Metabolism marker related to idiopathic male infertility in urine, and detection method and application thereof

A technology for metabolic markers and male infertility, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve the problems of prolonging the experimental cycle, long detection time, and limiting applications

Active Publication Date: 2016-12-07
NANJING HANWEI PUBLIC HEALTH RES INST CO LTD
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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 separat

Method used

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  • Metabolism marker related to idiopathic male infertility in urine, and detection method and application thereof
  • Metabolism marker related to idiopathic male infertility in urine, and detection method and application thereof
  • Metabolism marker related to idiopathic male infertility in urine, and detection method and application thereof

Examples

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 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. 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 and...

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 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, T...

Embodiment 3

[0158] Example 3: Stability analysis of L-leucine and L-palmitoylcarnitine in urine

[0159] The stability of the levels of L-leucine and L-palmitoylcarnitine in urine was evaluated by the method of Example 2 (the interval was 2 weeks). The results showed that the levels of L-leucine and L-palmitoylcarnitine in urine were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the field of analytic chemistry and clinical medicine, and discloses a metabolism marker related to idiopathic male infertility in urine, and a detection method and application thereof. The metabolism marker is L-leucine and/or L-palmitoyl carnitine, is detected by adopting UPLC-Qexactive MS method, can be used for auxiliary diagnosis and monitoring of idiopathic male infertility, has higher sensitivity and specificity, and has a clinical spread value.

Description

field of invention [0001] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to metabolic markers L-leucine and / or L-palmitoylcarnitine in urine related to idiopathic male infertility and its detection based on UPLC-Q exactive MS methods and applications. Background technique [0002] Currently, about 10-15% of couples of reproductive age worldwide suffer from fertility disorders. Due to the large population base in my country, the number of infertility patients among newlyweds is far more than one million, and the infertility caused by male factors is higher than 50%. There are still about 40%-75% of male infertility whose cause cannot be found, which is called idiopathic male infertility. Idiopathic male infertility poses a disproportionately high disease burden. [0003] In fact, the diagnosis of idiopathic male infertility is very difficult. The WHO diagnostic criteria for male infertility are couples who have lived together...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 陈敏健肖甜陈婷陈道帧周作民张婷王玉邦胡艳辉环飞陆春城吴炜夏彦恺王心如
Owner NANJING HANWEI PUBLIC HEALTH RES INST CO LTD
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