1. Bucharest - September 2010 University of Bucharest, ROMANIA Iulia Gabriela David, Vasile David, Department of Analytical Chemistry, Faculty of Chemistry Marius Matache, Centre for Environmental Research and Impact Studies

2. Bucharest - September 2010  essential elements Fe, Se, Cu, Zn   toxic metals Pb, Cd, Hg, As

3. Bucharest - September 2010 Obtain &preserve representative sample Transform the sample to an analyzable form Calculate the result (Input) (Black box) (Output)

4. Bucharest - September 2010 Ialomita River Prut River Seasonal variations in trace metals concentrations Quantification of xenobiotics bioaccumulation in wetland food chains aim technique ICP-AES ASV

5. (filtration)

8. Bucharest - September 2010 Hydrographical basin of the Ialomiţa river with indications of the sampling points. 1 Pietroşiţa – upstream of any pollution source - natural background 2 Pucioasa – downstream from the Pucioasa reservoir lock -> contribution of 2 economic operators – the cement factory + light sources factory Fieni . 3 Ciolpani – characterises the region Pucioasa reservoir lock and DN 1 Buc harest – Ploieşti , possible pollution sources : Pucioasa city , thermo - electric p ower station Doiceşti, Târgovişte city with the special steel aggregate works 4 Dridu – downstream from the Dridu reservoir lock – characterises region of agricultural activities; 5 Albeşti – Urziceni town , live-stock farms Căzăneşti, contribution of the Prahova river; 6 Bucu – Slobozia twon with chemical fertilizers aggregate works, 7 Vlădeni – Ţăndărei town Doiecesti  Targoviste 

9. Bucharest - September 2010

10. Bucharest - September 2010 Model l for the disturbance of ecological balance in wetlands (CCMESI, 2008) Bioaccumulation of heavy metals and/or pescticides along food chains Domestic, industrial and agricultural waste waters; organic substances; nutrients Excessive exploitation of fisheries resources Toxic atmospheric inputs: NOX, SX, heavy metals Uncontrolled hunting; Poaching Uncontrolled deforestation Excess of nutrients for fisheries

11. Bucharest - September 2010 Sampling sessions: Spring - april 2009 Summer - july 2009 Sampling: 6 sampling places along the Romanian side of Prut River from the axis of maximum turbulence of the river, from the water-sediment interface Sample preservation: - in Teflon bottles - concentrated HNO 3 added to avoid analytes losses

12. 1. Upstream of the M aţa-Rădeanu complex water quality at the entrance of the Prut river into the Lower Prut Floodplain Natural Park 2. Downstream of the Rogojeni village in fluence of some pollution sources: Maţa-Rădeanu complex, Pochina lake , Cacia and Leahu pools , Broscarului and Teleajen lakes and the localities Vădeni and Rogojeni 3. Downstream of Vlădeşti and Măicaşu lakes impact of the two lakes, of Şovârca pool and localities Oancea, Slobozia-Oancea and Vlădeşti 4.Downstream of Vlăşcuţa lake influence of lakes Brăneşti , Vlăşcuţa, and of Manta lake on the left side of Prut ( Moldova Rep ), and localities Brăneşti and Măstăcani 5.Downstream of Beleu, at Tuluceşt i co vers a region including the loop Cotul Hiului and some localities on the left river side and Beleu lake 6. Upstream of Prut ’s run into the Danube final sampling point-----  influence of Brateş lake, of agricultural fields -gives an image of pollutants concentrations transferred by Prut into the Danube Bucharest - September 2010

13. Bucharest - September 2010

14. Bucharest - September 2010 SCHEMATIC OF AN ICP-AESpectrometer Nebuliser Ar excited atoms h  Detector Polychromator (Ar+sample aerosole) (T=8000K h=12 cm) (cooling, 12 L/min ) (0.8 L/min) (1 L/min) (Frequency 27.12 MHz Power adjustable 800 -1,600 W) (165–210 nm; 210-580 nm) Plasma torch Sample (0.002 l/min)

