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AS245: Bachelor in Science Applied Chemistry

Title: Analysis of plant tissue (AAS)


Course: Spectrochemical Methods of Analysis
(CHM580)
Name: Mohamad Nor Amirul Azhar b. Kamis
Matrix no:

2014647344 (AS245 3S)

Date of experiment: 25th March 2015


Date of submission: 8th April 2015
Group members: 1. Nurnailah bt. Noorazlan
2. Nor Amirah bt. Ahmad Azuan
3. Noramira bt. Saad
It is my responsibility as a student of UiTM to adhere
to truthfulness and avoid dishonesty, fraud or deceit
of any type in connection with write up and conduct
of this experiment.
Signature:__________
Date:

TITLE:

Determination of iron (Fe) and cadmium (Cd) in plant tissue (spinach).

ABSTRACT:
The experiment was done to determine the cadmium (Cd) and iron (Fe) in spinach
by using wet digestion method for the sample preparation. The amount of those
metals presence in the spinach was determined by using the atomic absorption
spectroscopy (AAS) instrument. The since the concentration of analytes in sample
are in small amount, the standard addition method was applied. From the calibration
curve plotted, the concentration of analytes is known by extrapolating the straight
line until the x-intercept where the absorbance is zero. The concentration of
cadmium in spinach sample is 1.17M while the concentration of iron in spinach
sample is1.47M.

INTRODUCTION:
Generally the atomic absorption spectroscopy is an instrument that used for
quantification of metal elements that present in the sample. The concentration
measured is based on the absorption of the radiation either from line source such as
hollow cathode lamp or continuum source. Each element will absorb radiation at their
specific wavelength from radiation source. The sample introduction is normally in
liquid form for flame atomizer. The sample is converted into the form of free atoms
before it can absorb the radiation from the radiation source. Atomic absorption
Spectroscopy (AAS) instrumentation consists of radiation source, wavelength
selector, sample container, detector and the signal processor and readout.
Sample used was spinach tissue that contains cadmium and iron. Cadmium is not
regarded as essential to human life, different with iron that is an essential mineral
that helps transporting oxygen throughout body. Excess cadmium exposure
produces adverse health effects on human beings. Wet digestion method is used to
extract the analyte out from the sample by treating with concentrated acid. The
standard addition method is used when the concentration of analyte in the sample is
very low while matrix is high. By using the standard addition method, the

concentration of the sample is added with known concentration solution in order to


increase the signal meanwhile accounting with matrix effect.
OBJECTIVE:
Determination of cadmium and iron in spinach by using wet digestion method and
also to know the amount of the metals (cadmium and iron) present in the plant
tissue.

EXPERIMENTAL:
A. Preparation of cadmium (Cd) standard:
1. 5mL of 100ppm of Cd stock solution was pipetted into 50mL volumetric
flask and was added with de-ionized water until the calibration mark to
make 10ppm Cd solution.
C1V1= C2V2
100V1 = 10(50)
V1 = 5mL
2. 10ppm Cd solution was pipetted into 6 different 50mL volumetric flasks to
make the following concentration:
Standard concentration

Volume of Cd 10ppm,

(mL)
X + 0.0
0.0
X + 0.2
1.0
X + 0.4
2.0
X + 0.6
3.0
X + 0.8
4.0
X + 1.0
5.0
Sample calculation: standard 1

Blank
Standard 1
Standard 2
Standard 3
Standard 4
Standard 5

C1V1= C2V2
10V1 = 0.2(50)
V1 = 1.0mL
3. Each of the volumetric flasks was mark up with de-ionized water until the
50mL calibration mark.
B. Preparation of iron (Fe) standard:

1. 5mL of 100ppm of Fe stock solution was pipetted into 50mL volumetric


flask and was added with de-ionized water until the calibration mark to
make 10ppm Cd solution.
C1V1= C2V2
100V1 = 10(50)
V1 = 5mL
2. 10ppm Fe solution was pipetted into 6 different 50mL volumetric flasks to
make the following concentration:
Standard

Volume of Cd 10ppm,

concentration
X + 0.0
X + 1.0
X + 1.5
X + 2.0
X + 2.5
X + 3.0

(mL)
0.0
5.0
7.5
10.0
12.5
15.0

Blank
Standard 1
Standard 2
Standard 3
Standard 4
Standard 5

Sample calculation: standard 1


C1V1= C2V2
10V1 = 1.0(50)
V1 = 5.0mL
3. Each of the volumetric flasks was mark up with de-ionized water until the
50mL calibration mark.
C. Preparation of sample:
1. Day 1:
a. The spinach was cut to smaller size and was dried in oven at 100C
overnight to remove water moisture in the plant tissue.

