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Recommendations on the quantitative analysis of pheophorbides, photosensitizers present in algal biomass intended as food supplement
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SYSNO ASEP 0544796 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Recommendations on the quantitative analysis of pheophorbides, photosensitizers present in algal biomass intended as food supplement Author(s) Činčárová, Dominika (MBU-M)
Hájek, Jan (MBU-M) ORCID
Dobřichovský, M. (CZ)
Lukeš, Martin (MBU-M) ORCID
Hrouzek, Pavel (MBU-M) ORCIDNumber of authors 5 Article number 102298 Source Title Algal Research-Biomass Biofuels and Bioproducts. - : Elsevier - ISSN 2211-9264
Roč. 56, JUNE 2021 (2021)Number of pages 10 s. Language eng - English Country NL - Netherlands Keywords Pheophorbide a ; Analysis ; Extraction ; Chlorophyllase inactivation ; Algae ; Chlorophyll degradation Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects TN01000048 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) Method of publishing Limited access Institutional support MBU-M - RVO:61388971 UT WOS 000663410300004 EID SCOPUS 85105254433 DOI 10.1016/j.algal.2021.102298 Annotation Pheophorbide a is the major photosensitizer present in algal biomass and its presence in food supplement was linked to human health issues in the past. Although several pheophorbide a quantification methods have been proposed including the United States Pharmacopeia-National Formulary protocol (USP-NFP), there are open methodological issues which make their results questionable. In the present study we have tested optimal extraction conditions for pheophorbide a from a disintegrated biomass of green algae Chlorella and Haematococcus, widely used as food supplement. Further we have compared the quantification of pheophorbide a in algal extracts using HPLC-HRMS with the spectrophotometric quantification after USP-NFP in Haematococcus biomass and oleoresin samples. We have demonstrated that USP-NFP provide considerably (1.7-2.4 times) higher pheophorbide contents compared to HPLC-HRMS quantification using a pheophorbide a analytical standard. This was mainly caused by the fact that USP-NFP integrates the concentration of all pheophorbide variants present in the samples. We have detected nine pheophorbide analogues in the original algal sample as well as in the final sample of USP-NFP, of which pheophorbide a represented only similar to 40%. However, this explains the values obtained by USP-NFP only partly. We also show that 60% acetone is the most suitable extraction solvent for pheophorbide a from various algal samples, however, we noted that such extracts might exhibit high activity of chlorophyllase generating pheophorbide a during the extraction process. This activity differed between studied Chlorella and Haematococcus biomass extracts. We have achieved the chlorophyllase inactivation by a short-term high-temperature treatment (90 degrees C, 5 min). Acceptable stability of pheophorbide a and only minor degradation of chlorophyll to pheophorbide a has been observed at these conditions. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2022 Electronic address https://www.sciencedirect.com/science/article/pii/S221192642100117X?via%3Dihub
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