Why Did That Happen?

Importance of the Topic

Liquid chromatography is fundamental for qualitative and quantitative analysis in pharmaceuticals, environmental monitoring, food safety and research. Robust troubleshooting ensures reliable data, reduces downtime and enhances method performance.

Objectives and Study Overview

This guide categorizes common LC issues into changes in system pressure, peak shape, separation and detection, complemented by sample related effects. It aims to provide systematic root cause analysis and corrective actions to maintain optimal chromatography workflows.

Methodology and Instrumentation

A structured approach aligns observed symptoms with potential causes and recommended actions. Preventive practices are highlighted alongside initial corrective measures. Sample cleanup techniques and various consumables are assessed for their contributions to performance improvements.

Used Instrumentation

• Agilent InfinityLab HPLC system with advanced diagnostics
• InfinityLab Quick Connect and Quick Turn fittings for minimal dead volume
• InfinityLab Quick Change inline filters and Stay Safe caps
• InfinityLab Poroshell 120 and ZORBAX UHPLC/HPLC columns
• Captiva syringe filters and filter vials
• Bond Elut solid phase extraction (Plexa, Silica), Chem Elut S supported liquid extraction and QuEChERS kits
• Online SPE housings and automation-friendly 96-well plates

Key Results and Discussion

System overpressure often stems from clogged frits or filters, while underpressure arises from leaks in fittings. Pressure fluctuations are linked to air, cavitation and pump head wear. Peak tailing, broadening, fronting and splitting can be traced to secondary interactions, injection volume, tubing dispersion and column damage. Shifts in retention time originate from mobile phase inconsistency, temperature variations and equilibration. Detection issues such as noisy baselines and reduced sensitivity are attributed to bubbles, contamination and lamp degradation. Sample-related chemical and physical effects are addressed via inline filtration, SPE, SLE and QuEChERS workflows to remove particulates, lipids, proteins and matrix interferences.

Benefits and Practical Applications

This comprehensive framework enhances routine troubleshooting in QA/QC, research laboratories and industrial environments. It extends component lifespan, ensures reproducible separations, improves sensitivity and streamlines sample preparation across diverse matrices.

Future Trends and Opportunities

Emerging developments include adoption of automated inline sample cleanup, RFID-enabled column tracking, software-driven preventive maintenance, biocompatible inert flow path materials and sustainable chromatography with reduced solvent consumption and advanced polymeric phases.

Conclusion

By translating complex chromatographic anomalies into clear diagnostic steps and targeted remedies, this guide empowers analysts to sustain high-performance chromatography, minimize downtime and achieve consistent, high-quality results.

References

• Javadi G Why Did That Happen? Common Chromatography Problems Agilent Technologies 2021
• Agilent Technologies InfinityLab Stay Safe Caps Operational Manual 5994-1507EN Technical Overview 5994-1504EN
• Agilent Technologies InfinityLab Poroshell 120 Column Specifications Poster 5991-9013EN
• Agilent Technologies ZORBAX Column Overview Poster 5994-2212EN
• Agilent Technologies LC Troubleshooting Poster 5994-0709EN