Standard procedure for the determ...
Standard procedure for the determination of chlorophyll a by spectroscopic methods Alain Aminot IFREMER Centre de Brest BP 70 F-29280 Plouzan�� France and Francisco Rey Institute of Marine Research P.O. Box 1870 Nordnes N-5817 Bergen Norway International Council for the Exploration of the Sea Conseil International pour l���Exploration de la Mer Palaegade 2���4 DK-1261 Copenhagen K Denmark March 2000 ISSN 0903-2606
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TABLE OF CONTENTS Section Page i 1 INTRODUCTION...............................................................................................................1 2 CONTEXT AND DIFFICULTIES OF CHLOROPHYLL a DETERMINATION............1 2.1 Interfering Chlorophylls and Degradation Products .................................................1 2.2 Occurrence and Significance of the Chlorophyll Group Pigments...........................2 2.3 Spectroscopic Characteristics and Resulting Measurement Methods.......................2 2.3.1 Spectrophotometry..........................................................................................2 2.3.2 Fluorometry ....................................................................................................3 2.4 Critical View of the Preparation of Pigment Extracts...............................................3 2.4.1 Sampling and sub-sampling............................................................................3 2.4.2 Filtration .........................................................................................................3 2.4.3 Storage of the filters........................................................................................4 2.4.4 Extraction of the filters...................................................................................5 2.5 Spectroscopic Measurements....................................................................................6 2.5.1 Choice of the method......................................................................................6 2.5.2 Spectrophotometry..........................................................................................7 2.5.3 Fluorometric measurement .............................................................................8 3 PROCEDURE ...................................................................................................................8 3.1 Sampling ...................................................................................................................8 3.2 Filtration ...................................................................................................................9 3.3 Storage Of The Filters...............................................................................................9 3.4 Pigment Extraction..................................................................................................10 3.5 Centrifugation .........................................................................................................10 3.6 Spectroscopic Measurement ...................................................................................11 3.6.1 Spectrophotometry: trichromatic method.....................................................11 3.6.2 Spectrophotometry: monochromatic method with acidification...................11 3.6.3 Fluorometry ..................................................................................................12 4 SUMMARY OF OPERATIONAL CONDITIONS..........................................................14 5 QUALITY CONTROL .....................................................................................................14 6 ALTERNATIVE PROCEDURES ....................................................................................15 7 ACKNOWLEDGEMENT ................................................................................................15 8 REFERENCES .................................................................................................................15
