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By Donald B. Nuzzio, Martial Tailjefert, S. Craig Cary, Anna Louise Reysenbach and George W

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By Donald B. Nuzzio, Martial Tailjefert, S. Craig Cary, Anna Louise Reysenbach and George W DELU-R 02-003 D EL-SG-06-02 IN SITU VOLTAMMETRY AT DEEP-SEA HYDROTHERMAL VENTS by Donald B. Nuzzio, Martial Tailjefert, S. Craig Cary, Anna Louise Reysenbach and George W. Luther, ill This work was sponsored, in qart, by the National Sea Grant College Program under Grant No. NA16RG0162-03 Project No. R/B-33!. Reprintedfrom American Chemical Society Symposmm Series ttt 1, EnvirOnmentalElectrochemistry: Analyses of TraceElement Universityot DelawareNewark, Delaware 19716 ACKNOWLEDGEMENTS This publication was supported,in part, by the National Sea Grant Coliege Program of the U.S. Department of Commerce's National Oceanic and Atmospheric Administration under NOAA Grant No. NA16FtG0162-03 Project No. R/B-33!. The views express herein do not necessarily reflect the views of any of those organizations. Chapter 3 ln SituVoltammetry at Deep-Sea Hydrothermal Vents DonaldB.Nuzzio'*, Martial Taillefert, S.Craig Cary', AnnaLouise Reysenbach, and George W. Luther,III' 'AnalyticalInstrument Systems, Inc.,P.O. Box 458, Flemington, IVJ08822 CollegeofMarine Studies, University ofDelaware, 700Pilottown, Road, Lewes, DE 19958 'DepartmentofBiology, Portland StateUniversity, Portland, OR97201 Thereis a needto buildinstrumentation andsensors that can measurein situ chemicalchanges in dynamicenvironments. Hydrothermalvents are arguably the most dynamic aqueous systemson earth. The orifice of a ventapproaches 360 'C and spewsvast quantities of dissolvedhydrogen sulfide and iron intoambient seawater at 2 'C.These chemical species fuel incredibledeep-sea micro!biological communities, which may bea modelfor life onother planets, Here we describe an in situ submersibleanalyzer and electrodes for themeasurement of aqueouschemical species found near hydrothermal vents. A standardthree-electrode arrangement is controlled by a voltarnmetricanalyzer that is deployedfrom the deep-sea submersible,Alvin. Real time measurements fora variety of redoxspecies under flow conditions were made with a I00Iim Au/Hgsolid-state working electrode ata depthof2500 m. The solid-stateworking electrode was used to detect dissolved 02, S -II!,Fe II! andFeS,q molecular clusters, Our in situ data showthat significant changes canoccur in chemical speciation 40 C 2002American Chemical Society andanalyte concentration when waters are sampledand then measuredaboard ship. Introduction Voltammetric micro!electrode techniques have been used in a varietyof geochemistryand marine chemistry applications. Recently, in situ voltammetric measurementshave received increasing attention !. Bothdirect -4! andon- line -11, flow cell! type arrangementshave been used for trace metal and inajor chemical speciesdeterminations. However, voltamrnetricinstrumentation has only been deployed in shallow waters to date. There is a need to inake measurementsin the deep-sea,and this is particularlytrue of deep-sea hydrothermal vents that have an mcredible microbial and macrofaunal communityfueling itself via chemosynthesis2,13!, Changesin temperature canbe dramaticin this environmentand electrodes must be ableto respond preciselyin watersof lowerpH, high pressure, high temperature and high water flow rates.When high temperature waters mix with low temperaturewaters, their chemistriescan differ dramatically . Because organisms live in thesedynainic andextreme conditions, it is criticalto understandhow that chemistry drives or influences biology, In this paper we describe i n siru electrochemical instruinentationand the initial deployment of'it at 9 'N EastPacific Rise EPR; at a 2500meter water depth; 250 atmof pressure!and at GuaymasBasin, Gulf of California,2000 meter water depth 00 atmof pressure!.A companion paper in this volume ll! demonstratesthat current-concentrationcurves are affected by waterflow ratesbut not by pressure.At highflow ratesand reasonable scan rates - I V s '!,the current-concentration curvesbecome independent of water flow rate for the targetredox species. Deep-seahydrothermal vents can havehigh concentrationsof iron and sulfide.In this paperwe demonstratethe use of voltamrnetryto ineasureFe and S species.Using a solid-stategold amalgam Au/Hg! working electrode, we showthe simultaneousdetection and quantificationof severalsulfur and iron dissolvedspecies. In previouswork, dissolvedchemical species >, H~O~, Sg03,S', HS, I, Fe II!,Mn II!,organically complexed Fe III!,and FeS clusters! have been simultaneouslymeasured in situ in sedimentsand natural waters , 14,15!. 