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High-sensitivity kit for quantifying tacrolimus in dried blood spot through LC-MS/MS method, and preparation method thereof

A technology of dry blood filter paper and liquid chromatography, applied in the field of tacrolimus quantitative detection kit in dry blood filter paper, to achieve the effect of solving the problems of monitoring cost and stability

Inactive Publication Date: 2019-02-26
刘丽宏 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The use of DBS for LC-MS / MS quantification of FK506 has not been reported

Method used

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  • High-sensitivity kit for quantifying tacrolimus in dried blood spot through LC-MS/MS method, and preparation method thereof
  • High-sensitivity kit for quantifying tacrolimus in dried blood spot through LC-MS/MS method, and preparation method thereof
  • High-sensitivity kit for quantifying tacrolimus in dried blood spot through LC-MS/MS method, and preparation method thereof

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Embodiment Construction

[0031] 1. Applicable instruments

[0032] Liquid chromatography-tandem mass spectrometer (LC-MS / MS), equipped with electrospray ionization source (ESI).

[0033] 2. Sample requirements

[0034] Collect venous whole blood samples, place them in anticoagulant tubes, and prepare blood spots on dried blood filter paper sheets. After completely drying, immediately place them in a refrigerator at 2-8°C for storage for testing.

[0035] 3. Inspection method

[0036] Dried blood filter paper sample pretreatment:

[0037]Use a hole puncher to punch the dried blood filter paper into a 1.5mL EP tube (1 hole with a hole punch with a radius of 6mm, and 3 holes with a hole punch with a radius of 3mm), add 20 μL of the sample treatment solution, and then add 60 μL of the internal standard solution, Vortex for 1 min, let stand for 10 min, vortex for 1 min, centrifuge for 5 min (12000 rpm), and inject 5 μL of the supernatant for LC-MS / MS quantitative analysis.

[0038] Mass Spectrometry Co...

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Abstract

The invention provides a high-sensitivity kit for quantifying tacrolimus in dried blood spot through a LC-MS / MS method, and a preparation method thereof, wherein the high-sensitivity kit comprises standard curve dried blood spots with specified concentrations, a quality control sample dried blood spot, a sample treatment solution, an internal standard solution and a mobile phase adjustment solution. The use method of the kit comprises: punching a dried blood spot, sampling, treating with a zinc sulfate solution, precipitating with acetonitrile by using ascomycin as an internal standard, carrying out reversed phase chromatography gradient separation by using a C18 bonded phase silica gel as a stationary phase and using acetonitrile-1 mM ammonium acetate as a mobile phase, and quantitativelydetecting by a mass spectrometer, wherein the ion selection channels respectively are m / z 821.4[arrow]m / z 768.4 (tacrolimus) and m / z 809.6[arrow]m / z 756.7 (internal standard ascomycin). According tothe present invention, the kit has the test sensitivity of 1.0 ng / ml, and various methodological indicators can meet the needs for the monitoring of the blood concentration of tacrolimus in the driedblood spot.

Description

technical field [0001] The invention relates to a kit for quantitative detection of tacrolimus in dry blood filter paper developed based on dry blood filter paper and liquid chromatography tandem mass spectrometry technology, more specifically, it uses liquid chromatography mass spectrometry technology to detect tacrolimus in dry blood filter paper Method for accurate quantitative detection of tacrolimus. [0002] The present invention also relates to the production method and application of the kit of the present invention. Background technique [0003] Tacrolimus (FK506 for short) is a new macrolide immunosuppressant isolated from the culture of Streptomyces, which can effectively inhibit the activation of T lymphocytes. FK506 binds to endogenous cell receptors to form an immunophilin complex, thereby exerting pharmacological effects. At present, FK506 has been widely used in the treatment of liver, kidney, heart, pancreas and other organ transplants for anti-rejection a...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 刘丽宏李鹏飞
Owner 刘丽宏