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Development and validation of a LC method for the separation and determination of the anticancer‐active F e III (4‐methoxy‐salophene) using the new second‐generation monolith
Author(s) -
El Deeb Sami,
Ma Benjamin N.,
Gust Ronald
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200734
Subject(s) - monolith , chromatography , chemistry , analytical chemistry (journal) , biochemistry , catalysis
LC method with the newly introduced second‐generation monolithic silica RP ‐18e column has been developed for the separation of F e III (salophene) and four methoxy‐substituted F e III (salophene) complexes. The method has been validated for the quantitation of F e III (4‐ OM e‐salophene), a highly active anticancer substance in vitro , bound to serum albumin. Our routinely used high‐resolution continuum‐source atomic absorption spectroscopy method based on the determination of the central iron atom was unsuitable in this case because serum originally contains significant amounts of iron as revealed by a blank sample of serum albumin. The developed LC method depends on detecting the whole complex rather than the bound iron. Two morphologically different first‐ and second‐generation HPLC monolithic columns have been compared for this purpose. The newly introduced second‐generation monolithic silica column C hromolith® H igh R esolution RP ‐18e column (100 × 4.6 mm, M erck) separated the mixture successful within 13 min. A mobile phase consisting of 25 mM phosphate buffer pH 3/methanol (60:40, v/v) was used at a flow rate of 1 mL/min. The dynamic linear working range of the calibration curve for F e III (4‐ OM e‐salophene) was found to be between 1 and 200 μg/mL. Detection and quantitation limits were 0.3 and 1 μg/mL, respectively.