LAB MANUAL

CHM580

EXPERIMENT 1

DETERMINATION OF Cr2+ AND Cd2+ IN PLANT TISSUE SAMPLES USING ATOMIC

ABSORPTION SPECTROSCOPY (AAS)

OBJECTIVES:

1. To determine Cr2+ and Cd2+ in plant tissue sample.

2. To familiarize with spiking technique in unknown sample

INSTRUMENT:

Atomic Absorption Spectroscopy (AAS), Microwave Digestion

APPARATUS:

Beaker, Volumetric Flask, Filter paper, filter funnel and pipette.

CHEMICALS:

Cr2+ standard solutions of 1, 2, 3, 4 and 5 ppm

Cd2+ standard solutions of 1, 2, 3, 4 and 5 ppm

SAMPLE

Spinach plant

PROCEDURE:

Cut the sample into small pieces and put 50 g of the sample in
oven (110oC) to dry for 24 hours. Weigh 3.0 g of the sample
and
spike Cr2+ and Cd2+ standard from each 100 ppm standard
solution
into the sample. Divide the sample into 3 portions
ranging from
0.5 g to 1.0 g accurately and
place
in
microwave digestion
vessels separately. Add 7 mL of
concentrated HNO3 and 1 mL of
H2O2 into each vessel. Then
digest the samples in the microwave
for 20 minutes. Remember
to cool down the vessels to room
temperature opening the
lid. (It takes 1 hour to cool down)
Determine the Cr2+ and Cd2+ concentration in the sample by
using Atomic Absorption Spectroscopy (AAS)

QUESTIONS :

1.

Explain a little bit about the digestion method used in this

experiment.

EXPERIMENT 2

DETERMINATION OF CAFFEINE IN TEA SAMPLES BY USING ULTRAVIOLET

SPECTROSCOPY

OBJECTIVES:

1.

INSTRUMENT:

Instrument Model : Perkin Elmer Lambda 35

APPARATUS:

Beaker, Volumetric flask, hot plate, pipette, filter paper and filter
funnel

CHEMICALS:

1.
2.
3.

SAMPLE:

Dried Tea Leaves

PROCEDURE:

Dissolve 2 g dried tea sample in 100 mL distilled water and boil for
15 min. After boiling and cool , filter the sample and mark up
mL volumetric flask with distilled water (100ppm).
the diluted sample into 50 mL volumetric flask
mark (10 ppm).

to 250
Pipette 5 mL of
and top up to the

shake
at the
obtain
extraction

To quantify caffeine concentration in tea sample.

Caffeine standard solution (2, 4, 6, 8 and 10 ppm)

0.005 M NaOH
CHCl3

Take 10 mL of the tea sample solution and pour into a separating

funnel. Add 20 mL of CHCl3 into the separating funnel and
the mixture. The mixture will separate into 2, aqueous layer
bottom and organic layer at the top. Centrifuge the mixture to
layer separation. Collect the aqueous layer. Repeat the
procedure 3 times.

respective

Pipette 1 mL of the extract into 5 different of 10 mL volumetric

flask. Add 0.0, 0.4, 0.6, 0.8 and 1.0 mL of NaOH in
flasks. Add distilled water to the mark.
Run the UV-Vis and determine the concentration of caffeine in the
sample.

EXPERIMENT 3
DETERMINATION OF CAFFEINE AND ACETYL SALICYCLIC ACID IN AN
ANALGESIC TABLET BY ULTRAVIOLET SPECTROSCOPY

OBJECTIVES:

To quantify caffeine and acetyl salicyclic acid in analgesic tablet

INSTRUMENT:

Instrument Model : Perkin Elmer Lambda 35

APPARATUS:

Beaker, Volumetric flask and pipette.

CHEMICALS:

1.

Caffeine standard solution (2,4,6,8 and 10 ppm)

2.

Acetyl salicyclic acid standard solution (2,4,6,8, and

10 ppm)

SAMPLE:

Analgesic tablet

PROCEDURE:

1.

Prepare all the standard solution using deionized water.

2.

