HPLC Separation of Phthalic Acids using Hydrogen Bonding

  Phthalic acid, isophthalic acid and terephthalic acid are all isomers of each other. Being structurally similar, they can present difficulties to reverse-phase HPLC separation. Methods that require high organic concentrations in the mobile phase can cause dewetting in many reverse-phase columns. SHARC 1 column can be operated in anhydrous conditions and uses hydrogen bonding as the mechanism of separation. Here, phthalic acids were separated in pure acetonitrile (ACN), with the ability to adjust retention times by adding methanol (MeOH) to the mobile phase with formic acid and ammonium formate as buffer, making the method MS-compatible. Can also be UV detected at 270nm.


Column Sharc 1, 3.2x100 mm, 5 µm, 100A
Mobile Phase MeCN/MeOH
Buffer AmFm, Formic acid
Flow Rate 1.0 ml/min
Detection UV, 270 nm


Class of Compounds Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds Phthalic acid, Terephthalic acid, Isophthalic acid
Application Analytes:
Phthalic Acid