ACQUITY UPLC Columns
Increase speed, sensitivity, and resolution, without compromising data integrity
Increasing sample throughput, achieving unmatched resolution, and improving analyte sensitivity are priorities for your separation goals. With Waters ACQUITY UPLC Columns, you can meet all your method requirements and substantially reduce your analysis time and cost-per-sample.
Waters ACQUITY UPLC Columns’ sub-2-μm particle chemistries enable chromatographers to harness the separation efficiency improvements at increased flow rates of ultra-performance liquid chromatography (UPLC) technology, thereby improving analyte resolution and sensitivity as well as business productivity and profitability.
ACQUITY UPLC Columns
Mitigate unwanted interactions between analytes and metal ions associated with stainless-steel column hardware with ACQUITY Premier Columns.
Specifications
Overview
- Best-in-class column efficiency for superior separation speed, sensitivity, and resolution
- 8 UPLC particles of various substrate technologies, including Ethylene Bridged Hybrid (BEH), High Strength Silica (HSS), and Charged Surface Hybrid (CSH)
- 22 stationary phases, including C18, Phenyl-Hexyl, C8, Embedded-Polar, C4, HILIC, Amide, Diol, Cyano, and PFP
- 7 application-specific UPLC Column chemistries for SEC, proteins, peptides, oligonucleotides, glycan, and amino acid analysis
- More than 50 different column dimensions and configurations, with internal column diameters ranging from 1.0–4.6 mm and column lengths ranging from 30–300 mm
- Prolonged column lifetimes with optional VanGuard Cartridges specifically designed using ACQUITY stationary phases
Recommended Use: For improved speed, sensitivity, and resolution in methods development.
ACQUITY UPLC BEH Column Characteristics
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Universal C18 column -
Suitable for a diverse range of analytes
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High efficiency
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Industry-leading mobile phase pH 1-12 and temperature compatibility
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Lower hydrophobicity and retention than a C18 column
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Suitable for a diverse range of analytes
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High efficiency
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Industry-leading mobile phase pH 1-12 and temperature compatibility
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Contains an embedded polar group combined with the hydrophobicity of C18
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Complementary selectivity to a C18 column
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Enhanced peak shape for basic compounds
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Compatible with 100% aqueous mobile phases
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High efficiency
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Industry leading chemical stability
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Suitable for a diverse range of analytes
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High efficiency
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Complimentary selectivity to straight-chain alkyl phases
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Improves the retention of very polar basic analytes
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Complementary selectivity to reversed-phase
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High efficiency
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Exceptional chemical stability and peak shape compared to silica-based HILIC stationary phases
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Designed to retain highly polar analytes that are too polar to retain by reversed-phase
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Exceptional chemical stability at high pH and high temperature
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Ideal for sugar/carbohydrate analysis
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Exceptional retention of polar analytes spanning a wide range of polarity
ACQUITY UPLC CSH Column Characteristics
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Universal column choice for UV and MS applications
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Suitable for a broad range of compounds
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Alternate selectivity to BEH C18
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Exceptional peak shape and increased load capacity
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Suited for basic compounds at low pH
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Exceptional selectivity for positional isomers, halogenated compounds and polar compounds
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Enhanced retention of acidic compounds compared to traditional PFP-bonded stationary phases
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Complementary selectivity to straight-chain alkyl phases
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Exceptional peak shape for basic compounds under low- and high-pH conditions
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Increased retention for acidic compounds at low pH
ACQUITY UPLC HSS Column Characteristics
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Ideal for the enhanced retention of polar and non-polar compounds
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Compatible with 100% aqueous mobile phases
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Mechanically stable at elevated pressures
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General purpose silica-based C18 chemistry
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Suitable for a broad range of compounds
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Provides exceptional peak shapes and low pH stability
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Delivers increased retention compared to hybrid organic/inorganic C18 columns
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Low coverage silica-based C18 chemistry
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Provides alternate selectivity for compounds impacted by silanophilic interactions
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Increased silanol activity results in greater retention of basic compounds while reducing the retention of non-basic analytes
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Provides exceptional peak shape and imparts Selectivity for Bases at low pH (SB)
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Pentafluorophenyl (PFP) chemistry
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Industry-leading reproducibility and peak shape
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Ideally suited for planar aromatic compounds, positional isomers, halogenated compounds and polar analytes
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Provides low hydrophobicity and unique selectivity compared to straight-chain-alkyl columns
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Ultra-stable retention
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Exceptional peak shape and reproducibility under low- to mid-pH conditions
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Compatible with both reversed-phase and normal-phase techniques
ACQUITY UPLC Application-Specific Column Characteristics
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Suited for the reversed-phase analysis of hydrophobic and hydrophilic peptides at low and high pH
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Outstanding peak capacity and peak shape in TFA or FA when compared to 100% silica based C18 columns
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Two pore sizes (130 Å and 300 Å) for a wide range of peptides
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Outstanding peak capacities with formic acid for MS-based applications
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Excellent performance with TFA for optical based applications
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Less dependence on TFA or FA to achieve optimal peak capacity
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Accepts greater peptide mass loads than many competitive technologies for detection of low-level impurities
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At low pH with the positive charge on CSH, you get less secondary interactions with the particle
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Separations that require silica-based selectivity
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Separations that necessitate increased peptide retention
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Ideally suited for small, hydrophilic peptides
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Sub-2-μm particle columns for UPLC separations (2.5 μm for 450 Å)
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Pore size options include: 125 Å, 200 Å, and 450 Å
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Specifically designed and QC tested with BEH125, BEH200, or BEH450 protein standards to help ensure batch-to-batch consistency
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10x faster than traditional HPLC-based SEC
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Separates proteins of various sizes, hydrophobicities, and isoelectric points
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Tolerates extreme pH and temperature, and provides minimal secondary interactions
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Improves sensitivity for phosphorylated proteins and low-level intact and subunit mAb analysis
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High efficiency separations with 1.7 µm particles
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Suited for the analysis of detritylated oligonucleotides using ion-pair, reversed-phase chromatography
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Suited for the HILIC analysis of neutral and acidic glycans
Get superior peak shape for basic analytes
Reversed-phase bonded phases typically have poor peak shape for basic compounds when using formic acid, even at analytical mass loads. However, ACQUITY UPLC CSH Columns are the exception. These rugged UPLC Columns provide superior peak shape for basic analytes with formic acid or other low ionic-strength, acidic mobile phases. The controlled, low-level, positive surface charge bonded to the ethylene-bridged hybrid (BEH) particles provides excellent peak shape for bases—without the need for the use of ion-pairing reagents.
