The Comparison of Spectrophotometric Method and High-Performance Liquid Chromatography in Photosynthetic Pigments Analysis

Abstract

Problem statement: Spectrophotometric method and High-Performance Liquid Chromatography (HPLC) are the most useful tools to photosynthetic pigments analysis. The main objective of this reaserch was to compere the results obtained by these two independent methods in experiments under controlled laboratory conditions. Approach: Comparison between spectrophotometric and HPLC methods for pigment analysis was carried out on Baltic cyanobacterium Geitlerinema amphibium, which is distinguished by photoacclimation capacity and wide range of changes in pigment cell concentration depending on light and temperature conditions. Results: The degree of relationship between the methods of pigment measurement was described by correlation coefficients, which were 0.970, 0.978 and 0.997 for the results of chlorophyll a, carotenoids and carotenoids to chlorophyll a ratio, respectively. Conclusion: It was found that spectrophotometric method gave comparable results to HPLC method, especially when there were not degradation products of chlorophyll a in samples. In contrast with HPLC method it allows to determine the only total amount of carotenoid pigments. Estimation of the carotenoids to chlorophyll a ratio is useful to understand the reaction of organism to changing environmental light condition.