HPLC AND UHPLC - Application Highlights

Significance of the Topic

High-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) underpin modern analytical chemistry across pharmaceuticals, food safety, environmental monitoring, and life-science research. Achieving optimal throughput, resolution, and robustness depends on selecting the appropriate stationary phase, particle technology, and column configuration. Waters’ portfolio of BEH, CSH, HSS, and solid-core columns addresses the diverse separation challenges posed by acidic, basic, neutral, polar, and nonpolar analytes.

Objectives and Overview

This brochure guides chromatographers in choosing the right column technology and instrument pairing to maximize efficiency and reproducibility. It presents:

• An introduction to Waters particle technologies
• Column nomenclature matched to Nano/Micro, UPLC, UHPLC, HPLC, and preparative systems
• Instrument dispersion and bandspread considerations for seamless method transfers
• Representative applications showcasing diverse compound classes

Methodology and Instrumentation

Waters particle platforms deliver targeted performance:

• BEH Technology: Ethylene-bridged hybrid particles tolerate extremes of pH (1–12) and temperature, ideal for broad-scope separations and method robustness.
• CSH Technology: Charged surface hybrid phases optimize peak shape for basic analytes under low-ionic-strength mobile phases and facilitate rapid method development.
• HSS Particle Technology: High-strength pure silica offers enhanced retention and selectivity for polar species at pressures up to 18,000 psi.
• Solid-Core Technology (CORTECS): Superficially porous particles reduce backpressure and improve mass sensitivity and resolution, scalable from UHPLC to HPLC.

Column formats and nomenclature are aligned to instrument bandspread:

• Nano/Micro (ID 75–300 μm, <2 μm particles)
• UPLC (ID 1.0–2.1 mm, sub-2 μm particles)
• UHPLC (ID 2.1–3.0 mm, 2–3 μm particles)
• HPLC (ID 3.0–4.6 mm, 3–5 μm particles)
• Preparative (>7.8 mm, >5 μm particles)

Main Results and Discussion

Comparative evaluations highlight:

• Improved peak capacity and faster equilibration with solid-core columns versus fully porous media.
• Superior pH stability and universal applicability of BEH phases for pharmaceuticals, food additives, and environmental analytes.
• Exceptional retention of polar compounds on HSS and HILIC chemistries for metabolites and sugar analysis.
• Efficient separation of basic drugs, steroids, vitamins, and natural products across isocratic and gradient modes.

Benefits and Practical Applications

Laboratories benefit from:

• Scalable method transfer across platforms without reoptimization
• Reduced solvent consumption and analysis time
• Consistent peak shape and sensitivity for trace-level quantitation
• Flexibility to analyze diverse matrices—from drinking water to complex biologics

Future Trends and Opportunities

Emerging directions include:

• AI-driven column and method selection tools
• Integration of UHPLC with MS/MS workflows for high-throughput screening
• Development of novel hybrid and multi-modal stationary phases
• Advances in sub-2 μm and sub-1 μm particle manufacturing
• Green chromatography initiatives to minimize organic solvent usage

Conclusion

Waters’ comprehensive suite of column technologies and system-matched configurations empowers analysts to tackle the most demanding separation challenges. By understanding particle chemistries, instrument dispersion, and scalable formats, laboratories can achieve robust, reproducible, and high-throughput methods across a spectrum of applications.

Reference

No external references were provided in the source document.