ACQUITY UPC2 - FOOD APPLICATION NOTEBOOK

Significance of the Topic

Modern food and supplement testing laboratories face a growing need for rapid, sensitive and environmentally responsible methods to separate, identify and quantify a wide range of analytes, from fat-soluble vitamins and carotenoids to pesticides and flavor compounds. Waters® UltraPerformance Convergence Chromatography™ (UPC 2®) leverages sub-2 µm particle columns and compressed CO₂ mobile phase to combine the selectivity of normal-phase separations with efficiency commonly associated with reversed-phase LC. This approach significantly reduces organic solvent use and analysis times while improving resolution, making it especially valuable for green initiatives and high-throughput environments.

Objectives and Overview

● Demonstrate UPC 2 applications across key food and supplement matrices (vitamins, lipids, carotenoids, aroma compounds, dietary supplements, pesticides)
● Show method performance: speed, resolution, sensitivity, linearity, reproducibility, solvent savings
● Illustrate orthogonal separations complementing or exceeding HPLC and GC methods

Methodology and Instrumentation

● UPC 2 separations on ACQUITY UPC 2 BEH, HSS, BEH C18 SB and Trefoil chiral columns (1.7 µm sub-2 µm particles)
● Compressed CO₂ with polar co-solvents (e.g., isopropanol, ethanol, methanol) and additives for pH control
● Detection via PDA, ELSD, MS (Q-Tof, Xevo TQ-S) with RADAR and PIC scans for confident peak ID
● Sample prep: saponification for vitamins, simple extraction for lipids, direct injection for nonpolar analytes; QuEChERS and SPE for pesticides

Main Findings

● Fat-soluble vitamins A, D₃, K isomers resolved in < 10 min with excellent R² (> 0.999) and repeatability (< 6% RSD)
● Triacylglycerols and fatty acids profiled in oils—free fatty acids directly injected, no derivatization, < 9 min separations
● β-carotene and astaxanthin quantified in supplements in < 2 min, LOD < 100 ng/mL, R² > 0.999, solvent use − 85%
● Vanilla extract authenticity screening; detection of coumarin adulterant and natural phenolics in < 2 min
● Chiral resolution of flavor compounds (carvone, linalool, nerolidol) in < 3 min on Trefoil columns
● Rapid enantiomeric pesticide assays: tebuconazole, flutriafol, etc., at ppt levels in commodities; MRM provides selectivity with ease of MS detection

Benefits and Practical Impact

● Dramatic increases in sample throughput and productivity—analysis times up to 10× faster than HPLC/GC
● Significant reductions in organic solvent purchase and disposal costs (green chemistry)
● Simplified workflows, minimal sample prep and direct injection of organic extracts
● Orthogonal separations to complement existing LC or GC methods, improving confidence in complex matrices

Future Trends and Applications

● Broader adoption of UPC 2 and sub-2 µm CSPs for chiral and achiral assays in food, environmental and pharmaceutical labs
● Integration with advanced MS tools (high-resolution, ion mobility) for rapid “omics” profiling of lipids, vitamins and flavor molecules
● Continued method scale-up strategies to semi-prep and preparative SFC for natural product isolation and purification
● Enhanced data-driven optimization (AI-assisted method scouting) for faster route to robust separations

Conclusion

UPC 2 technology unifies the speed and resolution of modern UPLC with the selectivity and green credentials of supercritical fluid chromatography. Its versatility across diverse food, supplement and crop testing applications—from routine quantitation to advanced chiral analyses—offers laboratories a powerful tool for high-performance, sustainable separations.