EXPERIMENTAL ANALYTICAL CHEMISTRY

(2021-2022)

LW2. Atomic Spectrophotometric

Methods
(AAS)

Last updated: October 2021

Aims of these Lab Work:

In this Lab Work, the following sub-objectives are proposed:

• To gain more in-depth knowledge about calibration methodology and to

introduce the use of Certified Reference Materials (CRM).
• To experiment with atomic absorption methods.
• To understand the effects of different sample treatments.
• To schedule the main tasks of the Lab Work beforehand.
W2. Elemental analysis: metals in estuarine
sediments after different digestion methods

1.1 Theoretical background

1.1.1 Sample treatment

The analysis of solid samples is always a challenging task. In real situations, as shown in
Figure 1.1 (right), the determination of analytes directly from the sample, without the
need of any treatment, is almost impossible to experience. It rather common the need
of a pretreatment of the sample by designing sample dissolution or digestion protocols,
which provide the extraction of the target analytes in the extension that is required
(totally or partially). Therefore, to manage digestion process is capital to understand the
analytes recoveries and how to deal with them.

Figure 1.1: General view of the sample treatment operation in elemental analysis.

In this Lab-Work, the direct analysis of a target analyte in a solid sample is not possible.
Therefore, in order to analyse copper in the proposed sediment sample, by an atomic
technique, two digestion procedures are considered:
• A METHOD of digestion consisting on boiling a mixture of HNO3:HCl.
• B METHOD of digestion consisting on dilute acid and without heating.
The obtained extracts will be analyzed by Flame atomic absorption spectroscopy (FAAS)
using external calibration method.

1.1.2 Instrumental analysis

Elemental spectrophotometric analysis is broadly carried out by two approaches: atomic
emission (ICP and AES) and atomic absorption (AAS). In this laboratory practice, we are
going to use AAS.

Atomic absorption Spectroscopy (AAS)

AAS is a spectroanalytical procedure for the quantitative determination of chemical
elements using the absorption of optical radiation by free atoms in the gaseous state. In
order to measure that light absorption it is necessary to transform the ions present in
the sample to elements by means of a flame that is used as atomizer. Then, atoms are
irradiated with a specific light source to excite the elements present in the flame (hollow
cathode lamp). Finally, we need the optical elements to measure the intensity of light
that crosses the sample path. However, there are more than one way of doing these
measurements; the most common instrument is the atomic absorption
spectrophotometer as shown in Figure 1.2.

Figure 1.2: Scheme of an atomic absorption spectrophotometer.

1.1.3 Calibration
One of the key operations in many instrumental methods is the calibration step. This
operation has to fulfill the requirements of precision and accuracy of the analysis and,
additionally, the estimation of the recoveries (the fraction of analytes that might be lost
during the analytical process). In this sense, the use of Certified Reference Materials will
be introduced in this Lab-Work.
1.2 Operating procedure
Each couple will cover one of the digestion strategies mentioned above. However,
broadly speaking the procedure is equivalent. In all the cases, each couple should carry
out the digestion of three samples: one unknown sample, one CRM sample and one
procedural blank. In the following points, both procedures will be explained.
The unknown sample is a lyophilized estuarine sediment taken in Kadagua river
(Zorroza) which has similar CRM properties. CRM sample has a total concentration of
the target element (Zn) of 1138 mg Kg-1. 1
Once all the solutions are fully prepared, we have to measure them by AAS. The linear
range is rather short and it is element dependent. In this case, linear range for copper is
up to 5 mg L-1.
Since the digestion capabilities of the two sample treatments are expected to be
different, the ranges of the expected concentrations would be different too, and
therefore, the calibration should accommodate the measurements of the samples
coming from the different digestions. In case of the A METHOD of digestion, the
expected recoveries will be close to 100 %, while in the case of the B METHOD of
digestion is around 60 %. In the same way, the total acidity that will be remained in the
samples after the extraction can be assumed to be around 50 % of the initial total
amount in both treatments due to volatilization and neutralization. Since the
instrumental measurements should be carried out at constant acidities, in this case it
will be fixed at 1% value in both calibration solution sets. Therefore, the required
dilutions to fit both concentration and acidity should be considered.

• A METHOD of digestion
o Weigh precisely 0.5 g of sample (estuarine sediment) in a beaker.
o Add 5 mL of distillate water and add 1 mL of HNO3 (69%) and 5 mL of HCl
(35%). Cover the beaker with the watch glass.

