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Non-target biomarker high-throughput screening method based on pollutant metabolism disturbance

A technology for biomarkers and pollutants, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as low accuracy and insufficient screening throughput

Active Publication Date: 2020-08-07
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Aiming at the problem of insufficient screening throughput and low accuracy existing in metabolomics research in the prior art, the screening method of the present invention first performs non-target screening of potential metabolites, and then gradually narrows the range to obtain a small range of metabolites. The number of biomarkers can ensure more comprehensive screening, achieve high-throughput, accurate screening and identification of biomarkers, and provide a scientific basis for toxicity prediction and pollutant risk assessment and control

Method used

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  • Non-target biomarker high-throughput screening method based on pollutant metabolism disturbance
  • Non-target biomarker high-throughput screening method based on pollutant metabolism disturbance
  • Non-target biomarker high-throughput screening method based on pollutant metabolism disturbance

Examples

Experimental program
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Embodiment 1

[0081] This embodiment is a high-throughput screening method for non-target biomarkers of metabolic disturbance caused by perfluorinated pollutants in blood media, and the steps are as follows:

[0082] (1) Take 0.5mL serum sample in a 15mL centrifuge tube, add 0.26~0.28g of magnesium sulfate-sodium chloride mixture and 1.5mL of acetonitrile and vortex immediately, the sample is in a suspension state at this time, after the sample is ultrasonically extracted for 30min Centrifuge and transfer supernatant. The remaining residue was repeatedly extracted twice with 95% acetonitrile-water solution, and the extracts were combined. Nitrogen was blown to near dryness, transferred to a chromatographic vial and dilute to 100 μL with acetonitrile.

[0083] (2) On-board measurement: the extracted samples are analyzed and detected in full scan by high performance liquid chromatography-time-of-flight mass spectrometry. Its parameters are:

[0084] High performance liquid chromatography: ...

Embodiment 2

[0139] The operating steps of this embodiment are basically the same as those in Example 1, except that: before step (5), the two most widely studied perfluorinated compounds for perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) are added. Docking steps to make predictions about their metabolic perturbation capabilities.

[0140] The molecules of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) were optimized in SYBYL software: applying Tripos force field, Gasteiger-Huckel charge, and Powell (Powell) gradient method until the termination gradient drops to 0.001kcal / (mol· )the following.

[0141] After downloading the human serum albumin structure, the ligand was extracted in SYBYL software to form a docking pocket, and the crystal water was removed and protonated. The optimized perfluoroligand was docked with the protein pocket in SYBYL software, and the optimal conformation was selected as the docking result. The greater the total...

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Abstract

The invention discloses a non-target biomarker high-throughput screening method based on pollutant metabolism disturbance, and belongs to the field of environmental exposure and health, and the methodcomprises the following steps: (1) extracting to obtain a to-be-detected extracting solution; (2) performing chromatographic analysis to obtain a spectrogram containing chromatographic peaks; (3) identifying and labeling pollutant characteristic peaks, taking chromatographic peaks except the pollutant characteristic peaks as potential metabolite characteristic peaks, and carrying out non-target labeling on the potential metabolite characteristic peaks; (4) establishing a linear regression model by taking the peak area of the potential metabolite characteristic peak as a dependent variable andthe peak area of the pollutant characteristic peak as an independent variable; (5) operating the model, carrying out non-target screening on the biomarkers, and preliminarily obtaining related biomarkers; and (6) identifying the preliminarily obtained primary and secondary spectrograms of the biomarker, and identifying the biomarker related to pollutant exposure. According to the method disclosedby the invention, the accuracy of biomarker screening is remarkably improved, and meanwhile, the flux of biomarker screening is improved.

Description

technical field [0001] The invention belongs to the field of environmental exposure and health, and in particular relates to a high-throughput screening method for non-target biomarkers based on pollutant metabolism disturbance. Background technique [0002] With the development of science and technology, people's living standards continue to improve, and the pollutants produced are also increasing. These substances are released into the environment and accumulate in the environment. Organisms are exposed to various environmental media, and pollutants in the environment may enter organisms through various channels such as touching, breathing, and eating, resulting in a transformation from external exposure to internal exposure. Exogenous environmental pollutants that enter organisms pose potential threats to organisms, and there is substantial evidence that exposure to specific chemicals may cause disease. [0003] Biomarkers are signal indicators for abnormal changes obser...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/86
CPCG01N30/02G01N30/06G01N30/08G01N30/8679G01N30/8665G01N2030/065
Inventor 韦斯李昱茜于红霞于南洋
Owner NANJING UNIV
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