Spectrophotometric quantification of pigments in Anabaena flos-aquae: investigation of cell disruption methods

Articles in Press

Document Type : Research Article

Authors

1 Department of Bioengineering, School of Chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran

2 Department of Chemical Engineering, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran

Abstract

Chlorophyll a and phycobiliproteins are ubiquitous cyanobacterial secondary products of vast scientific and technological interest and their accurate measurement is necessary in many application fields. In this work, three different cell disruption methods of freeze-thawing, ball milling, and ultrasonication were extensively investigated for the extraction and spectrophotometric determination of Chlorophyll a and phycobiliproteins from the filamentous cyanobacterium Anabaena flos-aquae. The effect of the duration of the last two treatments was also studied. While all methods were found to return statistically equivalent results for Chlorophyll a under the test conditions (4.078±0.256 to 4.706±0.201 µg/mL), ultrasonication and ball milling for longer durations outperformed others. In addition, the interference of the overlapping Chlorophyll a peak during the phycobiliproteins quantification was analyzed and ultrasonication for extended duration was shown to offer not only a reasonably high yield but a low Chlorophyll a interference as well. Furthermore, common equation sets for the calculation of phycobiliprotein concentration from the spectral data were compared and potential discrepancies were highlighted. This study is believed to provide insights for future research and address the mistakes and inconsistencies regarding the extraction and spectrophotometric determination of cyanobacterial pigments.

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Iranian Journal of Chemistry and Chemical Engineering

Articles in Press, Accepted Manuscript
Available Online from 29 October 2023

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