A simple and sensitive assay for eflornithine quantification in rat brain using pre-column derivatization and...

Area of Science:

• Pharmacology
• Analytical Chemistry
• Neuroscience

Background:

• Eflornithine treats human African trypanosomiasis and is studied for cancer prevention.
• Accurate quantification of eflornithine in brain tissue is crucial for pharmacokinetic and pharmacodynamic studies.

Purpose of the Study:

• To develop and validate a novel ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay for eflornithine quantification in rat brain homogenate.
• To assess the assay's sensitivity, accuracy, precision, and stability for reliable eflornithine measurement.

Main Methods:

• Eflornithine derivatization using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.
• Separation via reverse-phase C18 UPLC column with a 6-min gradient.
• Quantification using tandem mass spectrometry (MS/MS) with methanol protein precipitation for extraction.

Main Results:

• The assay demonstrated reproducible extraction (~70% recovery) and stability of derivatized eflornithine for at least 24 hours.
• High sensitivity (LLOQ of 0.1 µm) and a wide dynamic range (5000-fold) were achieved.
• The validated assay successfully determined eflornithine distribution in various rat brain regions.

Conclusions:

• A sensitive and validated UPLC-MS/MS assay for eflornithine in rat brain was successfully developed.
• This assay enables precise quantification of eflornithine, supporting its study in neurological contexts and drug development.