In this experiment, the concentration of iron was determined with the use of a

single-beam spectrophotometer.

Metal complexes do not form in strongly acidic solution. On the other hand, most
metals form insoluble metal hydroxides in basic solutions. For these reasons the iron determination
using phenanthroline is carried out in a slightly acidic solution by the addition of concentrated sulfuric acid.

Sodium acetate was used as a buffer to maintain a slightly acidic pH.

In most water samples, iron exists in its oxidized form, Fe(III), due to the
presence of oxygen. Since it is the Fe(II) species that forms the complex with
orthophenanthroline, a reduction was carried out by the addition of hydroxylamine
hydrochloride:

4Fe3+ + 2NH2OHHCL 4Fe2+ + N2O + 4H+ + H2O

The absorbances of the prepared standards were measured using a single-

beam spectrophotometer at a wavelength of 508 nm where maximum absorbance
is observed. The concentration of the standards was computed and a curve was
plotted (absorbance vs concentration). A sample calculation in the determination of
the concentration of the standard is shown in Calculation (2). The plot is shown in
Figure 1.

The best fitting line was obtained with the use of least-squares method. The
equation of the best fitting line was used to determine the concentration of the
unknown and the water sample. The equation the best fitting line is shown in Figure
1.

The concentration of the unknown and the water sample was determined by
substituting the absorbance in the equation of the best fitting line. A sample
calculation is shown in Calculations (3) and (4).

The obtained concentrations in Calculations (3) and ( 4) were substituted in

the Calculations (6) and (5), respectively, to account for the dilution and to compute
for the actual concentrations.

The obtained concentration of the water sample was compared with the
established limits and the absolute error was calculated. The obtained concentration
of the water sample is 0.9 units higher than the secondary maximum contaminant
level (SMCL) value set by the US EPA. The obtained concentration of the water
sample is 0.2 units higher than the maximum limit set by the Philippine
government.

and the blank solution used in measuring the absorbances of the solutions.
The blank solution used in measuring the absorbances of the water sample and the
Unknown sample was not the same as the blank solution used in the measurement
of the absorbance of the standards. This was due to the late preparation of the
water and Unknown samples and the limited number of spectrophotometers in the
laboratory.

ANSWERS TO QUESTIONS

CONCLUSION

In this experiment Fe2+ is made to react with orthophenantrholine to form a

colored complex ion. The intensity of the colored species was measured using a
single-beam spectrophotometer. A calibration curve was constructed and the
concentrations of the Unknown solution and water sample were determined.

The equation of the constructed calibration curve is:

Absorbance = 0.2451Conc. - 0.0008136

The coefficient of determination (R2) of the constructed curve is 0.9997,

indicating an excellent linearity.

The concentration of the Unknown sample was calculated; the dilution

accounted, and found to be 9.14 mg/L.

The concentration of the water sample was calculated; the dilution

accounted, and found to be 1.1 mg/L. the calculated value is 0.8 units higher than
the limit established by the US EPA and 0.1 units higher than the limit established
by the Philippine government.

To eliminate the errors committed in the experiment that contributed to the

high concentration of iron in the water sample, de-ionized water should be used to
quantitatively clean the glasswares and to dilute the solutions and the blank
solution should be the same in the measurement of the absorbance of the
standards and of the samples.