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Chlorophyll‐derived porphyrins co‐chromatograph with phospholipids in high performance liquid chromatographic separations of plant lipid classes
Author(s) -
Moreau Robert A.,
Powell Michael J.,
Agnew John,
Young David H.
Publication year - 1998
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/(sici)1099-1565(199801/02)9:1<1::aid-pca383>3.0.co;2-p
Subject(s) - chemistry , chromatography , high performance liquid chromatography , hydrolysis , spinach , phospholipid , elution , hydrolysate , porphyrin , enzymatic hydrolysis , phytol , chlorophyll , alkaline hydrolysis , organic chemistry , biochemistry , membrane
Total lipid extracts were prepared from the leaves of potatoes, tomatoes, grapes and spinach, and each was analyzed by normal‐phase high performance liquid chromatography (HPLC). When these total lipid extracts were subjected to mild alkaline hydrolysis and the hydrolysates analyzed by normal‐phase HPLC, distinct peaks were detected in the region where common phospholipids typically elute (i.e. at 48, 50, and 68 min). The ultra violet‐visible spectra of these peak fractions revealed that each exhibited absorption maxima at 400 and 660 nm, suggesting that the peaks were porphyrins, most likely derived from chlorophylls. Mild alkaline hydrolysis apparently cleaved the ester bond of the chlorophylls and released the porphyrin and phytol components. This explanation was verified when commercially prepared chlorophylls a and b were subjected to the same alkaline hydrolysis conditions and identical peaks at 48 and 68 min were observed. Experiments with buffered (pH 6.0) aqueous homogenates of potato and tomato revealed that similar chlorophyll‐derived porphyrins were generated by endogenous enzymes. With the increasing popularity of HPLC as a tool for plant lipid analysis, users of this methodology should be cautioned as to the occurence of these non‐phospholipid peaks in the retention time region where phospholipids commonly elute. © 1998 John Wiley & Sons, Ltd.