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Quantification and profiling of lipophilic marine toxins in microalgae by UHPLC coupled to high-resolution orbitrap mass spectrometry

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Abstract
During the last decade, a significant increase in the occurrence of harmful algal blooms (HABs), linked to repet- itive cases of shellfish contamination has become a public health concern and therefore, accurate methods to detect marine toxins in different matrices are required. In this study, we developed a method for profiling lipophilic marine microalgal toxins based on ultra-high-performance liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (UHPLC-HR-Orbitrap MS). Extraction of selected toxins (okadaic acid (OA), dinophysistoxin-1 (DTX-1), pectenotoxin-2 (PTX-2), azaspiracid-1 (AZA-1), yessotoxin (YTX) and 13-desmethyl spirolide C (SPX-1)) was optimized using a Plackett-Burman design. Three key algal species, i.e., Prorocentrum lima, Protoceratium reticulatum and Alexandrium ostenfeldii were used to test the extraction efficiency of OA, YTXs and SPXs, respective- ly. Prorocentrum micans, fortified with certified reference so- lutions, was used for recovery studies. The quantitative and confirmatory performance of the method was evaluated according to CD 2002/657/EC. Limits of detection andquantification ranged between 0.006 and 0.050 ng mL−1 and 0.018 to 0.227 ng mL−1, respectively. The intra-laboratory reproducibility ranged from 6.8 to 11.7 %, repeatability from 6.41 to 11.5 % and mean corrected recoveries from 81.9 to 119.6 %. In addition, algae cultures were retrospectively screened for analogues and metabolites through a homemade database. Using the ToxID software programme, 18 toxin der- ivates were detected in the extract of three toxin producing microalgae species. In conclusion, the generic extraction and full-scan HRMS approach offers an excellent quantitative per- formance and simultaneously allows to profile analogues and metabolites of marine toxins in microalgae.
Keywords
Plackett-Burman, Ultra-high-performance liquid chromatography high-resolution Orbitrap mass spectrometry, PROROCENTRUM-LIMA, FOOD AVAILABILITY, ALGAL TOXINS, OKADAIC ACID, PROTOCERATIUM-RETICULATUM, ALEXANDRIUM-OSTENFELDII, SPIROLIDE-C, DINOPHYSIS-ACUMINATA, LIQUID-CHROMATOGRAPHY, Validation, Harmful algae, Yessotoxins, Spirolide, NORWEGIAN BLUE MUSSELS

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Chicago
Orellana Mancilla, Gabriel, Lieven Van Meulebroek, Sarah Van Vooren, Maarten De Rijcke, Michiel Vandegehuchte, Colin Janssen, and Lynn Vanhaecke. 2015. “Quantification and Profiling of Lipophilic Marine Toxins in Microalgae by UHPLC Coupled to High-resolution Orbitrap Mass Spectrometry.” Ed. Aldo Laganà. Analytical and Bioanalytical Chemistry 407 (21): 6345–6356.
APA
Orellana Mancilla, G., Van Meulebroek, L., Van Vooren, S., De Rijcke, M., Vandegehuchte, M., Janssen, C., & Vanhaecke, L. (2015). Quantification and profiling of lipophilic marine toxins in microalgae by UHPLC coupled to high-resolution orbitrap mass spectrometry. (A. Laganà, Ed.)ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 407(21), 6345–6356.
Vancouver
1.
Orellana Mancilla G, Van Meulebroek L, Van Vooren S, De Rijcke M, Vandegehuchte M, Janssen C, et al. Quantification and profiling of lipophilic marine toxins in microalgae by UHPLC coupled to high-resolution orbitrap mass spectrometry. Laganà A, editor. ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 2015;407(21):6345–56.
MLA
Orellana Mancilla, Gabriel, Lieven Van Meulebroek, Sarah Van Vooren, et al. “Quantification and Profiling of Lipophilic Marine Toxins in Microalgae by UHPLC Coupled to High-resolution Orbitrap Mass Spectrometry.” Ed. Aldo Laganà. ANALYTICAL AND BIOANALYTICAL CHEMISTRY 407.21 (2015): 6345–6356. Print.
@article{6925014,
  abstract     = {During the last decade, a significant increase in the occurrence of harmful algal blooms (HABs), linked to repet- itive cases of shellfish contamination has become a public health concern and therefore, accurate methods to detect marine toxins in different matrices are required. In this study, we developed a method for profiling lipophilic marine microalgal toxins based on ultra-high-performance liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (UHPLC-HR-Orbitrap MS). Extraction of selected toxins (okadaic acid (OA), dinophysistoxin-1 (DTX-1), pectenotoxin-2 (PTX-2), azaspiracid-1 (AZA-1), yessotoxin (YTX) and 13-desmethyl spirolide C (SPX-1)) was optimized using a Plackett-Burman design. Three key algal species, i.e., Prorocentrum lima, Protoceratium reticulatum and Alexandrium ostenfeldii were used to test the extraction efficiency of OA, YTXs and SPXs, respective- ly. Prorocentrum micans, fortified with certified reference so- lutions, was used for recovery studies. The quantitative and confirmatory performance of the method was evaluated according to CD 2002/657/EC. Limits of detection andquantification ranged between 0.006 and 0.050 ng mL\ensuremath{-}1 and 0.018 to 0.227 ng mL\ensuremath{-}1, respectively. The intra-laboratory reproducibility ranged from 6.8 to 11.7 \%, repeatability from 6.41 to 11.5 \% and mean corrected recoveries from 81.9 to 119.6 \%. In addition, algae cultures were retrospectively screened for analogues and metabolites through a homemade database. Using the ToxID software programme, 18 toxin der- ivates were detected in the extract of three toxin producing microalgae species. In conclusion, the generic extraction and full-scan HRMS approach offers an excellent quantitative per- formance and simultaneously allows to profile analogues and metabolites of marine toxins in microalgae.},
  author       = {Orellana Mancilla, Gabriel and Van Meulebroek, Lieven and Van Vooren, Sarah and De Rijcke, Maarten and Vandegehuchte, Michiel and Janssen, Colin and Vanhaecke, Lynn},
  editor       = {Lagan{\`a}, Aldo},
  issn         = {1618-2642},
  journal      = {ANALYTICAL AND BIOANALYTICAL CHEMISTRY},
  language     = {eng},
  number       = {21},
  pages        = {6345--6356},
  title        = {Quantification and profiling of lipophilic marine toxins in microalgae by UHPLC coupled to high-resolution orbitrap mass spectrometry},
  url          = {http://dx.doi.org/10.1007/s00216-015-8637-y},
  volume       = {407},
  year         = {2015},
}

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