Sample Preparation
IndustriesManufacturerWaters
Importance of the Topic
Bioanalytical sample preparation is essential for reliable quantitation of drugs in biological matrices. Consistent and efficient extraction workflows directly affect data quality, throughput, and laboratory productivity.
Objectives and Study Overview
This study aimed to fully automate the extraction of the anticoagulant apixaban from plasma using the Andrew+ Pipetting Robot with the Extraction+ Connected Device. Method performance across multiple preparation techniques was evaluated through recovery, matrix effect, accuracy, and precision assessments.
Methodology and Instrumentation
The automated workflows were developed and executed in OneLab software controlling the Andrew+ platform and Extraction+ device. Standard sample preparation methods included:
- Protein precipitation (PPT)
- PPT with phospholipid removal
- Solid‐supported liquid extraction (SSLE)
- Reversed‐phase SPE (RP‐SPE) and RP‐SPE with phospholipid removal
- Mixed‐mode SPE (MCX and WAX)
Apixaban and isotopically labeled internal standard stocks were prepared in methanol and spiked into plasma for calibration and QC samples. Automated liquid handling steps, vacuum settings, waste management, and elution steps were synchronized.
Key instrumentation:
- Andrew+ Pipetting Robot with Extraction+ Connected Device (Waters Corporation)
- OneLab web‐based control software
- Waters ACQUITY I‐Class PLUS UPLC with BEH C18 column
- Waters Xevo TQ‐XS tandem quadrupole mass spectrometer (ESI+)
Main Results and Discussion
All automated extraction techniques demonstrated excellent calibration linearity (R² > 0.99), accuracy within ±15%, and precision (RSD) below 15%. Recoveries varied by method:
- PPT and PPT with phospholipid removal: ~77–92% recovery
- SSLE: ~91% recovery with moderate matrix effects
- RP‐SPE: ~96% recovery
- RP‐SPE with phospholipid removal: ~81% recovery
- Mixed‐mode SPE (MCX/WAX): ~100% recovery with minimal matrix suppression or enhancement
The automated workflows minimized variability, eliminated manual handling errors, and matched or exceeded conventional manual protocols in cleanliness and sensitivity.
Benefits and Practical Applications
Automation of bioanalytical extractions offers:
- Improved throughput by parallel processing of 96‐well plates or cartridges
- Enhanced reproducibility across users and laboratories
- Reduced hands‐on time, freeing analysts for data review and method development
- Rapid deployment of new assays without extensive method revalidation
Future Trends and Opportunities
Ongoing advancements may include integration with real‐time process monitoring, expansion to additional sample types (e.g., tissues, urine), and incorporation of advanced sorbent chemistries. Machine‐learning–assisted protocol optimization could further reduce development time and improve extraction selectivity.
Conclusion
The combination of Andrew+ Pipetting Robot, Extraction+ Connected Device, and OneLab software streamlines bioanalytical sample preparation for LC‐MS assays. This fully automated approach ensures robust performance, high throughput, and consistent results, enhancing overall laboratory efficiency.
Reference
- Danaceau JP, Trudeau ME. Automation of Bioanalytical Sample Extraction Techniques Using Andrew+ Pipetting Robot with the Extraction+ Connected Device. Waters Corporation; 2023.
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