UNIVERSITI TEKNOLOGI MARA

FACULTY OF APPLIED SCIENCES

BACHELOR OF SCIENCE (HONS.)
ENVIRONMENTAL TECHNOLOGY

CHEMICAL LABORATORY

COURSE : BASIC INSTRUMENTAL ANALYSIS (CHM571)

EXPERIMENT : ATOMIC ABSORPTION SPECTROMETRY

NAME : AIN ATHIRAH BINTI RAHIMI

MATRIC NO : 2021124045

GROUP : AS2292A

DATE OF SUBMISSION : 2ND DECEMBER 2021

LECTURER’S NAME : DR NORAINI KASIM

 EXPERIMENT 1: ATOMIC ABSORPTION SPECTROMETRY

OBJECTIVE

1. To determine the concentration of calcium in an unknown calcium solution using a

standard calibration curve and standard addition method.

2. To determine the concentration of a Calcium in tap water and 1:1 ratio (tap water:
distilled water) with calibration method.

INTRODUCTION

AAS (Atomic Absorption Spectrometry) is an analysis technique that uses light absorption to
determine the concentration of trace metals in various samples. Today, the technique is used all
over the world as an important analysis tool in a variety of industries. AAS is widely used in the
medical field for blood and soft tissue analysis, as well as in pharmaceutical research and
production. AAS is also used extensively in the trace analysis of metals in water. AAS functions
as atomization. AAS detects the presence of gas-phase atoms by measuring light absorption.
Because analyte atoms or ions must be vaporized in a flame or graphite furnace because samples
are usually liquids or solids. The AAS is a device that measures the amount of radiation
absorption of energy by a ground-state atom in a flame during these energy transitions. The light
source for the atomic absorption apparatus is a hollow cathode lamp with a cathode made of the
same metal as the metal being interpreted, in this case calcium.
METHODOLOGY

The Calibration Curve Method is covered in Part I:

1. From standard 100 ppm stock solutions, a series of standard solutions with the following
concentrations were prepared: 2, 4, 6, 8, and 10 ppm.
2. Then, 5 mL of an unknown solution was divided evenly among six 100 mL volumetric
flasks. Before taking solution measurements, an empty water (deionized water) was run.
3. Following that, the absorption of a standard solution series of 2, 4, 6, 8, and 10 ppm was
determined.
4. The absorption of an unknown calcium solution was then determined. The absorption of
tap water was then measured. The absorption ratio of 1: 1 (tap water: distilled water) was
then determined.
5. Finally, the calcium concentration in an unknown calcium solution (tap water, 1: 1 tap
water to distilled water ratio) was calculated using the calibration curve (Absorption
versus standard calcium concentration).

The Standard Addition Method is used in Part II:

1. To begin, 5 mL of an unknown solution was divided evenly among six 100 mL

volumetric flasks. For standard addition methods, Thermos for a succession of solutions
were produced and labelled.
2. Then, in each of the six volumetric flasks at number 1, a standard calcium solution (100
ppm) was added by 0, 2, 4, 6, 8, 10 mL.
3. After that, each of the six volumetric flasks was filled to the mark with distilled water.
After that, the standard addition method's absorption of the solution series was measured.
4. Finally, the calcium content in an unknown calcium solution was determined.
DATA, RESULTS AND CALCULATION

I. Calibration Curve Method

Concentration of Calcium, Ca in Absorbance

standard solution, ppm
0 0.002
2 0.109
4 0.193
6 0.280
8 0.351
10 0.417
1:1 (tap water : distilled water) 0.753
Tap water 0.488
Unknown 0.300

0.8
0.753
0.7
0.6
ABSORBANCE

0.5 0.488
0.4 0.417
0.351
0.3 0.28 0.3
0.2 0.193
0.1 0.109
0 0.002
0 2 4 6 8 10 1:1 (tap Tap Unknown
water: Water
distilled
water)
CONCENTRATION OF CA SOLUTION IN PPM
Calibration curve method
Formula: y = m𝑥 + c
y = 0.0413x + 0.019
y = absorbance, m = gradient, c = y-intercept. 𝑥 = concentration
We need to find the value of 𝑥.

