Simultaneous determination of chlorophyll a and chlorophyll b by derivative spectrophotometry

Anal Bioanal Chem. 2004 Jul;379(5-6):803-11. doi: 10.1007/s00216-004-2637-7. Epub 2004 May 26.


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.

MeSH terms

  • Calibration
  • Chlorophyll / analysis*
  • Chlorophyll A
  • Plant Extracts / chemistry
  • Solvents
  • Species Specificity
  • Spectrophotometry / methods
  • Vegetables / chemistry


  • Plant Extracts
  • Solvents
  • Chlorophyll
  • chlorophyll b
  • Chlorophyll A