Analysis of β-agonists Using a Core Enhanced Technology Accucore Phenyl-Hexyl HPLC Column

Significance of the Topic

The accurate and rapid analysis of β-adrenergic agonists is critical for monitoring their misuse as growth promoters in food livestock and ensuring consumer safety.

Objectives and Study Overview

This application note presents a liquid chromatography–mass spectrometry (LC/MS) method using a Thermo Scientific Accucore Phenyl-Hexyl HPLC column to achieve the separation of six commonly used β-agonists within a 6-minute run time.

Methodology and Instrumentation

Core Enhanced Technology with 2.6 µm superficially porous particles combines a solid core with a porous outer layer, providing high efficiency at lower backpressures. The phenyl-hexyl stationary phase delivers mixed-mode interactions—hydrophobic retention via the C6 chain and π-π interactions via the phenyl ring. Sample preparation involved diluting a methanol/water (95:5, v/v) β-agonist standard to 1 µg/mL. Chromatographic conditions were: 100 × 2.1 mm column, 40 °C, 0.25 mL/min flow, gradient elution from 5 to 100 % acetonitrile over 10 minutes with 5 mM ammonium acetate buffer at pH 4. Detection employed positive electrospray MS between m/z 150–350.

Instrumentation

• Accucore Phenyl-Hexyl 2.6 µm, 100 × 2.1 mm column
• Accela UHPLC system (Thermo Scientific)
• MSQ single-quadrupole mass spectrometer with +ESI

Main Results and Discussion

Extracted ion chromatograms showed the six β-agonists—terbutaline, cimaterol, salbutamol, ractopamine, clorprenaline, and clenbuterol—separated in under 6 minutes with clear peak resolution. While baseline separation was not fully achieved for all analytes, the selectivity provided by the phenyl-hexyl phase and MS detection suffices for regulatory quantification at low µg/L levels. The method combines speed with robust retention and reproducibility.

Benefits and Practical Applications of the Method

• Rapid analysis suitable for high-throughput screening
• Reduced backpressure compared with sub-2 µm columns
• Enhanced selectivity from mixed-mode interactions
• Compliance with regulatory limits for β-agonist residues

Future Trends and Potential Applications

Advances may include coupling superficially porous columns with high-resolution MS for increased sensitivity and selectivity, expanding the method to other veterinary drugs, and integrating automated sample preparation for routine food safety testing.

Conclusion

The Accucore Phenyl-Hexyl column paired with LC/MS enables the efficient and reliable separation of six β-agonists in a sub-6 minute analysis, offering a robust tool for regulatory laboratories monitoring food safety.

References

• Pereira L. Thermo Scientific Application Note ANCCSCETBETAAG, 2011.