Separation and quantification of pigments from natural phototrophic microbial populations

Abstract A simple high-performance liquid chromatography (HPLC) method is described for the rapid (approx. 20 min) simultaneous separation and identification of the major chlorophylls and carotenoids from phytoplankton cells and phototrophic sulfur bacteria. Lyophilized samples were extracted with acetone in the dark at room temperature. Pigments were eluted from a silica column with a hexane-acetone mixture (80: 20, v/v). About 20 algal and bacterial chlorophyll and carotenoid pigments could be separated in one run. The method allowed for the detection of trace amounts of major carotenoids (> approx. 6 ng · 1^?1) and of chlorophylls and pheophytins (> approx. 200 ng · 1^?1) in natural samples. The method has been applied to samples from the metalimnion of Lake Vechten (The Netherlands) and has proved very useful in estimating algal and bacterial pigments simultaneously with respect to depth distribution and biomass changes of the microbial populations.