JoVE - Journal of Visualized Experiments
JoVE Visualize
Contact Us

Liquid Chromatography-Tandem Mass Spectrometry Method Development and Validation for the Determination of a New Mitochondrial Antioxidant in Mouse Liver and Cerebellum, Employing Advanced Chemometrics

  • 0Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece.
Molecules (basel, Switzerland) +

|

May 14, 2025

|

May 14, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

A novel antioxidant, triphenylphosphine-hydroxytyrosol (TPP-HT), was orally administered to mice. TPP-HT successfully crossed the blood-brain barrier, demonstrating its potential for mitochondrial targeting in vivo.

Area Of Science

  • Neuroscience
  • Pharmacology
  • Biochemistry

Background

  • Anxiety and stress disorders are globally prevalent, impacting all age groups.
  • Chronic stress and anxiety disrupt mitochondrial function, driving research into new therapeutic strategies.
  • Targeting mitochondria with antioxidant compounds presents an innovative approach for managing these conditions.

Purpose Of The Study

  • To synthesize and evaluate a novel antioxidant, triphenylphosphine-hydroxytyrosol (TPP-HT), for its efficacy in vivo.
  • To determine the tissue distribution and blood-brain barrier penetration of orally administered TPP-HT in mice.
  • To assess the feasibility of using TPP-HT as an orally administered agent for mitochondrial targeting.

Main Methods

  • A new antioxidant, TPP-HT, was synthesized, combining triphenylphosphine (TPP), an alkyl spacer, and hydroxytyrosol (HT).
  • TPP-HT was administered orally to mice through their drinking water.
  • Cerebellum and liver tissues were analyzed using ultra-high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) to quantify TPP-HT levels.

Main Results

  • The UHPLC-MS/MS method was validated for selectivity, linearity, accuracy, and sensitivity (LOD/LOQ).
  • Quantifiable levels of TPP-HT were detected in both cerebellum (11.5 ng g<sup>-1</sup>) and liver (4.8 ng g<sup>-1</sup>).
  • The results confirm that TPP-HT penetrates the blood-brain barrier.

Conclusions

  • Orally administered TPP-HT effectively reaches target tissues, including the brain.
  • TPP-HT demonstrates potential as an orally available therapeutic agent for mitochondrial targeting.
  • This study supports the development of TPP-HT for conditions associated with mitochondrial dysfunction due to stress and anxiety.

Keywords: