Chemistry 223: Experiment 12

Determination of lead and zinc in water samples by F- AAS

References: 1. C, Chapter. 20 .
2. H7, Chapter 21,
3. Kellner, Chap 8
4. SWH, Chaps. 24
5. Standard method for examination of water and waste water. EPA.
6. http://www.cee.vt.edu/ewr/environmental/teach/smprimer/aa/aa.html

I. Purpose of the experiment

Atomic line absorption spectra are used for analyzing various metals. Atomic absorption
spectrophotometry ("AAS") offers sensitivity, selectivity and simplicity and was developed
by CSIRO Melbourne scientists around 1955. "AAS" is an important tool in analytical
laboratories. Using this technique, students will be determine Pb and Zn in water samples by
flame atomic absorbance spectroscopy. Students will also be familiar with QA/QC for the
analytical procedure through the example of analyzing Pb and Zn in soluble species and total
concentrations in water samples.

II. Introduction

Flame atomic absorption spectroscopy (F-AAS) is a very common technique for detecting
metals and metalloids in solid and aqueous samples. It is very reliable and simple to use. The
technique is based on the fact that ground state metals absorb light at specific wavelengths.
Metal ions in a solution are converted to atomic state by means of a flame. Light of the
appropriate wavelength is supplied and the amount of light absorbed can be measured against
a standard curve.

AAS quantitatively measures the concentrations of elements present in a liquid sample. It

utilizes the principle that elements in the gas phase absorb light at very specific wavelengths
which gives the technique excellent specificity and detection limits.

The sample may be an aqueous or organic solution, indeed it may even be solid provided it
can be dissolved successfully. The liquid is drawn in to a flame where it is ionised in the gas
phase.

Light of a specific wavelength appropriate to the element being analyzed is shone through the
flame, the absorption is proportional to the concentration of the element. Quantification is
achieved by preparing standards of the element.

The technique of (FAAS) requires a liquid sample to be aspirated, aerosolized, and mixed
with combustible gases, such as acetylene and air or acetylene and nitrous oxide. The mixture
is ignited in a flame whose temperature ranges from 2100 to 2800 oC. During combustion,
atoms of the element of interest in the sample are reduced to free, unexcited ground state
atoms, which absorb light at characteristic wavelengths. The characteristic wavelengths are
element specific and accurate to 0.01-0.1nm. To provide element specific wavelengths, a light
beam from a lamp whose cathode is made of the element being determined is passed through
the flame. A device such as a photonmultiplier can detect the amount of reduction of the light
intensity due to absorption by the analyte, and this can be directly related to the amount of the
element in the sample.
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Flame atomic absorption hardware is divided into six fundamental groups that have two major
functions: generating atomic signals and signal processing. Signal processing is a growing
additional feature to be integrated or externally fitted to the instrument. The instrument parts
are shown in figure 1 and 2.

Fig. 1: Atomic Spectrometer novAA-400 AnalytikJena

Figure 2. Schematic of basic instrumental parts of atomic absorption spectrometer

A cathode lamp (1), shown in figure 3, is a stable light source, which is necessary to emit the
sharp characteristic spectrum of the element to be determined. A different cathode lamp is
needed for each element, although there are some lamps that can be used to determine three or
four different elements if the cathode contains all of them. Each time a lamp is changed,
proper alignment is needed in order to get as much light as possible through the flame, where
the analyte is being atomized, and into the monochromator.

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 Fig.3: Hollow cathode lamp

The atom cell (2), shown in figure 4, is the part with two major functions: nebulization of
sample solution into a fine aerosol solution, and dissociation of the analyte elements into free
gaseous ground state form. Not all the analyte goes through the flame, part of it is disposed as
shown in the figure.

Fig. 4: Atom cell

As the sample passes through the flame, the beam of light passes through it into the
monochromator (3). The monochromator isolates the specific spectrum line emitted by the

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light source through spectral dispersion, and focuses it upon a photomultiplier detector (4),
whose function is to convert the light signal into an electrical signal.

