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This method, based on the glucose oxidase enzymatic reaction, is highly
specific for glucose and required only 25 ul of serum. Replicate determinations
of each serum sample were performed. Total and esterified
cholesterol in serum was determined by a cholesterol oxidase assay
system which is both highly snesitive and specific.
For tissue-free amino acid analysis, muscle tissue was thawed,
weighed and homogenized in distilled water using a 2/1 ratio of distilled
water/wet weight. Homogenates were deproteinized with 12.5% trichloroacetic
acid and then centrifuged. The supernates were frozen, thawed,
and centrifuged again to remove additional TCA precipitates. The supernates
were then evaporated to dryness on a rotary evaporator and the
residue dissolved in 0.2 M Citrate buffer adjusted to pH 2.2. The
extracts were analyzed with a Beckman automatic amino acid analyzer.
The amino acid composition of the extract and the concentration of
individual amino acids in it were determined. Taurine/glycine molar
ratios were computed. Variations in amino acid compositions and concentrations
among fish and oysters from different sampling stations were
analyzed statistically.
Plaice liver was analyzed for ascorbic acid. Tissue samples were
thawed, weighed and homogenized in 3% metaphosphoric acid-8% acetic acid
solution. After centrifugation, the supernates were analyzed immediately
by the a,a-diperidyl technique of Zannoni et al. (1974).
Oysters and plaice samples for hydrocarbon analysis were taken at
the same times and places as samples for biochemical/histopathological
analysis. Ten to twelve whole oysters were pooled for each sample.
They were shucked and tissues were rinsed in distilled water, blotted
dry, wrapped in hexane-cleaned aluminum foil and frozen at -60°C until
analysis. For the April 1979 sample, whole fish were used. For subsequent
samples, pooled samples of liver and muscle from 5-10 fish were
used. Fish tissue samples were handled like oyster samples.
Hydrocarbon analyses were performed by Dr. Paul Boehm, ERCO,
Cambridge, Massachusetts using capillary gas chromatography /mass
spectrometry.
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