Idiopathic male infertility-related medium-chain fatty acid marker in serum and detection method and application thereof
A medium-chain fatty acid and male infertility technology, applied in the fields of analytical chemistry and clinical medicine, can solve problems such as low sensitivity and complicated experimental steps
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Embodiment 1
[0105] Embodiment 1: Research object selection and group basis
[0106] 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 content. 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. In the first stage, 113 idiopathic male infertility cases and 113 healthy controls were included; in the second stage, 15 idiopathic male infertility cases and 15 healthy controls ...
Embodiment 2
[0137] Example 2: UPLC-MS metabolomics biomarker screening for idiopathic male infertility
[0138] 1. Sample pretreatment
[0139] 1.1. Take 10 μL of serum, add 10 μL of internal standard A, add 10 μL of internal standard B, add 10 μL of internal standard C, add 40 μL of methanol (reagent A), and vortex for 30 seconds.
[0140] 1.2. 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.
[0141] 1.3. Reconstitute with 5 μL ultrapure water (reagent D) and wait for analysis.
[0142] 2. Instrument testing
[0143] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.
[0144] 2.2. Liquid phase conditions:
[0145] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column (100mm×2.1mm, particle size 1.9μm, Th...
Embodiment 3
[0158] Example 3: Stability analysis of capric acid and dodecanoic acid in serum
[0159] The method of Example 2 was used to evaluate the stability of capric acid and dodecanoic acid levels in serum (the interval was 2 weeks). The results showed that the measured levels of capric acid and dodecanoic acid in serum were stable ( figure 2 ), with properties as diagnostic / monitoring markers.
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