1. Concentration of Ca in 1:1 (tap water : distilled water)

y = m𝑥 + c
0.753 = 0.0413𝑥 + 0.019
𝑥 = (0.753 − 0.019)/0.0413
𝑥 = 17.7723 𝑝𝑝𝑚

2. Concentration of Ca in tap water

y = m𝑥 + c
0.488 = 0.0413𝑥 + 0.019
𝑥 = (0.488 − 0.019)/0.0413
𝑥 = 13.3559 𝑝𝑝𝑚

3. Concentration of Ca in unknown solution

y = m𝑥 + c
0.300 = 0.0413𝑥 + 0.019
𝑥 = (0.300 − 0.019)/0.0413
𝑥 = 6.8039 𝑝𝑝𝑚
I. Standard Addition Method

Flask Concentration of Volume of standard Sample Absorbance

standard (ppm) added (mL) Id

1 0 0 X+0 0.423

2 1 1 X+1 0.461

3 2 2 X+2 0.511

4 3 3 X+3 0.574

5 4 4 X+4 0.581

6 5 5 X+5 0.600

0.7

0.6 0.6
0.574 0.581
0.5 0.511
0.461
ABSORBANCE

0.4 0.423

0.3

0.2

0.1

0
0 1 2 3 4 5
VOLUME OF STANDARD ADDED (mL)
Standard addition method

By using formula:-

𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥

𝐶𝑥 = concentration of the unknown, b = y-intercept, m = gradient of the graph, 𝐶𝑠 =

volume added, 𝑉𝑥 = volume of unknown.
We need to find the concentration of the unknown solution, 𝐶𝑥.

i. For 1 mL volume of standard added

𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥
(𝑜. 4316 × 1)
𝐶𝑥 =
(0.0374 × 5)
𝐶𝑥 = 2.3080 𝑝𝑝𝑚

ii. For 2 mL volume of standard added

𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥
(𝑜. 4316 × 2)
𝐶𝑥 =
(0.0374 × 5)
𝐶𝑥 = 4.6160 𝑝𝑝𝑚

iii. For 3 mL volume of standard added

𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥
(𝑜. 4316 × 3)
𝐶𝑥 =
(0.0374 × 5)
𝐶𝑥 = 6.9241 𝑝𝑝𝑚
iv. For 4 mL volume of standard added
𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥
(𝑜. 4316 × 4)
𝐶𝑥 =
(0.0374 × 5)
𝐶𝑥 = 9.2321 𝑝𝑝𝑚

v. For 5 mL volume of standard added

𝑏𝐶𝑠
𝐶𝑥 =
𝑚𝑉𝑥
(𝑜. 4316 × 5)
𝐶𝑥 =
(0.0374 × 5)
𝐶𝑥 = 11.5401 𝑝𝑝𝑚
DISCUSSION

The value of the absorbance was calculated using the calibration curve method when the quantity
of calcium, Ca in standard solution, ppm is 0, 2, 4, 6, 8, 10, 1: 1 (tap water: distilled water), tap
water, and unknown are 0.0002, 0.193, 0.280, 0.351, 0.417, 0.753, 0.488, and 0.300,
respectively. Thus, a calibration curve technique graph might be created using these data. The
concentration can then be calculated using the formula y = mx + c, where x is the concentration.
Ca concentration in 1: 1 (tap water: distilled water) is 17.7723 parts per million, Ca
concentration in tap water is 13.3559 parts per million, and Ca content in unknown solution is
6.8039 parts per million. For the standard addition method, the data for standard concentration
(ppm) is equal to the volume of standard added (mL) starting at 0, 1, 2, 3, 4, and 5, with
absorbance values of 0.423, 0.461, 0.511, 0.574, 0.581, and 0.600, respectively. As a result, the
conventional addition technique graph has been created using this entire set of data. The
𝑏𝐶𝑠
concentration can then be calculated using the formula 𝐶𝑥 = 𝑚𝑉𝑥. The 𝐶𝑥 were 2.308 ppm,

4.616 ppm, 6.924 ppm, 9.232 ppm, and 11.5401 for 1 mL, 2 mL, 3 mL, 4 mL, and 5 mL standard
added, respectively. While conducting this experiment, there are some precautions that can be
taken. To begin, wear safety goggles while preparing and evaluating the samples in this
experiment to avoid any eye injuries. Next, inspect the entire gas system, including the hose lines
that connect the AAS to the gas cylinders, for flexibility, cracks, and holes before running the
experiment. Finally, look for excessive carbon buildup across the entire gas burner system.

