Purification of microcystins by DEAE and C(18) cartridge chromatography.
Microcystins (MCs) were purified by DEAE and C(18) cartridge chromatography. Addition of EtOH to the eluents (20%) in DEAE chromatography gave higher resolution than no addition of EtOH. The chromatogram showed three peaks: MC-LR; MC-LY and MC-LF; MC-LW. MC-LR and MC-LW were obtained by one step chromatography with purity of 96 and 88%, respectively. The separation of MC-LF and MC-LW with DEAE chromatography was better than that with reversed-phase chromatography. MC-LY and MC-LF were separated with C(18) cartridge. On the chromatogram, there were three peaks consisting of MC-LY (81% purity), MC-LF (86%), and an unknown compound which was considered as a MC variant judging from the results in HPLC/PDA, FAB-MS, and 1H NMR analyses, but the structure could not be determined. It is concluded that the combination of DEAE and C(18) cartridge chromatography would be a practical approach for the purification of various MCs.
Saito
Kazunori
K
School of Marine Science and Technology, Tokai University, 3-20-1 Orido, Shimizu, 424-8610, Shizuoka, Japan.
Ishii
Hiroshi
H
Nishida
Fumiko
F
Saito
Hiroshi
H
Abe
Toshihiko
T
Toyota
Yoshimasa
Y
D001427:Bacterial Toxins / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D000458:Cyanobacteria / Q000378:metabolism*; D000431:Ethanol / Q000737:chemistry; D066298:In Vitro Techniques; D052998:Microcystins; D010456:Peptides, Cyclic / Q000302:isolation & purification*
D001427:Bacterial Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D000431:Ethanol; C078588:microcystin