Experiment 6 Date: 10 October 2006

Title
Atomic Absorption Spectroscopy – Determination of Zinc in Water

Abstract
The main objective of this experiment is to determine the concentration of zinc (Zn) in a water sample using
atomic absorption spectroscopy (AAS) via two methods, linear calibration and standard addition. The former
utilizes a series of standard Zn solutions, of which a plot of absorbance versus standard concentration is
obtained and the sample concentration is determined from the plot. The latter incorporates the addition of
different Zn stock solution aliquots into equal amounts of water sample. Extrapolating the plot of absorbance
versus added concentration to the intercept of the horizontal axis gives the sample concentration. Because of
large differences between the final values from the two methods, taking into consideration of several factors,
the water sample is concluded to contain 0.167 ppm Zn, but with errors.

1.0 Introduction
1.1 Background Information of Method
Atomic absorption spectroscopy (AAS) is one of the methods used to determine the
concentration of metal elements in samples, such as iron (Fe), copper (Cu), aluminium (Al), lead
(Pb), calcium (Ca), zinc (Zn), cadmium (Cd) and 55 different other metals. Typical
concentration values are measured as low as in mg/L or ppm (parts per million), although ppb
(part per billion) values may be possible. First developed by a team of Australian chemists in the
50s, the technique makes use of a flame to atomize the sample. However, other atomizers such
as the graphite furnace, have since then been developed. Three major steps are involved in
converting the liquid sample into atomic gas, formerly desolvation (evaporation of solvent),
vaporization (conversion of dried solid into gas) and volatilization (breaking up of sample
compounds into atoms). The following is a schematic diagram of the atomic absorption (AA)
spectrometer:

Figure 1: Schematic Diagram of Atomic Absorption Spectrometer

The light source (hollow cathode lamp) emits a spectrum specific to the element of which it is
made, which is focused through the sample cell into the monochromator. At the sample cell
section, sample is being taken and undergoes a nebulization process into a fine aerosol solution
and dissociation of analyte elements into free gaseous ground state form by the flame in that
section. Only 10 percent of the analyte go through the flame. Usually, either air – acetylene or
nitrous oxide – acetylene combinations are used as fuel for the flame. The atoms then absorb
light energy from the source and are excited to the excited state. As the excited state is unstable,
the atoms return to ground state (decay process), emitting light in the process.

Figure 2: Atomic Absorption and Decay Process

 The light source must be electronically modulated or mechanically chopped to differentiate
between the light from the source and the emission from the sample cell. The monochromator
then disperses the light and the specific wavelength of light isolated passes to the detector,
usually a photomultiplier tube. An electrical current is produced depending on the light intensity
and processed by instrument electronics, which measure the amount of light attenuation in the
sample cell. A computer data system converts the intensity change into an absorbance reading.

According to Beer-Lambert’s Law, absorbance is linearly proportional to concentration for

sample of low concentration. Thus, the data provided will then be linearly calibrated. Deionized
water is taken as the blank, i.e. zero Zn concentration, to give zero absorbance value. The graph
of the data will be drawn as a straight line from the origin.

The linear calibration method, however, does not take into account of interferences that might
be present. Among these is the matrix interference, a result from high concentrations of
dissolved salts other than the analyte or acid, or when the sample and standards are at radically
different temperatures. Matrix interferences either suppress or enhance the absorbance signal,
causing errors. To eliminate such interference might prove difficult; therefore the standard
addition method is used, whereby aliquots of a standard are added in different portions to an
amount of sample. Such a way allows an accurate determination to be executed without
eliminating the interference as it will also affect the standard similarly. The linear graph
obtained will be extrapolated to intercept at the added concentration axis to determine the
analyte concentration of the sample. However, this method should not be of the highest priority
as this indirect method can produce incorrect answers with other types of interferences, such as
chemical and ionization interferences, background absorption, and spectral interference.

