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.
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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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