Tryptophanol and xanthanin in urine as diagnostic markers for idiopathic male infertility and their application
A male infertility, idiopathic technology, applied in the field of analytical chemistry and clinical medicine, can solve problems that have not received corresponding attention, and achieve the effect of preventing disease progression, strong correlation, and easy detection
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Embodiment 1
[0105] Example 1: Research object selection and grouping basis
[0106] The subjects of this part of the study were first-diagnosed cases of idiopathic male infertility and healthy reproductive controls from the Affiliated Hospital of Nanjing Medical University. The research content and informed consent form were approved by the Ethics Committee of Nanjing Medical University and met the requirements of relevant regulations. Cases and controls signed the informed consent after learning the content. All study subjects underwent a complete physical examination and completed a questionnaire that included basic personal information, living habits, occupational and environmental exposures, genetic risk factors, sexual and reproductive function, disease history, and physical activity. The first phase included 607 eligible cases of idiopathic male infertility and 430 healthy controls; the second phase included 15 eligible cases of idiopathic male infertility and 15 healthy controls a...
Embodiment 2
[0137] Example 2: UPLC-MS Metabolomics Biomarker Screening for Idiopathic Male Infertility
[0138] 1. Sample pretreatment
[0139] 1.1. 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 40 μL of methanol (reagent A), and vortex for 30 s.
[0140] 1.2. Centrifuge at 16000g at 4°C for 15min in a centrifuge, transfer the supernatant to a 1.5mL inlet EP tube, and concentrate the supernatant to dryness in a centrifugal concentration dryer at room temperature.
[0141] 1.3. Reconstitute with 5 μL of ultrapure water (reagent D) for analysis.
[0142] 2. Instrument testing
[0143] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high performance liquid chromatograph; Q-Exactive high resolution mass spectrometer.
[0144] 2.2. Liquid phase conditions:
[0145] 2.2.1 The liquid chromatographic column was a Hypersil GOLD C18 chromatographic column (100 mm×2.1 mm, particle size 1.9 μm, Thermo...
Embodiment 3
[0158] Example 3 Stability analysis of tryptophanol and xanthoside in urine
[0159] The stability of tryptophanol and xanthoside levels in urine was evaluated using the method of Example 2 (interval time was 2 weeks). The results showed that the measured levels of tryptophanol and xanthoside in urine were stable ( figure 2 ), possessing properties as diagnostic / monitoring markers.
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