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Method for purifying, enriching and detecting steroid hormones in blood

A technology of steroid hormones and detection methods, applied in the field of purification, enrichment and detection of steroid hormones in blood, can solve the problems of low hormone content, interference, etc., and achieve the effects of high sensitivity, high recovery rate and novel structure

Active Publication Date: 2021-04-06
大连博源医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some problems in the detection of steroid hormones, such as the content of hormones in the human body is often low, the problem of detection sensitivity; the interference of endogenous substances on hormone detection; the variability of results caused by matrix effects; the pretreatment of biological samples Technical stability and other issues

Method used

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  • Method for purifying, enriching and detecting steroid hormones in blood
  • Method for purifying, enriching and detecting steroid hormones in blood
  • Method for purifying, enriching and detecting steroid hormones in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Preparation of three-point bonded C18 packing

[0059] Add dry silica gel into toluene for stirring, then dropwise add silane (mainly dimethyloctadecylchlorosilane), heat to 120° C., and stir for 6 hours. After cooling down to room temperature, it was filtered, and washed with toluene, methanol, water, and methanol in sequence. Placed at 80°C and dried for 12 hours.

[0060] The C18 solid-phase extraction filler was identified by solid-state nuclear magnetic resonance as a bonding density of 1.6-2.2 μmol / m 2 , low-density three-point bonded C18 filler with a carbon content of 10%-12%. The particle size of silica gel is 10μm-100μm, and the pore size is The specific surface area is 100m2 / g-600m2 / g. The structure looks like this:

[0061]

Embodiment 2

[0063] Detection of Estrogen in Serum

[0064] 1. Fill the solid phase extraction column: fill with 100mg of filler and place it in a 1mL column tube (the bonding density of silica gel filled is 1.8μmol / m 2 , the carbon content is 10%, the particle size is 30μm, and the pore size is Specific surface area 300m 2 / g of three-point bonded C18 packing);

[0065] 2. Sample pretreatment process

[0066] (1) Preparation of standard working solution: Accurately prepare estrone, estradiol, estriol standard stock solution (1mg / mL), dilute step by step with 50% methanol to obtain mixed standard working solution, estrone, estradiol The concentration is 20ng / mL, 10ng / mL, 5ng / mL, 1ng / mL, 0.2ng / mL, 0.1ng / mL and 0.05ng / mL, the concentration of estriol is 200ng / mL, 100ng / mL, 50ng / mL, 10ng / mL, 2ng / mL, 1ng / mL and 0.5ng / mL, spare;

[0067] (2) Preparation of internal standard working solution: accurately prepare estrone, estradiol, estriol isotope internal standard stock solution (1mg / mL), ...

Embodiment 3

[0088] Detection of Androgen in Serum

[0089] 1. Filling the solid phase extraction column: the filling specifications and process are the same as in Example 1.

[0090] 2. Sample pretreatment process

[0091] (1) Preparation of standard working solution: accurately prepare testosterone, androstenedione, dihydrotestosterone, dehydroepiandrosterone, and dehydroepiandrosterone sulfate standard stock solution (1mg / mL), and use 50% methanol gradually The mixed standard working solution was obtained by serial dilution, the concentrations of testosterone and androstenedione were 150ng / mL, 75ng / mL, 15ng / mL, 6ng / mL, 1.5ng / mL, 0.6ng / mL and 0.3ng / mL, dihydrotestosterone The concentration is 250ng / mL, 125ng / mL, 25ng / mL, 10ng / mL, 2.5ng / mL, 1ng / mL and 0.5ng / mL, and the concentration of dehydroepiandrosterone is 2500ng / mL, 1250ng / mL, 250ng / mL , 100ng / mL, 25ng / mL, 10ng / mL and 5ng / mL, the concentration of dehydroepiandrosterone sulfate is 100μg / mL, 50μg / mL, 10μg / mL, 4μg / mL, 1μg / mL, 0.4μg / m...

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Abstract

The invention relates to a method for purifying, enriching and detecting steroid hormones in blood, and belongs to the technical field of medical examination and analysis. A sample containing steroid hormones is prepared from a low-density three-point bonded C18 filler taking silica gel as a matrix, and purification and enrichment of steroid hormones in the sample are realized by optimizing parameters such as solvent dosage and leacheate proportion in the preparation process. The prepared steroid hormone sample is analyzed through liquid chromatography-tandem mass spectrometry under certain conditions, and multiple steroid hormones can be detected at the same time at a time. The method has the characteristics of good stability, high sensitivity, high recovery rate, simplicity, convenience and controllability in operation, capability of realizing automatic operation and the like.

Description

technical field [0001] The invention belongs to the technical field of medical examination and analysis, and relates to the purification and enrichment of steroid hormones in blood and a detection method thereof. [0002] technical background [0003] Steroid hormones are a class of tetracyclic aliphatic hydrocarbon compounds with a cyclopentane polyhydrophenanthrene nucleus, all of which are derived from cholesterol, and regulate the metabolic process in organisms through the endocrine system. Progesterone is the precursor substance for all other steroid hormones. Corticosteroids are mainly produced by the adrenal glands and are mainly related to the metabolism of glucose and minerals. Sex hormones are divided into male hormones and female hormones, which can be transformed into each other, and there is a balance in animals of different sexes. Males are dominated by androgens, while females are dominated by estrogens. When the hormone levels in the body fluctuate abnormal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/08G01N30/88
CPCG01N30/08G01N30/88
Inventor 曹云峰张爽王爽孙晓宇张亚莲崔丽洪沫王琳
Owner 大连博源医学检验实验室有限公司
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