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Method and kit for detecting content of dehydroepiandrosterone in saliva

A technique for detecting DHEA and its detection method, which is applied in the field of detection method and kit for DHEA content in saliva, and can solve the problems of poor ionization efficiency of DHEA, insufficient sensitivity, deviation of detection results, etc. Achieve reliable quantitative detection method, improve detection sensitivity and strong specificity

Pending Publication Date: 2021-06-22
泊迈生物医学检测(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Traditional methods for detecting DHEA mainly include enzyme-linked immunoassay and chemiluminescence immunoassay, etc. These methods are prone to interference from DHEA structural analogues in the sample, resulting in deviation of the detection results
In recent years, high performance liquid chromatography tandem mass spectrometry has been developed to detect the content of DHEA in human body fluids. However, due to the molecular structure of DHEA, direct detection will find that the ionization efficiency of DHEA is not good, and its content in saliva is very low, so The sensitivity of direct detection is not up to the requirement

Method used

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  • Method and kit for detecting content of dehydroepiandrosterone in saliva
  • Method and kit for detecting content of dehydroepiandrosterone in saliva
  • Method and kit for detecting content of dehydroepiandrosterone in saliva

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Embodiment 1

[0039] A method for detecting the content of dehydroepiandrosterone in saliva, comprising:

[0040]Preparation of standard product: Accurately weigh 10 mg of DHEA standard product, dissolve it in acetonitrile and set the volume to 10 ml, prepare 1 mg / mL DHEA standard product mother solution, dilute with acetonitrile and set the volume to 100 ml to obtain 100ng / mL dehydroepiandrosterone standard solution; accurately weigh 10mgd6-dehydroepiandrosterone standard substance, dissolve it in acetonitrile to 10ml, prepare 1mg / mL internal standard mother solution, take 10μl internal standard Dilute the mother liquor with acetonitrile to 10ml to obtain a 1μg / mL internal standard solution, then take 25μl of the internal standard solution, dilute to 10ml with acetonitrile to prepare a 2.5ng / mL internal standard solution for the experiment.

[0041] Quality control product preparation: Take 2μl, 10μl, 50μl of 100ng / ml DHEA standard solution respectively, dilute to 10ml with blank saliva ma...

Embodiment 2

[0073] A kit for detecting DHEA content in saliva, comprising:

[0074] Mobile phase, the mobile phase comprises equal volumes of formic acid water and 100% acetonitrile in a volume ratio of 0.1%;

[0075] Dehydroepiandrosterone standard solution, the standard mother solution is a concentration of 100ng / ml dehydroepiandrosterone in acetonitrile;

[0076] Internal standard solution, internal standard solution is the acetonitrile solution that concentration is 2.5ng / ml d6-t dehydroepiandrosterone;

[0077] Diluent, diluent includes: 3g / L bovine serum albumin aqueous solution and acetonitrile;

[0078] The extract, the extract is methyl tert-butyl ether;

[0079] Quality control products, quality control products include dehydroepiandrosterone solutions with three concentration gradients of 20pg / mL, 100pg / mL, and 500pg / mL obtained by diluting to volume with saliva blank matrix.

[0080] The specific composition of the kit is shown in Table 6:

[0081]

[0082] Table 6 Kit ...

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Abstract

The invention relates to a method and a kit for detecting the content of dehydroepiandrosterone in saliva, and the method comprises the following steps: preparing a standard substance, preparing a quality control substance, treating the standard substance, analyzing and detecting by adopting a high performance liquid chromatography-tandem mass spectrometry technology, pre-treating a saliva sample, analyzing and detecting by adopting the high performance liquid chromatography-tandem mass spectrometry technology, drawing a calibration curve: establishing a standard curve by adopting an isotope internal standard quantification method by taking the concentration ratio of a standard substance to an internal standard substance as an X axis and the peak area ratio of the standard substance to the internal standard substance as a Y axis, and calculating the content of the dehydroepiandrosterone in the to-be-detected sample. According to the method, dehydroepiandrosterone and interferents are separated by adopting the high performance liquid chromatography-tandem mass spectrometry technology, then the content of dehydroepiandrosterone in saliva is measured by combining an isotope internal standard quantification method, and the dehydroepiandrosterone reacts with methoxyamine to generate oxime ester, so that the detection sensitivity is remarkably improved, and the detection time is shortened. The specificity of the detection method is ensured by adopting a reaction detection mode (MRM).

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method and a kit for detecting the content of dehydroepiandrosterone in saliva. Background technique [0002] Dehydroepiandrosterone (DHEA) is a steroid hormone secreted by the adrenal cortex. It is a precursor for the synthesis of androgens and estrogens, and has a wide range of physiological and pathological effects. DHEA is secreted into the blood, and most of it is metabolized to form bound DHEA. These bound DHEA have weak physiological activity, while the remaining DHEA is called free DHEA, which mainly exerts physiological activity. Free DHEA in blood diffuses passively into saliva through salivary gland epithelial cells, so the content of DHEA in saliva is considered to be used to evaluate the level of free DHEA in humans. [0003] Traditional methods for detecting DHEA mainly include enzyme-linked immunoassay and chemiluminescence immunoassay, etc. These m...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/045G01N2030/062G01N2030/067
Inventor 林高坤徐斌林鈺庭
Owner 泊迈生物医学检测(苏州)有限公司
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