Laboratory of Chemistry TASK 2.1 Analysis of the seaweed biomass produced in WP1 Laboratory of Chemistry Pedro Alexandre Reis pedroareis@ciimar.up.pt

Task 2.1 – Analysis of the seaweed biomass produced in WP1 LABORATORY OF CHEMISTRY - MARINNOVA : Atomic Absorption Spectrometer (AAS): Metals Skalar Segmented Flow Autoanalyzer (SFA): Nutrients Gas Chromatography – Mass Spectrometer (GC-MS): PAHs Liquid Chromatography – Mass Spectrometer (LC-MS): Microcystins and other organic compounds Isotope Ratio - Mass Spectrometer (IR-MS): 15N/14N and 13C/12C MORE INFORMATIONS: www.marinnova.pt

Task 2.1 – Analysis of the seaweed biomass produced in WP1 Direct partners: AlgaPlus (Red seaweed: Gracilaria sp.) CEVA (Green seaweed: Ulva sp.) CPS (Brown seaweed: Alaria esculenta) Elements: Metals and Nutrients Metals: Essential elements: Copper (Cu) Iron (Fe) Zinc (Zn) No-essential elements: Arsenic (As) Cadmium (Cd) Chromium (Cr) Lead (Pb) Mercury (Hg) Nutrients: Ammonium (NH3) Nitrites (NO2) Nitrates (NO3) Total-Nitrogen (N-Total) Phosphates (PO4) Total-Phosphorus (P-Total)

AAS – Flame Atomization AAS – Electrothermal Atomization Task 2.1 – Analysis of the seaweed biomass produced in WP1 Chemical analyses: Metals Analytical methods: Atomic Absorption Spectrometry (AAS) AAS – Flame Atomization AAS – Electrothermal Atomization

Task 2.1 – Analysis of the seaweed biomass produced in WP1 Chemical analyses: Nutrients Analytical methods: Skalar Segmented Flow Autoanalyzer (SFA) Skalar San Plus System

Task 2.1 – Analysis of the seaweed biomass produced in WP1 Collection Strategy: Latex gloves and steril plastic bags (material isent of metals or nutrients) Three replicates per sample (N=3) Minimum mass (fresh weight/per replicate): 50 grams Storage and Preservation Strategy: Correct identification of samples: name + date + laboratory Immediatly freezed at least -20 ºC Time interval between collection and analyses: No significant losses or degradation within 4 weeks

Dry sample + solution of HNO3 + two-steps of 340W during 4 min Task 2.1 – Analysis of the seaweed biomass produced in WP1 Pre-treatment of seaweed samples for metal and nutrients analyses: Liophylized during 2-3 days (until constant weight) Reis and Almeida (2008): Acid digestion with high-pressure microwave system Dry sample + solution of HNO3 + two-steps of 340W during 4 min Analytical methods for metals and nutrients: Adapted from certified: EPA methods Standard methods for the examination of water and wastewaters (20th edition) Our analytical methods acreditation: ISO/IEC 17025 (initial stage)

Task 2.1 – Analysis of the seaweed biomass produced in WP1 Data Quality Assurance: Evaluation of analyst performance for each method; Evaluation of instrument performance for each method: precision, limits of detection and quantification and accuracy; Analyses of blanks and samples with known additions of metals and nutrients (at each batch of 5% of total samples). Analyses of Certified Reference Materials (CRM) for metals and nutrients: DORM-2 (Dogfish Muscle); TORT-2 (Lobster Hepatopancreas); SRM 2976 (Mussel Tissue).

Pedro Alexandre Reis CIIMAR - Laboratory of Chemistry Rua dos Bragas, 289 4050-123 Porto Portugal Phone: +351 22 340 18 30 E-mail: pedroareis@ciimar.up.pt

Laboratory of Chemistry TASK 2.1 Analysis of the seaweed biomass produced in WP1 Laboratory of Chemistry Pedro Alexandre Reis pedroareis@ciimar.up.pt