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Method and kit for detecting progesterone and testosterone in saliva through high performance liquid chromatography-tandem mass spectrometry technique

A technology of high performance liquid chromatography and tandem mass spectrometry, which is applied in the field of detection of progesterone and testosterone in saliva by high performance liquid chromatography and tandem mass spectrometry, achieving the effect of simple operation, short analysis time and good reproducibility

Active Publication Date: 2016-06-15
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 progesterone and testosterone in saliva, and provide a method and kit for detecting progesterone and testosterone in saliva by high performance liquid chromatography tandem mass spectrometry

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  • Method and kit for detecting progesterone and testosterone in saliva through high performance liquid chromatography-tandem mass spectrometry technique
  • Method and kit for detecting progesterone and testosterone in saliva through high performance liquid chromatography-tandem mass spectrometry technique
  • Method and kit for detecting progesterone and testosterone in saliva through high performance liquid chromatography-tandem mass spectrometry technique

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[0083] 1. Instrument and parameter setting:

[0084] High-performance liquid chromatography (Agilent1100), chromatographic column (50 × 2.1mm, 2.6μm; Kinetex Phenomenes), mobile phase is 0.05% formic acid water and methanol, column temperature is 40°C, flow rate is 200μl·min -1 , the analysis time is 6.5min, and the liquid phase condition is gradient elution (0~1min, 20% methanol; 1~3.4min, 20%~80% methanol; 3.4~5.9min, 80% methanol; 5.9~6.1min, 80% %~20% methanol; 6.1~6.5min, 20% methanol).

[0085] A mass spectrometer (4000Qtrap, AB Company) uses an ESI source, under positive ion conditions, and the detection mode is MRM.

[0086] Detection of ion pairs (m / z): progesterone: 315.3→109.1; testosterone: 289.3→109.1; internal standard (d9-progesterone): 324.4→100.1. Curtain gas (CUR) is 10, collision gas (CAD) is 6, ion spray voltage (IS) is 5000V, temperature is 500°C, GS1 is 50, and GS2 is 50.

[0087] Mass Spectrometry Parameters for Progesterone and Testosterone and Inter...

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Abstract

The invention provides a method for detecting progesterone and testosterone in saliva through a high performance liquid chromatography-tandem mass spectrometry technique.The method comprises the steps that the progesterone and the testosterone in pretreated saliva are detected by adopting the high performance liquid chromatography-tandem mass spectrometry technique, the progesterone and the testosterone are separated from impurities through high performance liquid chromatography, quantification is performed through an internal standard method, an isotope of the progesterone is taken as an internal standard, a calibration curve is built by taking the concentration ratio of a standard product to the internal standard as an X axis and taking the peak area ratio of the standard product to the internal standard as a Y axis, and the content of the progesterone and the testosterone is calculated.The method is high in sensitivity and specificity, good in accuracy and simple in pretreatment process; the analysis time is short, analysis of the progesterone and the testosterone can be completed in 6.5 min, and the precision degree and the standard recovery rate meet the detection requirement.

Description

technical field [0001] The invention relates to the technical field of saliva detection, in particular to a method for detecting progesterone and testosterone in saliva by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] At present, the commonly used methods for detecting progesterone and testosterone 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 pro...

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/027G01N2030/062G01N2030/8813
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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