Determination of Iron in Caco-2 cells by ET-AAS.
An electrothermal atomic absorption spectrometric method (ET-AAS) was developed for the direct determination of iron in intestinal Caco-2 cells after studying cell viability and proliferation using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT test). Zeeman background correction and end-capped graphite tubes with L'vov platforms were used. Samples were dissolved in dimethylsulfoxide (DMSO) and pipetted directly into the graphite tube. The preashing, pretreatment and atomization steps were optimized. The temperatures selected were 600, 1200, and 2100 degrees C, respectively. Stability measurements were performed using iron standard solutions in DMSO on the one hand and acidified cell solutions on the other. Direct measurement and standard addition were compared in order to determine possible influences of the matrix. The low detection limit of the ET-AAS method (1.3 microg/L or 3.3 microg/g) combined with the small sample quantities required are ideal for the determination of iron in cells due to the low iron content and the limited growth area of the cells. The method was developed for iron uptake studies for toxicological purposes.
Zeiner
Michaela
M
Institute of Analytical Chemistry, University of Vienna, Währingerstr. 38, 1090, Vienna, Austria.
Zödl
Bettina
B
Steffan
Ilse
I
Marktl
Wolfgang
W
Ekmekcioglu
Cem
C
D018938:Caco-2 Cells; D002138:Calibration; D006801:Humans; D007501:Iron / Q000032:analysis* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013054:Spectrophotometry, Atomic / Q000379:methods*
D007501:Iron