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Volume 7, Issue 4 (Suppl) J Biotechnol Biomater, an open access journal
ISSN: 2155-952X Bio America 2017
October 19-20, 2017
October 19-20, 2017 | New York, USA
18th Biotechnology Congress
Determination of PUFA content in microalgae through Nile red fluorescence method
Ruben Darío Múnera-Soto, Mauricio López Trejos, Juan Martin Delgado Naranjo and Mariana Peñuela Vásquez
Universidad de Antioquia, Medellín, Colombia
Polyunsaturated Fatty Acids (PUFA’s) are essential in children development and healthy diets. Among them Docosahexaenoic
Acid (DHA) is linked to brain/eye development in children along heart disease prevention. DHA is usually obtained from fish-
capture oil. But, due to depletion of its natural sources, seasonal variability in its composition, and potential chemical contamination
alternative sources are becoming more relevant. Microalgae are some of the most suitable candidates to supply PUFA’s. One problem
is quantifying the amount of oils after fermentation. Currently, from the methods to measure PUFA’s within microalgae cultures, the
most popular are gravimetric and dying methods., Gravimetric methods are time-consuming, require larger samples, several steps,
and solvents, therefore, they become impractical. On the contrary, dying methods, though more complex analysis-wise, are a good
option when the samples are small and frequent. Nile Red Fluorescence (NRF) have proven to be successful to determine the content
of PUFA’s in microalgae, though it is not effective for microalgae with thick, rigid cell wall. NRF is a lipophilic dye, which properties
are determined by the polarity of the surroundings. The method is based on dying the cell in the presence of a transporting carrier to
make the membrane permeable to the Nile Red molecules. Dimethyl-Sulfoxide (DMSO) was our carrier. A spectrophotometer with
the capacity to read microplates allowed the development of the protocols with an enhanced performance towards the qualitative
analysis of lipid content. Excitation and emission wave lengths were measured as 535 nm, 660 nm respectively. After defining the
excitation and emission characteristics, we built a calibration curve for determining lipids. The results were validated by comparing
results fromBligh and Dyer method. NRF is a viable way to determine the content of lipid after the microalgae has reach the maximum
oil production, this way quantifying PUFA yield will not be so time-consuming
Biography
Ruben Darío Múnera-Soto is a Chemical Engineer graduated from Universidad de Antioquia, the education field has been his passion ever since he has been graduated
from college and he has worked with several Universities in his country in the areas of engineering and basic sciences. Currently, he is a student in the Chemical Engi-
neering Master’s Program at Universidad de Antioquia as in the field Biotechnology as a Member of the Bioprocess Research Group in the same university. As part of his
forming activities he has been participated in a project aiming to solve malnutrition issues in Colombian children, and improve dietary problems in the population at large.
Ruben Darío Múnera-Soto et al., J Biotechnol Biomater 2017, 7:4 (Suppl)
DOI: 10.4172/2155-952X-C1-079