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Related Experiment Videos

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

S H Hoke1, J A Tomlinson, R D Bolden

  • 1Health Care Research Center, The Procter & Gamble Company, Mason, Ohio 45040, USA. hoke.sh@pg.com

Analytical Chemistry
|July 27, 2001
PubMed
Summary

Packed-column supercritical fluid chromatography (pcSFC) offers faster analysis times for high-throughput bioanalysis. This method, coupled with tandem mass spectrometry (MS/MS), enables rapid quantification of drugs in plasma samples.

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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:

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  • 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.