HPLC Method Optimization and Validation for Determination of Lycopene in Tomato ( Lycopersicon esculentum, Mill.) Fruits

Abstract

The aim of this work was to develop efficient and accurate protocol that can measure lycopene in tomato. A total of ten tomato varieties loose and cluster types were selected for the study. All the tomatoes were harvested at commercial ripening stage and prepared after 3 days of storage at 18°C and relative humidity (RH) of 80%. For analysis purpose each tomato was cut, quartered, chopped and frozen in liquid nitrogen and then subjected to grinding. The finely ground tomato samples were immediately filled into air-tight plastic tube and stored in freezer (-80 °C) for about two weeks. Finally, the tomato samples were prepared in different composition of extraction solvent before subjected to HPLC analysis. Mobile phase composition Acetonitrile/Methanol (50:50, v/v) added with Triethylamine 9 μM; extraction solvents hexane/acetone/ethanol (50:25:25 v/v/v); re-dissolving residue in Tetrahydrofuran, followed Acetonitrile/Methanol (15:30:55, v/v/v); flow rate 0.6 mL/min and λdetection near to 472nm were showed most suitable for lycopene determination in tomato, achieving best characteristic spectral profile with precision (RSD<15), accuracy and recovery (≥81.7%), and sensitive detection limit (0.0156 μg/mL) within separation of ~21 minutes. The result showed that HPLC is most accepted and efficient method for determining lycopene in tomato.