Multidimensional biochemical detection of microcystins in liquid chromatography.
The coupling of antibody-, receptor-, or enzyme-based inhibition assays postcolumn to chromatographic systems provides biological detectors with extraordinary high sensitivity and specificity. Three monoclonal antibodies (MC10E7, AD4G2, M8H5) directed against microcystins and protein phosphatase 1 (PP1) were used as off-line detectors for the HPLC separation of microcystins and nodularin in comparison to UV detection. For HPLC/ELISA coupling using antibody MC10E7, a detection limit of 0.04 ng microcystin-LR was achieved. The provisional guideline value for microcystin-LR (1 microg/L, WHO) could be monitored without prior sample concentration, in contrast to UV detection. Quantification of microcystin-LR and two cross-reactants was demonstrated. Furthermore, cross-reactivity or enzyme inhibition of new microcystins, only available in small amounts, can be determined by this method. Using a cyanobacterial extract, HPLC/ELISA coupling was compared to HPLC/UV and electrospray ionization mass spectrometry (ESI-TOFMS).
Zeck
A
A
Institute of Hydrochemistry, Technical University of Munich, München, Germany.
Weller
M G
MG
Niessner
R
R
D000911:Antibodies, Monoclonal / Q000276:immunology; D001427:Bacterial Toxins / Q000032:analysis* / Q000276:immunology; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D000458:Cyanobacteria / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods; D052998:Microcystins; D010456:Peptides, Cyclic / Q000032:analysis* / Q000276:immunology; D010749:Phosphoprotein Phosphatases / Q000276:immunology; D054645:Protein Phosphatase 1
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; C078588:microcystin; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1