2. Day 2:
a. 2.9477g of dried spinach leaves was put into a beaker and 20-30mL of
concentrated nitric acid solution (65%) was added into it.
b. The beaker was left overnight.
3. Day 3:
a. The mixture was heated in the fume hood until the red fume (NO 2) was
released.
b. After the red fume was released, the mixture was let to cool in the fume
hood.
c. 2mL of hydrogen peroxide solution (30%) was added into the cooled
mixture.

d. The mixture was heated again allowing evaporating to a small volume.


e. The sample was filtered and transferred into a 250mL volumetric flask.
f. De-ionized water was added until the 250mL calibration mark (calculate
for the dilution factor).
*CAUTION: always add HNO3 to the tissue samples and allow this mixture to digest
before adding H2O2. H2O2 reacts explosively with untreated organic matter.
D. Operational instrument:
1. Turn on argon gas and compress air.
2. Turn on spectrometer.
3. Open software WinLab32 For AA.
4. Turn on lamp (idle).
5. Select method, new, write method name, select element to be analysed.
6. Select setting, time (3.0), delay (18-20).
7. Select sample, never.
8. Select calibration, equation (nonlinear through zero), unit (ppm).
9. Select standard concentration, fill with standard names.
10. Select file, save as, method, write method name.
11. Check lamp, close.
12. Turn on flame.
13. Select auto, analyse, rebuild list, analyse all.
RESULT:
A. Cadmium (Cd):
standards
X + 0.0
X + 0.2
X + 0.4
X + 0.6
X + 0.8
X + 1.0

Concentration (ppm)
0.120
0.138
0.162
0.182
0.209
0.217

Graph of Absorbance vs standards Cd


0.25
f(x) = 0.1x + 0.12
R = 0.99

0.2
0.15
Absorbance

0.1
0.05
0
0

0.2

0.4

0.6

0.8

1.2

(sample + x) standards

y = mx + c
y = 0.1026x + 0.12
When y = 0, x-intercept = ?
0.1026x = -0.12
X = -1.17
From graph, the concentration of cadmium in sample is 1.17M
B. Iron (Fe):
standards
X + 0.0
X + 1.0
X + 1.5
X + 2.0
X + 2.5
X + 3.0

Concentration (ppm)
0.021
0.045
0.055
0.062
0.068
0.072

Graph of Absorbance vs standards Fe


0.08
f(x) = 0.02x + 0.03
R = 0.96

0.07
0.06
0.05
Absorbance

0.04
0.03
0.02
0.01
0
0

0.5

1.5

2.5

3.5

(sample + x) standards

y = mx + c
y = 0.017x + 0.0255
When y = 0, x-intercept = ?
0.017x = -0.0255
X = -1.5
From graph, the concentration of iron in sample is 1.47M.

DISCUSSION:
The standard addition method is used because the matrix effect is too high, so that
the sample is spiked with the known standard concentration. After plotting the
calibration curve, the concentration of the analyte in the sample can be determine by
extrapolating until the x-intercept. The x-intercept is at which the volume of standard
reagent equivalent to the amount of analyte in the sample. References stated that
spinach is a type of vegetable that rich with iron about 3.6mg in 100g. For this
experiment, the concentration of iron is 1.47M in 50mL sample. The actual

concentration is known when the diluted concentration is multiply with the dilution
factor. The dilution factor is crucial to get the actual concentration of the analyte. The
error that occurs in this experiment is when the dilution factor was not recorded, so
that the actual concentration of the sample is unknown. The concentration of
cadmium in the diluted sample is 1.17M. The safe amount of cadmium consume by
human is about lower than 2mg/kg. Higher amount of cadmium in human body can
bring to health problem.
From the calibration curve plotted, the R 2 values are nearly to 1 for both calibration
curves. The nearer the R2 value to 1, the straight line is better best fit.
When preparing the standard solution, make sure to use the calibrated apparatus
that is most important, the pipette. If the volume transferred is not correct, the
concentration of solution made will be wrong. Wet digestion method must be handled
with care because it used the concentration solution that harming.

CONCLUSION:
The concentration of cadmium in the spinach sample is 1.17M while the
concentration of iron the spinach sample is 1.47M.

REFERENCES:
1. J. Dolan, HPLC Solutions 89 Standard Additions, Retrieved April 3, 2015, from
http://www.sepscience.com/Techniques/LC/Articles/216-/HPLC-Solutions-89Standard-Additions.
2. F. J. Holler, D. A. Skoog, S. R. Crouch (2007), Principles of Instrumental
Analysis 6th ed.), United States, Brooks/Cole Cengage Learning.
3. S. Watson, Vitamins and Supplements Lifestyle Guide, Retrieved April 3, from
http://www.webmd.com/vitamins-and-supplements/lifestyle-guide-11/ironsupplements.

4. S. K. Naik, T. K. Pandit, P. K. Patra & D. K. Das, Effects of Graded Levels of


Cadmium on Spinach and Cabbage Grown in an Inceptisol, Retrieved April 3,
from http://www.tandfonline.com/doi/abs/10.1080/00103624.2013.768264.
5. Top
10
Foods
Highest
in
Iron,
Retrieved
April
3,
from
http://www.healthaliciousness.com/articles/food-sources-of-iron.php.
6. Cadmium exposure and human health, Retrieved April 3,
http://www.cadmium.org/pg_n.php?id_menu=5.

from