RECENT ADVANCES AND FUTURE TRENDS IN FLOW INJECTION AND THEIR APPLICATION TO CHEMICAL OCEANOGRAPHY
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1 RECENT ADVANCES AND FUTURE TRENDS IN FLOW INJECTION AND THEIR APPLICATION TO CHEMICAL OCEANOGRAPHY António O. S. S. Rangel 1, Raquel B. R. Mesquita 1, Collaborative on Oceanographic Chemical Analysis (COCA) Jarda Ruzicka 2
2 COCA - Collaborative on Oceanographic Chemical Analysis 2 Flow analysis as an efficient monitoring tool Analysis automation Real-time determinations On-line monitoring Low consumption values Multi-parametric determinations Speciation Multiparametric assessment Flow Analysis Water dynamic systems
3 COCA - Collaborative on Oceanographic Chemical Analysis 3 Natural waters Wide concentration ranges Variable salinity Colour and/or turbidity Chemical forms Spatial and temporal variability Complex matrix Real time assessment Speciation Interferences
4 COCA - Collaborative on Oceanographic Chemical Analysis 4 Flow methods vs Classical methods Classical methodologies High reagent consumption High sample volumes High reaction time Higher contact between the analyst and the reagents Sample pretreatments often required Flow methodologies Reduced reagent and sample consumption Lack of physical-chemical equilibrium Reduction in the reaction time Automation increased security of the analyst Possibility of in-line sample pretreatments
5 COCA - Collaborative on Oceanographic Chemical Analysis 5 Flow Injection Analysis - FIA Described in 1975 No physical nor chemical equilibrium Controlled dispersion same conditions for standards and samples Continuous reagent flow One manifold per determination Sequential Injection Analysis (SIA)
6 COCA - Collaborative on Oceanographic Chemical Analysis 6 Sequential Injection Analysis - SIA Described in 1990 Non continuous flow Minimization of reagent/sample consumption Multi-parametric determinations Higher degree of automation
7 COCA - Collaborative on Oceanographic Chemical Analysis 7 Sequential Injection Analysis - SIA Detection system: Optical Electrochemical Unit Separation units: Gas diffusion SPE Pre-treatment units: Digestion Dilution
8 COCA - Collaborative on Oceanographic Chemical Analysis 8 Sequential injection Analysis - SIA The configuration of the basic SI instrument can be expanded by a variety of peripherals that fall into two categories: detectors and solution handling devices.
9 COCA - Collaborative on Oceanographic Chemical Analysis 9 Water monitoring In situ measurements ph Temperature Salinity Turbidity Dissolved Oxigen Routine determinations Nitrogen (NO x, NH x, N total ) Phosphorus (phosphate) Carbon (DIC, DOC, TC)
10 COCA - Collaborative on Oceanographic Chemical Analysis 10 SIA for transistional waters quality assessment Estuary of Cávado km 2 rural and disperse urban area Estuary of Ave km 2 North west of Portugal - Porto (>1 million pop.) - Viana do Castelo - Braga farm and industrial area Estuary of Douro km 2 urban area (vol. Discharge > 500 m 3 /s)
11 COCA - Collaborative on Oceanographic Chemical Analysis 11 Monitored esturies Cávado (41.5ºN, 08.7ºW) Douro (41.1ºN, 08.6ºW) Ave (41.3ªN, 08.7ªW)
12 COCA - Collaborative on Oceanographic Chemical Analysis 12 Nutrients monitorization (NO x ) Simultaneous determination of nitrite and nitrate CR CR, colour reagent (sulfanilamide, 20 g/l; N1NED, 2 g/l; Phosphoric ac., 0,5 M); S, sample/standard; EDTA, 0.4 g/l and amonium chloride 20 g/l; Col., Cd column P S H 2 O HC SV Col. W 1 cm 2 cm EDTA R 1 Mesquita et al. (2009) Anal. Methods, 2009, 1, R 2 l W
