Quantitative analysis of extracted phycobilin pigments in cyanobacteria—an assessment of spectrophotometric and spectrofluorometric methods

Abstract

Phycobilins are an important group of pigments that through complementary chromatic adaptation optimize the light-harvesting process in phytoplankton cells, exhibiting great potential as cyanobacteria species biomarkers. In their extracted form, concentrations of these water-soluble molecules are not easily determined using the chromatographic methods well suited to solvent-soluble pigments. Insights regarding the quantitative spectroscopic analysis of extracted phycobilins also remain limited. Here, we present an in-depth study of two methods that utilize the spectral properties of phycobilins in aqueous extracts. The technical work was carried out using high-purity standards of phycocyanin, phycoerythrin, and allophycocyanin. Calibration parameters for the spectrofluorometer and spectrophotometer were established. This analysis indicated the possibility of detecting pigments in concentrations ranging from 0.001 to 10 μg cm−3. Fluorescence data revealed a reproducibility of 95 %. The differences in detection limits between the two methods enable the presence of phycobilins to be investigated and their amounts to be monitored from oligotrophic to eutrophic aquatic environments.