Student Research Projects

Extraction of antibacterial substance from Cinachyra alloclada

Student Britton Hipple, '16
Department Chemistry
Course CHEM 489: Independent research in the Natural Science

Abstract

\" Sponges have a unique chemical environment that provides a
natural defense against predators and to control the symbiont bacteria that
grow with the sponge. In this study, organic compounds were extracted from
samples of the common yellow ball sponge, Cinachyra alloclada, to be tested
for the presence of substances with antibacterial or antifungal capabilities. C.
alloclada, purchased from Gulf Specimens, were collected from the Gulf of
Mexico off the coast of Florida. Organic compounds were isolated using five
different extraction methods to optimize the separation of components. The
most productive method was a serial extraction of the macerated sponge in a
3.5% saline solution with solvents of increasing polarity: dichloromethane,
butanol, and ethyl acetate. Each extract was analyzed by gravimetric
analysis, thin layer chromatography, and bioassay. The microbial species
used in bioassays were Escherichia coli (DH5α), Pseudomonas flourecens,
Bacillus megaterium and Penicillium notatum. A substance that showed
antibacterial capabilities was observed in all three methods that extracted
directly from the macerated sponge samples, but not when the sponges were
dehydrated for 24 hours prior to extraction. The optimized extraction
procedure isolated a single component with antibacterial activity in the
dichloromethane extract. The bioactive extracts showed dose dependent
inhibition for both Gram-positive and Gram-negative bacterial strains, but no
inhibition of fungal strains. The isolated bioactive substance was
characterized by chemical classification tests, qualitative thin layer
chromatography staining, IR spectroscopy, and gas chromatography-mass
spectroscopy, which indicated the substance is aromatic with an aldehyde
and an amine functional group\"