Extraction of chlorophyll a from freshwater phytoplankton for spectrophotometric analysis

The efficiency of pigment extraction forms the crux of the spectrophotometric analysis of chlorophyll a. The alcoholic solvents, methanol and ethanol, proved to be superior to acetone and acetone with DMSO. Homogenisation and sonication did not improve the extraction in the alcoholic solvents. Boiling at 100°C had an adverse effect whereas complete extraction of the pigments was obtained at the solvents boiling point and allowing the samples to stand for 24 h in the dark.

[1]  A. Marker.  The measurement of photosynthetic pigments in freshwaters and standardization of methods : conclusions and recommendations , 1980 .

[2]  R. Daley,et al.  A Quantitative, Semiroutine Method for Determining Algal and Sedimentary Chlorophyll Derivatives , 1973 .

[3]  H. Robinson.  Principles and Procedures of Statistics , 1961 .

[4]  M. Holden,et al.  Changes in chlorophylls resulting from various chemical and physical treatments of leaves and leaf extracts , 1967 .

[5]  J. Wintermans,et al.  Spectrophotometric characteristics of chlorophylls a and b and their phenophytins in ethanol , 1965 .

[6]  W. Thomas Shoaf,et al.  Improved extraction of chlorophyll a and b from algae using dimethyl sulfoxide , 1976 .

[7]  D. J. Nelson,et al.  Improved Chlorophyll Extraction Method , 1960, Science.

[8]  G. W. Snedecor.  Statistical Methods , 1964 .

[9]  G. Hallegraeff,et al.  Some Problems in the Estimation of Chlorophyll-a and Phaeopigments from Pre- and Post-acidification Spectrophotometrie Measurements , 1978 .

[10]  C. Lorenzen,et al.  DETERMINATION OF CHLOROPHYLL AND PHEO‐PIGMENTS: SPECTROPHOTOMETRIC EQUATIONS1 , 1967 .

[11]  T. Parsons,et al.  Discussion of Spectrophotometric Determination of Marine-plant Pigments , with Revised Equations for Ascertaining Chlorophylls and Carotenoids , 2015 .

[12]  J. P. Riley,et al.  A thin-layer chromatographic method for the determination of plant pigments in sea water and cultures. , 1969, Analytica chimica acta.

[13]  B. Burnison,et al.  Modified Dimethyl Sulfoxide (DMSO) Extraction for Chlorophyll Analysis of Phytoplankton , 1980 .

[14]  A. Marker,et al.  The use of acetone and methanol in the estimation of chlorophyll in the presence of phaeophytin , 1972 .