Automated sample preparation using membrane microtiter extraction for bioanalytical mass spectrometry

Area of Science:

• Analytical Chemistry
• Pharmacology
• Biotechnology

Background:

• Traditional drug analysis in biological fluids is often time-consuming.
• Current methods like solvent extraction or single cartridge SPE lack high-throughput capabilities.
• Automated sample preparation is crucial for modern drug discovery workflows.

Purpose of the Study:

• To develop and apply an automated membrane solid phase extraction (SPE) method for drug analysis in biological fluids.
• To enhance the speed and efficiency of sample preparation for high-performance liquid chromatography-mass spectrometry (HPLC/MS).
• To demonstrate the utility of a 96-well membrane SPE format for drug discovery support.

Main Methods:

• Development of an automated membrane SPE protocol using a modified programmable liquid handling station (Quadra 96).
• Utilized a novel rapid succession protocol for condition, load, and wash steps to prevent SPE bed drying and improve precision.
• Employed a 96-well microtiter plate format for simultaneous sample processing.
• Quantitative analysis of Ziprasidone in plasma as a test case.

Main Results:

• Achieved automated extraction of 96 samples in 10 minutes, a 30-fold increase in speed compared to manual methods.
• Enabled direct HPLC injection due to small elution solvent volumes, eliminating solvent evaporation.
• Demonstrated favorable quantitative results for Ziprasidone analysis compared to traditional methods.
• Successfully applied the method to analyze numerous drug prototypes in biological fluids.

Conclusions:

• Automated 96-well membrane SPE offers a significant advancement in sample preparation speed and efficiency for drug analysis.
• The developed method is compatible with high-throughput HPLC/MS, accelerating drug discovery efforts.
• This technique provides a robust and rapid solution for analyzing drugs in biological matrices.