title: Laboratory-scale purification of microcystins using flash chromatography and reversed-phase high-performance liquid chromatography.
abstract: Microcystins were extracted from 7 l (equivalent to 313 g dry weight) of cyanobacterial scum collected from Rutland Water in Leicestershire, UK in 1989. The resulting aqueous extract was rapidly concentrated on a C18 flash chromatography cartridge and microcystins were eluted using a step gradient. Fractions were collected manually and monitored by UV spectrophotometer and analytical HPLC. Fractions containing microcystins of similar polarity were pooled to give three fractions. Simple isocratic methods for separating each fraction were developed on an analytical column and scaled up to a 15 x 7.5 cm I.D. column. Closed-loop recycling was used to maximise yield and purity of two hydrophobic microcystins.
authors.affiliation: Biotage Ltd., Hertford, UK.
authors.forename: C
authors.initials: C
authors.lastname: Edwards
authors.forename: L A
authors.initials: LA
authors.lastname: Lawton
authors.forename: S M
authors.initials: SM
authors.lastname: Coyle
authors.forename: P
authors.initials: P
authors.lastname: Ross
mesh_terms: D000595:Amino Acid Sequence; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000458:Cyanobacteria / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000302:isolation & purification; D052998:Microcystins; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry / Q000302:isolation & purification*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors
chemical_list: D004791:Enzyme Inhibitors; D052998:Microcystins; D010456:Peptides, Cyclic; C078588:microcystin; D010749:Phosphoprotein Phosphatases