Weigh to the nearest 1 g the sample, analgesic tablet

provided. Transfer the weighed solid into 250 mL
flask. Add about 200 mL of deionized water to
sample and fill the flask to the mark with

3.

Transfer accurately 0.5 mL of the sample solution into each

of three 50 mL volumetric flasks. Dilute each flask to
mark with water.

4.

Get ready to record UV spectra and determine the

concentration of caffeine and acetyl salicyclic acid in
sample.

volumetric
dissolve the
deionized water.

the

the

EXPERIMENT 4
DETERMINATION OF RIBOFLAVIN IN ENERGY DRINKS BY FLUORESCENCE
SPECTROPHOTOMETRY

OBJECTIVES:

To determine the mass of Riboflavin in some energy drinks by

Fluorescence spectrophotometry

INSTRUMENT:

Fluorescence Spectophotometer

APPARATUS:

Beaker, Volumetric flask and pipette.

CHEMICALS:
SAMPLE:

1.
Riboflavin standard solution (0.2,0.4,0.6, 0.8 and 1 ppm)
2.
5% Acetic acid
Energy drink

PROCEDURE:

A.

Chemical Preparation
a) Preparation of 100 ppm stock solution of Riboflavin
standards:
Prepare a 100 ppm riboflavin stock solution by accurately
weighing about 50 mg riboflavin, transferring to a 500 mL
volumetric flask, and diluting to volume with 5% (v/v) acetic
acid, This should be stored in a cool, dark place.
b). Preparation of 10 ppm stock solution of Riboflavin
standards and standard calibration curve:
Dilute an aliquot of the stock solution, 1:10 to obtain 10 ppm
working standard solution. Dilute aliquots of this with 5%
acetic acid to prepare standards of 0 (blank), 0.2, 0.4, 0.6,
0.8 and 1.0 ppm riboflavin.(All of the solution need to be
prepared in dark bottle and the bottle must be closed with
aluminium foil).

B)

Sample preparation
Pour 0.2 mL of energy drink into 50 mL of volumetric flask.
Dilute the sample with 5% acetic acid. Prepare triplicate
samples. These samples are ready for analysis using
fluorescence spectrophotometry.

C)

Plot of Calibration Curve

Using the standard calibration concentrations (x axis) and
their fluorescence values (y axis), plot a calibration curve
and from this curve, determine the amount of riboflavin in
sample.

EXPERIMENT 5
ANALYSIS OF ASPIRIN AND CAFFEINE IN COMMERCIAL APC TABLET USING
FTIR

OBJECTIVES:

To determine the aspirin (acetylsalicyclic acid) and caffeine content

in APC tablet.

INSTRUMENT:

Instrument Model : Perkin Elmer Spectrum One (FTIR)

APPARATUS:

Beaker, Volumetric flask and pipette.

SAMPLE:

APC tablet

PROCEDURE:

Solid Samples to prepare:

1.
Mixture of sample and KBr
Remove the agate mortar and pestle from the dessicator. Grind
0.001 g sample in agate mortar into powder. Add 0.080 g KBr into
the sample powder. Mix them using the pestle. Scrap and heap the
mixturein the center of the mortar and grind again for 1 minute.
Keep the KBr back into the dessicator after use.

2.
KBr pellets
Take one fourth of the KBr mixture and transfer into the collar of the
handpress.Place the anvil along with the longer die pin so that it
comes into contact with the samples. Lift the die set carefully
by
holding the lower anvil. Make sure the collar stays in places.
Open
the handle of the handpress slowly and insert the die set into
the
handpress.Close the handle. Rotate the dial pressure until
the
upper ram of the handpress slightly touches the upper anvil
on the
die assembly. Tilt back the unit in order to hold the die
set from
falling off. Open the handle. Rotate the pressure dial
clockwise in
one half turn.Compress the mixture slowly while
closing the handle
in 2 minutes. Tilt back the unit, open the handle
and remove the die
set from the unit carefully.Weigh and inspect the
pellet. These
samples are ready for analysis to obtain an IR
spectrum.
Get the calibration curve for standard ASA & Caffeine