Bruna Ferreira Silva Chemistry Institute Federal University of Goiás Environmental Health and Safety 2016 Conference Analysis of some chemical elements in marine microalgae for biodiesel production and other uses Bruna Ferreira Silva Chemistry Institute Federal University of Goiás Valencia - Spain October, 2016

Outline Introduction Objective Materials and Methods Microalgae cultivation and sampling Light microscopy Biomass Digestion ICP-OES analysis Chemometric analysis Results and Discussion Chemical elements in marine microalgae Conclusion Comments & Questions

Introduction Microalgae Base of aquatic food chain Variable uses Plants similarities Variable uses Food industry Bioremediation Bioproducts Biofuels Variety of aquatic environments Different microalgae metal profile

Introduction Biochemical and mineral content Elements uptake for growth (bioaccumulation) Cell Adsorption (biosorption) Influence in microalgae use Marine Microalgae for biodiesel Lipids and fatty acid productivity (↑) Replace diesel, petroleum, natural gas and coal More resistance to environmental changes Large seawater availability Do not compete with crops

Objetives Determine 26 chemical elements found in the biomass of marine microalgae, with potential to produce biodiesel, mainly by direct conversion of the biomass into biofuel; Identify promising species, regarding their chemical element profile; Find relations between taxonomic group and biosorption or bioaccumulation ability of species; Find relations between biomass elements content and cultivation medium;

Microalgae cultivation and sampling; Experimental Methods Microalgae cultivation and sampling; Light microscopy; Biomass digestion; ICP-OES analysis; Chemometric analysis.

Microalgae cultivation medium (Conway) Microalgae Species (No floculant; no washing process; lyophilization) Cyanobacteria Aphanothece sp. Planktolyngbia limnetica Planktothrix isothrix Romeria gracilis Synechococcus sp.(1) Synechococcus sp. (2) Synechococcus sp.(3) Synechocystis aquatilis (1) Synechocystis aquatilis (2) Synechocystis aquatilis (3) Synechocystis sp. (1) Synechocystis sp. (2) Chlorophyceae Chlorophyceae N. I. (1) Chlorophyceae N. I. (2) Chlorophyceae N. I. (3) Dynoflagellate Amphidinium carterae Diatom Achnanthes sp. (1) Achnanthes sp. (2) Amphora sp. (1) Amphora sp. (2) Biddulphia sp. Biddulphia sp. cf longicruris Biddulphia sp. cf. aurita Chaetoceros simplex Cylindrotheca closterium (1) Cylindrotheca closterium (2) Diatom N. I. Entomoneis alata Navicula sp. (1) Navicula sp. (2) Navicula sp (3) Phaeodactylum tricornutum Thalassiosira sp. (1) Thalassiosira sp. (2) Thalassiosira sp. (3) Thalassiosira sp. (4)

Light Microscopy Biomass Digestion Samples selected at random; 100 mg biomass Digestor Volumetric Flask 25 mL Precipitate in Eppendorf Solution in Bottles 3 mL HNO3 70% metal ultrapure Programação S1: 330W – 5’ S2: 800W – 8’ S3: - 0W – 7’ Samples selected at random; Zeiss Optics Microscope, model Axioskop2 – triocular; Dryed biomass and rehydrated biomass with 0,85% NaCl solution. Image capture of both conditions.

Chemical Elements Analysis by ICP-OES ICP-OES Thermo Fisher Scientific model iCAP 6300 Calibration curve - 8 standard LOD and LOQ calculated Parameter Condition Auxiliary gas (argon) 0,5 L/min Radiofrequency (RF) Power 1250 W Pump rate 50 RPM Nebulizer gas (argon) 0,18 MPa

Chemical Elements Analysis by ICP-OES Emission Lines (nm)

Chemometric Analysis PCA performed on Octave v. 3.6.1 Software for Windows; Data autoescaled Correlation Analysis performed on Microsoft Excel 2010, with suplement Data Analysis.

Results and Discussion Method LOQs (0,04 to 2,64, except Bi, Na and P >5,9 mg/Kg) r (0,99813 a 0,99999) Sensitivity and Linearity  Analysis time Chemical elements profile Cellular affinity Seawater quality

Results and Discussion

Results and Discussion Adicionado Quantificado Al   X B Ba Be Bi Ca Cd Co Cr Cu Fe K Li Mg Mn Mo Na Ni P Pb Sn Sr Ti Tl V Zn Results and Discussion Microalgae as bioremediators Bioindicators of contamined environments? Water analysis x Microalgae analysis Presence x Quantity

Results and Discussion Range: 12,9 a 36,3% Same gender Na: 50 a 88% Strains: Amphora sp. (1) Biddulphia sp. Navicula sp. (3) P.limnetica Synechococcs sp. (1)

Results and Discussion Cristals whithin the cels Pearson’s correlation coeficients Fe-Ti 0,96 Fe-Al 0,86 Al-Ti 0,85 Al-P 0,75 B-Mg 0,73 P-Pb 0,72 Ni-V 0,70 Na-P -0,71

Conclusions Microalgae biomass showed high inorganic content for a biodiesel production feedstock, (12.9% to 36.3% mass of analyzed elements per dry biomass); Then the use of microalgae as bioindicators of contaminated environments is indicated; Navicula species showed toxic elements in their biomasses, such as Ba and Pb, which point their use in wastewater treatment; most of the microalgae elements profiled were macroelements, Na >> Mg > K ≥ Ca > P, which are natural seawater components.

Questions?

Contact: Bruna Ferreira Silva Thank You! Agradecimentos Contact: Bruna Ferreira Silva brufesil@hotmail.com +55 62 98144-8467