An analytical technique is a way that is applied to determine the concentration of a

chemical compound or chemical element. There a numerous type of technique applied for
analysis, from gravimetric analysis which involves simple weighing to titrimetric which
involves titration and also to very advanced techniques which involves the application of
highly specialized instrumentation.

Controlling and assessment of water pollution has become an extremely critical area
of study due to its direct consequences on the aquatic life and human beings. The
contamination of water surface due to the presence of heavy metals is a critical ecological
problem because some of them such as Hg and Pd are poisonous even at low concentration,
non-biodegradable and they might be bio-accumulated through food chain. Some metals like
Fe, Cu and Zn are crucial micronutrients, however, they can be harmful to the physiology of
living organisms at high concentration. Thus, there are a wide variety of instrumental
methods applied in the determination of heavy metals in natural water, mostly with
preconcentration due to the insufficient sensitivity of direct analysis.

It is difficult to determine the concentration of trace heavy elements in environmental

samples and the results obtained always differ according to the choice of analytical technique
(Soylak et al., 2002; Zhang et al., 2002; Ko¨klu¨ et al., 2000; Rauf et al., 2002; Tsikrizis et al.,
2002; Orescanin et al., 2002; Zucchi et al., 2002). Atomic absorption spectroscopy (AAS) is a
crucial instrumental technique applied for determining trace heavy metal ion due to its low
cost and short analysis time (Tu¨tem et al., 2001; Dogan et al., 2002).

The existence of metal in water can be in several different forms (Kim, 1995). The
extent of complexation and the oxidation state are the features which determine the form of
metal ion. Those metals which pass through a 0.45µm membrane filter are known as
dissolved (filterable) metals; Those metals which are retained by a 0.45µm membrane filter
are known as suspended (non-filterable) metals; Those metals with its concentration in
solution after vital digestion of an unfiltered sample are known as total metals; Those metals
with its concentration in solution after treatment of an unfiltered sample by hot dilute mineral
acid are known as acid-extractable metals.

Analytical method for the determination of heavy metals is composed of 3 main

stages. At sampling and sample preparation stage, several samples are collected from
different points of the targeted river at the study area. Sampling procedure is completed with
collection of water samples from different depths of the river. It is important to collect a
statistically sufficient number of samples from the representative area in order to make sure
that the real error of the mean result of samples is lower than the error of the analysis method
(Taraškevičius, 1998). Next, treatment of the collected samples such as acidification, acid
digestion, filtration and preconcentration are carried out before entering the analysis stage.
Lastly, some analytical techniques are employed to further analyse the samples.

Most metals exist in water in the dissolved state, as well as those in the other three
forms. Atomic absorption spectrophotometry can be easily carried out to determine the heavy
metals in river water after some suitable treatment to form them into dissolved state. The
outline of this technique is basically the study of the absorption of radiant energy such as UV
and visible regions by neutral atoms in the gaseous state. In an atomic absorption analysis,
the targeted element to be determined must undergo reduction to the elemental state,
vaporization and imposition in the beam of radiation from the light source. This process
involves the withdrawal of a solution of the sample in the form of fine mist into a suitable
flame. The flame has the similar function as the cell and solution in conventional absorption
spectrophotometry. Measurement of the absorption is made at a specified wavelength which
is characteristic of each individual element. The absorbance measured is proportional to the
concentration. Analysis is done by doing comparison of these absorbance with reference
samples of known composition. Flame atomic absorption spectrophotometry (FAAS) is a
simple, fast, precise and highly specific technique which is free from interferences. FAAS
consists the following components:

(a) A stable light source for emission of sharp resonance line of the element to be
determined.
(b) A flame system applied to convert metal ion in solution to atomic state through
atomization.
(c) A monochromator used for separation of resonance line and focusing of selected
wavelength upon a photomultiplier.
(d) A photomultiplier used for detection of the intensity of light energy striking on it.
Simplification and readout can be done with the aid of facilities.
 Figure 1: Structure of atomic absorption spectroscopy.

There are three types of atomic absorption spectroscopy applicable for the
determination of heavy metals.

Direct flame absorption is one of the atomic absorption techniques applied for
determining heavy metals in river. This technique is used to determine most metals in routine
water samples (arsenic, selenium and mercury are exceptions). Usually, an air-acetylene
flame is used, except for the case which involves barium since it can form heat-resistant
compounds with oxygen in the flame. The higher temperature nitrous oxide-acetylene flame
is favoured in determining these metals. Sometimes, the presence of elements may be at
concentrations below the normal detection limit of the selected technique. Thus, several
methods are employed to increases the concentration of these elements such as evaporation
and solvent extraction (chelation). For evaporation, the samples are evaporated to a specified
smaller volume before being analysed. However, this method is time consuming and may be
problematic caused my matrix or background interferences. For chelation, concentration of
the element to be determined can be increases and the inherent analytical sensitivity may be
increases due to the presence of the sample in the organic solvent instead of in water.
Designation of chelation procedures are generally made for simultaneous extraction of
several metals so the determination of all the metals may be done in the same organic solvent
solution. Ammonium pyrrolidine dithiocarbamate (APDC) is commonly used as chelating
agent for most of the metals.