16. Bucharest - September 2010 -HMDE -MFE -Bare C, Au, etc. 10 -4 -10 -5 Hg +2 for co-metal deposition. A cathodic or reducing potential is applied for a fixed time interval reducing M n+ Potential is scanned in anodic or oxidizing direction to strip out M o

17. Bucharest - September 2010 • Sensitive and reproducible (RSD<5%) method for trace metal ion analysis in aqueous media. Accuracy is proportionate to the way of sample calibration: <5% when calibrated directly via the method of standard additions. 10% when a calibration curve is built before measuring 20% - 40% when operating uncalibrated • Concentration limits of detection for many metals are in the low ppb to high ppt range (S/N=3) ppm - instantaneous ppb < 30 seconds or less ppt - several minutes  compares favorably with AAS or ICP analysis.

21. Bucharest - September 2010 Variation of Zn and Mo concentration in water samples from the Ialomiţa River

22. Bucharest - September 2010 Technique : Differential Pulse Anodic Stripping Voltammetry (DP-ASV) Working electrode : Mercury film deposited on a glassy carbon electrode (MF-GCE) Optimum conditions : t a c = 120 s, E ac = -1,1 V; v = 20 mV/s, Pulse amplitude = 50 mV; Sampling width = 20 ms; Pulse width = 40 ms; Pulse periode = 300 s Concentration evaluation method : Standard addition Analyte : Cu(II), Pb(II), Cd(II) from Prut River water samples and mollusks

23. Bucharest - September 2010 DP-anodic stripping voltammograms recorded in HNO 3 0,1 M on MF-GCE for sample 6 collected upstream of Prut’s run in the Danube: (6b-5)=water sample 6; (6b-6)= water sample 6 + 0,1 mL standard solution; (6b-7)= water sample 6 + 0,2 mL standard solution containing Cu(II) = Cd(II)= Pb(II)= 8 10 -3 g/L.

24. Bucharest - September 2010 Heavy metals concentration in Prut River Water Samples Me(II) sample Cu(II) (g/L) Pb(II) (g/L) Cd(II) (g/L) april july april july april july pr1 3.20 10 -6 1.30 10 -5 3.00 10 -6 <LOD 2.10 10 -5 <LOD pr2 6.74 10 -5 1.60 10 -5 5.66 10 -7 2.60 10 -4 1.61 10 -5 <LOD pr3 1.93 10 -5 4.10 10 -5 1,83 10 -5 1.80 10 -5 3.30 10 -5 1.20 10 -4 pr4 3.88 10 -5 <LOD <LOD <LOD 1.48 10 -4 <LOD pr5 9.20 10 -6 9.50 10 -6 <LOD <LOD 5.15 10 -5 <LOD pr6 6.70 10 -6 2.50 10 -5 8.60 10 -6 <LOD 1.09 10 -4 <LOD

25. Bucharest - September 2010 9 Separation Shell Soft part Washing Weighing Adding 5 ml HNO3 (65%) 5 ml HCl (35-37%) 5 ml HClO4 Heating to dryness Adding 5 ml HNO3 (65%) 5 ml HCl (35-37%) Heating Filtering Diluting with MilliQ H2O to the mark of a 25 ml volumetric flask Lymnaea stagnalis , ( sample X 5 col l ect ed on 29.07.2009 , at sampling point 6, where Prut runs into Danube ).

26. Bucharest - September 2010 DP-anodic stripping voltammograms recorded in HNO 3 0,1 M on MF-GCE for dissoluted mollusk sample X4 ( Lymnaea stagnalis) collected upstream of Prut’s run in the Danube: (X4)=dissoluted mollusk sample; (X41)= dissoluted mollusk sample + 0,1 mL standard solution; (X42)= dissoluted mollusk sample + 0,2 mL standard solution containing Cu(II) = Cd(II)= Pb(II)= 8 10 -3 g/L.

27. Bucharest - September 2010 Acknowledgement Financial support is acknowledged from the PN-II- project BIOXEN 32111-2008.

28. Bucharest - September 2010