Column chromatography and verification of phytosiderophores by phenylisothiocyanate derivatization and UV detection
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abstract
A high-performance liquid chromatographic method was developed for the separation of phytosiderophores and for verification of the Fe3+-complexing ability of chromatographic separates. The chromatographic procedure involves separation on a C18 reversed-phase column by linear gradient elution with sodium acetate (pH 6.4) and methanol with UV detection, following pre-column derivatization of the phytosiderophores with phenylisothiocyanate (PITC). A modification of the procedure involving pretreatment of the sample with Fe3+, which blocks the derivatization by PITC, was used to confirm whether a given chromatographic peak was attributable to an Fe3+-complexing compound. The practical detection limit was approximately 100 pmol of phytosiderophore. Copyright (C) 1999 Elsevier Science B.V.