Besides, hydride generation is also used to determine arsenic, selenium and other
metallic elements which form volatile covalent hydrides under suitable chemical conditions.
Hydride generation system can be utilized. Treatment of an acid solution of the sample to be
analysed to done with sodium borohydride and the volatile hydride releases is delivered into a
quartz tube heated in an air-acetylene flame. By using this technique, detection limit of below
5µg/l is achievable.
 Lastly, in cold vapour generation, the detection limit for mercury is about 200µg/L
with conventional flame absorption methods. In water analysis, the detection limit is below
1µg/L for routine mercury determination. Mercury is special since its elemental form has
appreciable vapour pressure at room temperature, thus, determination of its concentration in
water can be done without conventional burner system. Numerous highly sensitive flameless
systems have been developed to determine mercury according to the system first established
by Hatch and Ott. This technique involves chemical reduction of mercury in the sample to its
elemental state and aeration of mercury from solution in a closed system. The mercury
vapour goes through a cell placed in the light path of the atomic absorption
spectrophotometer. Measurement of the absorbance determines the mercury concentration.

Figure 2: Instrumental conditions for determination of heavy metals by AAS.

Ion chromatography is one of the capable analytical techniques for the analysis of
natural water. It is the best method of determining anions in aqueous solutions. It is also a
suitable method to determine heavy metals. Dual-column techniques which is highly sensitive
have been established for determining metals in different types of water. It involves the
separation according to both the cation- and the anion-exchange mechanism. In the first case,
elution has been used to separate heavy metals such as Cd, Cu, Mn, Ni, Pb, Sr and Zn with
the aid of ethylenediammonium chloride or ethylendiammonium tartrate solutions and
conductometric detector is used for determination. The detection limits an amount of 0.3-6.0
µg/ml. Relative standard deviation equal to 0.015-0.050 is used to characterize the precision
of determining 20 µg/ml of these metals. For determinations by anion-exchange mechanism,
the metals: As, Cr, Mo, Se, Te, V and W are first converted into oxianions, followed by
separation conducted on columns with “Dionex” low-capacity anion-exchange resin; the
elution is carried out with sodium carbonate solution. For an example, chromatogram of a
mixture of SeO42-, MoO42-, AsO42-, WO42- and CrO42- is shown in Figure 3. The linear range of
determined content is 0.05-500µg/mL. The mean relative standard deviation within this
interval is not more than 0.05. Even without the aid of a concentrating column, the minimal
determined concentrations of the investigated metals do not exceed their maximum allowable
concentrations in water. The use of a concentrating column can lower the detection limit
value by another two orders of magnitude. The method has been applied for determining
tungsten, molybdenum and chromium in waste water (Figure 4). The results obtained are
compatible with the data of atomic emission (w), atomic emission with inductively couples
plasma (Mo) and spectrophotometric (Cr) methods.

When determining selenium in the form of SeO2-, it is possible to lower the detection
limit 10 times with the use of a concentrating column. The minimal determined selenium
concentration is 10µg/L without enrichment, thus, its can be reduced to 0.01 µg/L with the
use of concentrating column, which makes it applicable to determine selenium in natural
water. For an example, selenium was determined in the Moskva River water.

Figure 3: Chromatogram of a mixture of selenium(IV), molybdenum (VI), arsenic (V),

tungsten (VI) and chromium (VI) oxoanions. Separating brine column 3 x 500mm;
suppressing column eluent supply rate 230ml/h.
Figure 4: Determination of chromium, molybdenum and tungsten in waste water (number of
determination = 5; γ = 0.95.

Furthermore, analytical technique using a combination of solvent extraction and thin

layer chromatography is used in the determination of heavy metals in river water. TLC which
combined with preliminary solvent extraction concentration is applicable to analyse surface
water, which was established by determining heavy metals in river water samples. The
outline of analysis consists: solvent extraction preconcentration of metals with the aid of
sodium diethyldithiocarbamate, separation of chelates on commercially available plates with
a thin silica gel layer, and visual or densitometric determination of the separated complexes
directly in the chromatographic zones. When preparing the outline, some theoretical aspects
of the chromatographic of chelates was studied. An investigation regarding the influence of
the structure of chelates on their behaviour in the conditions of liquid adsorption
chromatography was carried out. Suggestions have been provided on the choice of a
convenient chelate-forming reagent (class of chelates) depending on the solvent extraction
recovery of a group of metals and their selective chromatographic separation. The absolute
detection limits of metals in the form of diethyldithiocarbamates is within the range 1-10 ng,
allowing their quantitative determination down to 0.05 µg/mL. The relative standard
deviation for determining Co, Cu, Cr, Fe, Ni, Pb and Zn at a level of their maximum
allowable concentration within 0.1.

Figure 5: Radiochromatogram of the extract of lead, cadmium and mercury 3H-

dipropyldithiophosphates (Silufol/toluene)
 It is possible to achieve their radiometric determination in a thin layer with much
lower absolute detection limits by converting the determined metals into their tritium-labelled
complexes. A radio-chromatogram of an extract obtained after the extraction of cadmium,
lead and mercury with 3H-dipropyldithiophosphate solution in CCl4 was shown in Figure 5.
With the application of a liquid scintillation counter, the detection limits for these metals
range from 9 x 10-7 – 2 x 10-6 M. A linear calibration plots on the average from 1 x 10-7 to 2 x
10-6 M. Thus, the methods demonstrated can solve the problem in concentrating and
determining heavy metals in natural water.

According to Journal of Environmental Analytical Chemistry, determination of Mn,

Pb, Cr and Cd was carried out in Kulufo River water, Arba Minch and Gamo Gofa. FAAS
technique was employed for the determination of the concentrations of these heavy metals.
From this work, it was showed that the highest concentration of these heavy metals was
found from the work areas. From the results obtained, Kulufo river water was contaminated
by these metals concentration, thus, treatment is required to improve this condition due to the
increasing of transport flows and operation of factory.
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