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DISSOLVED INORGANIC CARBON

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     SUMMARY: Seawater samples are collected at discrete depths 
     using CTD-rosette sampling protocols.  Subsamples for 
     dissolved inorganic carbon are collected, immediately 
     preserved with HgCl2 and stored for subsequent analysis 
     in the laboratory using a commercial CO2 coulometer.
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1.  Principle

    The accurate and precise determination of dissolved inorganic 
carbon (DIC) concentrations over annual and interannual time scales 
is central to the achievement of GOFS objectives.  In the central 
ocean basins, DIC concentration in surface seawater is believed to 
be controlled by air-sea exchange reactions.  However, physical 
processes and biological activity also influence the concentration 
of DIC in surface waters.  Beneath the mixed layer the concentration 
of DIC increases as a result of the decomposition of organic material.
 
    DIC concentrations are determined using a commercial CO2 coulometer.  
The coulometric determination of carbon dioxide has the unique distinction 
of performing with a high degree of both precision and accuracy while 
maintaining relatively high sample throughput.  The coulometric 
determination of carbon dioxide involves the stripping of an acidified 
seawater sample with a carbon dioxide-free air stream and subsequent 
absorption of the carbon dioxide by a solution of ethanolamine.  
The weak acid generated by carbon dioxide absorbed in ethanolamine is 
titrated by a strong base produced electrolytically.  The equivalence 
point is detected photometrically with thymolphthalein as the indicator.  
The number of coulombs required to reach the end-point is proportional 
to the quantity of carbon dioxide evolved from the sample.  


2.  Precautions

    DIC samples should be the first samples taken from the water bottle 
unless dissolved oxygen (DO) or pH is also sampled from the same 
hydrocast.  In this case, DIC samples are collected immediately after 
the DO and/or pH samples.

    Careful subsampling is important for all dissolved gases.  Samples 
should be taken as soon as possible and in the same manner as DO samples 
(i.e., no bubbles, low turbulence with adequate flushing; see Chapter 5).


3.  Water Sampling

3.1. Drawing the sample

  3.1.1. Samples are drawn into clean 300 ml glass reagent bottles as 
         soon as the rosette arrives on deck.

  3.1.2. The drawing tube is completely filled with sample by raising 
         the end of the drawing tube.  Bubbles are simultaneously 
         dislodged by manipulation of the tube.  The drawing tube is 
         flushed and inserted to the bottom of the sample bottle. 

  3.1.3. The sample bottle is overflowed with at least two volumes of 
         sample.  
  
  3.1.4. The tube is slowly withdrawn from the bottle while the sample 
         is flowing so that the bottle remains brimful when the tube 
         is completely withdrawn.


4.  Preserving the Sample

4.1. Some of the sample is removed from the reagent bottle using a 
     plastic pipette equipped with a rubber bulb.  Enough water is 
     removed so that approximately 1 ml of air is contained in the 
     bottle when the glass stopper is inserted.

4.2. 100 µl of saturated HgCl2 is added to each sample.  The tapered 
     ground glass bottle neck is dried with a Kimwipe wrapped on an 
     applicator stick.  The bottle is sealed with a ground glass 
     stopper coated with a light covering of Apiezon grease.  The 
     stopper is pressed firmly into the bottle to make a good seal.  
     The stopper is secured with polyethylene tape or a large rubber 
     band.  

4.3. The samples are stored in a cool location, in the dark.


5.  Coulometric Determination of DIC

5.1. Maintenance of extraction and analysis system and temperature control

  5.1.1. The glassware used in the extraction system is combusted (450°C, 
         3 hours) on a regular basis in order to prevent the buildup of 
         organic films within the extraction system.

  5.1.2. The titration cell and rubber stopper are dried overnight at 55°C 
         before use.

  5.1.3. The air stream leaving the extractor is passed through a condensor 
         and then through a Balston air filter.                       

  5.1.4. The temperature of the titration cell is maintained at 25°C with 
         circulating water.


5.2.Analysis

  5.2.1. A 50 ml plastic syringe is rinsed with sample and then filled and 
         weighed on a microbalance.   

  5.2.2. Five ml of 6% phosphoric acid is added to the extractor and the 
         acid is purged for 2-5 minutes with carbon dioxide-free carrier gas.  