13 COCA - Collaborative on Oceanographic Chemical Analysis 13 Nutrients monitorization (NH x ) Determination of ammonium and ammonia Reagent recirculation no consumption Det ector l GDU NaOH Waste PP 2 BTB S NaOH Waste SV Holding coil PP 1 BTB, 180 µm; S, sample/standard; NaOH 25 mm; Ppi, peristaltics pumps; SV, selection valve; GDU, gas difusion unit H2O Segundo et al. (2009) Anal. Methods, 2011, 3,
14 COCA - Collaborative on Oceanographic Chemical Analysis 14 Nutrients monitorization (NO x, NH x e PO 4 3- ) Bi parametric determination with inline nitrate reduction Elimination of salinity interference using a multireflective cell Separation of the analyte from the sample with a GDU and no reagent consumption recirculation mode
15 COCA - Collaborative on Oceanographic Chemical Analysis 15 Parameter Quantification range Calibration curves LOD (µm) LOQ (µm) Determination rate (h -1 ) NH x mm A = ± [NH 4+ ] ± Phosphate µm H c = ± [PO 4-3 ] ± Nitrite M A = 5.06x10-2 ± 1.02x10-3 [NO 2- ] 1.24x10-4 ± 4.68x Nitrate M A = 3.17x10-2 ± 7.57x10-5 [NO 3- ] 8.38x10-2 ± 1.19x ID ph Conductance (ms/cm) Salinity PO 4 3- (µm±sd) NO 2 - (µm±sd) NO 3 - (µm±sd) NH 4 + (µm±sd) Estuário 1 7,89 49,9 36,6 42,9 ± 2,5 1,75 ± 0,16 77,1 ± 13,9 2,78 ± 0,41 Estuário 2 7,89 0,227 < 2 28,2 ± 0,0 1,16 ± 0, ± 67 4,14 ± 0,12 Estuário 3 6,21 1,455 < 2 1,22 ± 0,06 1,07 ± ± 6 24,5 ± 4,3 Estuário 4 6,31 0,276 < 2 < LQ 1,12 ± ± 4 10,3 ± 2,9 Estuário 5 6,27 11,67 7,4 < LQ 0,959 ± 0,067 23,4 ± 0,8 44,8 ± 4,0 Estuário 6 6,59 1,393 < 2 1,28 ± 0,17 4,06 ± ± 2 9,8 ± 0,8
16 COCA - Collaborative on Oceanographic Chemical Analysis 16 Nutrients monitorization - carbon speciation Determination of DIC, TC (DOC), CO 2 and alkalinity
17 COCA - Collaborative on Oceanographic Chemical Analysis 17 Micro SIA Lab on valve (LOV) Described in 2000 Typical sample and reagent volumes: 5 to 25 µl/assay Consumption of carrier solution: 100 to 250 µl
18 COCA - Collaborative on Oceanographic Chemical Analysis 18 Solid Phase Extraction - SPE Objectives Sample clean-up Analyte pre-concentration Pure & Appl. Chem., Vol. 66, No. 2, pp , 1994
19 COCA - Collaborative on Oceanographic Chemical Analysis 19 Solid Phase Extraction - SPE Non-Automated format time consuming low reproducibility Automation Solid Phase Micro-Extraction (Needle trap, in-tube ) Flow-based methods (FIA, SIA) SPE FLOW-BASED FORMAT Reusable Renewable (bead injection)
20 COCA - Collaborative on Oceanographic Chemical Analysis 20 Reusable SPE - SIA Determination of enzymatic activity (alkaline phosphatase) NTA resin Zn 2+ AP pnpp pnp + Pi p-nitrophenyl phosphate + H 2 O p-nitrophenol + Pi
21 COCA - Collaborative on Oceanographic Chemical Analysis 21 Reusable SPE - SIA
22 COCA - Collaborative on Oceanographic Chemical Analysis 22 Bead injection renewable solid phase spectrometry in micro SI format
23 COCA - Collaborative on Oceanographic Chemical Analysis 23 Bead injection renewable SPS
24 COCA - Collaborative on Oceanographic Chemical Analysis 24 Renewable SPE using microsi-lov
25 COCA - Collaborative on Oceanographic Chemical Analysis 25 Renewable SPE using microsi-lov Determination of total protein content in white wine Protein Freshly prepared beads A) NTA resin HC W Cu 2+ Detector FCr B) ST Detector OF Beads after one working day SP W 1 Coloring 6reaction FC PP (Lowry 2 method) W P 2 FC Bs H 2 O Light Source Cu 2+ Protein sample Bead Suspension Light Source
