DETERMINATION OF DISSOLVED OXYGEN IN SEAWATER BY WINKLER TITRATION USING THE AMPEROMETRIC TECHNIQUE
1. SCOPE AND FIELD OF APPLICATION
This paper describes procedures to be used for the determination of dissolved oxygen in discrete samples of seawater. These procedures are based on the modified Winkler titration method (Carpenter 1965). These modifications reduced the loss of I2 during the titration due to volatilization by optimizing the concentrations of the “pickling” reagents to encourage the formation of the more stable triiodide complex I3- and by adopting the whole-bottle titration method which eliminates the loss of I2 during transfer of sample aliquots. This procedure is suitable for the measurement of the full range of oceanic oxygen concentrations (0-400 μmol kg-1) in uncontaminated seawater. The typical precision that can be achieved using automated amperometric endpoint detection systems is ±0.15 μmol kg-1. Carpenter (1965) established the accuracy of the method as <0.1% or ±0.3 μmol kg-1. This procedure is unsuitable for seawater containing hydrogen sulfide (H2S). In oxygen-deficient regions (<5 μmol kg-1) a high concentration of nitrite (NO2-) may cause a positive oxygen bias. Polluted waters may contain reducing substances that will react with the liberated I2 resulting in a negative oxygen bias.
2. INTRODUCTION
Dissolved oxygen measurement is commonly a standard part of most hydrographic studies. The data is of interest to physical, chemical and biological oceanographers. Physical oceanographers use it to characterize water masses. Chemical oceanographers use it study to study the production and destruction of organic matter. Biological oceanographers use it determine rates of photosynthesis and respiration. Climate scientists are finding it to be a sensitive indicator of climate-related changes in the ocean circulation and ventilation of intermediate and deep water. It is also used in the quantification of the uptake of anthropogenic CO2 by the ocean. The performance of oxygen sensors have dramatically improved in recent years; however, for the most precise work they still need to be calibrated frequently to correct for drift and temperature and pressure influences. The chemical titration method first described by Winkler (1888) remains the method of choice for the analysis of discrete water samples. The method is fast and inexpensive. The development of computer-controlled titrators has eliminated the tedium and operator variance of manual titration and has pushed the routine shipboard precision of the method to 0.06% or ± 0.15 μmol kg-1. In 1991, a report was published (Culberson,et al., 1991) describing an inter-comparison experiment between oxygen measuring groups at four institutions. The experiment compared three groups using automated endpoint detection methods (two amperometric, one photometric) with one group using the manual method with starch endpoint detection. The systematic differences between methods and groups was encouragingly small; i.e., worst case 0.6% and on average 0.3%. However, because the systematic differences were large relative to the precisions attained by the different groups (0.06-0.15%), it was recognized that there was significant room for improvement. After considering possible causes of the systematic differences between groups the report came out with a set of recommendations for improving the accuracy of the dissolved oxygen method:
1) Calibrate the volume of all pipettes, volumetric flasks, burettes and oxygen flasks and include correction for buoyancy.
2) Correct for effect of thermal expansion on the masses of thiosulfate and iodate dispensed.
3) Measure the temperature of the seawater sample at the time of pickling in order that it’s mass can be accurately determined.
4) Standardize the set of equations used to compute oxygen concentration and correct for the thermal expansion of the sample and solutions.
5) Determine the oxygen content of the pickling reagents as a function of temperature.
6) Perform intercomparison study at low oxygen concentration.
7) A study should be made of the seawater blank in coastal and open ocean waters at surface, oxygen minimum, nutrient maximum and bottom depths. Until more is known about the magnitude and variability of the seawater blank, only the pure water or reagent blank should be measured. While this will result in small errors in computed oxygen concentrations of -0.5 to + 0.8 μmol kg-1 they will be internally consistent between groups.
