Organic compounds in sludges 237
10.8 MISCELLANEOUS ORGANIC COMPOUNDS
10.8.1 Cobolamin
Beck and Brink et al [188] have described a sensitive method for the routine assay of
cobalamins in activated sewage sludge. The method involves extraction with benzyl
alcohol, removal of interfering substances using a combination of gel filtration and
chromatography on alumina, concentration of the extract by lyophilisation, and direct
determination of total cobalamin by high performance liquid chromatography.
10.8.2 Nucleic acids
Seibert and Dahn et al [189] used precipitation titration with the cationic detergent
N-cetyl-N-N, N-trimethylammonium bromide to quantify the nucleic acid content
in activated sludge. With the introduction of toxins into the influent of a sewage
treatment plant the ribonucleic acid content of the activated sludge fell sharply, and
it was possible to correlate the deterioration of effective purification with a reduction
in the ribonucleic acid fraction precipitate by N-cetyl-N, N, N-trimethylammonium
bromide. Advantages of the method are simplicity and speed. Deoxyribonucleic acid
and ribonucleic acid can be determined in one operation and the process is not affected
by the presence of dyes or compounds absorbing ultraviolet light.
10.8.3 Adenosine phosphate ether
Smirnova et al [190] have described a simple non-enzymatic method of quantitative determination of adenosine triphosphate in activated sludge from aeration tanks.
Extraction of the nucleotides in boiling distilled water was followed by removal of
the protein impurities by acidification. Barium salts of di- and triphosphates of the
nucleotides were precipitated and the precipitate was washed and dissolved in acid.
The quantity of adenosine triphosphate was determined quantitatively by determining
inorganic phosphorus in the liquid over the precipitate before and after acid hydrolysis, and by ultraviolet absorption spectraoscopy The method was tested in activated
sludge from operational sewage works. There was good agreement between the adenosine triphosphate content determined spectrophotometrically and by determination of
phosphorus, despite the presence of small quantities of secondary impurities.
Luminescence methods
Patterson et al [191] carried out determination of adenosine triphosphate in activated sludge. The method involved the use of firefly lantern extract. Kucnerowcz
and Verstraete et al [192] carried out direct measurements of microbial adenosine-5
-
triphosphate in activated sludge samples. The sample was diluted with triethanolamine
buffer, homogenised, mixed with adenosine-5
-triphosphate releasing agent and
the luminescence of the mixture measured after addition of luciferase. Results
obtained were in agreement with literature data for adenosine-5
triphosphate in activated sludge. Relationships established between adenosine-5
-triphosphate content of
sludges and other sludge parameters indicate that adenosine-5
-triphosphate determination could be used as a method of monitoring activated-sludge treatment processes.
10.8 MISCELLANEOUS ORGANIC COMPOUNDS
10.8.1 Cobolamin
Beck and Brink et al [188] have described a sensitive method for the routine assay of
cobalamins in activated sewage sludge. The method involves extraction with benzyl
alcohol, removal of interfering substances using a combination of gel filtration and
chromatography on alumina, concentration of the extract by lyophilisation, and direct
determination of total cobalamin by high performance liquid chromatography.
10.8.2 Nucleic acids
Seibert and Dahn et al [189] used precipitation titration with the cationic detergent
N-cetyl-N-N, N-trimethylammonium bromide to quantify the nucleic acid content
in activated sludge. With the introduction of toxins into the influent of a sewage
treatment plant the ribonucleic acid content of the activated sludge fell sharply, and
it was possible to correlate the deterioration of effective purification with a reduction
in the ribonucleic acid fraction precipitate by N-cetyl-N, N, N-trimethylammonium
bromide. Advantages of the method are simplicity and speed. Deoxyribonucleic acid
and ribonucleic acid can be determined in one operation and the process is not affected
by the presence of dyes or compounds absorbing ultraviolet light.
10.8.3 Adenosine phosphate ether
Smirnova et al [190] have described a simple non-enzymatic method of quantitative determination of adenosine triphosphate in activated sludge from aeration tanks.
Extraction of the nucleotides in boiling distilled water was followed by removal of
the protein impurities by acidification. Barium salts of di- and triphosphates of the
nucleotides were precipitated and the precipitate was washed and dissolved in acid.
The quantity of adenosine triphosphate was determined quantitatively by determining
inorganic phosphorus in the liquid over the precipitate before and after acid hydrolysis, and by ultraviolet absorption spectraoscopy The method was tested in activated
sludge from operational sewage works. There was good agreement between the adenosine triphosphate content determined spectrophotometrically and by determination of
phosphorus, despite the presence of small quantities of secondary impurities.
Luminescence methods
Patterson et al [191] carried out determination of adenosine triphosphate in activated sludge. The method involved the use of firefly lantern extract. Kucnerowcz
and Verstraete et al [192] carried out direct measurements of microbial adenosine-5
-
triphosphate in activated sludge samples. The sample was diluted with triethanolamine
buffer, homogenised, mixed with adenosine-5
-triphosphate releasing agent and
the luminescence of the mixture measured after addition of luciferase. Results
obtained were in agreement with literature data for adenosine-5
triphosphate in activated sludge. Relationships established between adenosine-5
-triphosphate content of
sludges and other sludge parameters indicate that adenosine-5
-triphosphate determination could be used as a method of monitoring activated-sludge treatment processes.