Validated RP-HPLC-UV Method for the Quantification of Trans-Resveratrol in Plasma: Application to Comprehensive...

Area of Science:

• Pharmacokinetics and Bioanalysis
• Green Chemistry in Analytical Methods
• Drug Development and Formulation

Background:

• Trans-resveratrol (trans-RES) exhibits therapeutic promise but suffers from poor bioavailability and photolability, hindering its clinical application.
• Accurate quantification of trans-RES in biological matrices is crucial for pharmacokinetic studies and formulation development.
• Existing analytical methods may not adequately address the stability and bioavailability challenges of trans-RES.

Purpose of the Study:

• To develop and validate a robust, ICH M10 compliant RP-HPLC method for quantifying trans-RES in rat plasma.
• To assess the photostability of trans-RES and establish necessary controls.
• To conduct a comprehensive pharmacokinetic study comparing five administration routes of trans-RES in rats.

Main Methods:

• Development and validation of a reversed-phase high-performance liquid chromatography (RP-HPLC) method with liquid-liquid extraction and isocratic elution.
• Quantification of trans-RES in plasma using UV detection, adhering to ICH M10 guidelines.
• Evaluation of method linearity, precision, accuracy, sensitivity, and stability, including photostability.
• Assessment of the method's environmental impact using the Analytical GREEnness (AGREE) metric.
• Pharmacokinetic analysis following administration via five different routes in rats.

Main Results:

• The RP-HPLC method achieved excellent separation of trans- and cis-isomers, demonstrating high linearity (R² > 0.999), precision (%CV < 5%), accuracy (93.90-98.19%), and sensitivity (LLOQ 0.025 μg/mL).
• Stability studies confirmed trans-RES's susceptibility to UV-induced isomerization, necessitating photostability controls.
• Intraperitoneal administration resulted in the highest bioavailability (84.92 ± 4.87%), significantly higher than oral administration (4.91 ± 0.22%).
• Flip-flop kinetics were observed for non-intravenous routes, indicating absorption-limited disposition.
• The method achieved a favorable AGREE score of 0.7, indicating alignment with green chemistry principles.

Conclusions:

• A validated, reliable, and sustainable RP-HPLC bioanalytical method for trans-RES quantification in plasma was successfully developed.
• Pharmacokinetic data highlight the critical need for strategic formulation to enhance trans-RES bioavailability and stability.
• The findings support the advancement of trans-RES's translational potential through improved drug delivery strategies.