Metrosep Carb 2 (Column manual)

Significance of the Topic

The analysis of carbohydrates and related anhydrosugars under alkaline conditions is crucial for quality control in food, beverage, environmental monitoring and clinical research. Metrosep Carb 2 columns offer targeted separation of monosaccharides, disaccharides, sugar alcohols and anhydrosugars with high resolution and reproducibility using pulsed amperometric detection.

Objectives and Overview

This manual provides a comprehensive guide to the Metrosep Carb 2 (6.1090.XX0 / 6.01090.XX0) separation columns, including:

• Ordering information and technical specifications for 4 mm and 2 mm formats
• Key handling and storage recommendations
• Eluent preparation and degassing procedures
• Step-by-step start-up, conditioning and troubleshooting protocols
• Representative application examples across diverse sample matrices

Methodology and Instrumentation

Instrumentation and consumables:

• High-pressure pump with CO₂-scrubbed eluent bottle and inline degasser
• Metrosep Carb 2 separation column with PEEK iColumn chip
• Appropriate guard column (Carb 2 Guard) and trap columns (CO₃²– or BO₃³– Trap 1)
• Pulsation absorber, 0.18 mm PEEK capillaries to minimize dead volume
• Pulsed amperometric detector (Au working electrode, Pd reference electrode)

Method steps:

• Prepare 100 mmol/L NaOH + 10 mmol/L sodium acetate eluent; degas and protect with inert gas
• Connect and rinse guard and separation columns at recommended flow and temperature
• Condition system until baseline stability is achieved
• Run analyses in PAD mode with defined potential sequence (0.05 V, 0.55 V, –0.10 V)

Main Results and Discussion

Performance studies demonstrate:

• Baseline chromatogram resolving eight representative sugars with sharp peaks
• Retention time modulation by flow rate (0.13–1.2 mL/min), column temperature (20–60 °C) and eluent strength
• Effects of organic additives and salts on sensitivity and separation order
• Application cases: apple juice sugars, levoglucosan in air particulates, cosmetic extractables, lactose in milk, yogurt, seawater anions, chromium speciation by ICP-MS

Benefits and Practical Applications

Metrosep Carb 2 columns deliver:

• Wide pH tolerance (0–14) and stable performance under constant chemical conditions
• Long service life when combined with guard columns and proper eluent care
• Flexibility for inline sample preparation (MISP techniques) to remove particles and matrix interferences
• Compatibility with pulsed amperometric, UV/VIS and mass spectrometric detectors

Future Trends and Opportunities

Emerging directions include:

• Integration of automated online sample prep and multidimensional separations
• Coupling with high-resolution mass spectrometry for comprehensive carbohydrate profiling
• Miniaturization for point-of-care and field applications
• AI-driven method development and real-time process monitoring
• Development of greener eluents and sustainable consumables

Conclusion

Metrosep Carb 2 columns represent a robust solution for selective, high-resolution carbohydrate analysis in diverse industries. Adhering to recommended handling, eluent preparation and maintenance protocols ensures reproducible results and maximizes column lifetime.

References

• Application Note P-54: Anhydrosugars besides sugar alcohols and sugars on Metrosep Carb 2 - 150/4.0
• Application Note P-55: Lactose in lactose-free milk applying Metrosep Carb 2 - 150/4.0
• Brochure 8.107.5002: Metrosep Carb 2 column overview
• Application Note P-60: Ethylene and propylene glycol by PAD
• Application Note P-61: Sugar analysis in honey by PAD
• Application Note P-62: Sugars in apple drinks by PAD
• Application Note P-63: Mannitol, rhamnose, lactulose and lactose in serum by PAD
• Application Note P-64: Sugars and sugar acids by low-pressure gradient
• Application Note P-65: Sugars including sucrose and cellobiose
• Application Note P-67: Sorbitol and sucrose in soap by PAD
• Application Note U-71: Nitrite, bromide and nitrate in artificial seawater by UV/VIS
• Info MI-2015-2-AP-1: Are you made of sugar? (Chromatography Today)