1.2 Rationale / Purpose of Experiment

This experiment is conducted to determine the concentration of zinc (Zn) in a water sample in
terms of ppm via linear calibration and standard addition methods using atomic absorption
spectroscopy.

1.3 Background Information of Analyte

The analyte in this experiment is Zn. Having an atomic number of 30, it is one of the transition
metals alongside copper, nickel, iron, manganese, etc. In ionic form, it usually possesses a
charge of +2 and does produce any colour in solution form. In its oxide form, it displays
amphoteric properties, meaning it can display characteristics of both acid and base.

For zinc, standard atomic absorption conditions are as follow:

Table 1: Standard Atomic Absorption Conditions for Zinc

Light Source Chopper Characteristic

Flame
Wavelength (nm) Slit (nm) Concentration (ppm)
213.9 0.7 Air – acetylene 0.018
213.9 0.2 Nitrous oxide – acetylene 0.084
307.6 0.7 Air – acetylene 79.0

2.0 Materials and Methods

2.1 Apparatus
Perkin-Elmer AAnalyst 400 atomic absorption (AA) spectrometer, 50 mL volumetric flask, 100
mL volumetric flask,beaker, 1 mL pipette, 10 mL pipette, 20 mL pipette, dropper.

2.2 Chemicals and Reagents

Zinc sulphate (ZnSO4) stock solution (1000 ppm, parts per million), deionized water, water
sample.

2.3 Experimental Procedures

A. Preparation of Intermediate 10 ppm Zn Stock Solution
A stock solution of Zn (1000 ppm) is provided for the use of all laboratory groups.
Approximately 2 mL of the stock solution is obtained from the stock solution with a beaker.
A 10 ppm Zn stock solution is prepared in a 50 mL volumetric flask using the formula
below:

M1V1 = M2V2
M1 = original concentration M2 = desired concentration
V1 = volume required from original concentration V2 = desired volume

1000 ppm  V1 = 10 ppm  50 mL

V1 = 0.5 mL

An amount of 0.5 mL of the 1000 ppm stock solution is pipetted into the 50 mL volumetric
flask. The solution is then diluted to the mark.

B. Linear Calibration Method

A series of five standard Zn solutions with concentrations of 0.2 ppm, 0.4 ppm, 0.6 ppm, 0.8
ppm and 1.0 ppm are prepared using 50 mL volumetric flasks by means of serial dilution.
The amounts required from the 10 ppm stock solution to prepare the standards are listed as
below:

Table 2: Amount Required from 10 ppm Zn Stock Solution

to Prepare Standard Solution Series

Amount required from Amount required from

ppm ppm
10 ppm stock solution 10 ppm stock solution
0.2 ppm  50 mL 0.8 ppm  50 mL
=1 =4
0.2 10 ppm 0.8 10 ppm
mL mL
0.4 ppm  50 mL 1.0 ppm  50 mL
=2 =5
0.4 10 ppm 1.0 10 ppm
mL mL
0.6 ppm  50 mL
=3
0.6 10 ppm
mL

A sample of deionized water is used as a blank (0 ppm). With the assistance of the
laboratory assistant, the blank and the standards are sampled with the AA spectrometer and
the absorbance reading of the blank and each standard are recorded in the computer. A
relevant amount of water sample and the sampling procedure is repeated again for the water
sample. The results are then printed out. Based on the results, the absorbance reading is
plotted against concentration (ppm) using Microsoft Excel. The concentration of Zn in the
water sample is determined from the graph plotted.

