HPLC Separation of Methyl Paraben, Benzonitrile, Propyl Paraben, and Toluene on Mixed-Mode and Reverse Phase Columns

Parabens are common preservatives in pharmaceutical and cosmetic industries. They are esters of p-hydroxybenzoic acid. Method for separation of methyl paraben, propyl paraben, benzonitrile and toluene was developed on a Obelisc R column. All four compounds are neutral and are retained by reverse-phase mechanism. In case of reversed-phase stationary phase, no effect of pH is observed. Retention time for all four compounds changes on an Obelisc R column when pH is changed. pH of the mobile phase affects ionization state of stationary phase. Obelisc R column has C12 carbon chain and carboxylic acid with pKa of 4. At lower pH (pH 2, TFA), carboxylic acid of stationary phase is not ionized and thus adds hydrophobicity to stationary phase. Obelisc R column can be used for analysis of basic, acidic and neutral compounds with suitable detection techniques - UV, ELSD, CAD, LC/MS.


Column Obelisc R, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmAc, TFA
Flow Rate 1.0 ml/min
Detection UV, 270 nm


Class of Compounds Preservatives, Neutral
Analyzing Compounds Methylparaben, Benzonitrile, Propyl paraben, Toluene
Application Analytes: