Determination of Multiclass Cyanotoxins in Blue-Green Algae (BGA) Dietary Supplements Using Hydrophilic Interaction Liquid Chromatography-Tandem Mass Spectrometry

In recent years, the consumption of blue-green algae (BGA) dietary supplements is increasing because of their health benefits. However, cyanobacteria can produce cyanotoxins, which present serious health risks. In this work we propose hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) to determine cyanotoxins in BGA dietary supplements. Target toxins, including microcystin-leucine-arginine (MC-LR) and microcystin-arginine-arginine (MC-RR), nodularin, anatoxin-a and three non-protein amino acids, β-N-methylamino-L-alanine (BMAA), 2,4-diaminobutyric acid (DAB) and N-(2-aminoethyl)glycine (AEG), were separated using a SeQuant ZIC-HILIC column. Cyanotoxin extraction was based on solid–liquid extraction (SLE) followed by a tandem-solid phase extraction (SPE) procedure using Strata-X and mixed-mode cation-exchange (MCX) cartridges. The method was validated for BGA dietary supplements obtaining quantification limits from 60 to 300 µg·kg−1. Nine different commercial supplements were analyzed, and DAB, AEG, and MCs were found in some samples, highlighting the relevance of monitoring these substances as precaution measures for the safe consumption of these products

Implementing the Use of Collision Cross Section Database for Phycotoxin Screening Analysis

: The increased consumption of blue-green algae (BGA)-based dietary supplements has raised concern about their food safety, especially about cyanotoxin presence. The hyphenation of liquid chromatography with ion mobility mass spectrometry represents a relevant tool to screen several compounds in a large variety of food matrices. In this work, ultrahigh-performance liquid chromatography coupled to traveling wave ion mobility spectrometry/quadrupole time-of-flight mass spectrometry (UHPLC-TWIMS-QTOF) was employed to establish the first comprehensive TWIMS-derived collision cross section database (TWCCSN2) for phycotoxins. The database included 20 cyanotoxins and 1 marine toxin. Accurate m/z, retention times, and TWCCSN2 values were obtained for 81 adducts in positive and negative electrospray (ESI+/ESI-) modes. Reproducibility and robustness of the TWCCSN2 measurements were determined to be independent of the matrix. A screening was carried out on 19 commercial BGA dietary supplements of different composition. Cyanotoxins were confidently identified in five samples based on retention time, m/z, and TWCCSN2

Monitoring of cyanotoxins in water from hypersaline microalgae colonies by ultra high performance liquid chromatography with diode array and tandem mass spectrometry detection following salting-out liquid-liquid extraction

In this study two different analytical approaches have been developed to determine the presence of sev-eral cyanotoxins in saline water samples from a continental salt marsh. A salting-out assisted liquid-liquidextraction (SALLE) has been used in combination with ultra-high performance liquid chromatography-tandem mass spectrometry and UV-diode array detection (UHPLC-MS/MS and UHPLC-DAD). The targetanalytes are eight microcystins named MC-RR, MC-YR, MC-LR, MC-WR, MC-LA, MC-LY, MC-LW, MC-LFand nodularin (NOD), covering a wide range of polarities. The separation was achieved using a ZorbaxEclipse Plus RRHD C18 column (50 × 2.1 mm, 1.8 m) in less than 7.5 and 5.5 min for UV and MS/MSdetection, respectively. The mobile phase used consisted of water (solvent A) and acetonitrile (MeCN)(solvent B), both containing 0.01% of formic acid for DAD and 0.4% of formic acid for MS/MS detection,at a flow rate of 0.4 mL min−1. The temperature of the column was set at 25◦C and 20 L of samplewere injected. The main parameters affecting the SALLE procedure were studied and the following opti-mum values were obtained: neutral pH, 2 mL of acetonitrile as extraction solvent and 1.2 g of ammoniumsulfate as salting-out agent for 4 mL of water sample. The validation protocols for both methods wereaccomplished with real water samples obtaining LODs ranging from 1.0 to 3.4 g L−1and 0.02 to 0.11 gL−1for DAD and MS/MS respectively. Although the SALLE-UHPLC-DAD methodology is easier and cheaperthan UHPLC-MS/MS significantly better detection limits were achieved with tandem mass spectrometryas well as allowing for unambiguous identification. Extraction recoveries were higher than 77.0% (exceptfor MC-RR and NOD which were 53.2% and 54.3, respectively) with satisfactory inter-day and intra-dayprecisions (RSD below 13.3%).Departamento de Química Analítica, Grupo FQM 302Ministerio de Ciencia, Innovación y Universidades (ProjectRef.: RTI2018-097043-B-I00)Ministry of Science, Research, and Technology of Ira