Reverse Phase HPLC Columns – High-Efficiency C18 and C8 Columns for Reliable Separations

Product Line Overview

What Is Reverse Phase HPLC Chromatography?

Reverse phase HPLC (RP-HPLC) is the most widely used separation technique in liquid chromatography, relying on hydrophobic interactions between the stationary phase and analyte molecules.

In this mode, the stationary phase is non-polar while the mobile phase is relatively polar—typically a mixture of water and organic solvents such as methanol or acetonitrile. This reversed polarity allows for controlled retention of analytes based on their hydrophobic character.

GL Sciences reverse phase columns are manufactured using high-purity silica chemically bonded with non-polar ligands such as C18, C8, C4, Phenyl, and Cyano, offering stable and predictable retention for compounds across a broad polarity range.

Through precise surface modification and end-capping, these columns minimize secondary interactions, ensuring sharp peak shapes and reproducible retention even for basic or polar analytes.

By adjusting the ratio of organic solvent in the mobile phase—or by modifying buffer pH and ionic strength—analysts can fine-tune separation selectivity, elution strength, and analysis time.

This flexibility enables high-resolution separations of complex mixtures such as pharmaceuticals, metabolites, and biomolecules without sacrificing robustness or reproducibility.

Available in multiple particle sizes (from sub-2 µm to 5 µm) and column formats, GL Sciences columns ensure compatibility with both HPLC and UHPLC systems, providing consistent performance across a wide pH range (1–10) and extended column lifetime under demanding conditions.

Retention Control and Selectivity

Reverse phase selectivity depends strongly on the chemical structure of the bonded phase and the nature of the analyte.

GL Sciences offers a broad lineup of bonded phases—C18, C8, C4, Phenyl, and Cyano—each providing distinct separation characteristics.

• C18 (ODS Series): Delivers strong hydrophobic retention and balanced selectivity for general-purpose use.
• C8: Offers faster elution and shorter analysis times for moderately hydrophobic compounds.
• C4: Suited for large biomolecules like peptides and proteins, ensuring better recovery and peak shape.
• Phenyl: Provides π–π interaction selectivity for aromatic and conjugated analytes.
• Cyano: Offers intermediate polarity and alternative selectivity for polar compounds.

These options allow analysts to fine-tune method performance according to compound polarity, functional group chemistry, and target application.

Wide pH Range and Column Durability

All GL Sciences reverse phase columns are designed for stable operation across a pH range of 1–10.

High-purity silica combined with optimized surface bonding ensures resistance to hydrolysis and phase collapse under both acidic and basic conditions.

This chemical robustness extends column lifetime, enabling repeated analyses without changes in retention or selectivity.

Especially the InertSustain Series, featuring enhanced surface shielding and end-capping, demonstrates superior long-term stability even in high-aqueous or high-pH mobile phases.

Peak Shape and Reproducibility

Reproducibility is a key performance factor in analytical chromatography.

GL Sciences ensures exceptional batch-to-batch consistency through strict manufacturing control of silica purity, bonding density, and end-capping process.

As a result, all reverse phase columns deliver sharp, symmetrical peaks with minimal tailing—even for basic analytes that typically show poor peak shapes.

This precision manufacturing supports reliable quantitative analysis, regulatory compliance, and long-term reproducibility in validated workflows.

Application Versatility & Use Cases

GL Sciences reverse phase HPLC columns are widely used in analytical laboratories across diverse fields:

• Pharmaceutical Analysis: Assay, impurity profiling, stability studies
• Biochemical Research: Peptide, protein, and metabolite analysis
• Environmental Testing: Pesticide and organic contaminant detection
• Food and Beverage Analysis: Vitamins, amino acids, and additives

Their broad selectivity, chemical stability, and reproducible retention make them the first choice for method development, routine analysis, and regulatory quality control.

Technical Resources & Support