A simplified and completely automated workflow for regulated LC-MS/MS bioanalysis using cap-piercing direct sampling...

Area of Science:

• Analytical Chemistry
• Bioanalysis
• Pharmacokinetics

Background:

• Automated sample extraction for regulated bioanalysis using liquid chromatography/tandem mass spectrometry (LC-MS/MS) faces significant challenges.
• Current methods often require manual intervention, increasing risks and potential for errors.

Purpose of the Study:

• To develop a simplified, fully automated sample preparation workflow for regulated bioanalysis.
• To eliminate manual handling and reduce bioanalyst exposure to infectious biofluids.

Main Methods:

• A novel methodology employing cap piercing for direct biofluid transfer and evaporation-free solid phase extraction (SPE).
• Utilized pierceable cap sample tubes and a robotic liquid handler for automated sampling without uncapping.
• Employed a mobile phase-compatible elution solvent and sample dilution to eliminate evaporation steps prior to LC-MS/MS analysis.

Main Results:

• Successfully validated three LC-MS/MS assays for CNS drug development programs in various plasma matrices (dog, monkey, human).
• Assays demonstrated linearity from 1.00 to 1000 or 2.00 to 2000 ng/mL with excellent accuracy, precision, and reproducibility.
• Achieved fully automated sample preparation, meeting acceptance criteria for regulated bioanalysis and application to study samples.

Conclusions:

• The developed methodology fully automates sample preparation without compromising assay performance.
• This innovative approach enhances bioanalyst safety by eliminating direct contact with biofluids.
• The method is robust and suitable for supporting regulated bioanalysis in drug development.