Sviluppo e validazione del metodo analitico tramite HPLC per la quantificazione di fitormoni in campioni di foglie di pomodoro. Development and validation of HPLC method to quantify phythormones in tomato leaf samples

The project I took part in at the Faculty of Pharmacy of University of Porto consisted in the set-up of a rapid high-performance liquid chromatography (HPLC-UV) method for the simultaneous detection of seven phytohormones (abscisic acid-ABA, gibberellic acid-GA, indoleacetic acid-IAA), jasmonic acid-JA, methyl-jasmonate acid-MeJA, salicylic acid-SA, 6-benzylaminopurine-BAP) in tomato leafs. The chromatographic separation was achieved with a C18 reversed-phase column using a gradient elution of acid formic 0.1% and methanol (60:40 v/v) and acid formic 0.1% and methanol (40:60 v/v) with the detector wavelengths set at 254 nm and 206 nm. The method was also validated according to International Council for Harmonisation (ICH) guidelines, evaluating linear range, linearity, the limit of detection (LOD), the limit of quantitation (LOQ), precision, and accuracy.