Urine fatty acid metabolite marker related to idiopathic male infertility, as well as detection method and application thereof

A technology for fatty acid metabolism and male infertility, applied in the fields of analytical chemistry and clinical medicine, which can solve the problems of low sensitivity and complicated experimental steps.

Active Publication Date: 2017-01-04
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

NMR technology is characterized by no damage to the components to be measured, simple sample pretreatment, but low sensitivity; gas chromatography-mass spectrometry has good sensitivity and reproducibility, but derivatization methods are generally used for sample pretreatment , making the experimental steps more complicated

Method used

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  • Urine fatty acid metabolite marker related to idiopathic male infertility, as well as detection method and application thereof
  • Urine fatty acid metabolite marker related to idiopathic male infertility, as well as detection method and application thereof
  • Urine fatty acid metabolite marker related to idiopathic male infertility, as well as detection method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Embodiment 1 Research Object Selection and Grouping Basis

[0109] 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 basic personal 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

[0140] Example 2 UPLC-MS metabolomics screening of idiopathic male infertility biomarkers

[0141] 1. Sample pretreatment

[0142] 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.

[0143] 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.

[0144] 3. Reconstitute with 10 μL ultrapure water (reagent D) and wait for analysis.

[0145] 2. Instrument testing

[0146] 1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0147] 2. Liquid phase conditions:

[0148] 2.1. The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column (100 mm×2.1 mm, particle size 1.9 μm, Thermo S...

Embodiment 3

[0161] Example 3 Stability Analysis of Urine Fatty Acid Metabolites

[0162] The method of Example 2 was used to evaluate the stability of urine tetradecanedioic acid and hexadecanedioic acid levels (the interval was 2 weeks). The results showed that the levels of tetradecanedioic acid and hexadecanedioic acid in urine were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the fields of analytical chemistry and clinical medicine, and discloses a urine fatty acid metabolite marker related to idiopathic male infertility, as well as a detection method and application thereof. The marker is urine fatty acid metabolites, namely, tetradecanedioic acid and / or hexadecanedioic acid, and is detected by adopting a UPLC-Q exactive MS method; the marker can be used for auxiliary diagnosis and monitoring of the idiopathic male infertility, has high sensitivity and specificity, and has a clinical popularization value.

Description

[0001] field of invention [0002] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to a urine fatty acid metabolite marker related to idiopathic male infertility and a detection method and application thereof based on UPLC-Q exactive MS. Background technique [0003] About 15% of couples of childbearing age in the world suffer from infertility. Due to the large population base in my country, the number of infertility patients among newly married couples 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. [0004] However, the diagnosis of idiopathic male infertility is very difficult. The WHO standard for male infertility is that the husband and wife have lived together for more than 1 year after marriage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/62G01N30/72
CPCG01N30/02G01N30/62G01N30/7253G01N30/7266G01N2030/027
Inventor 陈敏健胡艳辉陈婷肖甜周作民陈道帧徐菠周昆夏彦恺陆春城吴炜韩秀梅吉晓明王玉邦环飞王心如
Owner NANJING MEDICAL UNIV
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