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Urinary estrogen metabolite markers related to idiopathic male infertility and its detection method and application

A technology for estrogen metabolism and male infertility, which is applied in the field of analytical chemistry and clinical medicine, and can solve the problems that the detection range cannot cover the metabolic spectrum, the pretreatment method is complicated, and the occupancy rate of the instrument is increased.

Active Publication Date: 2019-03-01
NANJING MEDICAL UNIV
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Problems solved by technology

However, gas chromatography tandem mass spectrometry is limited to the detection of volatile substances, and the detection range cannot cover the requirements of metabolic profiling
In order to make up for this defect, gas chromatography tandem mass spectrometry often uses a derivatization method to detect chemicals, which makes the pretreatment method complicated and introduces more errors
At the same time, due to the limitation of separation technology, gas chromatography tandem mass spectrometry detection generally takes a long time, which prolongs the experimental period and increases the occupancy rate of the instrument, which limits its application in the detection of large sample populations.
Capillary electrophoresis tandem mass spectrometry has advantages in the separation and detection of polar compounds, but the problem of instrument stability is difficult to solve

Method used

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  • Urinary estrogen metabolite markers related to idiopathic male infertility and its detection method and application
  • Urinary estrogen metabolite markers related to idiopathic male infertility and its detection method and application
  • Urinary estrogen metabolite markers related to idiopathic male infertility and its detection method and application

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Embodiment 1

[0106] Embodiment 1 Research Object Selection and Grouping Basis

[0107] The subjects of this part of the study are from the first-diagnosed idiopathic male infertility cases and healthy childbearing controls in the Affiliated Hospital of Nanjing Medical University. The research content and the informed consent form were approved by the Ethics Committee of Nanjing Medical University and complied with the requirements of relevant laws and regulations. Cases and controls signed the informed consent after understanding the contents. All subjects underwent a complete physical examination and completed a questionnaire including personal basic information, living habits, occupational and environmental exposures, genetic risk factors, sexual and reproductive function, disease history, and physical activity. The first stage included 607 idiopathic male infertility cases and 430 healthy controls who met the requirements; the second stage included 15 idiopathic male infertility cases ...

Embodiment 2

[0138] Example 2: UPLC-MS metabolomics biomarker screening for idiopathic male infertility

[0139] 1. Sample pretreatment

[0140] 1. Take 300 μL of urine, add 10 μL of internal standard A, add 10 μL of internal standard B, add 10 μL of internal standard C, add 900 μL of methanol (reagent A), and vortex for 30 seconds.

[0141] 2. Centrifuge at 16000g at 4°C for 15min in a centrifuge, transfer the supernatant to a 1.5mL imported EP tube, and concentrate the supernatant to dryness in a centrifugal concentrator at room temperature.

[0142] 3. Reconstitute with 10 μL ultrapure water (reagent D) and wait for analysis.

[0143] 2. Instrument testing

[0144] 1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0145] 2. Liquid phase conditions:

[0146] 2.1. The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column (100 mm×2.1 mm, particle size 1.9 μm, Thermo ...

Embodiment 3

[0159] Example 3 Stability Analysis of Urine Estrogen Metabolites

[0160] The stability of urine 2-methoxyestradiol and androstenedione levels was evaluated by the method of Example 2 (the interval was 2 weeks). The results showed that the levels of 2-methoxyestradiol and androstenedione in urine were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the field of analytical chemistry and clinical medicine and discloses the urine estrogen metabolite markers related to idiopathic male infertility and the detection methods and application. The marker is the urine estrogen metabolite 2-methoxyestradiol and / or androstenedione and detected by UPLC Q exactive MS method. The urine estrogen metabolite markers related to idiopathic male infertility can be used for auxiliary diagnosis and monitoring of idiopathic male infertility, and has high sensitivity, specificity, and clinical spread value.

Description

field of invention [0001] The invention belongs to the fields of analytical chemistry and clinical medicine, and relates to urinary estrogen metabolite markers related to idiopathic male infertility and a detection method and application thereof based on UPLC-Q exactive MS. Background technique [0002] Currently, about 10-15% of couples of reproductive age worldwide suffer from fertility disorders. Due to the large population base in my country, the number of infertility patients among newlyweds is far more than one million, and the infertility caused by male factors is higher than 50%. About 40% to 75% of male infertility still cannot find the cause, which is called idiopathic male infertility. Idiopathic male infertility poses a disproportionately high disease burden. [0003] In fact, the diagnosis of idiopathic male infertility is very difficult. The WHO diagnostic criteria for male infertility are couples who have lived together for more than 1 year after marriage w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 陈敏健胡艳辉陈婷王笑笑肖甜周作民陈道帧夏彦恺陆春城吴炜韩秀梅吴笛杜桂珍王玉邦环飞王心如
Owner NANJING MEDICAL UNIV
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