42 Experimental Methods Chemicals and solutions Chemicalsused in laboratoryexperiments were analyticalgrade from Fisher Scientific Co, Milli-Q quality Millipore! was usedfor all reagents.Laboratory measureinents were carried out in a 0.55 M NaC1 solution or in seawater, Mn II!, Fe II! and S -II! standardswere preparedfrom MnC12~ 4 H,O, ferrous ammoniuinsulfate, and NaqS9HqO. The mercuryplating solution was prepared as 0.1 N Hg NO> in 0.05N HNO,. Electrodes Goldamalgam PEEK" electrodes were made as described by Lutheret al ! by fixing 100 pm-diameter Au wire soldered to the conductor wire of a BNC cablewithin a bodyof 0.125"-diameterPEEK" tubing, which is commercially available as standardHPLC high-pressuretubing. The metal is fixed within the tubing with West System 105 epoxy resin and 206 hardener, A portion of the black outercoat andbraid of the BNC wire arereinoved to exposethe teflon shield and Cu conductor wire so that the Au wire soldered onto the Cu conductor canbe inserted into the PEEK ". Theepoxy is injectedinto the PEEK" tubing which containsthe gold wire that was previouslysoldered to the conductorwire of theBNC cable. Then the teflon is insertedinto the PEEK" tubing until the black coating of the BNC wire fits againstthe PEEK tubing, and the assembly is held with epoxy, which hasa moderatesetting time -1 hr!, and doesnot drain out the lower open side. On setting,the epoxyseals the tip andthe lower endcan be refilled with epoxy if necessary,Then the top end is coatedwith Scotchkote M! electricalcoating and Scotchfil M! electricalinsulation putty. PEEK" and high-purity epoxy fill permit the determinationof metal concentrations without risk of contamination,and at temperaturesas high as 150 'C. Pt counter and solid Ag/AgCI reference electrodes were made similarly but 500 lim diameterwire was usedfor each.These PEEK" electrodescould be usedas is or mated with standardHPLC fittings from Vpchurch,Inc for insertion into a flow cell //!. Once constructedthe working electrode Au! surfacewas sanded,polished and plated with Hg by reducingHg II! from a 0,1N Hg / 0.05 N HNO3solution, for 4 minutesat a potentialof -0.1 V, while purging with N2, The mercury/gold amalgam interface was conditioned using a 90-second -9 V polarization procedurein a 1 N NaOH solution 6!, The electrodewas then run in linear 43 sweep mode &orn -0.05 to -1.8 V versus a Saturated Calomel Electrode SCE! or Ag/AgCl electrodeseveral tiines in oxygenatedseawater to obtain a reproducible 02 signal. For DSVAlvin work, four Au/Hg electrodes can be controlled by the analyzer. The reference electrode was Ag/AgCl and the counter electrode was Pt wire, both of which weremounted on the basketof DSV Alvin so that they would not enter sulfidic waters 5!. For hydrothermal vent work, the Ag/AgCl reference was silver wire, which was oxidized in seawater at +9 V for 10 sec to form a AgCl coating. This electrode was used as a solid-state electrode in the seawatermedium =0.7! so that no pressureeffects on filling solutionswould hinder electrodeperformance. Peakpotentials of the analytesmeasured in situ and aboard ship were the same and similar to those for a saturated caloinel electrode SCE!. All laboratory and shipboardanalyses were carried out using an Analytical InstrumentSystems AIS! DLK-100A potentiostatcontrolled by a microcomputerusing software provided by the manufacturer.A DLK-SUB-I was used for allinsituwork see below!. Field Experiments For hydrothermal vent work, two working electrodes as well as a therinocouplesensor and tubing that lead into a flow cell inlet and a discrete syringesampler system were placedin a sensoror wandpackage 5! that canbe held by a manipulator arm! of Alvin, The wand was held over the vent orifice and areas along the length of vent chimneys. These latter areas are termed diffuse flow becausewater temperaturescan rangefrom 8 to 125 'C and do not emanate&om the vent orifice. A flow cell 1,15! was fixed in the submersible's basket that was bathed by waters at 2 'C. A submersible electrochemical analyzer DLK-SUB I! from Analytical InstrumentSystems, Inc. was used for data collection see below!. The electrochemicalpackage was deployed during cruises to 9 'N East Pacific Rise May, 1999! and GuaymasBasin, Gulf of California January 2000!. Separatediscrete sampleswere taken with a gas tight syringe sampler 1,16! from the samewaters for comparisonwith the flow cell measurements. The discrete sampleswere measuredaboard ship by voltammetry 1!. We typically madethree to five replicatemeasurements per samplewith the flow cell system,Electrodes were calibrated at different temperature7! as well as for flow 1!. 44 Voltammetry The three-electrodeconfiguration working, reference and counter! was used to determine the concentrationof the speciespresent in natural waters. Linear sweepvoltammetry LSV!, cyclic voltammetry CV! and squarewave voltammetry SQW! were used for analyses, The following conditions were generallyapplied during the LSV and CV scans:scan rate = 200, 500 or 1000 mVs ',scan range = -0,1to -1.75V, equilibrationtime = 5 s, Squarewave
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