Separation and determination of minor photosynthetic pigments by reversed-phase HPLC with minimal alteration of chlorophylls.
Reversed-phase HPLC conditions for separation of chlorophyll (Chl) a, Chl a' (the C132-epimer of Chl a), pheophytin (Pheo) a (the primary electron acceptor of photosystem (PS) II), and phylloquinone (PhQ) (the secondary electron acceptor of PS 1), have been developed. Pigment extraction conditions were optimized in terms of pigment alteration and extraction efficiency. Pigment composition analysis of light-harvesting complex II, which would not contain Chl a' nor Pheo a, showed the Chl a'/Chl a ratio of 3-4 x 10(-4) and the Pheo a/Chl a ratio of 4-5 x 10(-4), showing that the conditions developed here were sufficiently inert for Chl analysis. Preliminary analysis of thylakoid membranes with this analytical system gave the PhQ/Chl a' ratio of 0.58 +/- 0.03 (n = 4), in line with the stoichiometry of one molecule of Chl a' per PS I.
Nakamura
A
A
Institute of Industrial Science, The University of Tokyo, Komaba, Meguro, Japan.
Watanabe
T
T
D002734:Chlorophyll / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000077194:Chlorophyll A; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008027:Light; D045342:Light-Harvesting Protein Complexes; D010674:Pheophytins / Q000302:isolation & purification; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000302:isolation & purification; D018724:Spinacia oleracea / Q000737:chemistry; D020524:Thylakoids / Q000737:chemistry; D013997:Time Factors; D010837:Vitamin K 1 / Q000302:isolation & purification
D045342:Light-Harvesting Protein Complexes; D010674:Pheophytins; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins; D002734:Chlorophyll; C085815:chlorophyll a'; C001785:chlorophyll P 700; C061694:pheophytin a; D010837:Vitamin K 1; D000077194:Chlorophyll A