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 ƒÊM; 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 ƒÉdetection 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
ƒÊg/mL) within separation of ~21 minutes. The result showed that HPLC is most accepted and efficient method for determining lycopene in tomato.