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).