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Idiopathic male infertility-related seminal plasma metabolic small molecular marker as well as detection method and application thereof

A technique for metabolizing small molecules and male infertility, which is applied in the fields of analytical chemistry and clinical medicine, and can solve the problems of complex experimental steps and low sensitivity

Active Publication Date: 2017-02-22
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

NMR technology is characterized by no damage to the components to be measured, simple sample pretreatment, but low sensitivity; gas chromatography-mass spectrometry has good sensitivity and reproducibility, but derivatization methods are generally used for sample pretreatment , making the experimental steps more complicated

Method used

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  • Idiopathic male infertility-related seminal plasma metabolic small molecular marker as well as detection method and application thereof
  • Idiopathic male infertility-related seminal plasma metabolic small molecular marker as well as detection method and application thereof
  • Idiopathic male infertility-related seminal plasma metabolic small molecular marker as well as detection method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Embodiment 1: Research object selection and group basis

[0108] 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. 148 eligible idiopathic male infertility cases and 148 healthy controls were included in the first stage; 15 idiopathic male infertility cases and 15 healthy controls were include...

Embodiment 2

[0139] Example 2: UPLC-MS metabolomics biomarker screening for idiopathic male infertility

[0140] 1. Sample pretreatment

[0141] 1.1. Take 10 μL of seminal plasma, 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), vortex for 30 seconds, and precipitate protein.

[0142] 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.

[0143] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0144] 2. Instrument testing

[0145] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0146] 2.2. Liquid phase conditions:

[0147] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column (10...

Embodiment 3

[0160] Example 3 Stability Analysis of Small Molecules in Seminal Plasma Metabolism

[0161] The method of Example 2 was used to evaluate the stability of seminal plasma acetylcarnitine and tryptophan levels (the interval was 2 weeks). The results showed that the measured levels of acetylcarnitine and tryptophan in seminal plasma were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the fields of analytical chemistry and clinical medicine, and discloses an idiopathic male infertility-related seminal plasma metabolic small molecular marker as well as a detection method and application thereof. The marker is prepared from seminal plasma metabolic small molecular acetylcarnitine and / or tryptophan, and is detected by adopting a UPLC-Q exactive MS method; the marker can be used for auxiliary diagnosis and monitoring of the idiopathic male infertility, and has higher sensitivity and specificity, thus having a clinical popularization value.

Description

[0001] field of invention [0002] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to a seminal plasma metabolic small molecule marker related to idiopathic male infertility and a detection method and application based on UPLC-Q exactive MS. Background technique [0003] According to WHO statistics, about 15% of couples of childbearing age in the world suffer from infertility. Due to the large population base in my country, the number of infertility patients among newly married couples is far more than one million, and the infertility caused by male factors is higher than 50%. About 40% to 75% of male infertility still cannot find the cause, which is called idiopathic male infertility. Idiopathic male infertility poses a disproportionately high disease burden. [0004] In fact, the diagnosis of idiopathic male infertility is very difficult. The WHO standard for male infertility is that the husband and wife have lived together fo...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02G01N2800/34
Inventor 陈敏健孙娴陈婷高嘉伟肖甜王逸桐胡艳辉陆春城吴炜张婷周昆徐菠夏彦恺陈道帧周作民王心如
Owner NANJING MEDICAL UNIV
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