Optimization of methods for the extraction of ciguatoxins from Lionfish (Pterois volitans) tissue
Kosar, Brittney N
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Ciguatera Fish Poisoning (CFP) is a severe human illness that results from the consumption of finfish containing ciguatoxins (CTXs). To protect human health in ciguatera endemic areas, monitoring agencies must be able to differentiate between toxic and nontoxic fish. Toxin extraction is required before toxins in fish tissue can be quantified. This study addressed the critical need for optimization of extraction protocols to improve recovery of CTX from fish tissue. Invasive lionfish (Pterois volitans) tissue was used to assess the efficiency of various extraction techniques. optimizing Liquid-Liquid (L-Ll partitioning steps and Solid Phase Extraction (SPE) steps. This research found that toxin concentrations were highest after wet tissue was processed by initial extraction steps followed by chloroform L-L solvent partitioning. Further processing with SPE cartridges reduced the concentration of toxin remaining in the extracts. This research demonstrated how lyophilization adversely affects extraction of Caribbean toxins. UPLC-TOF and the Neuro-2a bioassay were used to determine efficiency of extraction methods. The optimized method analyzed 1 g samples after chloroform L-L extraction; this method was capable of toxin detection at subnanogram concentration. The use of a highly sensitive UPLC-TOF system greatly reduces the processing time and reduces solvent cost. Results may enable future monitoring programs to be implemented using similar extraction protocols.
A thesis Submitted in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE in MARINE BIOLOGY from Texas A&M University-Corpus Christi in Corpus Christi, Texas.