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Method for taking tryptosol and xanthosine in urine as idiopathic male infertility diagnosis markers and application thereof

A male infertility, idiopathic technology, applied in the fields of analytical chemistry and clinical medicine, can solve problems such as lack of corresponding attention

Active Publication Date: 2019-01-11
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of metabolomics analysis of urinary metabolites in the diagnostic monitoring of idiopathic male infertility has not received corresponding attention

Method used

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  • Method for taking tryptosol and xanthosine in urine as idiopathic male infertility diagnosis markers and application thereof
  • Method for taking tryptosol and xanthosine in urine as idiopathic male infertility diagnosis markers and application thereof
  • Method for taking tryptosol and xanthosine in urine as idiopathic male infertility diagnosis markers and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Embodiment 1: Research object selection and group basis

[0105] 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 an...

Embodiment 2

[0136] Example 2: UPLC-MS metabolomics biomarker screening for idiopathic male infertility

[0137] 1. Sample pretreatment

[0138] 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 seconds.

[0139] 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.

[0140] 1.3. Reconstitute with 5 μL ultrapure water (reagent D) and wait for analysis.

[0141] 2. Instrument testing

[0142] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0143] 2.2. Liquid phase conditions:

[0144] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column (100mm×2.1mm, particle size 1.9μm, T...

Embodiment 3

[0157] Stability Analysis of Tryptophanol and Xanthanin in Example 3 Urine

[0158] The method of Example 2 was used to evaluate the stability of the levels of tryptophanol and xanthanin in urine (the interval was 2 weeks). The results showed that the levels of tryptophanol and xanthoside in urine were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the field of the analytical chemistry and the clinical medicine, and discloses a method for taking tryptosol and xanthosine in urine as idiopathic male infertility diagnosis markers and an application of the method. The markers are tryptosol and xanthosine which have relatively high sensitivity and specificity in the diagnosis of the idiopathic male infertility, and can beused for preparing the reagent for idiopathic male infertility diagnosis or monitoring.

Description

field of invention [0001] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to tryptophanol and xanthanin in urine related to idiopathic male infertility as biomarkers and a detection method and application thereof based on UPLC-Q exactive MS. Background technique [0002] At present, the number of infertile patients among newly married couples in my country is far more than one million. Current studies suggest that genetic factors such as sequence changes in the AZF gene region of the Y chromosome, DAZ gene copy number changes, and sex hormone receptor gene sequence changes, as well as environmental factors such as exposure to harmful factors, nutrition, and living habits, are all related to male infertility. It is worth noting that a considerable part of male infertility manifests as unexplained etiology, that is, idiopathic male infertility, which makes the diagnosis of idiopathic male infertility extremely difficult and delays ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 陈敏健丁兴旺管昱盛张怡云周昆徐菠庄婷钰张晓玲韩秀梅胡艳辉
Owner NANJING MEDICAL UNIV
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