Chromatographic separation parameters in pigment analysis

In the present study, chromatographic separation parameters used in pigment analysis phytoplankton size classes depending on pigments from December 2014 to August 2015 were seasonally investigated along the south-eastern coasts (Giresun-Artvin) of the Black Sea. Along the 12 stations, seasonal samplings were collected from surface waters. Based on the HPLC-DAD conditions, an appropriate separation of 16 pigment standard was achieved within 38 minutes. The separation parameters and the resolution values showed that the peaks were close to 100%. The used method was validated for pigment analysis. Contribution of phytoplankton size classes derived from pigments to total phytoplankton in the study area revealed significant changes and ranged from 3 to 60%; 1 to 60%; 8 and 88% for picophytoplankton, nanophytoplankton and microphytoplankton, respectively. Surface mean chlorophyll-a concentrations ranged from 0.37-2.67 µg/L, 0.16-2.02 µg/L and 0.17-1.79 µg/L for 2, 8 and 15 miles, respectively. Besides chlorophyll-a, peridinin, 19-hexanoloxyfucoxanthine, fucoxanthine, zeaxanthin, chlorophyll-b and alloxanthine were the most outstangin marker pigments.