Advancing Liquid Chromatography (LC) Separation Science

Ultra-Performance Liquid Chromatography (UPLC) Technology works at higher efficiencies with a much wider range of linear velocities, flow rates, and backpressures. Higher flow rates and increased throughput result in superior resolution and sensitivity. UPLC substantially reduces time and cost-per-sample analysis, while improving the quality of decision making results.  Waters’ ACQUITY UPLC Columns’ patented sub-2-μm hybrid particle chemistry enable chromatographers to harness the separation efficiency improvement s this separation technology delivers, thereby improving their business’ productivity and profitability. The ACQUITY UPLC Column family continues to evolve and expand with the addition of particles and stationary phases for the analyses of both small molecules as well as more complex biopharmaceuticals.

  • Description


BEH (Ethylene Bridged Hybrid) Technology

In 2004, Waters revolutionized liquid chromatography with the introduction of the ACQUITY UPLC System. At the heart of the system was Waters second generation hybrid, the Ethylene Bridged Hybrid [BEH] particle.

In 2005, BEH Technology preparative particle sizes (2.5, 3.5, and 5 μm) with the introduction of XBridge between HPLC and UPLC Technology platforms.

BEH Technology particles offer excellent peak shape and efficiency for basic analytes, a rational array of chromatographic selectivity and improvements in chemical stability at mobile phase extremes, particularly at elevated pH.

CSH (Charged Surface Hybrid) Technology

CSH particles incorporate a low level surface charge that improves sample loadability and peak asymmetry in low-ionic-strength mobile phases, while maintaining the mechanical and chemical stability inherent to BEH particle technology.

HSS (High Strength Silica) Technology

Silica particles designed for high mechanical stability and appropriate morphology provide long column lifetimes and UPLC efficiencies at high pressures.

VanGuard Column Protection

Many complex sample matrices encountered in pharmaceutical, natural product, environmental and industrial chemical analysis prematurely shorten analytical column lifetime and degrade chromatographic performance. Waters VanGuard Pre-columns help improve UPLC Column performance by removing particulate and chemical contamination that may be present in the mobile-phase stream without compromising chromatographic performance.

Columns for Glycan Analysis

ACQUITY UPLC Glycan BEH Columns utilize hybrid-silica BEH Technology particles functionalized with a stable, amide-containing species. The high resolving power of these columns is due in part to fully-porous particles with an optimal concentration of amide ligands for associated applications. These columns are optimized for use with ACQUITY UPLC Systems with fluorescence (FLR) detection for separating the released and 2-aminobenzamide (2-AB) labeled N-linked glycans from various biotherapeutics. Available and well documented methods are used to achieve HILIC-based separations of both neutral and charged labeled glycan species.

Columns for Oligonucleotide Separations

ACQUITY UPLC Oligo C18 Columns are well suited for the analysis of detritylated oligonucleotides using ion-pair, reversed-phase (RP) chromatography. Component resolution is equivalent to separations obtained using traditional capillary gel electrophoresis (CGE) methods. However, Waters UPLC Technology significantly reduces analysis times. Columns packed with sub-2-μm BEH particles, functionalized with an optimal concentration of C18 ligands, can deliver N from N-1 resolution of synthetic oligonucleotide sequences up to approximated 50 nucleotides in length. In addition, mass spectrometry-compatible separation eluents enable users to obtain molecular weights of the separated oligonucleotides using defined hyphenated UPLC-MS methods.

Peptide Analysis

ACQUITY UPLC Peptide BEH C18, 130Å or 300Å Columns offer a wide usable pH range and outstanding  temperature stability for applications ranging from proteomics to peptide mapping.   ACQUITY UPLC CSH C18, 130Å Peptide Columns with a low-positive charged surface can be used with standard TFA-containing eluents or with a weaker acid modifier such as formic acid, eliminating the need to compromise between a reversed-phase eluent that delivers sharp peaks or one that minimizes reduction of the MS signal.

Protein Separations

Waters ethylene-bridged hybrid (BEH) SEC and RP Protein Separation Columns are specifically designed and QC tested with BEH protein standards, ensuring unmatched batch-to-batch consistency for increased confidence in validated methods. The SEC-based offerings are ideally suited for the accurate determination of low levels of protein aggregates contained in therapeutic monoclonal antibodies up to 10X faster than traditional HPLC-based SEC. By comparison (data not shown), the BEH C4, 300Å UPLC Columns separate proteins of various sizes, hydrophobicities, and isoelectric points while maximizing recovery and minimizing protein carryover.

Amino Acid Analysis

AccQTag Ultra C18, 1.7 µm Columns combined with Waters ACQUITY UPLC System provides a total application solution optimized for accurate, reliable, and reproducible analysis of free amino acids (AAs). These columns deliver enhanced resolution of derivatized amino acids to ensure accurate and precise qualitative and quantitative results. These columns utilize Waters AccQ•Tag Ultra Chemistry to deliver assured AA determination performance as part of protein characterization, cell culture monitoring, and food and feed analysis.