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Method and kit for testing cortisol in saliva by efficient liquid chromatogram and tandem mass spectrometry combination technology

A high-performance liquid chromatography and tandem mass spectrometry technology is applied in the field of high-performance liquid chromatography-tandem mass spectrometry technology for detecting cortisol in saliva, and achieves the effects of good reproducibility, simple operation, and accurate and reliable results.

Inactive Publication Date: 2016-06-22
北京洛奇医学检验实验室股份有限公司
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Problems solved by technology

[0003] The purpose of the present invention is to solve some problems existing in the existing detection method of cortisol in saliva, and provide a method and kit for detecting cortisol in saliva by high performance liquid chromatography tandem mass spectrometry

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  • Method and kit for testing cortisol in saliva by efficient liquid chromatogram and tandem mass spectrometry combination technology
  • Method and kit for testing cortisol in saliva by efficient liquid chromatogram and tandem mass spectrometry combination technology
  • Method and kit for testing cortisol in saliva by efficient liquid chromatogram and tandem mass spectrometry combination technology

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specific Embodiment

[0079] 1. Instrument and parameter setting:

[0080] High-performance liquid chromatography (Agilent1100), chromatographic column (50 × 2.1mm, 2.6μm; Kinetex Phenomenes), the mobile phase is 0.05% formic acid water and methanol in the volume ratio, the column temperature is 40°C, and the flow rate is 200μl·min -1 , the analysis time is 5min, and the liquid phase conditions are gradient elution (0~0.2min, 10% methanol; 0.2~1min, 10%~90% methanol; 1~4min, 90% methanol; 4~4.2min, 90%~ 10% methanol; 4.2 ~ 5min, 10% methanol).

[0081] A mass spectrometer (4000Qtrap, AB Company) uses an ESI source, under positive ion conditions, and the detection mode is MRM.

[0082] Detection ion pair (m / z): cortisol: 363.2→121.1; internal standard (d4-cortisol): 367.3→121.1;

[0083] Curtain gas (CUR) is 10, collision gas (CAD) is 3, ion spray voltage (IS) is 5000V, temperature is 500°C, GS1 is 70, and GS2 is 70.

[0084] Mass Spectrometry Parameters of Cortisol and Internal Standard

[0085...

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Abstract

The invention provides a method for testing cortisol in saliva by the efficient liquid chromatogram and tandem mass spectrometry combination technology. Specifically, cortisol in saliva which is preprocessed is tested by the efficient liquid chromatogram and tandem mass spectrometry combination technology, cortisol is separated from impurity by means of the efficient liquid chromatogram and is quantified by an internal standard method; isotope of cortisol is taken as the internal standard, concentration ratio of a standard to the internal standard is taken as X axis, peak area ratio of the standard to the internal standard is taken as Y axis, a standard curve is established, and finally content of cortisol is calculated. The method has the advantages of high flexibility, specificity and accuracy and simple preprocessing; besides, the method needs short time for analysis, which can complete analysis of cortisol within five minutes; and precision and standard recovery rate both meet test requirements.

Description

technical field [0001] The invention relates to the technical field of saliva detection, in particular to a method for detecting cortisol in saliva by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] Currently, commonly used methods for detecting cortisol in saliva include enzyme-linked immunoassay, electrochemiluminescence immunoassay, etc. These methods are affected by various objective conditions, such as temperature, pH value, ionic strength, and reagent immune activity. Immunoassay is unavoidable due to the interference of structurally similar substances in the saliva matrix, and the sensitivity is low. The test results of kits produced by different manufacturers have large deviations, and the lack of uniform standards leads to large deviations in test results and low reference value. Contents of the invention [0003] The purpose of the present invention is to solve some problems existing in the existing detection method...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/027G01N2030/8813
Inventor 刘胜敏何健宋晓涛
Owner 北京洛奇医学检验实验室股份有限公司
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