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Development and evaluation of a liquid chromatography-tandem mass spectrometry method for simultaneous measurement of toxic aldehydes from brain tissue. | LitMetric

AI Article Synopsis

  • Reactive aldehydes are important low molecular weight compounds that affect health and are linked to diseases like cardiovascular and neurodegenerative disorders, but quantifying them in brain tissue is challenging.
  • This study created and validated a new LC-MS/MS method using 3-nitrophenylhydrazine, which proved to be more sensitive than other reagents for measuring four specific reactive aldehydes associated with lipid peroxidation.
  • The method showed that in a mouse model of traumatic brain injury, there was a significant increase in specific aldehyde levels 28 days after injury, highlighting the method's effectiveness in studying brain trauma progression.

Article Abstract

Reactive aldehydes are a class of electrophilic low molecular weight compounds that play an essential role in physiological function and lipid peroxidation. These molecules are implicated in many diseases, especially cardiovascular and neurodegenerative diseases, and are potential endogenous markers of lipid peroxidation. However, there are limited options to accurately quantify multiple reactive aldehydes in brain tissue. This study developed and validated a 3-nitrophenylhydrazine derivatization-based LC-MS/MS method to quantify four reactive aldehydes: malondialdehyde, acrolein, 4-hydroxy-2-hexenal and 4-hydroxy-2-nonenal. Method development involved comparing the sensitivity of detection between widely used derivatization reagents: 2,4-dinitrophenylhydrazine and 3-nitrophenylhydrazine. Our data showed that 3-nitrophenylhydrazine resulted in greater sensitivity. Additional method development included evaluation of hydrolysis sample pretreatment, selection of protein precipitation reagent, and optimization of derivatization conditions. The optimized conditions included no hydrolysis and use of 20 % trichloroacetic acid as the protein precipitation reagent. The optimized derivatization condition was 25 mM 3-nitrophenylhydrazine reacted at 20 °C for 30 min. The chromatographic conditions were optimized to reduce matrix effects, ion suppression, and efficient analysis time (<7-minute analytical run). The four aldehyde species were accurately quantified in brain tissue using stable-labeled internal standards. Application of this assay to a traumatic brain injury mouse model revealed significant accumulation of acrolein, 4-hydroxy-2-hexenal, and 4-hydroxy-2-nonenal at 28 days post injury. Overall, a validated method was developed to rapidly quantify the most prominent reactive aldehydes associated with lipid peroxidation during injury progression following acute brain trauma.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11227393PMC
http://dx.doi.org/10.1016/j.jchromb.2024.124208DOI Listing

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