CHM260 LABORATORY REPORT

EXPERIMENT 4
FLAME ATOMIC ABSORPTION SPECTROSCOPY

NAME AND STUDENT ID: NURUL SHAFIAH BINTI SUPIAN ZORI

(2020482028)
ATHIRAH ABD HALIM
(2020845588)
UMIE MAISARA BINTI CHE MAZLAN
(2020895798)
NURAIN FASIHAH BINTI JUNID
(2020895282)
FACULTY: APPLIED SCIENCE FACULTY

GROUP: A4AS120517

DATE OF SUBMISSION: 11/11/2022

LECTURE’S NAME: DR FAIQAH RAMLI

1.0 INTRODUCTION
Atomic absorption spectroscopy, AAS is an analytical technique used to determine how much
of certain elements are in a sample. It uses the principle that atoms (and ions) can absorb light
at a specific, unique wavelength. In the flame, the liquid/solvent is evaporated into droplets
and the metal is vaporized mainly as atoms. Alternatively, an atomic vapour can be produced
by a rapid electrochemical heating of a graphite rod or tube in which a drop of the sample has
been placed. When the sample contains this specific wavelength of light, the energy (light) is
absorbed by the atom. Electrons in the atom move from the ground state to an excited state.
When the atoms make their transitions, they absorb some of the light from the beam such as
the radiation of the particular wavelength that is produced by a special source made from that
element. The amount of light absorbed is measured and the concentration of the element in
the sample can be calculated. The more concentrated the solution, the lighter energy is
absorbed.

2.0 OBJECTIVE:

• To learn how to operate a flame AAS instrument.

• To plot a standard calibration curve for determination of Ca in a simple.
• To determine the amount of Ca in a sample using the standard calibration curve.

3.0 APPARATUS:

• Beaker
• Dropper
• Pipette
• Volumetric flask, 50 mL

4.0 CHEMICALS:

• Deionized water
• 1000 ppm Ca stock solution
• Nitric acid, HNO3 (1%)
• Mineral water
• Drinking water
• Tap water
5.0 PROCEDURE:

In this experiment, the liquid sample was introduced to the flame by aspirating it into the air
supplied to the flame. The place was taken at the nebulizer chamber where the sample
solution was broken up into a very fine mist of droplets (aerosol) before entering the flame.
In the flame, the solvent evaporates and most of the element is converted into free atoms.

The Ca was analysed in a liquid sample.

A. Preparation of Standard Solutions

1) 5.00 mL of 1000 ppm stock solution was pipetted. It was transferred into a 50.0 mL
volumetric flask and was diluted with deionized water until the mark. The solution
was labelled as ‘100 ppm’.
2) The 100 ppm solution was used to prepare a series of standard solutions with
concentration of 1 ppm,2 ppm,3 ppm,4 ppm,5 ppm by using 100 ml volumetric flask.
3) Unknown solution was obtained.
4) The absorbance of standard solution in (2) was measured, followed by the unknown.

B. Operating of the AAS

Instrument: Perkin Elmer AAnalyst 700

Safety information

1) Do not leave flammable material near AA

2) Do not let acetylene tank level drop below 70 psi
3) Do not stare into the lamp

Igniting and optimizing the Flame (Air / Acetylene operation)

1) The burner height was adjusted by turning the horizontal adjustment knob until the
light beam is centred over the burner slot.
2) The vertical positioning of the burner head was adjusted by rotating the knob until the
light was near but not hitting the top of the burner head.
3) The acetylene tank was opened and checked the flow, which to be from 85-100 kPa
4) The air valve was open and the flow was checked to be from 50-65 psi
5) The Oxidant switch was turned on to Air
6) The ignite button was pressed
 7) The blank (water) was aspirated for 60-90 seconds. [A/Z] was pressed.
8) A mid-concentration standard was aspirated. The horizontal and vertical rotations
were adjusted to obtain maximum absorbance.
9) A blank was aspirated. The aspiration hose was left in the water.

Measurement of Atomic absorbance

1) The blank was aspirated by pressing [Cont],[A/Z]

2) The standards were aspirated and the absorbance was recorded for each one.
3) The samples was aspirated and the absorbance was recorded for each one

Shut down procedure

1) The water was aspirated for a few minutes to clean the burner head.
2) The gas control switch on the pneumatics control panel was turned off
3) All the gas sources valves was closed
4) The gas control switch was turned to Air to allow the gasses to bleed. The gas
control was turned off.
5) The instrument was turned off.

