Instrumental Methods of Analysis -2

Activity 1

Spectrophotometric Determination of Fe in Dietary Supplements

Introduction
The reaction between iron(II) ion and 1,10-phenanthroline to form a red complex
serves as a good sensitive method for determining iron. The molar absorptivity of the
complex [(C12H8N2)3Fe]2+, is 11,000 at 508 nm. The intensity of the color is independent of pH
in the range 2 to 9. The complex is very stable and the color intensity does not change
appreciably over very long period of time. Beer’s law is obeyed.
The iron must be in the +2 oxidation state, and hence a reducing agent is added
before the color is developed. Hydroxylamine, as its hydrochloride, can be used, the
reaction being
2Fe3+ + 2NH2OH  2Fe2+ +N2 + H2O
The pH is adjusted to a value between 6 and 9 by addition of ammonia or sodium acetate.
An excellent discussion of interferences and of applications of this method is given by
Sandyll.
OBJECTIVE:

The goal of this experiment is to use the quantitative technique of spectrophotometry to

determine the mass of iron (in mg) contained in a commercially available supplement tablet.

SAFETY AND DISPOSAL:

Any solution containing the red Fe–1,10-phenanthroline complex should be disposed of in

the appropriately labeled container. All other waste can be disposed of down the drain

Reagents:
a. 0.2 % of 1,10- phenanthroline; Dissolve 0.1g of 1,10-phenanthroline in 100mL
distilled water
b. 6 M hydrochloric acid; Transfer 100ml 12M HCl to 100mL distilled water and stir to
mix. Transfer the resulting solution to a clean and labelled reagent bottle
c. 10 % sodium acetate; Dissolve 10 grams of sodium acetate in 100mL water
10 % hydroxylamine hydrochloride; Dissolve 10g hydroxylamine hydrochloride in
100mL distilled water.
d. Ferrous ammonium sulfate hexahydrate, 100mgFe/Li
Weigh accurately about 0.0700g of pure ferrous ammonium sulfate, dissolve in
water, and transfer the solution to a 100-mL volumetric flask. Add 2.5mL sulfuric acid
and dilute it o the mark.

Procedure
 1. Purchase two brands (5 tablets per type) of iron supplementary tablets.
2. Weigh the 5 tablets (each type) in an analytical balance and record the mass in your
data notebook.
3. Manually ground the tablets using an agate Mortar and pestle and pass the ground
materials in 200 mesh filter to obtain fine powder. Place the fine powder in a clean
and labelled vial and store it in a desiccator.
4. In three replicates, transfer 0.10xx g of the fine powder into 100mL beaker and boil
gently in a fume hood with 25mL of 6M HCl for about 15 minutes. Cover the beaker
with a watch glass (Atkins 1975).
5. When necessary, add distilled water (or deionized water) to keep a constant volume
of 15mL.
6. When digestion is finished, cool the beaker containing the digested sample. Rinsed
the watch glass with distilled water, and then filter the solution using Whatman-40
filter paper. Collect the filtrate in 250mL volumetric flask.
7. Rinse the residue remaining in the filter paper using deionized water and complete
the volume using distilled water.
8. Label the solution as “Stock sample solution”.

Preparation of Fe Standard solution, 100mg/Li

1. Weigh accurately about 0.0700g of pure ferrous ammonium sulfate, dissolve in
water, and transfer the solution to a 100-mL volumetric flask. Add 2.5mL sulfuric acid
and dilute it to the mark.
2. Into 5-10mL volumetric flask, 10uL, 50uL, 100uL, 200uL and 250uL portions of the
100ppm Fe standard solution.
3. Transfer 1000uL of deionized water into another 10-mL volumetric flask to serve as
blank.
4. To each flask, add 100uL hydroxylamine hydrochloride solution, 1000uL of the 1,10-
phenanthroline solution, and 800uL of the sodium acetate solution. Then dilute all
solutions to the 10-mL marks and allow them to stand for 10 minutes.
5. Using blank and the 2.50ppm Fe standard solution, scan the solution in the UV-Vis
Spectrophotometer at wavelength range of 400 to 800nm to obtain the ƛ max. Record
the ƛmax in your data notebook.
6. Measure the absorbance of each standard at the ƛmax that will be obtained.
7. Using Excell, plot the absorbance (y-axis) against concentration of Fe standard (X-
axix). The linear curve with corresponding line equation will be generated.
Determination of Fe in the sample solution.
1. Transfer into 10-mL volumetric flask 200uL of the original stock solution of sample.
Add100uL hydroxylamine hydrochloride, 1000uL 1,10-phenanthroline solution and
800uL ammonium acetate solution. Dilute to final volume and allow them to stand
for minutes.
2. Measure the absorbance of the solution at the obtained ƛmax.
Calculations
1. Give the line equation and r2
2. Calculate the average mass of Fe in each tablet and report as mgFe/Tablet.
3. Calculate the standard deviation, CV and RSD in ppt
4. Report the result of analysis in mg Fe /tablet.