HPLC Analysis of Etidronic Acid, Citric Acid and Mannitol on Amaze HD Mixed-Mode Column

Application description

Aromatic organic acids are used as building blocks in chemistry, additives in various formulations and counter-ions in drug production. In most of the cases, these acids are hydrophilic and acidic in nature with limited retention in reversed-phase chromatography. We have developed a simple, robust and isocratic method to separate mandelic, gallic, phthalic and 4-hydroxybenzoic acids on the Amaze TR HPLC column. All four acids are retained and separated by combination of weak reversed-phase and weak anion-exchange mechanisms. The baseline separation is achieved within 5 minutes. This method can be used for the analysis of hydrophilic and hydrophobic organic and inorganic acids with LC/MS-compatible conditions.

Conditions of Experiment
Column: Amaze HD
Separation Modes: HILIC, anion-exchange
Column Dimenstions: 4.6x50 mm, 5 um, 100A
Mobile Phase: 80% ACN with 20 mM AmFm pH 3
Detection: ELSD, 50°C
Sample: 2 mg/ml
Injection: 2 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Drug, Organic acid, Preservative, Sugar
Nature of compounds: Acidic, Hydrophilic, Neutral, Polar
Compounds: Citric acid, Etidronic acid, Mannitol