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Separation of microcystins by capillary electrochromatography in monolithic columns
Authors:Zeisbergerová Marta  Kost'ál Vratislav  Srámková Markéta  Babica Pavel  Bláha Ludek  Glatz Zdenek  Kahle Vladislav
Institution:Masaryk University, Faculty of Science, Department of Biochemistry, Kamenice 5, 625 00 Brno, Czech Republic. zeisbergerova@iach.cz
Abstract:Contribution on microcystin variant analysis by capillary electrochromatography (CEC) with easily affordable spectrophotometric detection is presented. Two types of reversed-phase capillary columns formed by inorganic or organic polymer monoliths were prepared for this purpose. The analyses were performed isocratically by means of tris(hydroxymethyl) aminomethane (TRIS) buffers of mildly alkaline pH containing 30% (v/v) acetonitrile as the mobile phases. The samples were injected electrokinetically and the analyses were done at the same separation field strength of 500 V/cm. Microcystins were detected at 238 nm. Although both column types differ not only in monolith quality (inorganic versus organic) but also in the length of the aliphatic moiety (C8 versus C12) similar results were achieved. The on-column preconcentration as the encouraging prospect of electrochromatographic technique was also tested. Consequently 5% of column volume was injected in contrast with 0.5% at standard injection scheme resulting in the six times enrichment of the low concentrated cyanobacterial extract at the top of the separation column. From these preliminary results can be seen that the CEC method is fully applicable for rapid microcystin screening.
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