Separation of Generic Compounds by HPLC on Zodiac HST OG

Mixed-mode HPLC columns offer the unique capability to separate compounds with widely differing chemical properties in a single run. The Zodiac HST OG column enables simultaneous analysis of acidic, basic, and neutral compounds using either isocratic or gradient elution.

In this application, a diverse set of analytes is separated using a gradient of acetonitrile (ACN), including:

• Neutral hydrophilic compounds: uracil, phenol, hydroquinone

• Neutral hydrophobic compound: toluene

• Hydrophilic acidic compound: benzoic acid

• Hydrophilic basic compound: lutidine

Neutral compounds are retained primarily through reversed-phase interactions. The hydrophilic acidic compound (benzoic acid) becomes more hydrophobic at low pH, enhancing its retention by the reversed-phase mechanism. Basic compounds are retained via cation-exchange interactions, while hydrophobic basic compounds are influenced by both reversed-phase and ion-exchange mechanisms.

All compounds are effectively separated within 14 minutes using a short column format, demonstrating high efficiency and versatility. This method is suitable for the analysis of a broad range of polar and nonpolar compounds on a single column and within one chromatographic run.

The Zodiac HST OG column operates in one or more of the following modes: reversed-phase, ion-exchange, ion-exclusion, and HILIC, making it a powerful general-purpose column for mixed-mode separations across a wide array of analyte types.