Objective

1. To determine the amount of calcium in a sample and in tap water by calibration

curve.
2. To determine the amount of calcium in a sample by standard addition method.

Introduction

Atomic absorption spectrometry (AAS) is an analytical technique that measures the

concentrations of elements. Atomic absorption is so sensitive that it can measure down to
parts per billion of a gram (µg dm–3) in a sample. The quantity of interest in atomic
absorption measurements is the amount of light at the resonant wavelength which is
absorbed as the light passes through a cloud of atoms. As the number of atoms in the light
path increases, the amount of light absorbed increases in a predictable way. By measuring
the amount of light absorbed, a quantitative determination of the amount of analyte element
present can be made. The use of special light sources and careful selection of wavelength
allow the specific quantitative determination of individual elements in the presence of others.

The atom cloud required for atomic absorption measurements is produced by supplying
enough thermal energy to the sample to dissociate the chemical compounds into free atoms.
Aspirating a solution of the sample into a flame aligned in the light beam serves this
purpose. Under the proper flame conditions, most of the atoms will remain in the ground
state form and are capable of absorbing light at the analytical wavelength from a source
lamp. The ease and speed at which precise and accurate determinations can be made with
this technique have made atomic absorption one of the most popular methods for the
determination of metals.

Instrument

PYE UNICAM SP9 ATOMIC ABSORPTION SPECTROPHOTOMETER

Materials

1. Standard Ca++ solution, 100ppm

2. Unknown solution
3. Tap water
4. Distilled water
Experimental procedures
A series of standards solution was
prepared by diluting stock solution
to the concentration of 2ppm,
4ppm, 6ppm, 8ppm and 10ppm.
A series of solution for
standard addition method
was prepared and labeled as
X+0, X+1, X+2, X+3, X+4, and
X+5.
The absorbance of standard
solution in step 1, 3, 4 and 5
were measured
The sample was
measured by using
pipette and diluted
until its calibration
mark (CM).
A 5mL of unknown A sample of tap water
sample, X, is placed and a solution of tap
into 100 mL of water: distilled water
volumetric flask and in ratio of 1:1 were
diluted until its CM. prepared in a 100mL
volumetric flask.
Standard calibration curves of
absorbance vs concentration of
standard were plotted and the
calcium concentration in samples
were determined
Results:

Wavelength of the measurement: 422.7 nm

1) Standard solution of Ca++ solution

Sample Concentration, ppm Absorbance

Blank 0 0.00
2ppm 2 0.08
4ppm 4 0.14
6ppm 6 0.20
8ppm 8 0.26
10ppm 10 0.31

2) Unknown solution

Sample Concentration of standard Absorbance

solution, ppm
X+0 0 0.33
X+1 1 0.47
X+2 2 0.59
X+3 3 0.71
X+4 4 0.82
X+5 5 0.91

3) Unknown, X, tap water and ratio of tap water : dH2O ( 1:1 )

Sample Concentration, ppm Absorbance

Unknown 4.03 0.13
Tap water 11.80 0.38
Tap water : dh2o ( 1:1 ) 5.629 0.18
Discussion:

The basic principle of AAS

Every absorption spectrometer must have components which fulfill the three basic
requirements shown in Figure 1. There must be:

1. A light source
2. A sample cell
3. A means of specific light measurement.

Figure 1 Requirements for a spectrometer

There are five basic components of an atomic absorption instrument:

1. The light source that emits the spectrum of the element of interest

2. An "absorption cell" in which atoms of the sample are produced (flame, graphite
furnace, MHS cell, FIAS cell, FIMS cell)

3. A monochromator for light dispersion

4. A detector, which measures the light intensity and amplifies the signal

5. A display that shows the reading after it has been processed by the instrument
electronics

Figure 2 Basic principle of Atomic Absorption Spectrometer.

There are two basic types of atomic absorption instruments: single-beam and
double-beam.

Single-Beam Photometers

The instrument diagrammed in Figure 5 represents a fully functional ‘‘single-beam’’

atomic absorption spectrometer. It is called ‘‘single-beam’’ because all
measurements are based on the varying intensity of a single beam of light in a single
optical path.

Figure 3 A single-beam AA spectrometer

The primary advantage of a single-beam configuration:

a. It has fewer components and is less complicated than alternative designs. It is

therefore easier to construct and less expensive than other types of photometers.
b. With a single light path and a minimum number of optical components, single-
beam systems typically provide very high light throughput.

The primary limitation of the singlebeam photometer:

a. It provides no means to compensate for instrumental variations during an

analysis, such as changes in source intensity.

Double-Beam Photometers

In the double-beam system as in Figure 6, additional optics is used to divide the light
from the lamp into a ‘‘sample beam’’ which directed through the sample cell and a
‘‘reference beam’’ which directed around the sample cell. The reference beam
serves as a monitor of lamp intensity and the response characteristics of common
electronic circuitry.
Figure 4 A double-beam AA spectrometer.

The primary advantages of double-beam AA spectrometer:

a. Double-beam is automatically compensates for source and common electronics

drift which allows these instruments to change lamps and begin an analysis
immediately with little or no lamp warm-up for most elements. This reduces
analysis time and prolongs lamp life, since lamp warm-up time is eliminated.

