SLAS 2022: AUTOMATED SAMPLE PREPARATION PLATFORM FOR BIOANALYTICAL SPE METHOD DEVELOPMENT & OPTIMIZATION USING ANDREW+™ PIPETTING ROBOT

Importance of the topic

The reliable extraction of drugs and metabolites from biological fluids underpins critical bioanalytical workflows in drug discovery, clinical diagnostics, and forensic toxicology. Automated sample preparation addresses challenges in throughput, reproducibility, and operator variability, ensuring robust and efficient analyses.

Objectives and overview of the study

This work demonstrates a fully automated reversed‐phase solid‐phase extraction (RP‐SPE) method development strategy using the Andrew+ pipetting robot and OneLab software. Key goals include:

• Implementing a sequential elution scouting approach to optimize pH, solvent polarity, and organic composition.
• Evaluating analyte recovery and matrix effects for carbamazepine in neat and plasma samples.
• Streamlining sample preparation protocols to reduce manual workload and improve reproducibility.

Methodology and instrumentation

An integrated automation platform was utilized, featuring:

• Andrew+ pipetting robot controlled via cloud‐native OneLab software for graphical protocol design and execution.
• Oasis HLB 96‐well SPE plates for reversed‐phase extraction.
• Waters ACQUITY I‐Class UPLC with HSS T3 C18 column (2.1×50 mm, 1.7 µm) and water/acetonitrile gradient (0.5% formic acid).
• Waters Xevo TQ-S mass spectrometer in positive ESI mode with MRM transitions (237.1→179.1 and 237.1→194.1) for carbamazepine quantification.

The 3×3 elution profile scouting evaluated methanol, acetonitrile, and acetone at varying concentrations and pH levels. Top performing conditions were then applied to rat plasma spiked at 10 ng/mL for method evaluation.

Main results and discussion

Sequential elution screening revealed broad analyte elution across solvents and pH values, highlighting acetone’s superior performance. Final method assessment of the top three conditions yielded:

• Optimal wash: 30% methanol at pH 7.
• Optimal elution: 30% acetone at pH 10.
• Carbamazepine recovery of 84% and matrix effects limited to 3% in rat plasma.

Automation reduced variability and accelerated method optimization compared to manual workflows.

Benefits and practical applications

• Standardized, high‐throughput processing in 96‐well format enhances laboratory efficiency.
• Improved reproducibility across days, users, and laboratories.
• Minimized human error and freed analysts for data interpretation.
• Versatile application to diverse bioanalytical targets and matrices.

Future trends and potential applications

• Integration of AI‐driven optimization for real‐time protocol refinement.
• Expansion to multidimensional separations and alternative extraction chemistries.
• Large‐scale pharmacokinetic and toxicology screening with high‐throughput automation.
• Seamless connectivity with LIMS for end‐to‐end laboratory automation.

Conclusion

The Andrew+ pipetting robot coupled with OneLab software provides a flexible and fully automated platform for rapid SPE method development. Sequential elution profiling enables efficient optimization of recovery and matrix effects, supporting robust and reproducible bioanalytical workflows.

Reference

1. Novel Extraction Techniques with ACQUITY UPLC and 2D Technology: Part I Pesticide Screening in Drinking Water (720006588EN)