Quantitative HPLC/ETAAS hybrid method with an on-line metal scavenger for studying the protein binding and speciation of aluminum and iron.
Anal Chem
; 66(2): 216-22, 1994 Jan 15.
Article
em En
| MEDLINE
| ID: mdl-8116878
ABSTRACT
A quantitative metal-free high-performance liquid chromatographic/electrothermal atomic absorption spectrometric (HPLC/ETAAS) technique allowing an adequate separation of the proteins and of the inorganic/organic metal species of interest was developed. A silica-based scavenger was placed proximal to the injection valve retaining any remaining Al and Fe originating from buffer solutions and recipients. ETAAS instrumental conditions and matrix modifiers were carefully selected to eliminate interferences occurring due to the salt gradient elution. The postelution Al recovery (143-1428 micrograms L-1; 100-microL sample volume) was found to be 100.1 +/- 15.3% (N = 50) whereas that of Fe was 97.3 +/- 2.0% (N = 20; 442-2653 micrograms L-1). The protein recovery varied between 95 and 105%. The HPLC/ETAAS intraassay CV (N = 3) was 8.0% for Al (eluted amount of Al bound to transferrin (Tf), 7.3 ng) whereas that of Fe was 1.4% (eluted amount of Fe bound to Tf, 124.8 ng). The intraassay CV of the HPLC Tf determination was 1.6%. The proposed method is sensitive and accurate, allowing for the first time quantitative evaluation of the competition between Al and Fe for protein binding at clinical relevant concentrations. The technique will also be used for studying the protein binding of elements other than Al and Fe.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Compostos de Ferro
/
Compostos de Alumínio
Idioma:
En
Revista:
Anal Chem
Ano de publicação:
1994
Tipo de documento:
Article
País de afiliação:
Bélgica