Rational design of linear calibration experiments for the quantitative estimation of chlorophyll a using high-performance liquid chromatography, atomic absorption spectrometry and electronic absorption spectrometry

Abstract

There is considerable difference in published molar absorption coefficients for chlorophyll a. This paper involves estimating the molar absorption coefficient in acetone by calibrating against measured Mg using AAS. Chlorophyll can then be routinely estimated using peak areas in diode array detector HPLC. In order to account for the main errors in the estimates, designed linear calibration experiments are required, involving 14 measurements at five levels, with four instrumental and five dilution replicates. Three errors namely, lack-of-fit, dilution and instrumental errors can be calculated for each type of measurement, guiding the experimenter as to where problems may arise in the experimental procedure.