The Use of High Pressure Liquid Chromatography with Fluorimetric Detection for Chlorophyll A Determination in Natural Extracts of Chloropigments and their Degradation Products

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Abstract
To realize chlorophyll a analysis, a method is proposed that combines the sensitivity of fluorimetric determinations and the inherent selectivity and accuracy of chromatographic separations. A partial application of the overlapping resolution mapping (O.R.M.) method led to the use of a methanol:acetone:water (75:22:3, vol/vol) mixture for the isocratic separation by high pressure liquid chromatography on a reserved phase C-18 column of at least 16 chloropigments and degradation products in a natural extract. Ten replicate measurements of a 50 .mu.g .cntdot. l-1 standard showed a reproducibility of 3%, while a detection limit of 84 pg chlorophyll a was estimated. [Chlorophyll a is widely used as a parameter for living phytoplankton in natural waters.].