The processing of electrical signal is fulfilled by a signal amplifier (5). The signal could be
displayed for readout (6), or further fed into a data station (7) for printout by the requested
format.

F-AAS is also based on transmitting light through the sample and measuring the fraction of
light absorbed. Mathematically, the measurement can be expressed as:

T(λ) = P(λ) / P0(λ) (1)

In Eq. 1, T is termed the transmittance, P denotes the light intensity transmitted through the

sample, and P0 is the light intensity incident on the sample. The P0 value is measured by use
of a blank solution that contains the sample matrix but contains no analyte. The values T, P,
and P0 are all a function of λ, the wavelength of the light. The measured transmittance can be

related to the analyte concentration through the Beer-Lambert law:

A(λ) = -log10(T(λ)) = εbc (2)

In Eq. 2, A is termed the absorbance, c is the analyte concentration, and ε (absorptivity) and b
(optical path length) are constants that define the linear relationship between A and c.

Absorptivity is an intrinsic property of the analyte, and the path length is a characteristic of
the instrumental measurement. The value of εb is determined by a calibration procedure that
employs a set of known standards that bracket the desired concentration range.

III. Reagents, apparatus and materials.

Apparatus: - All sample plastic containers must be prewashed with detergents, acids, and
water. Both plastic and glass containers are suitable.

- Atomic Spectrometer novAA-400 AnalytikJena.

- Watch glasses or equivalent.

Provided Reagents: - Reagent Water shall be interference free. All references to water in the
method refer to reagent water unless otherwise specified.

- Nitric acid (concentrated), HNO3. Acid should be analyzed to determine level of impurities.
If method blank is < MDL, then acid can be used.

- Hydrochloric acid (concentrated), HCl. Acid should be analyzed to determine level of

impurities. If method blank is < MDL, then acid can be used.

- Water Sampling:

+Total recoverable metals - All samples must be acidified at the time of collection with
HNO3 (5 mL/L).

+ Dissolved metals - All samples must be filtered through a 0.45-m filter and then
acidified at the time of collection with HNO3 (5 mL/L).

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To be prepared:

Concentrations of the metal ion standard solutions need to cover a range of 0 to 25 mg/L (ie. 0
to 25 ppm). Some metal ion stock solutions (250 mg/L or 1000 mg/L) will already have been
prepared for you and simply require dilution with 0.002M to the appropriate final
concentrations.

Note: High levels of silicon, copper, or phosphate may interfere to the determination of Zn.
Addition of strontium (1,500 mg/L) removes the copper and phosphate interference. Zinc is a
universal contaminant, and great care should be taken to avoid contamination.

Sample preparation:

* Dissolved metals - The sample is filtered through a 0.45-m filter at the time of collection
and the liquid phase is then acidified at the time of collection with nitric acid. Samples for
dissolved metals do not need to be digested as long as the acid concentrations have been
adjusted to the same concentration as in the standards.

* Total recoverable metals: The entire sample is acidified at the time of collection with nitric
acid. At the time of analysis the sample is heated with acid and substantially reduced in
volume. The digestate is filtered and diluted to volume, and is then ready for analysis.

Interferences: The analyst should be cautioned that this digestion procedure may not be
sufficiently vigorous to destroy some metal complexes.

Precipitation will cause a lowering of the silver concentration and therefore an inaccurate
analysis.
Together with unknown sample, add the amount of several ppm of mixture containing heavy
metals to the samples and digest follow the above procedure to evaluate the recovery rate.

IV. Experimental Procedure

1. Sample preparation for F- AAS analysis

- Transfer a 100-mL aliquot of well-mixed sample to a beaker.

- For metals that are to be analyzed, add 2 mL of concentrated HNO3 and 5 mL of

concentrated HCl. The sample is covered with a ribbed watch glass or other suitable covers
and heated on a steam bath, hot plate or other heating source at 90 to 950C until the volume
has been reduced to 15-20 mL.

