Toxic Cyanopeptides Monitoring in Thermal Spring Water by Capillary Electrophoresis Tandem Mass Spectrometry

Cyanobacteria are an ancient group of prokaryotes capable of oxygenic photosynthesis. Recently, thermal crises symptoms in hot springs have been associated with acute cyanopeptides poisoning. The aim of this work is to develop a fast, easy and reliable method to monitor the presence of toxic cyanopeptides in geothermal waters. The analytical method based on capillary zone electrophoresis coupled with tandem mass spectrometry (CZE-MS/MS) was developed for the simultaneous determination of 14 cyanopeptides in less than 7.5 min. A basic 50 mM ammonium acetate buffer at pH 10.2 was selected as the background electrolyte, positive electrospray ionization (ESI+) was employed for all compounds, and a salting-out assisted liquid-liquid extraction (SALLE) protocol with acetonitrile as an extraction solvent and MgSO₄ as an auxiliary salting-out agent was optimized as sample treatment. Six natural hot springs in the province of Granada (Andalucía, Spain) were sampled at the beginning of the summer season (June) and at the end (September). Biomass collected at two sample points (Santa Fe and Zújar) contained cyanobacteria cells from the genera Phormidium, Leptolyngbya, and Spirulina. Nevertheless, cyanotoxins covered by this work were not found in any of the water samples analyzed. The greenness and transferability of the method was evaluated highlighting its sustainability and applicability.