Separation of Benzoic and Acetylbenzoic acid in Hydrogen-Bonding Mode on Zodiac HST HB Column

A robust and sensitive method for analyzing a broad range of organic acids was developed using the Zodiac HST HB column, which employs hydrogen-bonding as the primary retention mechanism. This approach is compatible with LC/MS detection in negative ion mode, enabling trace-level quantification of organic acids across diverse sample matrices.

The method utilizes a mobile phase composed of acetonitrile (ACN), methanol (MeOH), and a low concentration of ammonia, ensuring MS compatibility. In hydrogen-bonding chromatography, acetonitrile acts as a weaker solvent, while alcohols like methanol serve as stronger eluents, allowing fine-tuning of analyte retention. Both gradient and isocratic conditions can be optimized based on the application.