Al-Quds University

Faculty of Health Professions

Department of Medical Laboratory Sciences
Clinical Laboratory Instrumentation
Lab (3): Determination of iron with 1, 10 phenanthroline
Section: 1
Instructor: Ibrahim Ghannam.
Students’ Names: Shahd abu sneineh 22111938
Noor saefan
Date: 18/3/2023
Introduction
A single beam UV-Visible spectrophotometer was used in this lab, as it is a quantitative technique
used to measure used to measure how much a chemical substance absorbs light. In this lab, the
analyte don’t absorb in this region.

So, It is accomplished by causing them to react with a substance as an indicator (1,10- phenanthroline)
that produces a product that readily absorbs in the area, by creating a complex species(colored
solution) that absorbs light in the visible spectrum.
This property provides an excellent and sensitive method for determining iron ion in aqueous
solution.

Calibration plot (absorbance- concentration plot) is created when used measurements of the
absorbance of the standard solutions at known concentrations.

Figure1.a: represent calibration plot

Methods and materials

A variety of volumes (1, 2, 3, 5, 10) ml of the standard iron solution were taken,
and each volume was put in a different beaker. Moreover, 5 ml of
hydroxylamine hydrochloride and 1, 10-phenathroline solution were added to
each individual beaker, as a reducing agent from ferric(Fe+3) to ferrous(Fe+2).as
the following equation:

2 Fe+3 + 2 NH2OH + 2 OH− → 2 Fe+2 + N2 + 4 H2O

The PH level was measured as sodium acetate calibrated it to be 4.0. the
solution has been diluted with D.W after the adjusted solution has been
transferred to a 100ml volumetric flask. After thoroughly mixing, it was put in a
cuvette, and readings at 490 nm and 510 nm wavelengths were taken.
Moreover, the unknown sample followed the same procedures.

Figure 1.b: dilution with distilled water

results
Sample concentation Absorbance at Molar Absorbance at Molar
number 490nm absorbtivity at 510nm absorbbtivity
490nm at 510nm
1 0.1 0.195 1.95 0.206 2.06
2 0.2 0.316 1.58 0,333 1.665
3 0.3 0.540 1.8 0.580 1.933
4 0.5 0.891 1.782 0.957 1.914
5 1 1.654 1.654 1.776 1.776
unknown ? 0.776 1.526 0.833 1.638

Calibration curve at 490nm:

Calibration curve at 510nm:

Calculations for the unknown

At 490 nm, the linear equation of the calibration curve:

Y= 1.6415 x + 0.0298
0.776= 1.6415 x + 0.0298
0.746= 1.6415 x
x= 0.454 mg/ml
At 510 nm, the linear equation of the calibration curve :
Y= 1.7674 x + 0.0281
0.833= 1.7674 x + 0.0281
0.805= 1.7674 x
x= 0.454 mg/ml
Discussion
The calibration curve must be linear, but when it was drawn,
it appeared to be slightly tilted from the straight degradation.
This indicates that there was a mistake made in the practical
work as a result of a flaw in the device, a mistake made when
the solutions were made, or an interference that occurred in
the sample inside the cuvit.