Micro-method for the qualitative and quantitative analysis of photosynthetic pigments using high-performance liquid chromatography

Abstract4haracteristic chlorophyll (Chl) u aggregates formed in aqueous dimethyl sulfoxide (DMSO) were investigated spectroscopically. Four chlorophyll forms were found with increasing DMSO concentration; they are called A-672, A-683, A-695 and A-665 according to the wavelengths of their absorption maxima. Transformation occurred only in this order. Reverse transformation could not be realized. A-683 and A-695 were apparently formed by the interaction of Chl a with DMSO in the linear dimer and linear polymer arrangements, respectively. Coordination of the Mg atom with a DMSO 0 atom and interaction between the S atom of one DMSO molecule and the 0 atom of an other DMSO molecule should lead to formation of a sandwich-type complex of partially overlapping chlorophyll macrocycles (Chl a-DMSO),. A472 and A-665 were assigned to Chl u micelles and to dissolved monomeric Chl (I in DMSO, respectively. Fluorescence spectra showed that the A-683 was highly fluorescent, while the A-695 was less fluorescent. Energy migration within the A495 form to a trap with a low fluorescence yield might be responsible for this difference in the emission intensity.

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“…Purity of our samples was higher than 99.5% by HPLC analysis (cf. Iriyama et al, 1978). No contamination by carotenoids was found.…”

Section: Methodsmentioning

confidence: 86%

SPECTROSCOPIC STUDIES OF CHLOROPHYLL a AGGREGATES FORMED BY AQUEOUS DIMETHYL SULFOXIDE

Uehara

Mimuro

Tanaka

1991

Photochem & Photobiology

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“…In this context, an isocratic eluent system is favorable. In 1978, a simultaneous separation of Chls and Phes by normal-phase HPLC was attained by an isocratic procedure (Iriyama et al 1978). In 1984, the isocratic normalphase HPLC was established as a powerful tool for chlorophyll analysis ).…”

Section: Hplcmentioning

confidence: 99%

Physicochemical Properties of Chlorophylls in Oxygenic Photosynthesis — Succession of Co-Factors from Anoxygenic to Oxygenic Photosynthesis

Kobayashi¹,

Akutsu²,

Fujinuma³

et al. 2013

Photosynthesis

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“…Procedures for HPLC measurements of photosynthetic pigments are well established. 3,[19][20][21] The conditions used here have been described above. Three chromatograms characteristic of these measurements are given in Figure 3.…”

Section: B Measurements By Hplcmentioning

confidence: 99%

Stability of Chlorophyll a at the Gas−Water Interface in Pure and Mixed Monolayers. An Evaluation of Interfacial pH

1996

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The stability of chlorophyll a, Chl a, in neat and mixed lipid monolayers at the nitrogen-water interface has been measured using Langmuir trough and HPLC techniques. In neat monolayers, at subphase pH less than 8, Chl a degrades rapidly to produce pheophytin a, Phe a. Above this pH, Chl a appears to hydrolyze, giving a new product, Chl a hyd , spectroscopically similar to the parent compound. This latter process appears to involve alteration at the pentanone ring peripheral to the porphyrin structure. For mixed monolayer studies, four oleoyl-bearing lipids with headgroups differing in charge have been used to explore the influence of such headgroups on these two pH dependent processes. Consistent with a mechanism by which a negatively charged headgroup may enhance H 3 O + concentration in the monolayer region, it has been shown that L-Rdioleoylphosphatidyl-DL-glycerol markedly increases pheophytinization while eliminating the formation of Chl a hyd . By contrast, positively charged dioleoyl-N-(3-trimethylammoniopropyl)carbamoylglycerol serves to generate formation of Chl a hyd with no production of Phe a. In neutral but polar 1,2-dioleoyl-sn-glycerol monolayers, product yields were observed that are consistent with the larger acidity in the region of Chl a. Finally, L-R-dioleoylphosphatidylcholine, which bears a zwitterionic headgroup, significantly enhanced apparent Chl a hydrolysis. These results are interpreted in terms of alterations by the host lipid in pH local to Chl a.

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Micro-method for the qualitative and quantitative analysis of photosynthetic pigments using high-performance liquid chromatography

Cited by 48 publications

References 13 publications

SPECTROSCOPIC STUDIES OF CHLOROPHYLL a AGGREGATES FORMED BY AQUEOUS DIMETHYL SULFOXIDE

SPECTROSCOPIC STUDIES OF CHLOROPHYLL a AGGREGATES FORMED BY AQUEOUS DIMETHYL SULFOXIDE

Physicochemical Properties of Chlorophylls in Oxygenic Photosynthesis — Succession of Co-Factors from Anoxygenic to Oxygenic Photosynthesis

Stability of Chlorophyll a at the Gas−Water Interface in Pure and Mixed Monolayers. An Evaluation of Interfacial pH

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