Simultaneous determination of chlorophyll a and chlorophyll b by derivative spectrophotometry

Emel Ergun, Birsen Demirata*, Gulcin Gumus, Reşat Apak

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)


Chlorophyll a (Chl a) and chlorophyll b (Chl b) plant pigments, which are important in the food industry and are beneficial as environmental pollution indicators, have been extracted with a novel solvent mixture (1:1 v/v acetone-propanol) not containing chloroform and simultaneously determined by first-derivative spectrophotometry. The results were statistically compared to those obtained by the ordinary absorption spectrophotometric reference utilizing the principle of additivity of absorbances. The testing of the developed method in synthetic mixtures of Chl a and Chl b and in real plant material samples (grass, spinach, chard, purslane, black cabbage, crisp lettuce, rocket, dill and seaweed) proved successful in that the developed extractive derivative spectrophotometric method was both rapid and precise, and was not dependent on the Chl a/b ratio in contrast to the reference method which was adversely affected by the latter parameter.

Original languageEnglish
Pages (from-to)803-811
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Issue number5-6
Publication statusPublished - Jul 2004


  • Chlorophyll a
  • Chlorophyll b
  • Derivative spectrophotometry
  • Extraction
  • Simultaneous determination


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