Simultaneous bioanalysis of a phosphate prodrug and its parent compound using a multiplexed LC-MS method

Area of Science:

• Analytical Chemistry
• Pharmacology
• Biochemistry

Background:

• Drug discovery requires complex bioanalysis of drugs, prodrugs, and metabolites.
• Varying physiochemical properties necessitate optimized, yet challenging, liquid chromatography-mass spectrometry (LC-MS) conditions.
• Compromised single methods or manual condition switching hinder efficient quantitative bioanalysis.

Purpose of the Study:

• To address challenges in simultaneous quantitative bioanalysis of multiple analytes with differential LC-MS requirements.
• To develop a multiplexed approach for overcoming limitations in current bioanalytical methods.

Main Methods:

• A multiplexed approach employing a mixed mode of turbulent- and laminar-flow chromatography.
• Simultaneous quantitative bioanalysis of multiple analytes within the same biological sample.

Main Results:

• Demonstrated a method for simultaneous quantitative bioanalysis of multiple analytes.
• Successfully addressed the challenge of differential analyte optimization in LC-MS.

Conclusions:

• The developed approach enables individually optimized LC-MS conditions for multiple analytes in a single run.
• Illustrated with the quantitation of a lipophilic drug and its hydrophilic prodrug in a biological matrix.