Immunoaffinity purification method for detection and quantification of microcystins in lake water.
We have developed a new clean-up method, which consisted of solid-phase extraction on a Sep-Pak PS-2 (styrene-divinylbenzene copolymer) or Excelpak SPE-GLF (polymethacrylate) cartridge instead of conventional ODS silica gel and silica gel together with following immunoaffinity purification using anti-microcystin-LR monoclonal antibodies. This newly developed method was demonstrated to eliminate co-existing substances and to concentrate microcystins in the lake water. The recoveries from lake water (1 liter) spiked with 100 ng each of microcystins-RR, -YR and -LR were 85.5, 89.2 and 92.2%, respectively, with coefficients of variation of 3.3-7.6%. Only 3 h were required to complete the total procedures starting from the microcystin extraction, the immunoaffinity purification, and the quantification using HPLC. The detection limits for all of the 3 microcystins in lake water were 0.005 microg/l. Applicability of this method has been demonstrated by measuring the concentrations of microcystins in water samples collected from lakes where water blooms occurred, which turned out to be 0.012-0.177 microg/l of total microcystins.
Kondo
F
F
Aichi Prefectural Institute of Public health, Kita, Nagoya, Japan. fumio_2_kondou@mail.pref.aichi.jp
Matsumoto
H
H
Yamada
S
S
Tsuji
K
K
Ueno
Y
Y
Harada
K
K
D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D000458:Cyanobacteria / Q000378:metabolism; D005618:Fresh Water; D007475:Ion Exchange Resins; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic / Q000276:immunology / Q000302:isolation & purification*; D011109:Polymethacrylic Acids; D011137:Polystyrenes; D018508:Water Purification / Q000379:methods*
D000911:Antibodies, Monoclonal; D007475:Ion Exchange Resins; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D011109:Polymethacrylic Acids; D011137:Polystyrenes; C065685:microcystin YR; C030613:polymethacrylic acid; C003771:Amberlite XAD-2 resin; C063855:microcystin RR; C057862:cyanoginosin LR