  5.2.3. The contents of the syringe are injected into the extractor through 
         a septum.  

  5.2.4. The syringe is weighed again and the mass of the extracted sample 
         determined.                 

  5.2.5. The acidified sample is purged with carrier gas.  Successive 
         coulometer readings are integrated at 1 minute intervals until 
         they differ by less than 0.01%


6.  Determination of the Coulometer Blank

    The coulometer blank is determined at intervals throughout each 
day by allowing the coulometer to titrate a CO2-free air stream.  The 
blank is taken as the µg C per minute value detected by the coulometer 
when a steady-state reading is achieved.


7.  Calibration

    Although the digital coulometer output is fairly accurate, the 
coulometer response per unit carbon may vary with time.  In order to
achieve maximum accuracy, it is necessary to calibrate the coulometer 
with samples containing known quantities of inorganic carbon.  We are 
presently using anhydrous sodium carbonate standards.  The dried (270°C 
for 3 hours) reagent is carefully weighed on a microbalance to the 
nearest 0.1 µg and introduced into a degassed acidified sample solution 
in a combusted aluminum boat through a port in the side of the extractor.  
Recoveries are generally slightly less than 100%.  


8.  Data Reduction and Calculations

    In order to compute the absolute concentration of DIC in a water 
sample, the integrated reading given at the titration endpoint must 
be corrected for both the coulometer blank and the recovery of NaCO3
standards.  These corrections are made by multiplying the blank µg C 
min-1 by the time taken to reach the endpoint and subtracting this 
value from the integrated reading.  This value is then corrected for 
the recovery of standards by dividing by the average percentage 
recovery of known standards run on the day of analysis.


9.  Precision and Accuracy

    Three replicate samples from a single Niskin bottle generally yield 
a coefficient of variation of less than 0.1% (+2 µmol/kg).  The 
precision of our replicate determinations of liquid standards averages
approximately 0.5 µmol/kg over periods of months.  The accuracy of our 
determinations is within 1 µmol/kg as determined by the routine analysis 
of liquid standards provided by Dr. Andrew Dickson of Scripps Institution 
of Oceanography under the U.S. Department of Energy Global Change 
Research Program.


10.  Quality Control

     As a safeguard on the quality of our results, we maintain a set 
of secondary seawater standards.  These are made from a large surface 
seawater sample, which is preserved with saturated HgCl2 and subdivided 
into 300 ml bottles.  These are sealed as described above and stored 
in the dark.  These secondary standards are analyzed relative to a 
primary seawater standard provided by Dr. Andrew Dickson. 


11.  Equipment/Supplies

     5011 Coulometer (UIC Inc.) and modified glassware
     300 ml ground glass stoppered reagent bottles
     kimwipes and applicator sticks
     large plastic pipette
     Apiezon grease
     Cahn microbalance
     volumetric flasks
     analytical balance / pan balance
     muffle furnace
     carbon dioxide-free carrier gas
     data acquisition system
     water bath

12.  Reagents

     distilled deionized water
     high purity sodium carbonate
     potassium iodide
     coulometer cathode solution (UIC Inc.)
     coulometer anode solution (UIC Inc.)
     orthophosphoric acid


13.  References

     Johnson, K. M., A. E. King and J. McN. Sieburth.  1985.  
     Coulometric TCO2 analysis for marine studies; An introduction. 
     Marine Chemistry, 16, 61-82.    

     Johnson, K. M., J. McN. Sieburth, P. J. LeB. Williams and L. 
     Brandstrom.  1987.  Coulometric total carbon dioxide analysis 
     for marine studies:  Automation and calibration.  Marine 
     Chemistry, 21, 117-133.                                      

     Lindberg, A. O.  1978.  Automatic coulometric titration with 
     photometric end-point detection. Part II. Coulometric 
     determination of nanomole amounts of carbon dioxide by non-
     aqueous titration.  Analytica Chimica Acta, 96, 327-333.      

     Robinson, C. and P. J. LeB. Williams.  1991.  Development 
     and assessment of an analytical system for the accurate and 
     continual measurement of total inorganic carbon. Marine 
     Chemistry, 34, 157-175.