26 COCA - Collaborative on Oceanographic Chemical Analysis 26 Dear Colleagues, It is our pleasure to announce and invite you to participate in the: 18 th International Conference on Flow Injection Analysis ICFIA, September 15-20, 2013, Porto, Portugal 18 th ICFIA Contact icfia18@gmail.com
27 COCA - Collaborative on Oceanographic Chemical Analysis 28 GREEN CHEMISTRY APPROACH FOR IRON DETERMINATION IN BATHING WATERS Exploiting the lab on valve concept to study the 3,4- HPO chelator as non-toxic reagent for the determination of iron in coastal and inland bathing waters
28 COCA - Collaborative on Oceanographic Chemical Analysis 29 Coastal and inland balnear waters Important to monitor some aesthetic parameters High impact waters More often quality assessment Public health
29 COCA - Collaborative on Oceanographic Chemical Analysis 30 3,4-HPO Ligands 3,4-HPO Ligand Structure Chemical formula Molecular weight pka 3-Hydroxy-2- methyl-4- pyridinone 3-Hydroxy-1, 2- methyl-4-pyridinone 2-Ethyl-3-hydroxy- 4-pyridinone 2-Ethyl-3-hydroxy- 1-methyl-4- pyridinone Hmpp Hdmpp Hetpp Hempp O O O O N H OH CH C 6 H 7 O 2 N C 7 H 9 O 2 N C 7 H 9 O 2 N C 8 H 11 O 2 N pka 1 = 3.62 ± 0.05 pka 2 = 9.48 ± N CH 3 OH CH pka 1 = 3.69 ± 0.01 pka 2 = 9.61 ± N H OH pka 1 = 3.63 ± 0.04 pka 2 = 9.62 ± 0.05 OH C 2 H 5 N C 2 H 5 CH 3 pka 1 = 3.53 ± 0.02 pka 2 = 9.46 ± 0.05
30 31 Detector HC µsi-lov manifold for iron determination with 3,4-HPO ligand Hmpp SP W W 1 FC 2 3 SV Sample / standard SP, syringe pump; HC, holding coil; OF, optical fibers H 2 O Light Source 3,4-HPO ligand Buffer
31 Parameters for iron quantification with 3,4- HPO ligand Hmpp 32 Parameter Volumes Buffer composition Flow rate for determination Ligand 40 µl Buffer 5 µl Sample 50 µl Used value [HCO 3 - ] =0.25 M ph= µl/s Slope (Lmg-1) intercept The decrease of ligand concentration results in slope decrease and intercept increase Concentration of ligand solution 12 mg/l Dilution factor
32 33 Interference assessement Major ions possibly present in water and bivalent (and trivalent) metal ions
33 34 Features of the developed µsi-lov method Dynamic range (mg/l) Calibration curve A = m.[fe +3 ] + b LOD (µg/l) LOQ (µg/l) Quantification rate (h-1) Reagent/ sample consumption Effluent production (µl) y = (±0.0008) [Fe +3 ] (±0.0004) R² = mg Hmpp 0.11 mg NaHCO mg HNO 3 50 µl 350 Lower dynamic range Iron preconcentration Decrease the quantification limit Limitations for coastal waters application Use of NTA Superflow resin Solid phase spectrometry (SPS)
34 35 µsi-lov for iron determination with 3,4-HPO ligand with preconcentration in NTA resin SP HC W Detector FC 2 W 1 3 SV ,4 HPO ligand /buffer HNO 3 Sample / standard H 2 O 2 SP, syringe pump; HC, holding coil; OF, optical fibers NTA resin (B) for Light Iron preconcentration; Source HMeasurement 2 O based on solid phase spectrometry SPS 3,4-HPO ligand Buffer
35 Salinity interference minimization with the SPS method µsi-lov for iron determination slope (L/mg) salinity added µsi-lov-sps 36 for iron speciation slope (L/mg) salinity added
36 Bathing waters 37
37 38 Coastal bathing waters locations Porto urban area 5 Km
38 39 µsi-lov [Fe 3+ ] µsi-lov = (± 0.140) [Fe 3+ ] RM (± 0.011) RM
39 COCA - Collaborative on Oceanographic Chemical Analysis 40 Acknowledgments Raquel B. R. Mesquita thanks to Fundação para a Ciência e a Tecnologia (FCT, Portugal) and Fundo Social Europeu (FSE) the grant SFRH/BPD/41859/2007 and to Luso American Foundation (FLAD). This work was supported by National Funds through FCT, projects PTDC/AAC- AMB/104882/2008 and Pest- C/EQB/LA0016/2011.
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