The ruggedness you want, the repeatability you need
ACQUITY UPLC BEH 1.7 μm fully porous columns provide superior analyte peak shape, efficiency, and chemical stability to any separation. ACQUITY UPLC BEH Columns are available in both reversed-phase and HILIC separation modes, with chemistries that provide selectivities for a wide range of compounds. With the ruggedness to operate at extreme pH conditions, ACQUITY UPLC BEH Columns enable separation scientists to use a wide pH range to influence retention, selectivity, and sensitivity for ionizable compounds.
Achieve flexibility in method development
Chromatographic laboratories need to quickly and easily develop robust methods that are compatible with all modern chromatographic detection modes and transferrable to laboratories and sites that operate different LC system platforms. With ACQUITY UPLC CSH and XSelect Columns, method development scientists in all application areas can achieve a wide range of selectivities without compromising performance across attributes, including superior peak shape for basic compounds, low column bleeds, excellent batch-to-batch reproducibility, and high efficiency.
Applications
Experience HILIC for large biomolecules, an alternative high-resolution technology
Hydrophilic Interaction Chromatography (HILIC) has been widely used to separate small polar compounds, yet its application to large biomolecules, other than released glycans, has been surprisingly limited. With Waters wide-pore ACQUITY UPLC Glycoprotein BEH Amide 300 Å, 1.7 μm Column and new ideas on separation methods, now you can use HILIC to glean previously unattainable information from intact proteins (with or without glycosylation), protein fragments, and complex, released glycans. The ACQUITY UPLC Glycoprotein BEH Amide 300 Å, 1.7 µm Column offers high resolution glycopeptide mapping without limitations due to peptide/glycan size or composition.
Robust LC or LC-MS analyses of biotherapeutic glycoproteins
The analysis of complex N- and 0-linked glycans composed of frequently similar and repeating sugar moieties requires a highly discriminating chromatographic method. Hydrophilic liquid interaction chromatography (HILIC) using UPLC, HPLC, or UHPLC columns containing Waters amide bonded, Ethylene Bridged Hybrid (BEH) Technology particles separates compounds based on differing hydrophilicity. ACQUITY UPLC and XBridge Glycan BEH Amide 130 Å Columns are best suited for the analysis of released, N-labeled glycans using pre-column labeling with 2-AB, 2-AA, or Waters innovative and enabling RapiFluor-MS reagent. Each batch of the Glycan BEH Amide 130 Å chemistry is specifically QC tested with a complex mixture of 2-AB labeled glycans from a mAb digest to help ensure consistent column performance in validated methods.
Orthogonal techniques for quantitation of sialic acids and monosaccharides
Evidence for reliable and consistent glycosylation of glycoprotein therapeutics is typically obtained through LC or LC-MS analysis of released N-linked glycans. However, drug discovery and development organizations, as well as regulatory agencies, frequently find value using orthogonal techniques to determine and/or confirm quantitative levels of sialic acids and monosaccharides contained in a therapeutic protein due to potential effect on biological activity and stability of the drug.
Waters offers UPLC, UHPLC, and HPLC-based C18 columns and defined application conditions that can assist you with the analysis and quantitation of these important glycan derived components.
Explore ACQUITY UPLC and XBridge BEH Columns for sialic & monosaccharide analyses
Reproducibly generate high quality reversed-phase peptide separations
ACQUITY UPLC Peptide BEH C18, 130 Å and 300 Å Columns offer a wide pH range and outstanding temperature stability for applications ranging from proteomics to peptide mapping. ACQUITY UPLC CSH C18, 130 Å 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. The Peptide HSS T3, 100% silica-based particle offering is the ideal choice for the separation of small, polar peptides since retentivity is greater than that obtained using either of Waters BEH or CSH, hybrid-based peptide separation columns.
Exceptional resolution of synthetic oligonucleotide mixtures
Chromatography of synthetic oligonucleotides and DNA/RNA species of less than approximately 12,000 base pairs can be performed using reversed-phase or ion-exchange gradient techniques capable of resolving sample components based on minor changes in size (i.e. charge). ACQUITY UPLC Oligonucleotide C18, 1.7 μm Columns and XBridge Oligonucleotide C18, 2.5 μm Columns are well suited for the analysis of detritylated oligonucleotides using ion-pair, reversed-phase chromatography. Waters Oligonucleotide columns are packed with BEH Technology particles, ensuring column longevity under demanding separation conditions while maintaining outstanding separation performance.
Explore ACQUITY UPLC and XBridge Oligonucleotide BEH Columns
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