ACQUITY UPLC Columns

ACQUITY UPLC Columns

Increase speed, sensitivity, and resolution, without compromising data integrity

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 (IVD) harness the potential of sub-2-µm particles for faster separations with higher resolution and sensitivities. ACQUITY UPLC Columns
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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

ACQUITY UPLC BEH Column Characteristics
ACQUITY UPLC BEH C18 Columns
  • Universal C18 column
  • Suitable for a diverse range of analytes
  • High efficiency
  • Industry-leading mobile phase pH 1-12 and temperature compatibility
ACQUITY UPLC BEH C8 Columns
  • Lower hydrophobicity and retention than a C18 column
  • Suitable for a diverse range of analytes
  • High efficiency
  • Industry-leading mobile phase pH 1-12 and temperature compatibility
ACQUITY UPLC BEH Shield RP18 Columns
  • Contains an embedded polar group combined with the hydrophobicity of C18
  • Complementary selectivity to a C18 column
  • Enhanced peak shape for basic compounds
  • Compatible with 100% aqueous mobile phases
  • High efficiency
ACQUITY UPLC BEH Phenyl Columns
  • Industry leading chemical stability
  • Suitable for a diverse range of analytes
  • High efficiency
  • Complimentary selectivity to straight-chain alkyl phases
ACQUITY UPLC BEH HILIC Columns
  • Improves the retention of very polar basic analytes
  • Complementary selectivity to reversed-phase
  • High efficiency
  • Exceptional chemical stability and peak shape compared to silica-based HILIC stationary phases
ACQUITY UPLC BEH Amide Columns
  • Designed to retain highly polar analytes that are too polar to retain by reversed-phase
  • Exceptional chemical stability at high pH and high temperature
  • Ideal for sugar/carbohydrate analysis
  • Exceptional retention of polar analytes spanning a wide range of polarity

ACQUITY UPLC CSH Column Characteristics

ACQUITY UPLC CSH Column Characteristics
ACQUITY UPLC CSH C18 Columns
  • Universal column choice for UV and MS applications
  • Suitable for a broad range of compounds
  • Alternate selectivity to BEH C18
  • Exceptional peak shape and increased load capacity
  • Suited for basic compounds at low pH
ACQUITY UPLC CSH Fluoro-Phenyl Columns
  • Exceptional selectivity for positional isomers, halogenated compounds and polar compounds
  • Enhanced retention of acidic compounds compared to traditional PFP-bonded stationary phases
ACQUITY UPLC CSH Phenyl-Hexyl Columns
  • Complementary selectivity to straight-chain alkyl phases
  • Exceptional peak shape for basic compounds under low- and high-pH conditions
  • Increased retention for acidic compounds at low pH

ACQUITY UPLC HSS Column Characteristics

ACQUITY UPLC HSS Column Characteristics
ACQUITY UPLC HSS T3 Columns
  • Ideal for the enhanced retention of polar and non-polar compounds
  • Compatible with 100% aqueous mobile phases
  • Mechanically stable at elevated pressures
ACQUITY UPLC HSS C18 Columns
  • General purpose silica-based C18 chemistry
  • Suitable for a broad range of compounds
  • Provides exceptional peak shapes and low pH stability
  • Delivers increased retention compared to hybrid organic/inorganic C18 columns
ACQUITY UPLC HSS C18 SB Columns
  • Low coverage silica-based C18 chemistry
  • Provides alternate selectivity for compounds impacted by silanophilic interactions
  • Increased silanol activity results in greater retention of basic compounds while reducing the retention of non-basic analytes
  • Provides exceptional peak shape and imparts Selectivity for Bases at low pH (SB)
ACQUITY UPLC HSS PFP Columns
  • Pentafluorophenyl (PFP) chemistry
  • Industry-leading reproducibility and peak shape
  • Ideally suited for planar aromatic compounds, positional isomers, halogenated compounds and polar analytes
ACQUITY UPLC HSS Cyano Columns
  • Provides low hydrophobicity and unique selectivity compared to straight-chain-alkyl columns
  • Ultra-stable retention
  • Exceptional peak shape and reproducibility under low- to mid-pH conditions
  • Compatible with both reversed-phase and normal-phase techniques

