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Development of an Optimized Two-Step Solid-Phase Extraction Method for Urinary Nucleic Acid Adductomics.

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Summary
This summary is machine-generated.

This study developed a novel method to extract and measure nucleic acid adducts in urine, crucial for understanding disease etiology. The optimized solid-phase extraction technique successfully identified hundreds of adducts in human and mouse samples.

Keywords:
DNA adductsDNA damageDNA repairRNA adductsadductomicsexposomemass spectrometryurine

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Area of Science:

  • Environmental Health Sciences
  • Toxicology
  • Analytical Chemistry

Background:

  • The exposome encompasses all exposures influencing health.
  • Nucleic acid adducts, formed by exposures, are implicated in diseases like cancer.
  • Urine analysis offers a non-invasive method to study the body's adductome.

Purpose of the Study:

  • To develop an optimized solid-phase extraction (SPE) method for maximal recovery of diverse nucleic acid adducts from urine.
  • To demonstrate the method's efficacy in identifying endogenous urinary adducts using untargeted high-resolution mass spectrometry.

Main Methods:

  • Evaluation of various SPE sorbent combinations for nucleic acid adduct extraction from urine.
  • Utilizing an ENV+ and PHE SPE column combination for enhanced adduct retention.
  • Employing untargeted high-resolution mass spectrometry with FeatureHunter 1.3 software for adduct identification.

Main Results:

  • The ENV+ and PHE SPE combination demonstrated superior recovery of a 20-adduct standard cocktail.
  • The method successfully recovered endogenous nucleic acid adducts from mouse and human urine.
  • Approximately 500 adducts were identified in both species, with isotopically labeled standards aiding in endogenous adduct characterization.

Conclusions:

  • An optimized SPE method coupled with high-resolution mass spectrometry enables comprehensive analysis of the urinary nucleic acid adductome.
  • This approach facilitates the study of exposome-related health effects and disease mechanisms.
  • The characterized adductome provides a foundation for understanding individual susceptibility and disease risk.