Spectrofluorimetric method for the determination of large chlorophyll a/b ratios

A practical spectrofluorimetric calibration method at room temperature is described for determining large chlorophyll a/b ratios on direct extracts from plant material of pigments in 80% aqueous acetone, dimethyl sulfoxide, and N,N'-dimethylformamide. The method is based on the work of Boardman...

Full description

Saved in:
Bibliographic Details
Published inPhotosynthesis research Vol. 53; no. 1; pp. 73 - 79
Main Authors Talbot, M.F.J, Sauer, K
Format Journal Article
LanguageEnglish
Published Dordrecht Springer Nature B.V 01.07.1997
Subjects
Analytical chemistry
biochemical techniques
calibration
Chlorophyll
leaves
plant extracts
ratios
solvents
spectral analysis
spectrofluorimetry
Spinacia oleracea
Online AccessGet full text

Cover

More Information
Summary:A practical spectrofluorimetric calibration method at room temperature is described for determining large chlorophyll a/b ratios on direct extracts from plant material of pigments in 80% aqueous acetone, dimethyl sulfoxide, and N,N'-dimethylformamide. The method is based on the work of Boardman and Thorne [Boardman NK and Thorne SW (1971) Biochim Biophys Acta 253: 222-231] who used diethyl ether as solvent. We repeated the calibration in diethyl ether and find significantly different parameters for the calibration curve. The range of standards in this work included solutions with chlorophyll a/b ratios of 10-125 in dimethyl sulfoxide, and of 10-220 in the other solvents. Fluorescence emission spectra were found to be a highly sensitive method for assessing chlorophyll purity. We determined the limits of sensitivity for each solvent from the calibration data. The empirically determined slope of the calibration curve was shown to be related to intrinsic properties of the chlorophylls in solution; this allows predictions of the performance of the method in other solvents.[PUBLICATION ABSTRACT]
ISSN:0166-8595
1573-5079
DOI:10.1023/A:1005809801266