How to Run the AAS

1) The air compressor, stock cock air, acetylene regulator and acetylene tank was
opened.
2) The compressed air must be between 5-6 kg/cm3 and the acetylene pressure is
100kPa.
3) Exhaust fan was switched on
4) Isolating switch was switched on
5) The aspiration hose should be in water or blank.
6) Instrument was turned on
7) The computer was turned on
8) WinLab 32 was double clicked until the calibrating equipment was ready.
9) The method was created by clicking on click file new method starting
condition : Element Ca
10) Method editor : untitled (type in the date)
a. Method description: To determine the concentration of Ca
b. Calibration equation: linear through zero
c. Change unit (for standard and unknown)
 11) Standard concentration
i. Blank : DI water
ii. Standard : std1 , std 2 , std 3 , std 4 and std 5
iii. Concentration : 1 ppm , 2 ppm , 3 ppm , 4 ppm , 5 ppm
12) File → save as → method
i. Name :
ii. Method :
13) File → new → sample into file →ok
Sample ID : unknown Ca
14) File→ save as→ sample info
File name : → save

15) Workspace ( workspace ) was clicked - whatever is in the box was highlighted →
open → type in name.
16) Burner switch was clicked→ analyzed standard→ analyze blank →analyze sample
unknown.
17) Burner was switched off.
18) Tools was clicked → calibration was edited
19) To print→ file was click → active window preview
20) To open the file, ‘file’ was clicked → open →workspace→ filename→ok→analyze→
Recall calibration

6.0 TABULATED DATA

Solution Concentration of Solution, ppm Absorbance

Standard 1 1 0.113

Standard 2 2 0.178

Standard 3 3 0.254

Standard 4 4 0.320

Standard 5 5 0.390

Mineral water
1.081 0.088
(1:10 dil)
 Drinking water 0.753 0.061

Tap water 1.1015 0.083

7.0 GRAPH

Absorbance vs Concentration of Standard (ppm)

0.5

0.4 y = 0.0767x + 0.014

Concentration of Standard (ppm)

R² = 0.9886

0.3

0.2

0.1

0
-2 -1 0 1 2 3 4 5 6

-0.1
Absorbance

Graph of absorbance vs Concentration of standard (ppm)

Volume of unknown solution:

Equation:
𝑌 = 0.0767𝑥 + 0.014

i) Mineral Water:

0.0767𝑥 + 0.014 = 0.088

0.0767𝑥 = 0.074

𝑥 = 0.9648 × 10(𝑑𝑖𝑙𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟)

𝑥 = 9.648𝑚𝑔/𝐿

ii) Drinking Water:

0.0767𝑥 + 0.014 = 0.061

0.0767𝑥 = 0.047

𝑥 = 0.612𝑚𝑔/𝐿

iii) Tap Water:

0.0767𝑥 + 0.014 = 0.083

0.0767𝑥 = 0.069

𝑥 = 0.900𝑚𝑔/𝐿

8.0 QUESTIONS:
In a standard addition method, you will prepare a series of solutions in which you add different
increments of standard solutions to fixed aliquots of an unknown sample X i.e., 10.00 mL of
X. All solutions were prepared using a 50.00 mL volumetric flask. Suppose that the analysis
of standard solutions of X gave the following results:

Solution Concentration of Added Standard, ppm Absorbance

Standard 1 0.00 0.201

Standard 2 2.44 0.292

Standard 3 4.88 0.378

Standard 4 7.32 0.467

Standard 5 9.76 0.554

Table 2
 1. Plot the standard addition graph.

2. Determine the concentration of X (in ppm) in the unknown sample.

𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑋 𝑖𝑛 𝑢𝑛𝑘𝑛𝑜𝑤𝑛 𝑠𝑎𝑚𝑝𝑙𝑒 = |𝑥 − 𝑖𝑛𝑡𝑒𝑟𝑐𝑒𝑝𝑡|

𝑥 − 𝑖𝑛𝑡𝑒𝑟𝑐𝑒𝑝𝑡, 𝑤ℎ𝑒𝑛 𝑦 = 0

𝑌 = 0.0361𝑋 + 0.2022

0.0361𝑋 + 0.2022 = 0

0.0361𝑋 = − 0.2022

𝑋 = | − 5.60|

𝑋 = 5.60 𝑝𝑝𝑚
9.0 DISCUSSION

Atomic spectroscopy is a class of spectroscopic method in which the species examined in the
spectrometer are in the form of atom. It is a technique for measuring the concentration of
metallic element in different material. Based on the principle that the ground state metals
absorb light at a specific wavelength. During the atomic absorption spectroscopy process,
these atoms will absorb electromagnetic radiation at a specific wavelength. This produces a
measurable signal. By looking at these signals, it is then possible to determine the parts per
million, or ppm, levels of specified metals in the material that is being tested. Within an atom,
there are electrons at various energy levels. During the spectroscopy process, the absorbing
of energy moves electrons to a more energetic level.

