228
Phytoplankton
chromatographic methods. However, although the results for chlorophyll a by the
polychromatic method were in reasonable accord with those derived chromatographically, many of those for the other chlorophylls showed a high discrepancy. Obviously, the polychromatic method is particularly unsatisfactory with respect to the
interference of chlorophyll degradation products, as these are nearly always present in
environmental samples.
Sartory [655] used a combination of high performance liquid chromatography and
spectrometry to determine algal pigments. He discussed sample cleanup procedures
which allowed determination of chlorophylls free from carotenoids, and a high performance liquid chromatography procedure with fluorescence detection which allowed separation of all chlorophyll pigments within 40 min, using a simple solvent
programme. The method had detection limits of 10 pg for chlorophyll a and phaeophytin b, 15 pg for chlorophyll b, and 20 pg for phaeophytin a. Comparative analyses
of carotenoid-free extracts by high performance liquid chromatography and several
spectrophotometric procedures tended to over-estimate chlorophyll a and phaeophytin a, and to underestimate chlorophyll b.
9.4.7
Humic Substances
Gadel and Brucket [659] applied pyrolysis-gas chromatography-mass spectrometry to
the characterization of humic substances resulting from the decay of aquatic algae and
macrophytes. The composition of humic substances from decaying algae and aquatic
macrophytes in a coastal Mediterranean lagoon and from sediments from different
sources, including a lake on the Greenland icecap, was investigated. The material was
also analysed by gas elemental analysis and by infrared spectroscopy. The humic
matter included varying proportions of carbohydrates, n-acetylamino sugars, proteinaceous material, and lignin derivatives: lesser amounts of phthalates and aliphatic
compounds were found in some of the samples. There were marked differences
between humic and fulvic fractions, with most of the carbohydrates being included in
the fulvic acids.
9.4.8
Adenosine Triphosphate
Shoaf and Lium [660] compared three extraction methods involving dimethyl sulphoxide, boiling tris buffer and butanol-octanol for the isolation of adenosine triphosphate from algae prior to its luminometric determination using luciferin-Iuciferase
assay. All were equally effective.
Martin [661] describes the chemistry and biology of adenosine triphosphate. The
extraction of adenosine necessitates rupturing cell envelopes, and inhibiting adenine
triphosphaterase and other intracellular enzymes. Adenosine triphosphate has up to
now been determined by bioluminescence techniques, but these have been subject to
interference problems. High performance liquid chromatography and nuclear magnetic resonance using phosphorus-31 showed promise in overcoming these problems.
The adenosine triphosphate content in cells of algae is discussed, and the relationship
Phytoplankton
chromatographic methods. However, although the results for chlorophyll a by the
polychromatic method were in reasonable accord with those derived chromatographically, many of those for the other chlorophylls showed a high discrepancy. Obviously, the polychromatic method is particularly unsatisfactory with respect to the
interference of chlorophyll degradation products, as these are nearly always present in
environmental samples.
Sartory [655] used a combination of high performance liquid chromatography and
spectrometry to determine algal pigments. He discussed sample cleanup procedures
which allowed determination of chlorophylls free from carotenoids, and a high performance liquid chromatography procedure with fluorescence detection which allowed separation of all chlorophyll pigments within 40 min, using a simple solvent
programme. The method had detection limits of 10 pg for chlorophyll a and phaeophytin b, 15 pg for chlorophyll b, and 20 pg for phaeophytin a. Comparative analyses
of carotenoid-free extracts by high performance liquid chromatography and several
spectrophotometric procedures tended to over-estimate chlorophyll a and phaeophytin a, and to underestimate chlorophyll b.
9.4.7
Humic Substances
Gadel and Brucket [659] applied pyrolysis-gas chromatography-mass spectrometry to
the characterization of humic substances resulting from the decay of aquatic algae and
macrophytes. The composition of humic substances from decaying algae and aquatic
macrophytes in a coastal Mediterranean lagoon and from sediments from different
sources, including a lake on the Greenland icecap, was investigated. The material was
also analysed by gas elemental analysis and by infrared spectroscopy. The humic
matter included varying proportions of carbohydrates, n-acetylamino sugars, proteinaceous material, and lignin derivatives: lesser amounts of phthalates and aliphatic
compounds were found in some of the samples. There were marked differences
between humic and fulvic fractions, with most of the carbohydrates being included in
the fulvic acids.
9.4.8
Adenosine Triphosphate
Shoaf and Lium [660] compared three extraction methods involving dimethyl sulphoxide, boiling tris buffer and butanol-octanol for the isolation of adenosine triphosphate from algae prior to its luminometric determination using luciferin-Iuciferase
assay. All were equally effective.
Martin [661] describes the chemistry and biology of adenosine triphosphate. The
extraction of adenosine necessitates rupturing cell envelopes, and inhibiting adenine
triphosphaterase and other intracellular enzymes. Adenosine triphosphate has up to
now been determined by bioluminescence techniques, but these have been subject to
interference problems. High performance liquid chromatography and nuclear magnetic resonance using phosphorus-31 showed promise in overcoming these problems.
The adenosine triphosphate content in cells of algae is discussed, and the relationship