Research methods
The Daily Water Sample program was started in 1978 to moniter the daily
nutrient cycling within the North Inlet Estuary System. The original sampling site
was at Town Creek. Later samples were taken at Clambank and Oyster Landing as well.
Samples were taken 365 days a year at the same time every day, approximately 10:00
AM EST. The samples were filtered at the Lab to separate the particulates from the
water. The filtered water was then run through a colorimeter and the nutrient value
peak heights derived were recorded on a ticker tape type printout. These values were
recorded into a master nutrient note book, and entered onto a CMS file by the Data
Management Department at the lab. The filters that were used in the filtering
process were first weighted, burnt off and cleaned, then they were weighted with the
particulates dried onto them. These filters were then cooked again for 24 hours at
450 degrees. This was done to determine the inorganic particulates that were left.
All data files have since been stored on a SUN workstation and backed up on 8mm
tapes at both the field lab and the data management office at the University of
South Carolina.
Analysis methods for the individual variables are described below:
Ortho-Phosphate
The basic method was the Technicon Industrial Method No. 155-71W (1973), which
was a modification of the Murphy and Riley (1962) single solution method. The method
depended on the formation of a phospomolybdate blue complex, the color of which is
read at a wavelength of 880 mm.
Sulfate
For this automated procedure for sulfate (Technicon Industrial Method No.
118-71W/B), the sample was first passed through a cation-exchange column to remove
interferences. The sample containing sulfate was then reacted with barium chloride
at a pH of 2.5-3.0 to form barium sulfate. Excess barium reacted with methylthmol
blue to form a blue-colored was gray; if it was all chelated with barium, the color
was blue. Initially, the barium chloride and methylthymol blue were equimolar and
equivalent to the highest concentration of sulfate ion expected; thus the amount of
uncomplexed methylthymol blue, measured at 460 mm, was equal to the sulfate
present.
Nitrate/Nitrite
The basic method was Technicon Industrial Method No. 158-71W/B, which utilized
the reaction in which nitrate was reduced to nitrite by a copper- cadmium reductor
column. The nitrite ion then reacted with sulfanilamide under acidic conditions to
form a diazo compound. This compound then coupled with N-1-napthylethylenediamine
dihydrochloride to form a reddish-purple azo dye.
Chlorides
The basic method was Technicon Industrial Method No. 99-70W/B. This automated
procedure for the determination of chloride depended upon the the liberation of
thiocyanate ion from mercuric thiocyanate by the formation of unionized but soluble
mercuric chloride. In the presence of ferric ion, the liberated thiocyanate formed a
highly colored ferric thiocyanate proportional to the original chloride
concentration.
Ammonia
This method was from "The Automated Analysis of Nutrients in Seawater: A Manual
of Techniques" by Gilbert and Loder (1977), a Woods Hole Oceanographic Institution
publication No. WHO-77-47. This method was dependent upon the Berthelot Reaction,
during which the formation of a blue colored compound closely related to indophenol
occurred when the solution of an ammonium salt was added to sodium phenoxide,
followed by the addition of sodium hypochlorite. A solution of potassium sodium
tartrate and sodium citrate was added to the sample stream to eliminate the
precipitation of the hydroxides of calcium and magnesium. Total Nitrogen - Total
Phosphorus The procedure consisted of an alkaline persulfate oxidation followed by
automated analysis for nitrogen (as nitrate) and phosphorus (as phosphate) on a
Technicon Auto Analyzer. With alkaline persulfate digestion, all nitrogen in the
sample was oxidized to nitrate, which was then reduced by cadmium and analyzed as
nitrite. The precision and recovery of this method compared favorably with a
Kjeldahl procedure (D'Elia et al., 1977). The main advantage of the persulfate
oxidation was the speed and convenience with which it was accomplished. Unlike the
Kjeldahl method, which recovered only organic and NH4 - nitrogen, the persulfate
oxidation also included NO3 and NO2 nitrogen. The procedure described by D'Elia
(1977) has been so the digest could be analyzed using an autoanalyzer (Gilbert et
al., 1977). This semi- automated procedure combined the persulfate oxidation with
automated nitrate and phosphate analysis to provide simultaneous analysis for total
persulfate nitrogen (TPN) and total persulfate phosphorus (TPP). Gilbert's procedure
was further modified to use a smaller sample size so that the digestion flask could
double as the sampler tray cup (Loder, 1978). Using this method, many samples could
be run quickly with a minimum of handling. Carbon (prior to September 1989)
Particulate Carbon 20 ml of daily water sample was filtered through a precombusted
(24 hr @ 450 C) Whatman GFF 0.7 micron glass filter. The filter was waved over
concentrated HCl to eliminate inorganic carbon, then placed in a precombusted ampule
and frozen. Batches were dried at 60 C for 48 hrs. 0.1 gr of precombusted (48 hr @
750 C) reagent grade CuO was added to each ampule, the ampule was then purged with
pure oxygen and sealed, using an Oceanigraphics International Purging and Sealing
Unit #524PS. Ampules were etched, then heated to 580 C for 4 hrs. Sorted ampules
were then analyzed on an Oceanigraphics International #524C TOC Analyzer by
comparing infrared absorption peak heights (Horiba PIR 2000) on an LDC strip chart
recorder with those of KHP (Potassium Hydrogen Phthalate) standards. Standards were
prepared by microsyringing a series of volumes of a KHP solution into ampules and
processing them as samples. Reagent blanks were subtracted and average concentration
of duplicate samples was calculated for each DWS site. Dissolved Organic Carbon The
daily water sample was filtered through a precombusted (24 hr @ 450 C) Whatman GFF
0.7 micron glass filter. An aliquot of filtrate was refrigerated. Samples were
purged of inorganic carbon by adding 25percent phosphoric acid to lower the pH to 2
and bubbling for 10 minutes with pure oxygen. The pH was then neutralized using
ammonium hydroxide to protect the oxidizing catalyst. Every 5 minutes, using a
Hamilton CR-700-200 spring loaded syringe, a 100 microliter sample was injected into
a Beckman 915A TOC Analyzer and infrared absorption peak heights were compared to
those of KHP standard solutions to calculate carbon concentration. Carbon (after
September 1989) Dissolved Organic Carbon Water samples were filtered through a
precombusted Whatman GFF 7.0 micron glass filter. Samples were purged of inorganic
carbon by adding 10 percent HCl and sparged with ultra zero grade air. A 40
microliter aliquot was injected, via a autosampler, into a Shimadzu TOC-500 organic
carbon analyzer. A non-dispersive infrared gas analyzer was utilized to measure the
carbon dioxide produced and samples were run in triplicate with means reported.
***NOTE*** A Whatman GFF 7.0 micron glass fiber filter was used throughout the LTER
Daily Water Sample analysis.