LIGHT SOURCES

An atom absorbs light at discrete wavelengths. In order to measure this narrow light
absorption with maximum sensitivity, it is necessary to use a line source, which emits
the specific wavelengths which can be absorbed by the atom. Narrow line sources
not only provide high sensitivity, but also make atomic absorption a very specific
analytical technique with little spectral interference. The two most common line
sources used in atomic absorption are the hollow cathode lamp and the
electrodeless discharge lamp.

Figure 5 Hollow cathode lamp Figure 6 Electrodeless discharge lamp

Burner System

In AAS, burner system functions as (i) sample holder, (ii) desolvation source, and (iii)
volatilization source. The dual option burner system can be operated either with or
without a flow spoiler for optimum operation under different analytical conditions.
However, for routine operation it is recommended that the burner system be
operated with a flow spoiler. The flow spoiler helps to remove large droplets from the
nebulizer aerosol and thus minimizes chemical interferences. A diagram of the dual
option burner system is shown in Figure 7.

Figure 7 Diagram of Dual-Option Burner System

Monochromator – Wavelength Separator

Detector

Photomultiplier tubes
The dilution calculations of each working solution

The preparation of series of standards solution by diluting the stock solution:

2 ppm: 4 ppm: 6 ppm:

MssVss = M2V2 MssVss = M2V2 MssVss = M2V2
(100)Vss = (2)(100) (100)Vss = (4)(100) (100)Vss = (6)(100)
Vss = 2 ml Vss = 4 ml Vss = 6 ml

8 ppm: 10 ppm:
MssVss = M2V2 MssVss = M2V2
(100)Vss = (8)(100) (100)Vss = (10)(100)
Vss = 8 ml Vss = 10 ml

Standard calibration of unknown solution, X

Gradient:

𝑌2 − 𝑌1
m=
𝑋2 − 𝑋1

0.82−0.47
m=
4−1

m = 0.1167

Concentration of unknown, X:

b x Cstd
Cx =
𝑚 𝑥 𝑉𝑥

0.35 x 100
Cx =
0.1167 𝑥 5

Cx = 59.98ppm

Amount of calcium in unknown solution, X:

59.98 mg/L x 0.005L = 0.3mg

Amount of calcium in tap water:

11.80 mg/L x 0.1L = 1.18mg

In this experiment, Flame Atomic Spectroscopy is used in order to determine the
concentration of calcium in tap water and unknown sample by using two methods. a
series of calcium solution is prepared from a stock solution of 100 ppm calcium
solution. The methods are calibration curve method and the standard addition
method. Firstly, we had used the calibration method and the concentration of calcium
in the unknown sample is determined by comparing it to a set of known
concentration samples. A graph of Calibration curve is plotted and a linear
relationship is produced in order to obey Beer’s law.

Based on the graph of calibration curve, we can determine the concentration of

calcium in the solution of tap water : distilled water in ratio 1 : 1 and tap water
solution. since the graph of concentration calcium is proportional to its absorbance,
we could say that as the concentration of tap water increases in solution, the
absorbance also increases. From the graph, the solution of tap water : distilled water
in ratio 1 : 1 and tap water has the concentration of calcium of 5.60ppm and
11.90ppm respectively. Thus, the amount of calcium in tap water is 1.18 mg.

Next is the standard addition method. It is a common method that had been used by
adding one or more increment of standard solution to sample aliquots of the same
size. This is also known as spiking the sample. A series of standard with 1, 2, 3, 4
and 5ppm were added to the 15mL of unknown in separate volumetric flask and was
diluted. The standard addition graph was plotted and the concentration of calcium in
the unknown sample, X, is determined which is 19.84ppm. Thus, the amount of
calcium in the unknown is 1.984mg

During the experiment was conducted, there could be errors that had occur. To avoid
errors, several precautions should be taken by students. To make sure that the
concentration of the standards are accurate during preparation of the solution, we
have to make sure that our eye is parallel with the volumetric flask mark and to use a
dropper when the solution almost reach the mark. This is so that we do not exceed
the mark. Besides that, when using the instrument for analysis, we must set the auto
sampler with correct location of vials so that there will be no incorrect sampling.
Conclusion:

Based on this experiment, we can conclude that the concentration of calcium in the
tap water and in the unknown sample is 11.90ppm and 19.84ppm respectively. The
concentration of calcium in tap water is determined by using the calibration curve
whereas the concentration of calcium in the unknown sample is determined by using
the standard addition method. Meanwhile, the amount of calcium in the tap water
and in the unknown sample is 1.18mg and 1.984mg respectively.

Reference

(n.d.). Retrieved November 11, 2017, from Chapter 9: Atomic Absorption Spectrometry:
http://www2.chemistry.msu.edu/courses/cem434/chap9atomabsspec.pdf

(1996). Analytical Methods for Atomic Absorption Spectroscopy. United States of America: The
Perkin-Elmer Corporation.

Beaty, R. D., & Kerber, J. D. (1993). Concepts, Instrumentation and Techniques in Atomic Absorption
Spectrophotometry. Norwalk, CT, U.S.A: The Perkin-Elmer Corporation.

Levinson, R. (n.d.). Royal Society of Chemistry. Burlington House, Piccadilly, London : The Education
Department, The Royal Society of Chemistry.
The calibration curve of tap water and tap water:distilled water
The calibration curve of standard solution
The calibration curve of the unknown solution