XBridge Columns

XBridge Columns for High pH and Method Development

XBridge HPLC and UHPLC columns, utilizing BEH Particle Technology, are built to withstand high pH, high temperatures , and other extreme operating conditions, including volatile eluents, for a wide range of small molecule and biomolecule LC-MS analysis. These columns are available in a broad range of sorbent particles and sizes. XP Columns (2.5 µm) for rapid analytical UHPLC separations, (3.5 and 5 µm) for reliable analytical method development and OBD Prep (5 and 10 µm) for purification chromatography.

High pH stability and chemical resistance for reliability and longevity
Maximum method development flexibility
Seamless method transfer of analytical to preparative LC

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XBridge HPLC Analytical and Preparative Columns

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Accelerated High pH Stability Test of Competitive Columns

HPLC and UHPLC pH Lifetime and Reliability

These HPLC and UHPLC columns offer mobile phase pH stability that is higher than any other commercially available chromatographic phase, making it ideal for method development. Unlike approaches that claim high pH resistance, the pH stability of these columns is created by utilizing proprietary surface modifications and is incorporated into the particle synthesis process.

The columns were tested under accelerated pH 10 stability test conditions against some of the most popular chromatographic phases that claim extended high pH stability. The C18 columns showed very little degradation in chromatographic performance, exceeding the performance of the best silica column by over 1000%.

General Use Chemistry Selection

Column	Performance Benefit	UHPLC	HPLC
C18	General purpose, reversed-phase chromatography, ideally suited for method development due to extreme pH stability and applicability to the broadest range of compound classes.	2.5 μm XP	Analytical to Prep 3.5, 5, 10 μm
C8	General purpose reversed-phase chromatography, ideally suited for method development due to extreme pH stability and applicability to the broadest range of compound classes.	2.5 μm XP	Analytical to Prep 3.5, 5, 10 μm
Shield RP18	Alternate selectivity compared to straight chain C18, particularly with phenolic analytes. Compatible with 100% aqueous-phase composition.	2.5 μm XP	Analytical to Prep 3.5, 5, 10 μm
Phenyl	Excellent for method development for alternate selectivity, particularly in regard to polyaromatic compounds. Unique level of pH stability for a phenyl bonded phase.	2.5 μm XP	Analytical to Prep 3.5, 5 μm
HILIC	Excellent for retention of very polar, basic, water soluble analytes. Specifically designed and tested for HILIC separations using mobile phases containing high concentrations of organic solvent.	2.5 μm XP	Analytical to Prep 3.5, 5 μm
Amide	Rugged HILIC stationary phase designed to separate a wide range of very polar compounds. Especially good at separating carbohydrates (saccharides) using high concentrations of organic modifier, elevated temperature, and high pH. Compatible with all modern detectors including MS, ELSD, UV, and fluorescence.	2.5 μm XP	Analytical to Prep 3.5, 5 μm

Method Development Flexibility with pH Range

These columns are designed to eliminate the compromise of sorbent selection and deliver the flexibility to work under any mobile phase, temperature and pH chromatography conditions, which is a requirement for method development. Even the most challenging sample mixtures can be quickly resolved with a sample method development protocol consisting of two mobile phase pH’s and two organic solvents. See below for examples of C18, Shield RP18 and Phenyl chromatography in low pH in Methanol, high pH in Methanol, in low pH in Acetonitrile, and in high pH in Acetonitrile.

Method Development XBridge Low pH in Methanol XBridge High pH in Methanol XBridge Low pH in Acetonitrile XBridge High pH in Acetonitrile

LC of Proteins, Peptides, Glycans, and Oligos

BEH HPLC/UHPLC Columns are available for application specific analysis. These are dedicated high-performance sorbents for isolation, analysis, and characterization of biomolecules. They support UHPL and HPLC for LC-MS analyses as well as lab scale purification of oligonucleotides, amino acids, peptides, proteins and associated glycans.

