneous determination of the concentrations of ticagrelor, active metabolites and endogenous adenosine in human plasma by liquid chromatography-mass spectrometry. The concentration of ticagrelor and its active metabolites and endogenous adenosine in the medium, the method includes: A. Simultaneously determine the concentrations of ticagrelor, its active metabolites and endogenous adenosine in plasma. B. All samples to be tested can be measured under the same chromatographic and mass spectrometry conditions. C. Simultaneous extraction of analytes of different polarities in the sample. Advantages: Accurate, simple and rapid evaluation of changes in plasma adenosine concentration in patients and its relationship with plasma ticagrelor and its active metabolite concentrations after taking the antiplatelet drug ticagrelor. In addition, the present invention optimizes the pretreatment and chromatographic and mass spectrometry conditions, so that the determination method is simple, rapid, sensitive, specific, and repeatable.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a method for simultaneous determination of ticagrelor, active metabolites and endogenous adenosine concentrations in human plasma by liquid chromatography-mass spectrometry. Background technique [0002] Ticagrelor has a dual mechanism of action on platelet aggregation. On the one hand, it can block adenosine phosphate (ADP)-induced platelet aggregation by inhibiting the P2Y12 receptor, and on the other hand, it can inhibit the reuptake of adenosine by ENT-1 on erythrocytes. inhibit platelet aggregation. In addition to the original drug itself, ticagrelor, its active metabolite AR-C124910XX, can also effectively inhibit platelet aggregation. These drug characteristics make ticagrelor in clinical use, compared with other antiplatelet drugs such as aspirin and clopidogrel, can significantly reduce cardiovascular mortality, as well as stroke and death caused by cardiovascul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 王卓张文静高丽红曹爱霖许小建仰先蜜毕娟杨云云丁楠邓敏
Owner SHANGHAI CHANGHAI HOSPITAL
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