HPLC Separation of Bile Acids Using a CORTECS™ Premier Phenyl Column

Significance of Topic

Accurate analysis of bile acids is critical for understanding lipid metabolism, monitoring human health, and assessing colon cancer risk associated with elevated plasma bile acid levels.

Study Objectives and Overview

This study evaluated the performance of a 2.1×50 mm, 2.7 µm CORTECS Premier Phenyl Column with MaxPeak Premier hardware for baseline separation and LC-MS quantification of six bile acids, including two isobaric pairs.

Methodology and Instrumentation

The analytes (50 µg/mL each) were prepared in 90:10 water:acetonitrile with 0.1% formic acid. Chromatographic separation employed a screening gradient from 5% to 95% acetonitrile over 6.86 minutes at 0.5 mL/min and 40°C. A Waters ACQUITY QDa Detector operated in negative electrospray ionization mode. The CORTECS solid-core particles provided high efficiency, while MaxPeak Premier surfaces minimized analyte–hardware interactions.

Results and Discussion

Baseline resolution was achieved for taurodeoxycholic acid/taurochenodeoxycholic acid and glycodeoxycholic acid/glycochenodeoxycholic acid, as confirmed by single ion recording plots. Peak shapes were sharp, with no tailing or area loss, demonstrating effective suppression of metal-induced interactions affecting sulfonated bile acids.

Benefits and Practical Applications

• Reliable baseline separation of isobaric bile acids for accurate quantification.
• High column efficiency supports rapid screening in clinical and quality control laboratories.
• Reduced peak tailing enhances sensitivity and data quality in LC-MS workflows.

Future Trends and Opportunities

Advances in stationary phase design and inert surface technologies will further improve separations of challenging analytes. Integration with high-resolution MS, automation, and AI-driven data analysis promises greater throughput and deeper metabolic insights.

Conclusion

The combination of CORTECS Premier Phenyl Columns with MaxPeak Premier hardware enables high-efficiency, inert-surface chromatography ideal for challenging bile acid analyses. This approach delivers baseline separation of isobaric compounds and robust LC-MS performance for health monitoring and research applications.

Reference

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