Peptide HPLC: ideally suited for reversed-phase chromatography of a wide range of peptides including large and small, acidic and basic, and hydrophilic and hydrophobic species while providing the ability to scale up to prep.
Protein SEC: deliver higher resolution protein aggregate from monomer separations in less time than many traditional SEC columns.
Protein: separate proteins of various sizes, hydrophobicities, and isoelectric points while maximizing recovery and minimizing protein carryover while providing the ability to scale up to prep.
Glycans: perfect for analyzing released and 2-aminobenzamide (2-AB) labeled N-linked glycans from various biotherapeutics.
Oligonucleotide: well-established ion-pair, reversed-phase chromatography method for separating detritylated synthetic oligonucleotide samples while providing the ability to scale up to prep.

Column	Performance Benefit	UHPLC	HPLC
Peptide C18, 300Å	For reversed-phase chromatography of peptides. Suitable for a low pH through high pH LC and outstanding temperature stability for applications ranging from proteomics to peptide mapping.	N/A	Analytical to Prep 3.5, 5, 10 μm
Peptide C18, 130Å	Ideally suited for reversed-phase chromatography of a wide range of peptides including large and small, acidic and basic, as well as hydrophilic and hydrophobic species. Stable at low through high temperatures.	N/A	Analytical to Prep 3.5, 5, 10 μm
Protein C4, 300Å	Well suited for HPLC-based protein and polypeptide characterization, pH and temperature stable.	N/A	Analytical to Prep 3.5, 5, 10 μm
Protein SEC, 125Å	Ideal HPLC based SEC separation of proteins ranging in size from 1,000 to 80,000 daltons.	N/A	3.5 μm
Protein SEC, 200Å	Ideal HPLC based SEC separation of proteins ranging in size from 10,000 to 450,000 daltons.3.5 µm BEH SEC particles are flow and pressure tolerant for increased sample throughput compared to use of silica-based, SEC columns containing >5 µm particles.	N/A	3.5 μm
Protein SEC, 450Å	Ideal HPLC based SEC separation of proteins ranging in size from 100,000 to 1.5 million daltons.	2.5 μm	3.5 μm
Oligo C₁₈, 130Å	pH and temperature stable, for synthetic DNA and RNA. Well suited for the analysis of detritylated oligonucleotides by means of ion-pair, reversed-phase chromatography.	2.5 μm	N/A
Glycan Amide, 130Å	Designed for the LC and LC-MS analysis of fluorescently labeled glycans. Specifically QC tested with 2-AB labeled human mAb IgG derived glycans.	2.5 μm XP	3.5 μm

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BEH Technology for extreme conditions in reverse phase, HILIC, C18, C8, SEC, silica, phenyl, Octyl, size exclusion, LCMS, and Amide stationary phases

Mechanical Strength and Chemical Stability

Ethylene Bridged Hybrid (BEH) Technology synthesis creates particles that ensure extreme performance and longer column lifetimes under all chromatographic conditions. Hybrid particle technology offers many advantages over conventional silica-based particles, including the ability to control surface silanol activity for better reproducibility, peak shape and retention time. The BEH particle (1.7, 2.5, 3.5, 5, and 10 μm) is prepared from two high purity monomers: tetraethoxysilane (TEOS) and bis(triethoxysilyl) ethane (BTEE), which results in a highly stable, pH resistant, and mechanically strong particle allowing seamless transfer from analytical to preparative separations.

Method Scaling and Transfer

All of these Columns are designed for stability to preserve separation efficiency and seamless method transfer throughout the available particle sizes 2.5, 3.5, 5, and 10 μm prep. These configurations and particle sizes meet the most demanding chromatographic challenges. Whether you require robust HPLC methods, or preparative scaling for product isolation.

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Maximum efficiency/30% lower backpressure

Preparative Separation Performance

Waters innovative Optimum Bed Density (OBD) Preparative design ensures direct scalability that is independent of particle size. The combination of the revolutionary OBD format with the BEH base particle delivers the most stable, efficient, and reproducible preparative LC columns available. XBridge OBD Preparative Columns for LC users benefit from high pH and temperature stability for increased mass loading for basic compounds.

Protect your Column without Compromise

Don’t take chances… Complex sample matrices encountered in pharmaceutical, natural product, environmental, and industrial chemical analysis can prematurely shorten analytical UHPLC/HPLC column lifetime and degrade chromatographic performance. Using guard cartridges like VanGuard Protection products improve analytical performance by removing particulate and chemical contamination that may be present in the mobile-phase stream—without compromising chromatographic performance.

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Minimal Chromatographic Effects with VanGuard Cartridges

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