Verification and quantification of saxitoxin from algal samples using fast and validated hydrophilic interaction liquid chromatography–tandem mass spectrometry method

Abstract Hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC–MS/MS) method was validated with algal samples for verification and quantification of saxitoxin (STX), a potent neurotoxin which is listed in the Chemical Weapons Convention (CWC) in Schedule 1A. Isocratic elution, conventional bore HILIC column and high flow rate together with accurate post-column splitter provided detection of STX in 6.5 min with total analysis time of 9 min per sample. STX analogue, gonyautoxin 1 (GTX 1) was used as an internal standard. Sample preparation of freeze-dried algae included liquid extraction and centrifugal filtering with mean recovery of 99.9% at concentration level of 10 ng/ml ( n  = 3). Retention times for STX and GTX 1 were 6.47 ± 0.03 min and 4.44 ± 0.01 min ( n  = 45), respectively. Four diagnostic product ions were used for reliable verification of saxitoxin. Method was found to be precise and linear ( R 2  = 0.9714 and R 2  = 0.9768) in concentration ranges of 5–50 ng/ml and 25–200 ng/ml, respectively. For saxitoxin, calculated LOD was 3 ng/ml and LLOQ 11 ng/ml. Validation was conducted using spiked algal matrix since this method is not only needed for verification analysis for the CWC but also for safety analysis of other environmental samples for presence of STX. Identification criteria for verification of STX with HILIC–MS/MS method are discussed.