Analysis of Catecholamines

Importance of the Topic

Catecholamines such as norepinephrine, epinephrine, and dopamine are crucial signaling molecules in physiological and pathological processes. Accurate quantitation in plasma supports research in neuroscience, endocrinology, and clinical diagnostics.

Study Objectives and Overview

This application note demonstrates a rapid, sensitive LC-MS method using a Shim-pack MAqC-ODS I reversed-phase column to separate and quantify three catecholamines in plasma samples.

Methodology and Instrumentation

• Column: Shim-pack MAqC-ODS I (150 mm × 2.0 mm I.D., 5 µm)
• Mobile Phase A: 0.1% formic acid in water
• Mobile Phase B: 0.05% formic acid in methanol
• Gradient: 1% B (0–0.5 min) to 50% B (3.0 min), 99% B (3.1–7.0 min), re-equilibration to 1% B (7.1–12.0 min)
• Flow Rate: 0.2 mL/min; Injection Volume: 5 µL; Post-column Addition: 0.2 mL/min of Phase B
• MS System: LCMS-8600 with ESI in positive MRM mode (interface voltage +0.6 kV; DL 250 °C; interface 400 °C; nebulizing 2.2 L/min; drying 3 L/min; heating 17 L/min)

Main Results and Discussion

Baseline resolution of norepinephrine, epinephrine, and dopamine was achieved within 5 minutes. Sensitivity was enhanced by post-column addition, providing clear MRM signals. The method showed reproducible retention times and peak shapes, supporting robust quantification.

Benefits and Practical Applications

• Fast analysis cycle enables high throughput.
• High sensitivity and selectivity for trace-level detection.
• Applicable to clinical and research laboratories for stress hormone profiling.

Future Trends and Possibilities

Miniaturized LC-MS interfaces and advanced columns promise faster separations with lower solvent consumption. Multiplexed MRM panels could expand analyte coverage for comprehensive neurochemical profiling.

Conclusion

The described method offers a rapid, reliable platform for catecholamine analysis in plasma, enhancing workflow efficiency and data quality in biomedical studies.

References

Application News C114 (JP), Shimadzu Corporation, First Edition Aug. 2023