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Steroid hormone markers related to idiopathic male infertility in serum and its detection method and application

A technology for steroid hormones and male infertility, which is applied in the fields of analytical chemistry and clinical medicine, and can solve the problems of limited application, complicated pretreatment methods, and difficult instrument stability.

Active Publication Date: 2021-03-16
NANJING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gas chromatography tandem mass spectrometry is limited to the detection of volatile substances, and the detection range cannot cover the requirements of metabolic profiling
In order to make up for this defect, gas chromatography tandem mass spectrometry often uses a derivatization method to detect chemicals, which makes the pretreatment method complicated and introduces more errors
At the same time, due to the limitation of separation technology, gas chromatography tandem mass spectrometry detection generally takes a long time, which prolongs the experimental period and increases the occupancy rate of the instrument, which limits its application in the detection of large sample populations.
Capillary electrophoresis tandem mass spectrometry has advantages in the separation and detection of polar compounds, but the problem of instrument stability is difficult to solve

Method used

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  • Steroid hormone markers related to idiopathic male infertility in serum and its detection method and application
  • Steroid hormone markers related to idiopathic male infertility in serum and its detection method and application
  • Steroid hormone markers related to idiopathic male infertility in serum and its detection method and application

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Experimental program
Comparison scheme
Effect test

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 contents. All subjects underwent a complete physical examination and completed a questionnaire including personal basic information, living habits, occupational and environmental exposures, genetic risk factors, sexual and reproductive function, disease history, and physical activity. The first phase included 113 idiopathic male infertility cases and 113 healthy controls who met the requirements; the second phase included 15 idiopathic male infertility cases an...

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 3000 system (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, T...

Embodiment 3

[0158] Example 3 Stability Analysis of Cortisol and DHEA in Serum

[0159] The method of Example 2 was used to evaluate the stability of cortisol and DHEA levels in serum (the interval was 2 weeks). The results showed that the measured levels of cortisol and DHEA in serum were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

Belonging to the field of analytical chemistry and clinical medicine, the invention discloses an idiopathic male infertility related steroid hormone marker in serum, a detection method and application thereof. The marker is cortisol and / or dehydroepiandrosterone, a UPLC-Q exactive MS method is employed for detection, the marker can be used for auxiliary diagnosis and monitoring of idiopathic male infertility, has high sensitivity and specificity, and has clinical promotion value.

Description

[0001] field of invention [0002] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to a steroid hormone marker related to idiopathic male infertility in serum and a detection method and application based on UPLC-Q exactive MS. Background technique [0003] Currently, about 10-15% of couples of reproductive age worldwide suffer from fertility disorders. The number of infertile couples in our country is far more than one million, and the infertility caused by male factors is higher than 50%. Idiopathic male infertility is male infertility with no clear cause, accounting for 40%-75% of infertility. The WHO diagnostic criteria for male infertility are couples who have lived together for more than 1 year after marriage without taking contraceptive measures, and the woman is infertile due to the man. However, the 1-year observation time of cohabitation has greatly delayed the time for early treatment and intervention of male infertilit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/88
CPCG01N30/02G01N30/88
Inventor 陈敏健周作民陈道桢肖甜夏彦恺王玉邦环飞胡艳辉吴笛王心如
Owner NANJING MEDICAL UNIV
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