EXPERIMENT 1: DETERMINATION OF IRON USING UV-VISIBLE (UV-VIS)

SPECTROPHOTOMETER

Objectives:

Upon completion of this experiment, students should be able to:

1. Prepare samples for UV-Visible analysis

2. Plot calibration curve for standard solution
3. Perform UV-Visible analysis on aqueous solution

Introduction:

A complex of iron(II) is formed with 1,10-phenanthroline (Fe(C12H8N2)32+) and the

absorbance of this coloured solution is measured with a UV-Visible spectrophotometer. The
spectrum is plotted to determine the absorption maximum. Hydroxylamine (as the
hydrochloride salt to increase solubility) is added to reduce any Fe3+ to Fe2+ and to maintain it
in that state. The formation of tris(1,10-phenanthroline) iron(II) complex is shown in equation
below:
+¿ +H O ¿
2
2+¿+2 NH 2OH → 4 Fe 2+ ¿+ N 2 O +4 H ¿
¿
4 Fe

Chemicals and Apparatus:

Chemicals:

Ferrous ammonium sulfate

Concentrated sulfuric acid

1, 10-phenanthroline

Hydroxylamine hydrochloride

Sodium acetate

Distilled water
Apparatus:

Beaker (100 mL)

Volumetric flask (100 mL and 1 L)

Pipette (1, 5, 10, 25 and 50 mL)

Measuring cylinder (10 mL)

Procedure:

Part A: preparation of solutions

1. Standard iron(II) solution: A standard iron solution was prepared by weighing

~0.0700 g ferrous ammonium sulfate (Fe(NH4)2(SO4)2.6H2O; RMM = 392.14) in 100
mL beaker and was dissolved in ~30 mL distilled water. Quantitatively, the solution
was transferred to a 1 L volumetric flask and sufficient distilled water was added to
dissolve the salt. 2.5 mL concentrated sulfuric acid was added, was diluted exactly to
the mark with distilled water and was mixed thoroughly.
2. 1, 10-phenanthroline solution: 0.1 g, 1, 10-phenanthroline monohydrate was dissolve
in 100 mL distilled water (beaker)
3. Hydroxylamine hydrochloride solution: 5 g hydroxylamine hydrochloride was
dissolved in 50 mL distilled water (beaker)
4. Sodium acetate solution: 10 g sodium acetate was dissolved in 100 mL distilled water
(beaker)

Part B: UV-Visible analysis

1. Into a series of 100 mL volumetric flasks, 1, 5, 10, 25 and 50 mL of the standard iron
solution was added with pipette.
2. Into another 100 mL volumetric flask, 50 mL distilled water was placed for a Blank.
3. 10 mL of solution X (unknown sample) was pipette into a 100 mL volumetric flask.
4. To each of the volumetric flask (including the blank and unknown), 1 mL of the
hydroxylamine hydrochloride solution, 10 mL of 1 ,10-phenanthroline solution and 8
mL of the sodium acetate solution was added. Distilled water was added to the mark.
 After adding the reagents the solution was allowed at least 15 minutes before making
absorbance measurements. Each solution was shaken, once in a while.
5. By using blank solution as the reference solution and one of the standard iron(II)
solutions prepared in procedure 1, absorbance, A was measured, for the iron(II)
solution chosen at wavelength of 400-600nm. Readings at 20-nm intervals was taken
except near the vicinity of the absorption maximum, where we should take readings at
5-nm intervals.
6. The absorbance against the wavelength graph was plotted and the wavelength of the
absorption maximum was selected.
7. The absorbance of each of the standard solutions of the wavelength of maximum
absorbance was measured and a calibration curve was prepared. The unknown was
measured in the same way.
8. The absorbance of the standards was plotted against concentration in ppm. From this
plot and the absorbance of the unknown, the concentration of iron was determined in
our unknown solution.

Results:

Part A: Preparation of Fe(II) standard solution

Weight of ferrous ammonium sulfate:

Concentration of Fe(II) standard solution in mg Fe mL-1:

Calculation:

mg Fe m L−1=¿

Part B: curve of absorbance of Fe-1, 10-phenanthrolinecomplex against wavelength

Concentration of Fe(II) standard solution used:

λ (nm) Absorbance (A)

400
420
440
460
480
500
505
 510
515
520
540
560
580
600

Plot graph of the absorbance (A) against λ (nm)

From graph, maximum λ:

Part C: Calibration Curve For The Determination of Fe(II).

Volume of Fe(II) stock Absorbance (A) at maximum

Concentration (mg Fe mL-1)
solution (mL) λ
1
5
10
15
25
50
Unknown

Plot graph of absorbance (A) against concentration (mg Fe mL-1)

From graph, concentration of unknown:

Concentration of solution X: