Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Nakamura, Akimasa; Watanabe, Tadashi

doi:10.2116/analsci.17.503

Cited by 16 publications

(29 citation statements)

References 28 publications

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“…1), PhQ (peak 1), Chl a (peak 2), Chl a′ (peak 3), and Phe a (peak 4) can be detected and clearly resolved on the HPLC traces as previously reported for spinach thylakoid membranes. 10 The Phe a peak area significantly decreased from thylakoid membranes to PS I-LHC I and TX-PS I. Most of the xanthophyll peaks, which are ingredients of LHC I, 11 at around 5 -10 min disappeared in going from PS I-LHC I to TX-PS I.…”

Section: Pigment Composition Of P Purpureummentioning

confidence: 90%

“…10 A pigment solution filtered with a 0.45 µm PTFE filter, Millex FH13 (Millipore), was directly injected to HPLC without removing the solvents. For P. purpureum, reversed-phase HPLC conditions were the same as in Ref.…”

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

“…For P. purpureum, reversed-phase HPLC conditions were the same as in Ref. 10. For C. caldarium, pigments were eluted with acetonitrile/2-propanol/water = 75/20/5 (v/v) through a polyetheretherketone (PEEK) column (4.6 mm × 150 mm) packed with Pegasil ODS (Senshu Science, Tokyo) at a flow rate of 1.2 mL min -1 .…”

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

“…The Chl a/PhQ and Chl a/Phe a ratios were deduced from a comparison of the peak areas corrected with factors determined from a calibration curve of each pigment. 10 …”

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

“…9 Recently, we developed such conditions which can deduce the Chl a′/PS I ratio on a single reversed-phase HPLC trace by the simultaneous detection of Chl a′ and phylloquinone (PhQ), the secondary electron acceptor of PS I. 10 Pigment composition analyses of two cyanobacteria, a green alga Chlamydomonas reinhardtii, and spinach, combined with spectrophotometric determination of the Chl a/P700 ratios showed that one Chl a′ molecule exists in the vicinity of P700 in these organisms. 38 The light-harvesting system exhibits a significant variety among oxygenic photosynthetic organisms.…”

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Reversed-phase HPLC Determination of Chlorophyll a′ and Naphthoquinones in Photosystem I of Red Algae: Existence of Two Menaquinone-4 Molecules in Photosystem I of Cyanidium caldarium

2003

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The light reaction of oxygenic photosynthetic organisms proceeds by the cooperation of hundreds of such cofactors as chlorophyll (Chl), quinones, and carotenoids, placed within large pigment-protein complexes, photosystem (PS) I and PS II, in thylakoid membranes. Most Chl molecules function as lightharvesting pigments to funnel the excitation energy to the primary electron donor, P700 in PS I and P680 in PS II (for review, see Ref. 1). In the electron-transfer chains of PS I and PS II, minor Chl a derivatives play crucial roles in light-induced charge separation. 2 PS II contains two molecules of demetallated Chl a, pheophytin (Phe) a, as the primary electron acceptor. 3 We have proposed the existence of one or two molecules of Chl a′, the C13 2 -epimer of Chl a, in PS I through HPLC analysis of pigments.4,5 Recent X-ray crystallography on the PS I trimer of a thermophilic cyanobacterium Synechococcus elongatus revealed that P700 is a heterodimer of Chl a′ and Chl a. However, whether other oxygenic photosynthetic organisms use one Chl a′ molecule in P700 is still unknown. Because Chl a′ has practically the same absorption and fluorescence spectra as Chl a, 7 but shows a different chromatographic behavior, 8 an HPLC analysis of pigment extracts is the sole reliable way for the precise determination of Chl a′ in PS I. For this purpose, the conditions for pigment extraction and HPLC must strictly ensure the molecular integrity of Chl a, since Chl a undergoes several alterations in organic solvents. 9 Recently, we developed such conditions which can deduce the Chl a′/PS I ratio on a single reversed-phase HPLC trace by the simultaneous detection of Chl a′ and phylloquinone (PhQ), the secondary electron acceptor of PS I. 10 Pigment composition analyses of two cyanobacteria, a green alga Chlamydomonas reinhardtii, and spinach, combined with spectrophotometric determination of the Chl a/P700 ratios showed that one Chl a′ molecule exists in the vicinity of P700 in these organisms. 38 The light-harvesting system exhibits a significant variety among oxygenic photosynthetic organisms. 11 Typical cyanobacteria use soluble membrane-extrinsic phycobiliproteins as a light-harvesting system. Green algae and higher plants utilize only membrane-intrinsic Chl a/b complexes, LHC I and LHC II. The light-harvesting systems of red algae are known to be a transitional state from cyanobacteria to green plants, 12,13 and contain both phycobiliprotein and membrane-intrinsic LHC I (containing Chl a alone as the chlorophyllous pigment). 14,15 Hence, the pigment composition of red algae is of much interest to examine the universality of the existence of one Chl a′ molecule in P700.In view of this, we extended the pigment composition analysis to two red algae, a marine alga Porphyridium purpureum employed for many biochemical studies [12][13][14] and an alga living in an extreme environment, Cyanidium caldarium. 16 Experimental Photosynthetic organismsPorphyridium purpureum IAM R-1 was grown in an artificial seawater medium 17 for 7 days at 25...

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Section: Pigment Composition Of P Purpureummentioning

confidence: 90%

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

“…The Chl a/PhQ and Chl a/Phe a ratios were deduced from a comparison of the peak areas corrected with factors determined from a calibration curve of each pigment. 10 …”

Section: Pigment Extraction and Hplcmentioning

confidence: 99%

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Reversed-phase HPLC Determination of Chlorophyll a′ and Naphthoquinones in Photosystem I of Red Algae: Existence of Two Menaquinone-4 Molecules in Photosystem I of Cyanidium caldarium

2003

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Lipophilic constituents from aerial and root parts of Mercurialis perennis L.

Lorenz¹,

Hradecky²,

Berger³

et al. 2009

Phytochemical Analysis

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Current Awareness

2001

Phytochemical Analysis

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In order to keep subscribers up‐to‐date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of phytochemical analysis. Each bibliography is divided into 13 sections: 1 Books, Reviews & Symposia; 2 General; 3 Nucleic Acids; 4 Amino Acids, Proteins & Enzymes; 5 Carbohydrates; 6 Lipids; 7 Secondary Products; 8 Growth Regulators; 9 Industrially‐Important Products; 10 Toxins/Allergens; 11 Pigments; 12 Vitamins; 13 Others. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted.

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Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Cited by 16 publications

References 28 publications

Reversed-phase HPLC Determination of Chlorophyll a′ and Naphthoquinones in Photosystem I of Red Algae: Existence of Two Menaquinone-4 Molecules in Photosystem I of Cyanidium caldarium

Reversed-phase HPLC Determination of Chlorophyll a′ and Naphthoquinones in Photosystem I of Red Algae: Existence of Two Menaquinone-4 Molecules in Photosystem I of Cyanidium caldarium

Lipophilic constituents from aerial and root parts of Mercurialis perennis L.

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