Increasing bioanalytical throughput using pcSFC-MS/MS: 10 minutes per 96-well plate

Area of Science:

• Analytical Chemistry
• Chromatography
• Mass Spectrometry

Background:

• Packed-column supercritical fluid chromatography (pcSFC) is gaining traction for high-throughput analyses.
• Compared to traditional liquid chromatography, pcSFC uses volatile mobile phases (like CO2) reducing viscosity for faster flow rates.
• The volatile nature of pcSFC mobile phases is compatible with mass spectrometry (MS), allowing direct effluent analysis.

Purpose of the Study:

• To demonstrate the utility of pcSFC coupled with tandem mass spectrometry (pcSFC-MS/MS) for high-throughput bioanalytical quantitation.
• To validate pcSFC-MS/MS for pharmacokinetic applications using dextromethorphan as a model compound.
• To assess the speed, accuracy, and precision of the pcSFC-MS/MS method for analyzing plasma samples.

Main Methods:

• Plasma samples were processed using automated liquid/liquid extraction in a 96-well format.
• Dextromethorphan was analyzed using packed-column supercritical fluid chromatography coupled with tandem mass spectrometry (pcSFC-MS/MS).
• Method validation included three days of accuracy and precision assessments for quality control samples at various concentrations.

Main Results:

• Quantification of dextromethorphan in 96-well plates was achieved at a rate of approximately 10 minutes per plate.
• The method demonstrated excellent intraday accuracy, with results within 9% of the true value.
• Daily relative standard deviations (RSDs) for quality control samples were consistently below 10% (for higher concentrations) and below 15% (for lower concentrations).

Conclusions:

• pcSFC-MS/MS is a highly efficient technique for high-throughput bioanalytical quantitation in pharmacokinetic studies.
• The method provides rapid analysis with high accuracy and precision, suitable for processing large numbers of samples.
• This approach facilitates faster drug development by enabling quicker analysis of patient samples.