Use of gas chromatography in the elucidation of carbohydrates in the bodily fluids of the gulf pipefish, Syngnathus scovelli

CHED 613

Jessica Rozelle1, Charlyn Partridge1, Anne Boettcher1, and David C. Forbes2. (1) Department of Biology, University of South Alabama, Mobile, AL 36688, (2) Department of Chemistry, University of South Alabama, Mobile, AL 36688-0002
The Gulf pipefish, Syngnathus scovelli, is known for its unique sex-role reversed reproduction. The reproductive mode of this family of fish, specifically the role that the male brood pouch plays, has stimulated considerable research and has been an area of debate for nearly 100 years. It has been theorized that the brood pouch functions as an osmoregulatory device and/or that it functions in the passage of nutrients by diffusion through the epithelium. The current study focuses on the nutritional role of the brood pouch. An area of interest is the presence and concentrations of carbohydrates in the blood and brood pouch fluid of the male pipefish. Our approach is to use gas chromatography (GC) to either confirm the presence or absence of specific analytes proposed to exist in the bodily fluids of the Gulf pipefish. Three carbohydrates, galactose, glucose and mannose, have been studied thus far. Authentic samples are first derivatized using N-methyl-bis(trifluoroacetamide) as acylating reagent and pyridine as solvent medium. Once acylated, the sugar derivatives are suitably modified for GC analysis. The results are then cross-referenced the test studies. The benefits with this approach are that more information can be gained from small sample sizes and that the limits of detection approach parts per million. Gulf killifish, Fundulus grandis, blood is used in preliminary assays for development of protocols. The results for F. grandis blood are then compared with those for pipefish blood and brood pouch fluid. Once the makeup of these bodily fluids is determined, a better understanding of Syngnathid reproduction as a whole can be gained.