Spectrophotometric determination of iron

A 0.25 mM ferrous ammonium sulphate stock solution was prepared from the salt
(Fe(NH4)2(SO4)2C6H2O, Aldrich) in purified water. Six standard solutions with iron
concentrations of 0.03, 0.05, 0.07, 0.09, 0.10 and 0.13 mM were prepared from the stock
solution. The unknown was provided by the instructor and a stock unknown solution was
prepared by dissolving 0.0815 g to 500.0 mL with purified water. Dilutions of 30, 50, 70, 90,
110 and 130 in 250 were all prepared from the unknown stock solution.
All final solutions, standards, unknowns and blank, contained 0.017 M hydroxylamine
hydrochloride (NH2OHCHCl, Fisher) to reduce any iron (III) to iron (II), 0.15 M sodium acetate
(CH3COONaC3H2O. Fisher) to maintain pH at ~3.5, and 0.6 mM 1,10-phenanthroline (Aldrich)
as complexing agent.
Absorbance readings at 508 nm were determined for all standard and unknown solutions using a
Biotek :Quant Microplate Spectrophotometer.