Statistical Optimization of Reverse Phase High Performance Liquid Chromatography for the Analysis of Caffeine Paracetamol and its Degradation Product p-aminophenol

Milkica Crevar,* Branka Ivkovic, Sote Vladimirov, Vesna Kuntica and Zorica Vujic

Department of Pharmaceutical Chemistry and Drug Analysis, Faculty of Pharmacy, Belgrade
aDepartment of Physical Chemistry, Faculty of Pharmacy, Belgrade
* Corresponding author: E-mail:

Paracetamol, analgoantipyretic and anti-inflammatory drug, is available in numerous pharmaceutical formulations in common combination with caffeine and some other drug substances. p-aminophenol is paracatamol’s process-related impurity that also may be present in formulations containing paracetamol. This paper presents a RP-HPLC method for simultaneous determination of caffeine, paracetamol and p-aminophenol, using chromatographic system Hewlett Packard 1100 series. In defining optimal RP-HPLC chromatographic conditions for the separation of these three compounds, experimental design was applied. Completely separation was achieved using Zorbax Extend C18 column (150 mm × 4.6 mm, 5 µm), with mobile phase consisting of methanol-phosphate buffer pH 6 (20:80 v/v), flow rate of 1 mL/min, and column temperature of 30 °C. UV detection was performed at 230 nm.

Keywords: RP-HPLC, caffeine, paracetamol, p-aminophenol, statistical optimization