The development of a fluorimetric method for the determination of chlorophyll a

Abstract

Klorofil a je zeleni pigment prisutan u biljkama, cijanobakterijama i algama te je ključan u procesu fotosinteze. U prehrambenoj industriji njegovi se derivati koriste kao prirodna bojila biljnoga podrijetla, a u posljednje vrijeme raste broj literaturnih primjera u kojima se klorofilu a pripisuje antikancerogena, antimutagena i antioksidativna aktivnost. Koncentracija klorofila a u biljnim ekstraktima u laboratoriju se rutinski određuje mjerenjem apsorbancije pri apsorpcijskom maksimumu klorofila a (660 nm) te primjenom empirijskih jednadžbi. Imajući u vidu veću osjetljivost i selektivnost fluorimetrije kao analitičke metode u odnosu na UV/ViS spektrofotometriju te dvojbenu pouzdanost empirijskih jednadžbi, cilj ovoga rada bio je razvoj metode za fluorimetrijsko određivanje koncentracije klorofila a u biljnim ekstraktima. Standard klorofila a izoliran je iz cijanobakterije Arthrospira platensis primjenom tankoslojne preparativne kromatografije.Chlorophyll a is a green pigment present in plants, cyanobacteria and algae and is crucial in the photosynthesis process. Besides the usage of its derivatives as natural dyes of plant origin in food industry, there is a growing number of literary examples in which anticancer, antimutagenic and antioxidant activity is attributed to chlorophyll a. Concentration of chlorophyll an in plant extracts is routinely determined in the laboratory by measuring the absorbance at the absorption maximum of chlorophyll a (660 nm) and using empirical equations. Having in mind greater sensitivity and selectivity of fluorimetry as analytical method compared to UV/ViS spectrophotometry and reliability of empirical equations, the aim of this study was to develop a method for fluorimetric determination of chlorophyll a concentration in plant extracts. Chlorophyll a used as standard was isolated from cyanobacteria Arthrospira platensis by preparative thin layer chromatography