CONCLUSION

Subsequently, the atomic absorption spectrometry experiment has been done successfully, and
the objectives of this experiment were met, which have been to determine the unknown
concentration of calcium in an unknown calcium solution using standard addition methods and
standard calibration curves. Using the calibration curve method, these experiments successfully
determined the calcium concentration in tap water and 1: 1 (tap water: distilled water). It has
been proved that the calibration curve method has a higher concentration than for the standard
addition method. Likewise, as the absorbance increases, so will the volume and the
concentration. As a byproduct, this hypothesis is accepted.
ANSWERS TO QUESTIONS (Pre-Lab Assignment)

1. Calculate and describe the preparation of standard solutions with concentrations of 2, 4,

6, 8 and 10 ppm in 100 mL volumetric flask from a standard stock solution (100 ppm).
𝐶1 = (0, 2, 4, 6, 8, 10)𝑝𝑝𝑚, 𝑉1 = 100 𝑚𝐿
𝐶2 = 100 𝑝𝑝𝑚, 𝑉2 =?
𝐵𝑦 𝑢𝑠𝑖𝑛𝑔 𝑓𝑜𝑟𝑚𝑢𝑙𝑎 ∶ 𝐶1 𝑉1 = 𝐶2 𝑉2

𝐹𝑜𝑟 𝐶1 = 0 𝑝𝑝𝑚 𝐹𝑜𝑟 𝐶1 = 6 𝑝𝑝𝑚

𝐶1 𝑉1 = 𝐶2 𝑉2 𝐶1 𝑉1 = 𝐶2 𝑉2
2(100) = 100𝑉2 6(100) = 100𝑉2
𝑉2 = 2 𝑚𝐿 𝑉2 = 6 𝑚𝐿

For C1 = 2 ppm For C1 = 8 ppm

(C1)(V1) = (C2)(V2) (C1)(V1) = (C2)(V2)

(2)(100) = (100)(V2) (8)(100) = (100)(V2)

V2 = 2 mL V2 = 8 mL

For C1 = 4 ppm For C1 = 10 ppm

(C1)(V1) = (C2)(V2) (C1)(V1) = (C2)(V2)

(4)(100) = (100)(V2) (10)(100) = (100)(V2)

V2 = 4 mL V2 = 10 mL
2. In the standard addition method, you will have to prepare a series of solution in which
you add increments of standard solution (100 ppm) to sample aliquots of same volume (to
a 5 mL of samples). The volumes of the added standards in the six 100 mL volumetric
flasks are 0, 2, 4, 6, 8 and 10 mL respectively, then dilute to mark. Calculate the
concentration of the added standard in each flask.

C1 = 100 ppm, V1 = 5 mL, C2 = ?, V2 = (0, 2, 4, 6, 8, 10) mL

By using formula: (C1)(V1) = (C2)(V2)

For V2 = 0 mL For V2 = 6 mL

(C1)(V1) = (C2)(V2) (C1)(V1) = (C2)(V2)

(100)(5) = (C2)(0) (100)(5) = (C2)(6)

C2 = 0 ppm C2 = 83.3 ppm

For V2 = 2 mL For C1 = 8 ppm

(C1)(V1) = (C2)(V2) (C1)(V1) = (C2)(V2)

(100)(5) = (C2)(2) (100)(5) = (C2)(8)

C2 = 250 ppm C2 = 62.5 ppm

For V2 = 4 mL For C1 = 10 ppm

(C1)(V1) = (C2)(V2) (C1)(V1) = (C2)(V2)

(100)(5) = (C2)(4) (100)(5) = (C2)(10)

C2 = 125 ppm C2 = 50 ppm

 3. How do you determine the concentration of an unknown form
a. standard calibration curve.
A calibration curve, also known as a standard curve, is a general method for determining
the concentration of a substance in an unknown sample by comparing it to a set of known
concentration standard samples. To evaluate the concentration of an unknown calcium
solution from this curve, take this value on the intensity axis and draw a line parallel to
the x-axis where it intersects the standard curve, then draw a line perpendicular to the
concentration axis, which gives the concentration.

b. standard addition method.

A quantitative analysis technique used to minimize matrix effects that interfere with
analyte measurement signals is known as a standard addition method. The intensity of the
light of the test solution is measured with reference to the blank to determine the
concentration of an unknown calcium solution that use this method. The element to be
determined is then added in greater concentrations to a few test solutions, and the
solutions are diluted to the same volume in each case.

REFERENCES

1) Walter Slavin, Glen R. Carnrick & S. Roy Koirtyohann (1988) Background Correction in
Atomic Absorption Spectroscopy (AAS), C R C Critical Reviews in Analytical
Chemistry, 19:2, 95-134
2) Akram, S., Najam, R., Rizwani, G.H. and Abbas, S.A., 2015. Determination of heavy
metal contents by atomic absorption spectroscopy (AAS) in some medicinal plants from
Pakistani and Malaysian origin. Pakistan journal of pharmaceutical sciences, 28(5).
3) Ali Bazzi, Bette Kreuz, and Jeffrey Fischer
Journal of Chemical Education 2004 81 (7), 1042