Publisher Summary This chapter presents detailed information on chlorophylls and carotenoids to give practical directions toward their quantitative isolation and determination in extracts from leaves, chloroplasts, thylakoid particles, and pigment proteins. The chapter focuses on the spectral characteristics and absorption coefficients of chlorophylls, pheophytins, and carotenoids, which are the basis for establishing equations to quantitatively determine them. Therefore, the specific absorption coefficients of the pigments are re-evaluated. This is achieved by using a two-beam spectrophotometer of the new generation, which allows programmed automatic recording and printing out of the proper wavelengths and absorbancy values. Several procedures have been developed for the separation of the photosynthetic pigments, including column (CC), paper (PC), and thin-layer chromatography (TLC) and high-pressure liquid chromatography (HPLC). All chloroplast carotenoids exhibit a typical absorption spectrum that is characterized by three absorption maxima (violaxanthin, neoxanthin) or two maxima with one shoulder (lutein and β-carotene) in the blue spectral region.

Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes

Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy

/pdf/laboratory-manual-for-physiological-studies-of-rice-37e0a0qv1v.pdf

Laboratory manual for physiological studies of rice

There is a rapidly growing body of literature covering the role of plant secondary metabolites in food and their potential effects on human health. Furthermore, consumers are increasingly aware of diet related health problems, therefore demanding natural ingredients which are expected to be safe and health-promoting. By-products of plant food processing represent a major disposal problem for the industry concerned, but they are also promising sources of compounds which may be used because of their favourable technological or nutritional properties. The purpose of this review is to highlight the potential of selected by-products as a source of functional compounds.

By-products of plant food processing as a source of functional compounds — recent developments

Over the last half century, the most frequently used assay for chlorophylls in higher plants and green algae, the Arnon assay [Arnon DI (1949) Plant Physiol 24: 1–15], employed simultaneous equations for determining the concentrations of chlorophylls a and b in aqueous 80% acetone extracts of chlorophyllous plant and algal materials. These equations, however, were developed using extinction coefficients for chlorophylls a and b derived from early inaccurate spectrophotometric data. Thus, Arnon’s equations give inaccurate chlorophyll a and b determinations and, therefore, inaccurate chlorophyll a/b ratios, which are always low. This paper describes how the ratios are increasingly and alarmingly low as the proportion of chlorophyll a increases. Accurate extinction coefficients for chlorophylls a and b, and the more reliable simultaneous equations derived from them, have been published subsequently by many research groups; these new post-Arnon equations, however, have been ignored by many researchers. This Minireview records the history of the development of accurate simultaneous equations and some difficulties and anomalies arising from the retention of Arnon’s seriously flawed equations.

/pdf/the-chequered-history-of-the-development-and-use-of-1ndjqtjezd.pdf

The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b

/pdf/modern-methods-of-plant-analysis-3t59z9gik3.pdf

I. Kawamura

Papers

Modern methods of plant analysis