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Method for detecting estrogens and estrogen metabolites in human urine

A detection method and technology for metabolites, applied in the field of estrogen metabolite detection, can solve the problems of increasing the error chance of DNA replication, increasing the risk of breast cancer, weak estrogen activity, etc., to meet the needs of large-scale laboratory testing, Efficient and accurate qualitative and quantitative analysis, high precision effect

Inactive Publication Date: 2018-12-28
成都益康谱科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The half-life of 4-OHE1 in the body is longer than that of 2-OHE1, and it is more likely to be oxidized into quinones, which binds to DNA and increases the chance of DNA replication errors. The change is more obvious in breast cancer patients, thereby increasing the risk of breast cancer. disease risk
Catechol-O-methyltransferase-mediated partially methylated estrogen metabolites such as 2-MeOE2 are thought to be detoxifying and even tumor suppressive, with low binding to hormone receptors ER and ER , so the estrogenic activity is also weak

Method used

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  • Method for detecting estrogens and estrogen metabolites in human urine
  • Method for detecting estrogens and estrogen metabolites in human urine
  • Method for detecting estrogens and estrogen metabolites in human urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Detection method establishment and optimization.

[0046] 1. Materials

[0047] The samples of the methodological research experiment came from the urine samples of some patients in the Department of Endocrinology, West China Hospital, Sichuan Province, from May to June 2018.

[0048] Instruments: Shimadzu 8050 triple quadrupole mass spectrometer (Shimadzu); Shimadzu liquid chromatography system (with automatic sampler); one-hundred-thousandth balance; chromatographic column (C18); nitrogen blowing instrument; high-speed benchtop Refrigerated centrifuges; freeze dryers; solid phase extraction columns; adjustable pipettes; glass instruments, beakers, graduated cylinders, etc.

[0049] Reagent consumables: chromatography pure methanol; acetonitrile; formic acid; ammonium acetate; ascorbic acid; sodium acetate; hydrolase Glucuronidase / Sulfatase; sodium bicarbonate; sodium hydroxide; acetone; dansyl chloride; glacial acetic acid; sodium hydroxide; activated carb...

Embodiment 2

[0059] Embodiment 2: method verification.

[0060] 1. Method specificity.

[0061] Take 2 parts of 0.5ml blank matrix and divide into 2 groups for treatment:

[0062] 1. Leave blank;

[0063] 2. Add 10μl working standard solution (4ng / ml) containing 14 EMs, and 10μl internal standard solution (50ng / ml).

[0064] The two groups of samples were vortex mixed for 10s and left for 30min. This was followed by solid phase extraction, nitrogen blowing and derivatization. After membrane filtration, the samples were injected for analysis.

[0065] Compare the two sets of chromatograms to analyze the interference of endogenous substances in urine on the tested compound and internal standard.

[0066] The specificity verification results showed that no obvious interference peaks appeared in the blank matrix samples at the peak position of the standard. The peak shapes of 14 estrogen and estrogen metabolite standards and urine samples are symmetrical, and there is basically no interf...

Embodiment 3

[0101] Example 3: Detection of patient samples.

[0102] The samples of the research experiment came from the urine samples of breast cancer patients in the Department of Breast Surgery of Huaxi Hospital, Sichuan Province.

[0103] Reagents and equipment are the same as in Example 1 in this experiment.

[0104] Chromatographic conditions and mass spectrometry detection conditions are the same as in Example 1 in this experiment.

[0105] The measurement results and data statistics are shown in the table below.

[0106]

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Abstract

The invention provides a method for detecting estrogens and estrogen metabolites in human urine, and relates to the technical field of estrogen and estrogen metabolite detection. According to the detection method, the human urine is mixed with an internal standard solution, and then a mixed solution is subjected to enzyme hydrolysis, organic solvent extraction, reversed phase medium solid-phase extraction purification and target product chemical derivatization in sequence. The full dissolution of the estrogens and the estrogen metabolites in the urine is guaranteed, and the interference of impurities in the urine is eliminated, so that the accuracy and stability of a detection result are ensured. Then, a liquid chromatography separation and mass spectrometry ion pair detection combined method is adopted for carrying out synchronous qualitative and quantitative analysis on the estrogens and the estrogen metabolites. The method is applicable to the synchronous batch detection of the estrogens and the estrogen metabolites with similar structures and chemical physical properties in the human urine, and has the advantages of high sensitivity, high precision, good repeatability, high detection speed and the like. The method fully meets the requirements of clinical examination, and has relatively high clinical application values.

Description

technical field [0001] The invention relates to the technical field of estrogen metabolite detection, in particular to a method for detecting estrogen metabolites in human urine. Background technique [0002] Estrogen is an important class of sex hormones in vertebrates, which plays an important role in the growth and development of the body and the maintenance of system functions. It plays a key role in reproductive system, endocrine system, nervous system and so on. [0003] Estrogens are a class of fat-soluble steroid small molecule compounds, which have two forms, free and conjugated. Among them, it mainly exists in a bound state in the body. In the blood, 70% is bound to sex hormone binding globulin (SHBG), 25% is bound to albumin, and 5% remains in a free state. Among them, the estrogen in the bound state has no physiological activity, while the estrogen in the free state can freely penetrate the target cells and bind to the corresponding receptors, regulate cell tra...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 邬智刚朱文漓叶尚宇
Owner 成都益康谱科技有限公司
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