Ja. Howe et al., Column chromatography and verification of phytosiderophores by phenylisothiocyanate derivatization and UV detection, J CHROMAT A, 841(2), 1999, pp. 155-164
A high-performance liquid chromatographic method was developed for the sepa
ration of phytosiderophores and for verification of the Fe3+-complexing abi
lity of chromatographic separates. The chromatographic procedure involves s
eparation on a C-18 reversed-phase column by linear gradient elution with s
odium 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 F
e3+, 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 phytosideroph
ore. (C) 1999 Elsevier Science B.V. All rights reserved.