An optimized analytical method based on C8 core-shell reverse phase chromatographic separation and
high resolution mass spectral (HRMS) detection is developed for a fast analysis of unbound phytoche-
latins (PCs) in plants. Its application to analysis of Clinopodium vulgare L. is demonstrated where proper
PCs liberatingandpreservationconditionswereemployedusingdithiotreitolintheextractionstep.A
baseline separationofglutathione(GSH)andphytochelatinsfrom2to5(PC2–PC5) for3minwas
achieved atconventionalHPLCbackpressure,withdetectionlimitsfrom3ppt(forGSH)to2.5ppb(for
PC5). Itisshown,thattheuseofHRMSwithtandemmassspectral(MS/MS)capabilitiespermitsaddi-
tional widerangescreeningabilityforiso-phytochelatinsandPCsimilarcompounds,basedonexact
mass andfragmentspectrainapostacquisitionmanner