ACQUITY UPLC Application-Specific Column Characteristics

ACQUITY UPLC Application-Specific Column Characteristics
ACQUITY UPLC Peptide BEH C18 Columns
  • Suited for the reversed-phase analysis of hydrophobic and hydrophilic peptides at low and high pH
  • Outstanding peak capacity and peak shape in TFA or FA when compared to 100% silica based C18 columns
  • Two pore sizes (130 Å and 300 Å) for a wide range of peptides

Try ACQUITY UPLC Peptide BEH C18 Columns

ACQUITY UPLC Peptide CSH C18 Columns
  • Outstanding peak capacities with formic acid for MS-based applications
  • Excellent performance with TFA for optical based applications
  • Less dependence on TFA or FA to achieve optimal peak capacity
  • Accepts greater peptide mass loads than many competitive technologies for detection of low-level impurities
  • At low pH with the positive charge on CSH, you get less secondary interactions with the particle

Try ACQUITY Premier Peptide CSH C18 Columns

ACQUITY UPLC Peptide HSS T3 Columns
  • Separations that require silica-based selectivity
  • Separations that necessitate increased peptide retention
  • Ideally suited for small, hydrophilic peptides

Try ACQUITY UPLC Peptide HSS T3 Columns

ACQUITY UPLC Protein SEC Columns
  • Sub-2-μm particle columns for UPLC separations (2.5 μm for 450 Å)
  • Pore size options include: 125 Å, 200 Å, and 450 Å
  • Specifically designed and QC tested with BEH125, BEH200, or BEH450 protein standards to help ensure batch-to-batch consistency
  • 10x faster than traditional HPLC-based SEC

Try ACQUITY UPLC Protein BEH SEC Guards and Columns

ACQUITY UPLC Protein BEH C4 Columns
  • Separates proteins of various sizes, hydrophobicities, and isoelectric points
  • Tolerates extreme pH and temperature, and provides minimal secondary interactions
  • Improves sensitivity for phosphorylated proteins and low-level intact and subunit mAb analysis
  • High efficiency separations with 1.7 µm particles

Try ACQUITY UPLC Protein BEH C4 Columns

ACQUITY UPLC Oligonucleotide Columns
  • Suited for the analysis of detritylated oligonucleotides using ion-pair, reversed-phase chromatography

Try ACQUITY UPLC Oligonucleotide Columns

ACQUITY UPLC Glycan Columns
  • Suited for the HILIC analysis of neutral and acidic glycans

Try ACQUITY UPLC Glycan Columns

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.

Explore ACQUITY UPLC Glycoprotein Columns

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.

Explore ACQUITY UPLC and XBridge Glycan BEH Amide Columns

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. 

Explore ACQUITY UPLC Peptide 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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ACQUITY Premier Columns
Achieve increases in sensitivity and analyte recovery while reducing column passivation time and mitigating unwanted analyte/surface interactions with ACQUITY Premier Columns.

ACQUITY Premier System
Experience the ultimate in sensitivity, reproducibility, and speed. The ACQUITY Premier LC-MS provides the best sensitivity and peak shape for analytical separations.

Bio LC Columns
Waters series of ion-exchange, size-exclusion, reversed-phase, hydrophobic-interaction, and hydrophilic-interaction chromatography columns offer you the diverse range of chromatographic selectivities needed to help characterize today's biotherapeutics and biosimilars. These columns support HPLC, UPLC, UHPLC, LC-MS analyses or lab-scale purification of oligonucleotides, amino acids, peptides, proteins and glycans.

VanGuard Column Protection
Prolong analytical column lifetime while maintaining chromatographic performance with VanGuard Pre-columns and cartridges.