CAUTION: Do not boil. Antimony is easily lost by volatilization from hydrochloric acid
media.

- Remove the beaker and allow to cool. Wash down the beaker walls and watch glass with
water and, when necessary, filter or centrifuge the sample to remove silicates and other
insoluble material that could clog the nebulizer.

Filtration should be done only if there is concern that insoluble materials may clog the
nebulizer; this additional step is liable to cause sample contamination unless the filter and
filtering apparatus are thoroughly cleaned and pre-rinsed with dilute HNO3.

- Adjust the final volume to 100 mL with reagent water.

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2. Quality control

- For each analytical batch of samples processed, blanks should be carried throughout the
entire sample preparation and analytical process. These blanks will be useful in determining if
samples are being contaminated.

- Replicate samples should be processed on a routine basis. A replicate sample is a sample

brought through the whole sample preparation and analytical process. Replicate samples will
be used to determine precision. The sample load will dictate the frequency, but 5% is
recommended.

- Spiked samples or standard reference materials should be employed to determine accuracy

by calculating recovery rate. A spiked sample should be included with each batch.

3. Determination of Pb and Zn (direct aspiration)

3.1. Instrument parameters (general):

Parameters Determination of Pb Determination of Zn

Lead hollow cathode lamp Zinc hollow cathode lamp
Fuel Acetylene.
Wavelength 283.3 nm 213.9 nm.
Fuel Acetylene Acetylene
Oxidant Air Air
Type of flame Oxidizing (fuel lean) Oxidizing (fuel lean)
Background correction Required Required

3.2. Calibration standards

Calibration standards are prepared by single or multiple dilutions of the stock metal solutions.
Prepare a reagent blank and at least 3 calibration standards in graduated amount in the
appropriate range of the linear part of the curve. The calibration standards must contain the
same acid concentration as in the samples following processing. For precipitation samples,
that would be 1% (v/v) HNO3 and for suspended particulate matter10% (v/v) HNO3. The
calibration standard should be transferred to polyethylene bottles.

- For determination of Pb: Optimum concentration range: 1-20 mg/L ;

Sensitivity: 0.5 mg/L.
Detection limit: 0.1 mg/L.

- For determination of Zn: Optimum concentration range: 0.05-1 mg/L

Sensitivity: 0.02 mg/L.

Detection limit: try to calculate from measuring the procedural
blank at least 8 times.

3.3. Blank solutions

Three different blank solutions are required; calibration blank, procedural blank and rinse
blank.

 Calibration blank is used for establishing the calibration curve. The calibration blank
consists of the same concentration(s) of the acid(s) used to prepare the final dilution of
the calibration standards. In addition, an appropriate concentration of internal standard
is added.

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  Procedural blank (or reagent blank) is used to monitor for possible contaminations
resulting from the sample preparation procedure. The procedural blank must be carried
through the same procedure and contain the same volume of reagents as the sample
solution. In addition an appropriate concentration of internal standard is added.

 Rinse blank is consist of 1-2 % (v/v) HNO3 and is used to flush the sample
introduction system between standards and samples.

3.4. Instrumental procedure

The operating procedure will vary between instrument brands, so the instrument manual
should be followed carefully. The position of observation and the fuel:oxidant ratio must be
optimised. Some general guidelines are outlined below:

 Light the hollow cathode lamp or electrode discharge lamp and D2-lamp if such
background correction is used. Set the lamp current to the value specified by the
manufacturer.

 Position the monocromator at wavelength 283.3 for Pb and 213.9 for Zn and choose
slit with 0.7 and slit height “high”.

 Carefully balance the intensity of the hollow cathode lamp and the D2-lamp if such
background correction is used.

 Align the burner head to assure that the centre of the light beam passes over the burner
slot.

 Light the flame and regulate the flow of fuel and oxidant to produce an oxidising
flame (lean blue).

 Aspirate calibration blank and establish a zero point.

 Aspirate standard solutions and construct a calibration curve.

