Determination of paralytic shellfish poisoning toxins in cultured microalgae by high-performance liquid chromatography with fluorescence detection

A novel method for the determination of paralytic shellfish poisoning (PSP) toxins using high-performance liquid chromatography with fluorescence detection was developed. The fluorescent derivates of neosaxitoxin (neoSTX), saxitoxin (STX), gonyautoxins 1 and 4 (GTX1+4), and gonyautoxins 2 and 3 (GTX2+3) were separated on a mu Bondapak NH2 column (300 mm x 3.9 mm, 10 mu m) using water and acetate buffer (pH 6.5) as the mobile phase (1.00 mL min(-1)) in gradient mode with fluorescence detection at 390 nm (excitation at 330 nm). The linear ranges of neoSTX, STX, GTX1+4 and GTX2+3 were 3.31-331, 0.952-95.2, 3.78-378 and 0.124-12.4 ng mL(-1), respectively. The detection limits of neoSTX, STX, GTX1+4 and GTX2+3 were 1.10, 0.32, 1.26 and 0.041 ng mL(-1), respectively. The method was successfully applied to the determination of PSP toxins in microalgae. The recoveries ranged from 88 +/- 2% to 107 +/- 4% and the relative standard deviations were 0.16% to 4.4%. The procedure is also environmentally friendly because no organic solvent is used in the mobile phase.