Quantification of Methylene Blue and Evaluation of Its Pharmacokinetics in ICR Mice by Liquid Chromatography‐Quadrupole Time‐of‐Flight Mass Spectrometry Using Difluoroacetic Acid
Year: 2025
Type: article
Abstract: ABSTRACT Methylene blue (MB), a phenothiazine derivative, is currently under clinical trials for Alzheimer's disease ( AD ) due to its potential to inhibit tau aggregation, a key pathological process in AD . In this study, we developed and qualified a rapid and reliable liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (LC‐qTOF‐MS) method for the quantification of MB in mouse plasma and brain samples. Chromatographic separation was achieved using a PolymerX RP‐1 100 Å (50 × 2 mm, 5 μm) column with a mobile phase consisting of water and methanol containing 0.5% difluoroacetic acid, delivered at a flow rate of 0.5 mL/min. Calibration curves were constructed using quadratic regression (weighted 1/concentration 2 ) over a range of 3.05–2222.22 ng/mL in both matrices. The method was successfully applied to characterize the pharmacokinetics of MB in male ICR mice, revealing a high systemic clearance (65.64 mL/min/kg) and substantial brain penetration, as indicated by a brain‐to‐plasma partition coefficient ( K p,brain ) of 23.50 following single intravenous bolus administration. These findings provide critical insights into MB's in vivo behavior and demonstrate the utility of this bioanalytical method for evaluating MB in preclinical studies.
Source: Biomedical Chromatography
DOI: 10.1002/bmc.70080
Institution: Chungnam National University
Language: English
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $3,710
OpenAlex ID: W4409609083