HPLC Separation of Carboxylic Acids

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Condition 1

Column Newcrom  B, 3.2x50 mm, 3 µm, 100A
Mobile Phase MeCN/H2O- 30/70%
Buffer H3PO4 - 0.5%
Flow Rate 0.5 ml/min
Detection UV, 275 nm
HPLC separation of carboxylic acids (nicotinic acid, benzoic acid and acetylbenzoic acid) on Primesep B2 column using an isocratic method by reverse-phase and anion-exchange mechanisms. The elution of carboxylic acids using the same isocratic method and mobile phase can be reversed by using a reverse-phase cation-exchange mechanisms of the Primesep 100 column. The mobile phase is water, acetonitrile (MeCN, ACN) and formic acid with a UV detector.

Condition 2

Column Primesep B2, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer Formic Acid
Flow Rate 1.0 ml/min
Detection UV, 250 nm


Class of Compounds Acid, Hydrophilic
Analyzing Compounds Nicotinic acid, Benzoic acid, Acetylbenzoic acid, 1.4-acetylbenzoic Acid
Application Analytes:
Acetylbenzoic Acid
Benzoic Acid
Nicotinic Acid/Niacin (3-pyridinecarboxylic acid)