1
More details about these values and how they were obtained can be found in Amigo J., Arana A.,Etxebarria N., Fernández L.A.,
Emerging needs for sustained production of laboratory reference materials, Trends in Analytical Chemistry, 23, 1, 80-84, 2004.
 o Put the mixture in the sand bath and keep it boiling for 10 min. The liquid
volume that is lost during the process can be replaced adding Milli Q
water in order to avoid the total elimination of the liquid.
o The sample should be filtered by a disk filter of 0.45 μm and then the
liquid should be transferred to a volumetric flask of 50 mL. At this
moment, it should be considered if this solution is the one that can be
measured or if a further dilution would be needed to determine the
concentration of copper and to perform the analysis in correct acidity
values.
o Data of the replicates of the unknown sample, CRM sample and
procedural blank, prepared by the different couples using this digestion
method should be compiled to estimate the concentration of the sample,
the recovery and the limits of detection of the process.
o From a 1000 mg L-1 standard solution, a set of calibration solutions should
be prepared for AAS measurements, taking into account both linear
range of the instrument and suspected recovery. In addition, 0.025, 0.05,
and 0.1 mg/L concentration solutions should also be prepared to
estimate the instrumental limit of detection.

• B METHOD of digestion
o Weigh precisely 0.5 g of sample (estuarine sediment) in a beaker.
o Add 10 mL of distillate water and add 0.6 mL of HNO3 (69 %) and 1 mL of
hydrogen peroxide (H2O2). 2 Cover the beaker with the watch glass.
o Put the mixture in a magnetic stirring plate and keep it stirring for 10 min.
If ultrasonic baths were available, the samples can be sonicated for 10
min.
o The sample should be filtered by a disk filter of 0.45 μm and then liquid
should be transferred to a volumetric flask of 50 mL. At this moment, it
should be consider if this solution is the one that can be measured to

2
In this case, the acid is to assure a slightly acid media in the initial stage and the hydrogen peroxide is to strengthen the oxidation
capacity of the nitric acid.
 estimate the concentration of copper or if a further dilution would be
needed.
o Data of the replicates of the unknown sample, CRM sample and
procedural blank, prepared by the different couples using this digestion
method will be compiled to estimate the concentration of the sample,
the recovery and the limits of detection of the process.
o The calibration step is carried out as explained in the previous case. Keep
in mind that the recovery of the process could be lower in this case and,
therefore, the calibration and sample preparation should consider this
fact.

1.2.1 Work schedule

In this Lab-Work you are supposed to deal with a unique analysis technique but two
sample treatments (A and B methods of digestion). In order to organize the duties at the
lab we will consider the following scenario with two groups in each session:
o Group A will be in charge of the A METHOD of digestion and the calibration curve
fitted to the concentration values suspected to be obtained with this extraction.
o Group B will be in charge of the B METHOD of digestion and the calibration curve
fitted to the concentration values suspected to be obtained with this extraction.
The analytical measurements will be carried out during the second day. Nonetheless, all
the solutions (calibrates and samples) should be prepared by the end of the first day.
Since it is not going to be known in advance which digestion method will be commended
to each couple, it is compulsory to prepare both procedures before entering to the lab.
This means that calculations agreed in the last seminar should be made and
conveniently reported in the lab notebook for both digestion types.

1.3 Results and reports

Assuming for granted some of the skills that were introduced in the first Lab-Work we
would like to initiate some new ones and to intensify those already acquired. Among the
known skills, this Lab-Work is the workbench for the external calibration, and calculation
of recoveries and different type of limits of detection are new.
Finally, since the same sample will be extracted by two digestion methods, it offers the
chance to integrate the results and to compare them accordingly, that is, it has to be
decided which is the best one and why.
Based on the data collected, you should report about the following items:
• Calibration:
o Range of Calibration
o Detection limits
• Sample preparation:
o Recoveries: Use the Certified Reference Material, with known
concentration of the analyte, to estimate the recovery and correct the
concentration of the unknown sample if necessary.
• Analysis:
o Final results (average, standard deviation, trueness and precision taking
into account the whole procedure and replicates of the group).
o Comparison of the procedures. Each of you should include the results
obtained by both methods and compare them in terms of accuracy
(trueness + precision) and try to give a general conclusion.