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Seminal plasma organic acid markers related to idiopathic male infertility and their detection methods and applications

A technology of male infertility and markers, which is applied in the field of analytical chemistry and clinical medicine, can solve the problems of complex experimental steps and low sensitivity, and achieve the effects of improved sensitivity and specificity, high sensitivity and specificity, and easy detection

Active Publication Date: 2021-07-20
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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  • Seminal plasma organic acid markers related to idiopathic male infertility and their detection methods and applications
  • Seminal plasma organic acid markers related to idiopathic male infertility and their detection methods and applications
  • Seminal plasma organic acid markers related to idiopathic male infertility and their detection methods and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Embodiment 1 Research Object Selection and Grouping Basis

[0085] 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. Specific sample classification criteria are as follows:

[0086] first stage screening

[0087] A total of 30 people were included in the study, including 15 patients with def...

Embodiment 2

[0116] Example 2 UPLC-MS metabolomics screening of idiopathic male infertility β-alanine and tetradecanedioic acid

[0117] 1. Sample pretreatment

[0118] 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.

[0119] 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.

[0120] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0121] 2. Instrument testing

[0122] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0123]2.2. Liquid phase conditions:

[0124] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chr...

Embodiment 3

[0137] Example 3 Stability analysis of seminal plasma β-alanine and tetradecanedioic acid

[0138] The stability of seminal plasma β-alanine and tetradecanedioic acid levels was evaluated by the method of Example 2 (the interval was 2 weeks). The results showed that the measured levels of β-alanine and tetradecanedioic acid 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 provides a method for detecting β-alanine and tetradecanedioic acid in seminal plasma related to idiopathic male infertility by UPLC-Q active MS and an application thereof. The present invention compares the metabolic small molecules in the seminal plasma of the normal control and idiopathic male infertility by using UPLC-Q exact MS, and finds that there are small molecules in the seminal plasma that can be used to evaluate whether there is idiopathic male infertility, and have diagnostic value. The combination of β-alanine and tetradecanedioic acid, and the application of UPLC-Q exact MS for the detection of β-alanine and tetradecanedioic acid, developed a diagnosis of idiopathic male infertility that can be easily applied clinically , Monitoring kit.

Description

[0001] field of invention [0002] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to the detection of β-alanine and tetradecanedioic acid in seminal plasma related to idiopathic male infertility based on UPLC-Q active MS and the application thereof. Background technique [0003] According to the statistics of the World Health Organization (WHO), 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. WHO's standard for male infertility is th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88G01N30/06G01N33/68
CPCG01N30/06G01N30/88G01N33/6848G01N2030/045G01N2030/8813
Inventor 陈敏健王洁胡艳辉周景景王玉邦陈道桢夏彦恺
Owner NANJING MEDICAL UNIV
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