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ACQUITY UPLC (Ultra-Performance Liquid Chromatography) Columns engineered to excel in the intricate processes of compound separation, analysis, and quantification across diverse sample types. These columns are meticulously crafted with state-of-the-art technologies to cater to the exacting demands of contemporary analytical laboratories. Below is an in-depth exploration of the principal features and advantages of ACQUITY UPLC Columns:

Sub-2-μm Particle Size: ACQUITY UPLC Columns harness sub-2-μm particle size technology, facilitating extraordinarily high-resolution separations. By leveraging these minute particles, the columns achieve optimal packing efficiency, resulting in narrow peak widths and heightened separation efficacy for compounds with closely similar elution profiles.

Efficiency: The fusion of sub-2-μm particle size and sophisticated column chemistry engenders unparalleled efficiency in peak resolution and chromatographic performance. This heightened efficiency empowers the separation of intricate mixtures with superior peak capacity, thereby enabling more precise compound identification and quantification.

Resolution: ACQUITY UPLC Columns has exceptional resolution capabilities, enabling the discrimination of closely related compounds that might co-elute on conventional chromatographic columns. Such augmented resolution proves pivotal in applications necessitating the detection and quantification of trace-level components within complex samples.

Sensitivity: The enhanced efficiency and resolution offered by ACQUITY UPLC Columns contribute to heightened sensitivity in detection. Sharper peak profiles and enhanced baseline separation facilitate more accurate measurements, even for analytes present at low concentrations.

Accuracy: ACQUITY UPLC Columns uphold analytical accuracy to the highest degree by ensuring reliable separation and quantification of target compounds. The consistent performance of these columns yields reproducible results across multiple analyses, mitigating variability and enhancing data reliability.

Speed: ACQUITY UPLC Columns facilitate rapid analysis times while preserving resolution and sensitivity. The efficient packing of particles and optimized column design expedite compound elution, thereby reducing overall analysis time and bolstering laboratory throughput.

Reliability: Engineered with robust materials and subject to stringent quality control measures, ACQUITY UPLC Columns epitomize high reliability and exhibit exceptional batch-to-batch reproducibility. This reliability guarantees consistent performance over time, minimizing instrument downtime and optimizing productivity within the laboratory.

ACQUITY UPLC Columns epitomize a state-of-the-art solution for high-performance liquid chromatography, delivering unmatched efficiency, resolution, sensitivity, accuracy, speed, and reliability across a broad spectrum of analytical applications.

FAQs About ACQUITY UPLC Columns

What types of stationary phases are available for ACQUITY UPLC columns?

ACQUITY UPLC columns offer a wide range of stationary phases to suit various analytical needs. These include C18, C8, phenyl, and BEH (Ethylene Bridged Hybrid) phases, among others. Each phase is designed to provide optimal performance for different types of compounds and separation challenges. For example, C18 and C8 phases are ideal for hydrophobic compounds, while phenyl phases are suited for aromatic compounds.

What are the key applications of ACQUITY UPLC columns?

ACQUITY UPLC columns are used in a wide range of applications due to their high resolution, sensitivity, and speed. Key applications include pharmaceutical analysis, where they are employed for drug development, quality control, and stability testing. They are also used in biopharmaceutical research for protein and peptide analysis, as well as in environmental testing for detecting pollutants and contaminants. Additionally, ACQUITY UPLC columns are valuable in food and beverage testing, clinical research, and metabolomics, providing precise and reliable results across various fields.

What is the typical lifespan of an ACQUITY UPLC column?

The typical lifespan of an ACQUITY UPLC column can vary depending on factors such as sample cleanliness, operating conditions, and maintenance practices. Generally, these columns can last from several months to over a year. Proper sample preparation, such as filtration, helps extend the column"s life by preventing clogging and contamination. Regular maintenance, including flushing with appropriate solvents and correct storage, is essential for maximizing column longevity.

How do ACQUITY UPLC columns improve analytical performance?

ACQUITY UPLC columns significantly enhance analytical performance by leveraging advanced technology and design. The use of sub-2 µm particles in these columns provides higher resolution, sensitivity, and faster separation compared to traditional HPLC columns. This allows for sharper peaks and more accurate quantification of analytes. The robust column design ensures stable and consistent performance under high pressure, enabling efficient handling of complex mixtures.