THE IDENTIFICATION OF CHLOROPHYLL AND ITS DERIVATIVES IN THE PIGMENT MIXTURES: HPLC-CHROMATOGRAPHY, VISIBLE AND MASS SPECTROSCOPY STUDIES

Abstract

This work represents an application of the Visible Spectrometry, High Pres­sure Liquid Cromatography and Electronspray Ionisation-Mass Spectrometry analysis for the identification of chlorophyll and its derivatives, pheophytin and chlorophyllide, in their purified mixtures (the chlorophyll, pheophytin and chlorophyllide fraction, respectively). The chlorophyll, pheophytin and chlorophyllide fractions were obtained by using a pigment extraction from the plant material (Spinacia Oleracea L), coupled with the column chromatography. The main components in the chlorophyll, pheophytin and chlorophyllide fractions were identified as chlorophyll a & b, pheophytin a & b and chlorophyllide a, respectively. Spectrophotometrically determined concentration ratios of the main identified pigments, chlorophyll a/b (in the chlorophyll fraction), pheophytin a/b (in the pheophytin fraction) and chlorophyllide a/b (in the chlorophyllide fraction) were: 4.95/1, 1.05/1 and 6.89/1, respectively.