It’s Not All About the Column: Sample Preparation and Containment

Importance of the topic

Sample preparation and containment are vital steps in liquid chromatography workflows, ensuring removal of particulates, proteins and lipids prior to analysis and preventing sample degradation and column fouling.
Appropriate sample handling enhances reproducibility, sensitivity and column lifetime, reducing ion suppression in LC-MS and minimizing the risk of contaminants entering the analytical system.

Objectives and overview

This article reviews best practices and product solutions for efficient sample preparation and secure containment in high-performance liquid chromatography and LC-MS applications.
Key goals include demonstrating filtration methods for particulate, protein, and lipid removal, and outlining vial and plate options that maintain sample integrity, compatibility and throughput.

Methodology

Filtration strategies covered comprise:

• Particulate removal via syringe filters, filter vials, and 96-well filter plates and cartridges with various membrane chemistries (PTFE, nylon, PES, glass fiber, depth filters).
• Comprehensive cleanup of lipids and proteins using EMR-Lipid sorbents in pass-through formats (1 mL, 3 mL, 6 mL cartridges and 96-well plates) to reduce matrix effects and enhance analyte sensitivity.

Containment solutions include:

• Deactivated, inert borosilicate and polypropylene vials certified for low adsorption and UV-invisible extractables.
• Precision screw-cap and crimp-cap closures with chemically compatible septa, including PFC-free options for PFAS analysis.
• Well plates and silicone sealing mats conforming to ANSI/SLAS standards for automated autosampler compatibility.

Used Instrumentation

• Captiva Premium Syringe Filters and Filter Vials
• Captiva 96-well Filter Plates and Cartridges
• Captiva EMR-Lipid Cleanup Cartridges and Plates
• Agilent A-Line Certified Deactivated Vials and Closures
• Agilent Electronic Crimper and Decapper
• InfinityLab Polypropylene Well Plates and Silicone Sealing Mats
• Vacuum, positive pressure and centrifuge manifolds for sample processing

Main results and discussion

Targeted EMR-Lipid cleanup achieved over 99 % phospholipid removal from biological fluids, dramatically reducing background noise in LC-MS analyses.
Comparisons show relative standard deviations below 3 % with EMR-Lipid versus over 25 % for protein precipitation alone, highlighting improved precision.
Efficient lipid removal also allowed significantly shorter LC gradients while maintaining peak shape and sensitivity, improving throughput and reducing solvent consumption.
Selecting appropriate vial materials and closures proved essential to prevent analyte adsorption, septum coring or sample evaporation during automated injections.

Benefits and practical applications

• Enhanced analytical reproducibility and sensitivity in complex matrices such as plasma, blood and food extracts.
• Extended column and instrument lifetime by minimizing fouling and carry-over.
• Increased laboratory throughput and reduced re-analysis rates.
• Compatibility with automated workflows in QA/QC, bioanalysis, environmental and PFAS monitoring.

Future trends and potential applications

Sample preparation is moving toward integrated, high-throughput and automated platforms, combining filtration, cleanup and direct introduction to LC-MS.
Emerging chemistries target broader classes of interferences, including polar contaminants and microplastics.
Advances in PFC-free consumables support stringent PFAS regulations and minimal blank levels.
Digital tools will guide optimal selection of vials, filters and plates based on sample type, autosampler models and analytical goals.

Conclusion

Effective sample preparation and secure containment are critical to achieving reliable, sensitive and reproducible chromatographic analyses.
A combination of targeted filtration chemistries and inert, well-matched vials and closures significantly improves data quality and laboratory efficiency.
Selecting the right consumables early in the workflow prevents common sources of error and supports a robust, high-performance analytical pipeline.

References

• Brochure: Agilent vials and sample containment solutions (5990-9022EN)
• White paper: An Agilent vial is not just a vial (5991-6769EN)
• White paper: An Agilent septum is not just a septum (5991-6770EN)
• Case study: Lower costs with Agilent A-Line vials (5991-7845EN)
• Poster: Influence of glass vial type upon trace level recovery rates (5991-7712EN)
• Poster: Agilent Vials (5991-6960EN)