Microwave oven and boiling waterbath extraction of hepatotoxins from cyanobacterial cells.
Low-cost, straightforward methods for the extraction of microcystins and nodularins from cyanobacterial cells were developed using a microwave oven and boiling waterbath. The use of organic solvents, such as methanol, which can interfere with sensitive analytical procedures, e.g. immunoassays, can thus be avoided. Analysis by protein phosphatase inhibition assay and high performance liquid chromatography indicated that purified microcystin-LR was unaffected by the microwave oven and boiling waterbath treatments. Four microcystins of differing hydrophobicities were successfully extracted from Microcystis PCC 7813 by both treatments at yields equivalent to those obtained by longer protocols using methanol. Assessment of the microwave oven and boiling waterbath extraction methods with laboratory strains and environmental samples of cyanobacteria showed good correlation with results from lyophilisation and methanol extraction, when extracts were analysed by high performance liquid chromatography with diode array detection (R(2)>/=0.92). The microwave and boiling waterbath extraction methods also sterilised the environmental bloom samples, as evidenced by the abolition of heterotrophic bacterial growth.
Metcalf
J S
JS
Department of Biological Sciences, University of Dundee, Dundee, UK.
Codd
G A
GA
D001427:Bacterial Toxins / Q000032:analysis / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D000458:Cyanobacteria / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D004783:Environmental Microbiology; D005612:Freeze Drying; D006358:Hot Temperature; D000432:Methanol / Q000737:chemistry; D052998:Microcystins; D008872:Microwaves; D010456:Peptides, Cyclic / Q000032:analysis / Q000302:isolation & purification*; D014867:Water
D001427:Bacterial Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D014867:Water; C078588:microcystin; D000432:Methanol