Implementation of the recommendations made by Culberson (1991) and Dickson (1995) with respect to the careful calibration of all glassware used in the method, the corrections for the effect of temperature in the lab on the mass of thiosulfate and iodate dispensed, and the effect of draw temperature on the mass of sample contained in the oxygen flask (i.e., recommendations 1-4) have contributed to a general improvement in the accuracy and precision of discrete dissolved oxygen measurements. It is important to note that recommendations 5-7 have still not been addressed. It would be wise in the not-too-distant future to conduct another intercomparison experiment both as an opportunity to judge how far systematic differences between groups have improved since 1991 and also to tackle the seawater blank and oxygen content of the reagents as a function of temperature.
In this paper I review the procedures that are followed to implement recommendations 1-4. Sample calculations are provided so that investigators coding the equations into their software can confirm that they are getting the correct values. New in this paper, I discuss the amperometric method of endpoint detection in detail and present the results of a side by side comparison of an amperometric and a photometric system and show that the oxygen concentrations obtained are identical to within the measurement uncertainties of 0.06% or ± 0.15 μmol kg-1. Also new in the paper, I compare the “standard curve” method of thiosulfate standardization and reagent blank determination that some groups around the US are using with the methods described in Carpenter (1965) and in the 1991 and 1995 Standard Operating Procedures by Culberson et al. and Dickson, respectively.
3. PRINCIPLE OF THE WINKLER METHOD
Manganous chloride (MnCl2) solution is added to a known quantity of seawater and is immediately followed by the addition of an alkaline sodium hydroxide-sodium iodide solution (NaOH/NaI). Manganous hydroxide (Mn(OH)2) precipitates and reacts with the dissolved oxygen in the water with the formation of a hydrated tetravalent oxide of manganese (MnO(OH)2).
Mn2+ + 2OH– → Mn(OH)2
2Mn(OH)2 + O2 → 2MnO(OH)2
Upon acidification, the manganese hydroxides dissolve. In the acid solution, the tetravalent manganese in MnO(OH)2 acts as an oxidizing agent and liberates iodine (I2) from the iodide ions (I-).
2MnO(OH)2 + 8H+ + 4I– → 2Mn2+ + 2I2 + 6H2O
Two moles of I2 are formed for each mole of O2 present in the sample. The amount of I2 in the solution is determined by titration with a standardized sodium thiosulfate (Na2S2O3) solution.
I2 + 2S2O32- → 2I– + S4O62-
Two moles of thiosulfate are required to titrate each mole of I2. Since two moles of I2 were formed for each mole of O2 the final stoichiometry is four moles of thiosulfate equals one mole of O2. By knowing the concentration of the thiosulfate solution and the volume required to titrate the liberated I2 the amount of the oxygen dissolved in the seawater sample can be easily computed.
4. AMPEROMETRIC ENDPOINT DETECTION
The method of titration in which the endpoint is detected by the sudden increase or decrease in current flow between two similar electrodes immersed in a solution due to polarization effects is known as the “dead-stop” endpoint amperometric method. Foulk and Bawden (1926) and Bradbury and Hambly (1952) investigated the advantages of using the amperometric method over the visual starch endpoint method in the titration of iodine with thiosulfate. The first report of the use of the amperometric method of endpoint detection applied to the Winkler titration is Truesdale and Knowles (1956). They described two methods, the first involved adding an excess of thiosulfate and then back-titrating with potassium iodate. The second method involved titrating directly with thiosulfate. It is the later method that is more widely used today (Culberson and Huang, 1987). The amperometric method involves applying a potential to an electrode placed in a solution so that an analyte is reduced and using the resulting current as an indicator of the concentration of that analyte. In the case of the Winkler titration the analyte is I2. A potential of 100 mV is applied between two platinum electrodes. In an acidified solution the iodine is reduced at the cathode to iodide (I2 + 2e- → 2I-) and the reverse reaction occurs at the anode (2I- →I2+2e-). The result is a stable current proportional to the concentration of I2 in solution. During the titration, thiosulfate (Na2S2O3) is added and it reacts with the I2 to form 2I-. As the I2 concentration decreases so does the current measured at the electrodes. The endpoint is detected as the point at which the current ceases to
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