title: Microanalytical method development for Fe, Cu and Zn determination in colorectal cancer cells.
abstract: Microanalytical methods suitable for the determination of Fe, Cu in HT-29 (human colon adenocarcinoma) cells treated with different iron compounds (Fe(II) sulfate, Fe(III) chloride, Fe(III) citrate and Fe(III) transferrin) and cultured in medium supplemented or not with 10% (v/v) fetal calf serum (FCS) by total reflection X-ray fluorescence spectrometry (TXRF) and simultaneous graphite furnace atomic absorption spectrometry (GF-AAS) were developed. The developed TXRF method was also suitable for Zn determination in the samples. The main advantage of the proposed methods is the execution of all sample preparation steps following incubation and prior to the elemental analysis in the same Eppendorf tubes. Sample preparation was performed at microscale (115 μL sample volume) with 65% nitric acid and 30% hydrogen peroxide. According to scanning electron microscopic measurements, the organic matrix of the cell samples could be eliminated to the extent that accurate results were obtained for Cu and Fe by analyzing the same samples by TXRF and GF-AAS. Concerning the iron uptake, HT-29 cells incubated in FCS-free medium contained Fe in cca. 5-50 times higher amounts compared to cells cultured in FCS supplemented medium. Pronounced differences in the iron uptake compared to the iron supply (inorganic vs. organic chelated as well as iron(II) vs. iron(III)) were observed in the case of cell lines incubated in FCS-free medium.
authors.affiliation: Laboratory of Environmental Chemistry and Bioanalytics, Department of Analytical Chemistry, Institute of Chemistry, Eötvös Loránd University, H-1518, P.O. Box 32, Budapest, Hungary.
authors.forename: Zsófia
authors.initials: Z
authors.lastname: Polgári
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Ajtony
authors.forename: Peter
authors.initials: P
authors.lastname: Kregsamer
authors.forename: Christina
authors.initials: C
authors.lastname: Streli
authors.forename: Victor G
authors.initials: VG
authors.lastname: Mihucz
authors.forename: Andrea
authors.initials: A
authors.lastname: Réti
authors.forename: Barna
authors.initials: B
authors.lastname: Budai
authors.forename: Judit
authors.initials: J
authors.lastname: Kralovánszky
authors.forename: Norbert
authors.initials: N
authors.lastname: Szoboszlai
authors.forename: Gyula
authors.initials: G
authors.lastname: Záray
mesh_terms: D015179:Colorectal Neoplasms / Q000473:pathology*; D003300:Copper / Q000032:analysis; D006108:Graphite / Q000737:chemistry; D019073:HT29 Cells; D006801:Humans; D007501:Iron / Q000032:analysis / Q000378:metabolism; D019216:Metals, Heavy / Q000032:analysis*; D013052:Spectrometry, X-Ray Emission / Q000379:methods*; D013054:Spectrophotometry, Atomic / Q000379:methods*; D013997:Time Factors; D015032:Zinc / Q000032:analysis
chemical_list: D019216:Metals, Heavy; D006108:Graphite; D003300:Copper; D007501:Iron; D015032:Zinc