Agilent Captiva EMR–Lipid Manual Plate Preconditioning

Importance of the Topic

Efficient sample preparation is fundamental for reliable LC/MS-based omics analyses. Preconditioning of solid-phase extraction devices helps stabilize sorbent performance, reduces carryover, and improves reproducibility when fractionating metabolites, lipids, and proteins from plasma or cell samples.

Objectives and Study Overview

This method guide presents a manual preconditioning protocol for Agilent Captiva EMR–Lipid 96-well plates. It details solvent composition, workflow steps, and equipment settings to ensure consistent plate performance prior to automated or manual omics sample preparation.

Methodology and Instrumentation

The protocol uses a 2:1:1 water:methanol:ethanol mixture as the preconditioning solvent. Key steps include:

• Prepare 180 mL of solvent per plate (use LC/MS-grade solvents and store up to four months).
• Add 800 µL of solvent to each well, then apply positive pressure to elute.
• Repeat the addition and elution for a second cycle, employing a low-flow setting (gradually increasing pressure) and finishing with a high-flow purge (10–20 psi) to clear sorbent frits.

Instrumentation Used

• Agilent Captiva EMR–Lipid 96-well plates (Part #5190-1001), pack of 5.
• Agilent 2-column reservoir (Part #203852-100), pack of 25.
• Agilent single-cavity reservoir (Part #201244-100), pack of 25.
• Agilent positive pressure manifold PPM-96 (Part #5191-4116).
• Milli-Q Integral water purification system with 0.22 µm filter cartridge.

Key Results and Discussion

Plate preconditioning with the specified solvent system and pressure settings yields uniformly conditioned sorbent beds. This process reduces nonspecific binding, ensures reproducible flow rates during subsequent sample loading, and minimizes risk of frit blockage.

Benefits and Practical Applications

• Enhanced reproducibility across wells and plates in lipid and metabolite extraction workflows.
• Reduced risk of clogging and carryover in high-throughput LC/MS omics pipelines.
• Compatibility with automated liquid-handling platforms for streamlined sample processing.

Future Trends and Potential Applications

Advancements may include integration of automated preconditioning steps into robotic workflows, optimization of solvent compositions for diverse sample types, and adaptation of high-pressure systems to accelerate throughput. Expanding EMR sorbent surfaces and novel plate formats could further improve selectivity and capacity.

Conclusion

The manual preconditioning of Agilent Captiva EMR–Lipid 96-well plates using a 2:1:1 water:methanol:ethanol solvent under controlled pressure significantly improves omics sample preparation. This straightforward protocol supports robust, reproducible fractionation of lipids and metabolites in LC/MS workflows.

References

1. Sartain M. et al. Enabling Automated, Low-Volume Plasma Metabolite Extraction with the Agilent Bravo Platform. Agilent Technologies Application Note 5994-2156EN (2020).
2. Van de Bittner G.C. et al. An Automated Dual Metabolite and Lipid Sample Preparation Workflow for Mammalian Cell Samples. Agilent Technologies Technical Overview 5994-5065EN (2022).
3. Van de Bittner G.C. et al. Automated Fractionation of Low-Volume Plasma Samples for LC/MS Multi-Omics. Agilent Technologies Technical Overview 5994-7357EN (2024).
4. Agilent Technologies. Captiva EMR–Lipid Manual Fractionation of Low-Volume Plasma Samples for LC/MS Multi-Omics. Method Guide 5994-7482EN (2024).