“…Cartridges were stored at -20°C prior to laboratory analysis. the main steps of the laboratory procedure included elution of SPE cartridges with approximately 40 mL of acetone, transfer into iso-octane by adding 1 mL of this solvent to the acetone extract previously rotor-evaporated to 3 mL, vortexing and transferring the more non-polar phase (top phase) into the new test tube, rinsing the acetone phase with hexane three times, reducing the volume of the solvent mixture to 1 mL under the nitrogen flow, and cleaning up the solution with 0.5 mL of concentrated sulphuric acid (Mukherjee and Gopal, 1996). Ice algae samples were filtered through 0.7 μm pre-combusted and pre-weighed GFF filters (Whatman) and stored at -20°C.…”