Analysis of cyanobacterial toxins by hydrophilic interaction liquid chromatography–mass spectrometry

• Published in: Journal of Chromatography A Vol. 1028; no. 1; pp. 155 - 164
• Main Authors: Dell’Aversano, Carmela, Eaglesham, Geoffrey K, Quilliam, Michael A
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 27.02.2004; Elsevier
• Subjects: Anatoxins; Bacteria; Bacterial Toxins - analysis; Bacteriology; Biological and medical sciences; Chromatography, Liquid - methods; Cyanobacteria - chemistry; Cyanotoxins; Cylindrospermopsin; Fundamental and applied biological sciences. Psychology; Hydrophilic interaction chromatography; Microbiology; Microcystins; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Saxitoxins; Spectrometry, Mass, Electrospray Ionization - methods; Toxins; Anatoxins; Hydrophilic interaction chromatography; Cyanotoxins; Cylindrospermopsin; Bacteria; Toxins; Saxitoxins; Microcystins; Chemical analysis; Angular nitrogen heterocycle; Nitrogen heterocycle; Coupled method; Cyclic peptides; Toxoid; Liquid chromatography; Tricyclic compound; Electrospray; Hydrophilic Interaction Liquid Chromatography; Toxin; Anabaena; Cyanobacteria; Bicyclic compound; Mass spectrometry
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2003.11.083

The combination of hydrophilic interaction liquid chromatography with electrospray mass spectrometry (HILIC–MS) has been investigated as a tool for the analysis of assorted toxins produced by cyanobacteria. Toxins examined included saxitoxin and its various analogues ( 1– 18), anatoxin-a (ATX-a, 19), cylindrospermopsin (CYN, 20), deoxycylindrospermopsin (doCYN, 21), and microcystins-LR ( 22) and -RR ( 23). The saxitoxins could be unequivocally detected in one isocratic analysis using a TSK gel Amide-80 column eluted with 65% B, where eluent A is water and B is a 95% acetonitrile/water solution, both containing 2.0 mM ammonium formate and 3.6 mM formic acid. The analysis of ATX-a, CYN and doCYN required 75% B isocratic. Simultaneous determination of 1– 21 was also possible by using gradient elution. HILIC proved to be suitable for the analysis of microcystins, but peak shape was not symmetric and it was concluded that these compounds are best analysed using existing reversed-phase methods. The HILIC–MS method was applied to the analysis of field and cultured samples of Anabaena circinalis and Cylindrospermopsis raciborskii. In general, the method proved quite robust with similar results obtained in two different laboratories using different instrumentation.