Oasis™ PRiME HLB Solid Phase Extraction for High Bioanalytical Plasma Analyte Recovery and Low Matrix Effects of Top-Selling Pharmaceuticals

Significance of the Topic

Robust and efficient sample preparation is critical for accurate quantification of pharmaceutical drugs in plasma matrices. Traditional methods often require extensive optimization to balance analyte recovery and matrix removal. A generic SPE approach that eliminates method development can accelerate bioanalytical workflows and improve data reliability.

Objectives and Study Overview

This application note evaluates a simple, generic solid phase extraction protocol using Oasis PRiME HLB to extract multiple top-selling small molecule therapeutics from plasma. The goals were to achieve high analyte recovery, minimal matrix effects, and rapid LC-MS/MS analysis without the need for individual method optimization.

Methodology and Instrumentation

A three-step reversed-phase SPE protocol was employed using Oasis PRiME HLB in a 96-well µElution format with load wash and elute steps completed in under 30 minutes

• Sample fortification at 0.5 to 1000 ng/mL in rat plasma for recovery and calibration curves
• LC conditions: Waters ACQUITY I-Class Plus UPLC system, ACQUITY HSS T3 column 2.1×30 mm 1.8 µm, 0.1 percent formic acid in water and acetonitrile, 3-minute gradient, 10 µL injection
• MS conditions: Xevo TQ-XS tandem quadrupole with electrospray ionization, optimized cone voltage and collision energies for each analyte in MRM mode
• Data acquisition and quantification performed with MassLynx v4.1 and TargetLynx software

Main Results and Discussion

The method delivered analyte recoveries greater than 80 percent and matrix effects between minus 15 and plus 5 percent. Endogenous phospholipids were reduced by over 90 percent relative to traditional SPE and protein precipitation, resulting in cleaner extracts and reduced ion suppression. Chromatographic separation of six diverse pharmaceuticals was achieved within 3 minutes and calibration curves showed linear fits above 0.99 with lower limits of quantification at 0.91 ng/mL and upper limits ranging from 250 to 1000 ng/mL depending on compound.

Benefits and Practical Applications

This generic SPE strategy offers significant advantages for bioanalytical laboratories:

• No method development required for a broad range of neutral and basic small molecules
• Fast 30-minute SPE extraction and 3-minute UPLC-MS/MS analysis per sample
• High reproducibility and quantitative performance within plus or minus 15 percent accuracy
• Effective removal of phospholipids to lower matrix effects and extend instrument uptime

Future Trends and Possibilities

Further integration of generic SPE sorbents with automated platforms can enhance throughput and consistency. Expanding applications to other biological matrices and emerging modalities such as peptide and oligonucleotide therapeutics may benefit from similar phospholipid-removal strategies. Coupling with high-resolution mass spectrometry and data-independent acquisition could widen the dynamic range and multiplexing capabilities.

Conclusion

The Oasis PRiME HLB SPE protocol delivers a straightforward, broadly applicable method for plasma sample preparation that combines high recovery, low matrix effects, and ultra-fast LC-MS/MS quantification. This approach streamlines bioanalytical workflows by eliminating extensive method development while maintaining rigorous quantitative standards.

References

1. Williams RE Leatherwood HM Top 200 Small Molecule Drugs by Retail Sales in 2023 2024
2. DrugBank Online go drugbank com drugs accessed June 2024
3. Zhang X Danaceau JP Chambers EE Improvements in Recovery Reproducibility and Matrix Effects with Oasis PRiME HLB Waters Application Note 2015
4. Danaceau JP Trudeau ME A Simple Broadly Applicable Automated Bioanalytical Sample Preparation Strategy for LC MS Quantification of Apixaban Waters Application Note 2023
5. Rahim F VanTran K Trudeau ME Simple Fast and Selective Bioanalytical Sample Extraction for Lenalidomide Using Oasis MCX SPE Waters Application Note 2023