 Aspirate samples.

Aspirate distilled water after each standard or sample.

3.5. Instrument performance

The “characteristic concentration” (sometimes called sensitivity) is defined as the

concentration of an element (mg l-1) that will absorb 1 % of the incoming radiation. This
equals a signal of 0.0044 absorbance units (AU). The “characteristic concentration” is
instrument dependent and is calculated as follows:

C = (S * 0.0044 AU) / measured absorbance

C: Characteristic concentration (mg l-1)
S: Concentration of measured standard (mg l-1)

Knowing the “characteristic concentration” allows the analyst to check if the instrument is
correctly optimised and performing up to specifications.

3.6. Sequence of analysis

 Aspirate calibration blank and establish a blank level

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  Aspirate calibration blank and standard solutions and construct a calibration curve.
Use at least 3 standard solutions in addition to the calibration blank to cover the linear
range. Every point at the calibration curve should, if possible, be based on replicate
analysis. Distilled water should be aspirated after each standard and sample.

 A quality control standard should be analysed to verify the calibration.

 A calibration blank should be analysed to check for memory effects.

 Aspirate unknown samples.

 Aspirate a quality control standard for every 10th sample to check for drift.

 Samples that are found to have concentration higher than the highest standard should
be diluted and reanalysed.

FOR YOUR REPORT

1. Report the experimental results. Summarize the calculations that produced your calibration
design.

2. Calculate the exact concentrations of your calibration standards. Be sure to use the
appropriate number of significant figures. data. Report values to the correct number of
significant figures.

4. Make calibration models for each set of data. Use least-squares procedures to determine the
slope and intercept of each calibration model. Report the value of r2 and the standard error of
estimate.

4. Include separate plots of absorbance (with error bars) vs. concentration (ppm) for each
calibration model.

5. Use your calibration model together with your dilution scheme to compute the weight
percent of iron in your original solid sample. Report as weight percent Fe2O3. Predict each
replicate of the unknown separately, then compute a mean, standard deviation, and 95%
confidence interval for the predicted amount. Repeat this calculation for both calibration
models.

6. The limit of detection associated with each calibration can be estimated by use of the
following procedure. The intercept of the calibration model is an estimate of the absorbance
when no metal is present in the sample (i.e., the blank signal). The standard error of estimate
is an estimate of the measurement noise (i.e., in the absence of systematic error, the variation
about the least-squares line should be due solely to instrumental noise). A widely used
definition of limit of detection is the concentration corresponding to an absorbance value
equal to the blank signal plus three times the noise level. Thus, ALOD = b + 3s (3)

where ALOD is the absorbance corresponding to the limit of detection, b is the intercept of the
calibration model, and s is the computed standard error of estimate. The limit of detection,
CLOD, can then be estimated as CLOD = (ALOD - b)/m (4)

where ALOD are as defined above and m is the computed least-squares slope of the calibration
model. Equations 3 and 4 can be combined to yield

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CLOD = 3s/m (5)

Use your calibration data to estimate the limit of detection for each method.

7. In your discussion section, compare the atomic and molecular spectroscopic methods

8. Tabulate the mean, standard deviation, and 95% confidence interval for your absorbance-
Report the data of the calibrations with linear regression.

- Calculate the detection limit of Zn determination.

- Calculate the Pb, Zn content in samples by using the calibration regression. Also report
uncertainty of the sample’s data from calibration curve.

ITEMS TO THINK ABOUT:

- Explain how metal atoms are produced and excited in HCL?

-Flame AA has poor sampling efficiency and sensitivity. Explain why?

- Spectral overlap is insignificant in AAS method. Explain!

- Discuss the major factors involved in the detection to use flame of electrothermal
atomization in AAS.

- F-AAS is known as a technique with few problems related to interference effects. Do you
know what are they?

- Why do hydride techniques for As and Se and the cold vapor techniques for Hg yield better
AA detection limit than conventional flame AAS with solution nebulization?