C. Standard Addition Method

To a series of five 100 mL volumetric flasks, water sample (20 mL) is pipetted into each
flask. Certain volumes of the 10 ppm Zn stock solution are pipetted into the flasks as well
and diluted to the mark to give the following concentrations:
 Table 3: Amount Required from 10 ppm Zn Stock Solution
to Prepare Solution Series with Added Standard

Amount required from Amount required from

ppm ppm
10 ppm stock solution 10 ppm stock solution
0.00 ppm  100 mL 0.75 ppm  100 mL
= 0.0 = 7.5
x + 0.00 10 ppm x + 0.75 10 ppm
mL mL
0.25 ppm  100 mL 1.00 ppm  100 mL
= 2.5 = 10.0
x + 0.25 10 ppm x + 1.00 10 ppm
mL mL
0.50 ppm  100 mL
= 5.0
x + 0.50 10 ppm
mL

The value of unknown x is the concentration of Zn in the water sample. A sample of

deionized water is used as a blank. With the assistance of the laboratory assistant, the blank
and the solution series are sampled with the AA spectrometer and the absorbance reading of
the blank and each solution are recorded in the computer. The results are then printed out.
Based on the results, the absorbance reading is plotted against the added concentrations
(0.00, 0.25, 0.50, 0.75, 1.00 ppm) using Microsoft Excel. The value of x is determined from
the negative intercept of the added concentration axis.

3.0 Results and Discussion

3.1 Experimental Results
The following results are obtained when the two methods mentioned above are carried out:

Table 4: Experimental Results of Linear Calibration Method

Concentration Mean absorbance

(ppm) signal
0.0 0.0000
0.2 0.2484
0.4 0.3169
0.6 0.4315
0.8 0.5099
1.0 0.6704
Water sample 0.210

Table 5: Experimental Results of Standard Addition Method

Concentration Mean absorbance

(ppm) signal
x + 0.00 0.0293
x + 0.25 0.2891
x + 0.50 0.4119
x + 0.75 0.5560
x + 1.00 0.6059

3.2 Discussion

A. Linear Calibration Method

 The graph shown in Figure 1 (Appendix page A6) is a representation of the response of the
AA spectrometer towards the solutions used in the linear calibration method. Based on the
graph, the linear equation given by Microsoft Excel is y = 0.688x, with y representing the
absorbance reading and x, the Zn concentration value in ppm. Analysis of the water sample
gives an absorbance reading of 0.210.

Therefore, 0.688x = 0.210

0.210
Concentration of Zn in water sample, x = ppm = 0.305 ppm
0.688

To determine whether the linear line is an acceptable fit of the data, the magnitude of the
squared value of the correlation coefficient, R2, is usually considered.

 xi  x yi  y  
Correlation coefficient, R =

 xi  x  yi  y
2
 2
 
0.0  0.2  0.4  0.6  0.8  1.0
Mean concentration value, x = = 0.5
6
0.0  0.2484  0.3169  0.4315
Mean absorbance value, y =  0.5099  0.6704 = 0.3629
6

Sum of products of deviations, x i 

 x yi  y 
=
0.0  0.5   0.0  0.3629    0.2  0.5   0.2484
  0.4  0.5   0.3169  0.3629    0.6  0.5   0
  0.8  0.5   0.5099  0.3629    1.0  0.5   0
5

= 0.4251

x  2
Sum of squared values of deviation of concentration values, i x
=  0.0  0.5 2   0.2  0.5 2   0.4  0.5 2   0.6  0.5 2   0.8  0.5 2
 1.0  0.5
2

= 0.7

 y  2
Sum of squared values of deviation of absorbance values, i y
=  0.0  0.3629 2   0.2484  0.3629 2   0.3169  0.3629 2
  0.4315  0.3629   0.5099  0.3629   0.6704  0.3629
2 2 2

= 0.2678

0.4251
R =
0.7  0.2678
= 0.9819, R2 = (0.9819)2 = 0.9641

B. Standard Addition Method

The graph shown in Figure 2 (Appendix page A7) is a representation of the response of the
AA spectrometer towards the solutions used in the standard addition method. Based on the
graph, the linear equation given by Microsoft Excel is y = 0.568x + 0.0944, with y
representing the absorbance reading and x, the added Zn concentration value in ppm.

