EXPERIMENT 1

Determination of Cu2+ and Pb2+ in Plant Tissue Samples Using Atomic

Absorption Spectroscopy (AAS)

NAME: MUHAMMAD IRFAN BIN AMIRUDDIN

ID: 2016709343

GROUP: AS2454B

NAME OF LECTURER: MADAM ROHAIZA MOHAMAD

DATE OF EXPERIMENT: 20/3/2018

DATE OF SUBMISSION: 4/4/2018

OBJECTIVES
a) To determine Cu2+ and Pb2+ in plant tissue sample.
b) To familiarize with spiking technique in unknown sample.

INTRODUCTION
Copper is an important element for growth and development in plants and its
absorption are achieved primarily as Cu2+. Copper has redox properties and also is a
structural and catalytic component for the involvement of protein and enzyme
molecules in the metabolic processes. Copper can penetrate the plant both through
its roots and leaves when assimilated by plants as divalent ions or as chelates.

Copper when present in excess can be very toxic to plants. The effect of the
toxicity of copper in plants can be seen by the discolouration of leaves and also
preventing the activity of enzyme and protein at cellular level.

AAS is an analytical technique that measures the concentration on elements.

Instruments for AAS consist of a radiation source, a sample holder, a wavelength
selector, a detector, a signal processor and readout. The sample holder in atomic
absorption instruments is the atomized cell that contains the gaseous atomized
sample.

INSTRUMENT:

Atomic Absorption Spectroscopy (AAS).

APPARATUS:

Beaker, Volumetric Flask, Filter Funnel and Pipette.

CHEMICAL:

Cu2+ standard solutions of 100, 5, 4, 3, 2 and 1 ppm.

Pb2+ standard solutions of 100, 5, 4, 3, 2 and 1 ppm.

SAMPLE:

Spinach plant.

PROCEDURE:

a) Sample Preparation:

The sample was cut into small pieces and put 50g of the sample in oven (110°C) to
dry for 24 hours. 3.0g of the dried sample was weighed and grind using mortar and
then divided equally into 3 portions (about 1g each). 7mL of concentrated HNO3 and
1mL of H2O2 were added to the sample. The sample is then were spiked with 50 µL
of 100 ppm Cu2+ and 50 µL of 100 ppm of Pb2+ standard solution. The spiked
sample was filtered using filter paper and put in 50mL volumetric flask. Diluted to the
mark using deionized water.

b) Preparation of standard solution. (Cu2+ and Pb2+)

All standard solutions, 100, 5, 4, 3, 2 and 1 ppm, were prepared using deionized
water from 1000 ppm stock solution.

For 100 ppm:

(1000ppm)(V1) = (100ppm)(50mL)

V1 = 5mL

For 5 ppm:

(100ppm)(V1) = (5ppm)(50mL)

V1 = 2.5mL

For 4 ppm:

(100ppm)(V1) = (4ppm)(50mL)

V1 = 2.0 mL
For 3 ppm:

(100ppm)(V1) = (3ppm)(50mL)

V1 = 1.5 mL

For 2 ppm:

(100ppm)(V1) = (2ppm)(50mL)

V1 = 1.0 mL

For 1 ppm:

(100ppm)(V1) = (1ppm)(50mL)

V1 = 0.5 mL

c) Determination of the Cu2+ and Pb2+ concentration by AAS.

The Atomic Absorption Spectroscopy was run to obtain curve and determine the
concentration of each cation in the sample.
DISCUSSIONS
Sample digestion is necessary to release the analyte of interest or for
determining trace metals in a sample. Two common sample digestion method which
are wet digestion and dry ashing. In wet digestion, acids is added to the sample in
order to dissolve it into solution until complete decomposition of the sample matrix.
For dry ashing digestion, high temperature is used in removal of organic sample
matrix, then the residue dissolved in a solvent. The temperature range for heating in
the muffle furnace for ashing is between 200 and 600 °C.

Atomic Absorption Spectroscopy uses a technique by which for measuring the

concentration of various elements in the sample through their absorption of light
(Absorbance). When a radiation is passed through a sample, the radiation is
absorbed depending upon the wavelength of the radiation. The absorption of
radiation would bring about an increase in the energy of the molecule. The energy
gained by the molecule is directly proportional to the wavelength of radiation. The
increase in the energy of the molecule leads to the electronic excitations where
electrons from lower energy level, jump to higher energy levels. The amount of
absorbance is directly proportional with concentration of analyte.

Based on the table above, the concentration of Copper in spinach is 0.46

mg/L while concentration of Lead is 1.05 mg/L. From the result, the average
concentration of Cu in the sample is 0.221 mg/L while the average concentration of
Pb is 0.572 mg/L. By comparing the experimental result with the table above, it was
determined that the concentration on Copper and Lead in the sample was
significantly lower than the table above.

CONCLUSION
In conclusion, the average concentration of three spinach samples for Cu2+ is
0.221mg/L and the average concentration for Pb2+ is 0.572 mg/L.
 REFERENCES

Guerra F., 2011. Scielo. Heavy metals in vegetables and potential risk
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http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-
90162012000100008

Horiba Scientific, N.D. Sample Preparation. Sample digestion. Retrieved

on April 4, 2018 from
http://www.horiba.com/scientific/products/sample-
preparation/sample-digestion/

Tutor Vista, N.D. Atomic Absorption Spectroscopy. Atomic spectroscopy

principle. Retrieved on April 4, 2018 from
https://chemistry.tutorvista.com/inorganic-chemistry/atomic-
absorption-spectroscopy.html