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Kit and detecting method for accurately measuring concentration of four immunosuppressant drugs in human whole blood

An immunosuppressant and accurate determination technology, which is applied in the field of medical testing, can solve the problems of difficult traceability of detection methods, and achieve the effects of small sample size, high throughput and high sensitivity

Inactive Publication Date: 2019-01-11
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, mass spectrometry clinical testing methods are all established by the laboratory (laboratory developed tests, LDT). , repeatability, and time comparability need further clinical verification

Method used

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  • Kit and detecting method for accurately measuring concentration of four immunosuppressant drugs in human whole blood
  • Kit and detecting method for accurately measuring concentration of four immunosuppressant drugs in human whole blood
  • Kit and detecting method for accurately measuring concentration of four immunosuppressant drugs in human whole blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] A kit for determining the concentrations of four immunosuppressant drugs in human whole blood, including

[0070] Contains calibrators with six concentration gradients, and the calibrators of each concentration gradient are methanol solutions of tacrolimus, sirolimus, everolimus and cyclosporine A;

[0071] Among them, the six concentration points of tacrolimus, sirolimus, and everolimus are the same, in order: 2ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 50ng / mL, 100ng / mL; The six concentrations of sporomycin A are 10ng / mL, 50ng / mL, 200ng / mL, 500ng / mL, 1000ng / mL, 2000ng / mL;

[0072] Internal standard: internal standard solution is containing tacrolimus- 13 Cd 2 , sirolimus-d 3 , everolimus-d 4 and cyclosporine A-d 4 methanol solution, in which tacrolimus- 13 Cd 2 , sirolimus-d 3 and everolimus-d 4 The concentration in the internal standard solution is 10ng / mL; cyclosporine A-d 4 The concentration in the internal standard solution is 200ng / mL;

[0073] Quality control p...

Embodiment 2

[0094] A kit for determining the concentrations of four immunosuppressant drugs in human whole blood, including

[0095] Contains calibrators with six concentration gradients, and the calibrators of each concentration gradient are methanol solutions of tacrolimus, sirolimus, everolimus and cyclosporine A;

[0096] Among them, the six concentration points of tacrolimus, sirolimus, and everolimus are the same, in order: 2ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 50ng / mL, 100ng / mL; The six concentrations of sporomycin A are 10ng / mL, 50ng / mL, 200ng / mL, 500ng / mL, 1000ng / mL, 2000ng / mL;

[0097] Internal standard: internal standard solution is containing tacrolimus- 13 Cd 2 , sirolimus-d 3 , everolimus-d 4 and cyclosporine A-d 4 methanol solution, in which tacrolimus- 13 Cd 2 , sirolimus-d 3 and everolimus-d 4 The concentration in the internal standard solution is 10ng / mL; cyclosporine A-d 4 The concentration in the internal standard solution is 200ng / mL;

[0098] Quality control p...

Embodiment 3

[0106] A kit for determining the concentrations of four immunosuppressant drugs in human whole blood, including

[0107] Contains calibrators with six concentration gradients, and the calibrators of each concentration gradient are methanol solutions of tacrolimus, sirolimus, everolimus and cyclosporine A;

[0108] Among them, the six concentration points of tacrolimus, sirolimus, and everolimus are the same, in order: 2ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 50ng / mL, 100ng / mL; The six concentrations of sporomycin A are 10ng / mL, 50ng / mL, 200ng / mL, 500ng / mL, 1000ng / mL, 2000ng / mL;

[0109] Internal standard: internal standard solution is containing tacrolimus- 13 Cd 2 , sirolimus-d 3 , everolimus-d 4 and cyclosporine A-d 4 methanol solution, in which tacrolimus- 13 Cd 2 , sirolimus-d 3 and everolimus-d 4 The concentration in the internal standard solution is 10ng / mL; cyclosporine A-d 4 The concentration in the internal standard solution is 200ng / mL;

[0110] Quality control p...

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Abstract

The invention provides a kit for accurately measuring the concentration of four immunosuppressant drugs in human whole blood. The kit comprises the calibrators of tacrolimus, sirolimus, everolimus andcyclosporine A, the internal standards of the tacrolimus-13Cd2, the sirolimus-d3, the everolimus-d4 and the cyclosporine A, the quality control serums of the tacrolimus, the sirolimus, the everolimusand the cyclosporine A, sample treatment liquid, a 96-well plate assembly, and a mobile phase. According to the kit and detecting method for accurately measuring the concentration of four immunosuppressant drugs in human whole blood, a clean treatment liquid can be obtained through a simple 96-well plate protein precipitation pretreatment method without complex purification steps, and the required sample amount is small and can be used together with a liquid chromatography-tandem mass spectrometry; and four different immunosuppressant drugs can be detected at the same time, that is, the single-sample multi-index synchronous detection can be realized, the detection specificity is good, the sensitivity is high, the whole detection time is short, the throughput is high, and the cost is low.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a mass spectrometry kit and a liquid chromatography tandem mass spectrometry detection method for measuring the concentration of immunosuppressant drugs in human whole blood. Background technique [0002] Immunosuppressants are mainly used clinically in anti-graft rejection, treatment of autoimmune diseases and allergic diseases, etc. These drugs have a narrow therapeutic window and large individual differences in pharmacokinetics, requiring therapeutic drug monitoring (TDM). In recent years, statistics from the World Health Organization (WHO) and the Adverse Drug Reaction Monitoring Center of the Ministry of Health of my country have shown that the number of deaths due to improper medication is much higher than the number of deaths from various infectious diseases in the same period. Among those who died from improper medication, most of them were caused by improper dos...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88
Inventor 邹继华沈敏杨晓东潘清清王琳李全乐邹炳德
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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