At the intercept of the added concentration axis, y = 0. Therefore, 0.568x + 0.0944 = 0

 0.0944
Concentration of Zn in water sample, x = ppm = 0.167 ppm
0.568
 The uncertainty of the negative intercept = – 0.167, or the standard deviation of the
unknown concentration (x), is determined as follow:
2
sy 1 y
Standard deviation of x = 
m n m  x x
2
i   2

Number of points, n = 5 Slope of graph, m = 0.568

0.00  0.25  0.50  0.75  1.00

Mean added concentration value, x =
5
= 0.50
Mean absorbance value, y
0.0293  0.2891  0.4119  0.5560  0.6059
=
5
= 0.37844

x  2
Sum of squared values of deviation of added concentration values, i x
=  0.00  0.50 2   0.25  0.50  2   0.50  0.50  2   0.75  0.50  2
 1.00  0.50 
2

= 0.625

 y  2
Sum of squared values of deviation of absorbance values, i y
=  0.0293  0.37844 2   0.2891  0.37844 2   0.4119  0.37844 2
  0.5560  0.37844    0.6059  0.37844
2 2

= 0.2143

 y  2
y 0.2143
Standard deviation of y, sy = i = = 0.2314
n 1 5 1
0.2314 1 0.37844 2
 Standard deviation of x =  = 0.3888
0.568 5 0.5682  0.625

The concentration values of Zn in the common water sample determined from the two methods
have a contrasting difference. In the linear calibration method, even without plotting the graph,
one would use the standard values to locate the concentration value range to determine where
the water sample absorbance reading (0.210) lies, that is between 0.0 ppm and 0.2 ppm. This
range coincides with where the concentration value determined from the standard addition
method is. However, the concentration value determined from the linear equation given by
Microsoft Excel (0.305 ppm) lies beyond that range. Furthermore, if the regression line in the
linear calibration method is an acceptable fit of the data, R2 should be greater than 0.999.
However, calculations show otherwise. In addition to that, the standard deviation of the sample
concentration value in the standard addition method is fairly larger than the value itself. This
suggests that the experiment was conducted with errors, as it is assumed that there should be no
problem with the AA spectrometer, which is designed to perform with high accuracy. Possible
errors might originate from inaccurate standard solutions that were prepared prior to the AA
analysis, or that the blank might contain interferences other than matrix interference, affecting
the absorbance readings. Nevertheless, since a conclusion needs to be reached in this
experiment, therefore the value obtained from the standard addition method will be taken as the
concentration of Zn in the water sample, i.e. 0.167 ppm Zn. This is because the standard
addition method is able to solve the matrix effect problem that might be occurring in the water
sample, as the water sample composition is not only unknown, but may be also very complex.

4.0 Results and Discussion

 The water sample in this experiment has a concentration of 0.167 ppm Zn. However, this is not
considered accurate as the final value obtain from two methods used in this experiment do not agree
with each other and that further numerical analysis suggests that the experiment could have errors.

5.0 Suggestions and Recommendations

The experiment should be run a few times to obtain an average of the final value. Interferences and
impurities in all solutions, including blanks, must be eliminated before conducting the experiment.

6.0 References

Mohd. Marsin Sanagi, Azli Sulaiman, Wan Aini Wan Ibrahim. 2004. Principles of Chemical Analysis.
Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia. Skudai. 32, 133, 136.

Fishman, Marvin J., Downs, Sanford C. 1966. Methods for Analysis of Selected Metals in Water by
Atomic Absorption. United States Department of the Interior. Washington D.C. 25 – 26.

The Perkin-Elmer Corporation. 1996. Analytical Methods for Atomic Absorption Spectroscopy.
Retrieved October 14, 2006, from http://perkin-elmer.com/AIMKT/lit.nsf/webfiles/AACKBK96.PDF/
.../AACKBK96.pdf

Wikipedia. 2006. Atomic Absorption Spectroscopy. Retrieved October 14, 2006, from
http://en.wikipedia.org/wiki/Atomic_absorption_spectroscopy

Wikipedia. 2006. Standard Addition. Retrieved October 14, 2006, from

http://en.wikipedia.org/wiki/Standard_addition