TITLE: UV SPECTROPHOTOMETRY

AIM: To determine the concentration of Fe 2+ in the stock solution

APPARATUS:
Helios Omega UV/Vis,
MATERIALS:
Hydroquinone, Trisodium citrate, 0- Phenanthroline, Standard Iron (II) Solution
METHOD:
1. One of the iron containing tablets was placed in a 125 mL flash or 100 mL beaker
and boiled gently ( in a fume hood) with 25 mL of 6 M HCL for 15 minutes. The
solution was then filtered directly into a 100 mL volumetric flask. The solution was
filtered directly into a 100 m L volumetric flask. The beaker was washed and
filtered several times with small portions of water to complete a quantitative
transfer. The solution was allowed to cool and diluted to the mark and mixed well.
5.00 mL of this solution was diluted to 100.0 mL in a fresh volumetric flask. If the
tablet contained < 15 mg of Fe, 10.00 mL was used instead of 5.00 mL.
2. 10.00 mL of the standard Fe solution was pipette into the beaker and the pH was
measured (with pH paper or a glass electrode). Sodium citrate solution was added
one drop at a time until a pH of ~3.5 was reached. The drops were counted if
needed (it required about 30 drops.).
3. A fresh 10.00 mL aliquot of Fe standard was pipetted into 100 mL volumetric and
the same number of drops of citrate solution as required in step 2 was added.
2.00mL of Hydroquinone solution and 3.00 mL of o-phenanthroline solution was
added and was diluted to the mark with water and mixed well.
4. Three more solution from 5.00, 2.00 and 1.00 mL of Fe standard and a blank
containing no Fe was prepared. Sodium citrate solution was used in proportion to
the volume of Fe solution. (If 10 mL of Fe required 30 drops of citrate solution, 5
mL of Fe required 15 drops of citrate solution.).
5. The amount of drops of Citrate solution that was needed to bring 10.00 mL of iron
tablet solution from step 1 to pH 3.5 was found out. This required about 3.5 or 7
mL of citrate, depending on whether 5 or 10 ml of unknown was diluted in the
second part of step 1.
6. 10.00 mL of solution from step 1 was transferred to a 100 mL volumetric flask. The
amount of citrate solution that was determined in step 5 was added, then 2.00 mL
of hydroquinone solution was added and 3.0 mL of o- phenanthroline solution was
added and diluted to the mark and mixed well.
7. The solution was allowed to stand for at least 10 minutes. The absorbance of each
was measured at 508 nm. (The colour is stable, so all solutions were prepared and
all the absorbance readings were measured at once.). Distilled water was used in
the reference cuvette and the absorbance of the blank was subtracted from the
absorbance of the standards.

8. A graph of absorbance versus micrograms of Fe in the standards was plotted. The

slope and intercept (and standard deviations) were determined by the method of
least squares.

9. The following were calculated;

The molarity of the Fe (o-phenanthroline)32+ in each solution
The average molar absorptivity ( in Beers Law) from the four absorbance
readings taken.
The number of milligrams of Fe in the tablets using the calibration curve.

OBSERVATIONS:
The more vivid coloured compounds had the higher amount of absorption of
radiation.

RESULTS:
TABLE SHOWING THE AMOUNT OF ABSORBANCE OF THE SOLUTIONS OF FE
[Fe]/ ppm
0.4
0.8
2
3
4
Unknown
0

Absorbance reading
0.071
0.151
0.389
0.585
0.775
0.382
0.001

CALCULATIONS:
1. Mass concentration of standard
0.2836 g in 1000mL
2. Mass concentration of iron in stock solution
[Fe stock] = 0.2836 * 14.24/100
= 0.04 g Fe/L or 40 mgFe/L
or 40 ppm
3. Sample concentration for making standards ( include Dilution factor) ( Show
working how to calculate concentration of standard)
1st Fe std
DF = 100/10 = 10
Hence conc. of [1st Fe std]
= 0.04gFe/L 10
= 0.004 gFe/L or 4mgFe/L or 4ppm

4.
5.
6.
7.

Table with data title and app data

Wavelength at maximum absorbance 508 nm
Name of instrument Helios Omega UV/Vis
Molar Absorptivity at lambda max from spectra
A= ECL
A/Cl= E
0.071/0.4= 0.1775 dm3cm-1mol-1
0.151/0.8= 0.18875
0.389/2=0.1945
0.585/3=0.195
0.775/4=0.19375
0.382/2=0.191

8. Molar absorptivity from calibration curve gradient

M= y1-y2/ x1-x2
0.85-0/4.5-0 =0.188888888
9. Equation for line y =mx+c
M= 0.188888888
C= 0
Y= 0.188888888X

0.9
0.8
0.7
0.6
0.5
Absorbance 0.4

Absorbance reading

0.3

Linear (Absorbance
reading)

0.2
0.1
0
0

Conc. of Fe/ppm

GRAPH SHOWING THE FE CALIBRATION CURVE FOR UV/VIS SPECTROSCOPY