The objective of this experiment was to learn how to operate flame AAS instrument, to
plot a standard calibration curve for determination of Ca sample and to determine the amount
of Ca in a sample using the standard calibration curve.In the beginning of the experiment, 5ml
of 1000ppm stock solution was pipet into 50 ml volumetric flask and it was diluted with deionize
water until the calibration mark. Then preparation of the series of the standard solution it must
using McVc=MdVd to find the volume of the dilution. Then the stock solution was diluted with
1% of HNO3. Besides that, it also used the mineral water, drinking water and tap water as a
standard solution. In this experiment, only 5ml mineral water should be diluted with the 1%
HNO3 until the calibration mark because of the high content of the mineral which can be
causing changes for obtaining the actual results. When light of the correct wavelength is
supplied, the amount of light absorbed is measured and a reading can be obtained. After all
the results was determine, the graph can be plotting absorbance versus concentration. By
plotting that, the calibration curve can be plotted. The intensity of the absorbed light is
proportional to the concentration of the element in the flame. In AAS instrument, the elements
in the gas phase absorb light at very specific wavelengths. From the results, the specific
wavelength of the Ca is 422.67. The atom is absorbing the visible light and make the transition
to higher electronic energy level. Then, the analyte concentration is determined from the
amount of absorption.

The standard AAS instrument consists of nebulizer, hollow cathode lamp,

monochromator, detector, and data processer. In this experiment the energy lamp for the AAS
instrument is 65. Firstly, the flame converts samples into free ground state atoms that can be
excited. In flame AAS, nebulizer will aspirate a sample into a flame. The flame is line up in a
beam of light of the appropriate wavelength. The flame causes the atom to undergo a transition
from the ground state to the first excited state. The hollow lamp must be perfectly aligned, so
the beam crosses the hottest part of the flame and travels into detector. Then, the detector
measures the intensity of the beam light. When some of the light is absorbed by a metal, the
beam’s intensity is reduced. The detector records that reduction as an absorption. The most
important part in AAS spectrometer is monochromator. It is used to separate all the thousand
lines to a single line. So, in ASS only one line spectrum will be form. The light selected by the
monochromator is directed onto a detector that is typically a photomultiplier tube, whose
function is to convert light signal into an electrical signal proportionally to the light intensity.
Then signal amplifier process of the electrical signal.

However, all measurements have some degree of uncertainty that may come from a
variety of sources. The process of evaluating the uncertainty associated with a measurement
result is often called uncertainty analysis or error analysis. From this experiment, there is
1.04% or error that may be coming from the possible error. There are many possible errors in
this experiment such as human error. Not labelling the 50ml volumetric flask according to the
5 series of the standard solution correctly that will maybe affect the result far from the original
due to the mismatched of the concentration of the standard solution. To avoid this
mismatched, the volumetric flask should be labelled properly. There are several precautions
should be acted during this experiment, which is the eyes must be perpendicular to the
meniscus or scale of the apparatus. It is because to avoid from affect the result. When handling
the sample of flame AAS instrument, the tip of the instrument must be paced into the solution
to enable the reading to appear correctly. Besides that, the switch of instrument must be turn
off after result was obtain. Furthermore, do not stare into the flame because it will deficit our
sightedness and in serious cases will cause blindness. Then, do not stare into the flame
because it will deficit our sightedness and in serious cases will cause blindness. Moreover, do
not leave flammable material near AA. Finally, do not let acetylene tank level drop below 70psi.

10.0 CONCLUSION:

To conclude, during this experiment and after this experiment was conducted, we were able
to learn on how to handle and operate Flame Atomic Absorption Spectroscopy instrument well
and at the same time was able to plot a standard calibration curve for determination of Ca in
a sample and determine the amount of Ca in a sample using the standard calibration curve. So
the objective for this experiment is achieve.
11.0 REFERENCE

Barron, P. M. (2022, August 29). 1.4: Introduction to Atomic Absorption Spectroscopy. Retrieved
November 6, 2022, from LibreTexts Chemistry:
https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Physical_Methods_in_Chemi
stry_and_Nano_Science_(Barron)/01%3A_Elemental_Analysis/1.04%3A_Introduction_to_At
omic_Absorption_Spectroscopy

Dancahill. (2022, October 13). What is Atomic Absorption Spectroscopy (AAS). Retrieved November
8, 2022, from SCIMED: https://www.scimed.co.uk/education/what-is-atomic-absorption-
spectroscopy-
aas/#:~:text=Atomic%20absorption%20spectroscopy%2C%20or%20AAS,will%20absorb%20t
hese%20wavelengths%20differently