Atomic

Spectroscopy
Absorption (AA)
Lab
By: Wesley Clair
Contents

Executive summary A brief overview of the series of experiments (slide 3)

Background information The Goal of the Analysis as well as some AA Instrumentation and Techniques
(slide 6)

Methods and process Brief run down of the basic steps taken for each experiment (slide 10)

Results The raw and processed data retrieved during the experiment (slide 13)

Discussion Written breakdown of what was retrieved from each experiment (slide 17)

Conclusion Overall findings and if it is trustworthy (slide 18)

Acknowledgements A thanks to all thoughts that helped me through the experiments. (slide 19)
 Executive summary
Objectives

The project will determine iron concentrations in soil, spinach, and an iron
vitamin tablet by atomic absorption spectrometry. This will be intellectually
important for understanding the iron content in environmental samples, plant-
based materials, and dietary supplements. The result of this analysis will also
verify the label claims accuracy for the iron vitamin tablet. Since this is of
great import both in environmental monitoring and nutritional analyses, and
into consumer product labeling, an iron content determination represents a
need in both scientific and regulatory considerations.
 Methods

The sample preparation involved weighing of soil and spinach and a crushed iron vitamin
tablet. Addition of concentrated nitric acid was added to the samples. These samples
were later on boiled, filtered, and diluted. Standards of iron were also prepared with
known concentrations to plot an appropriate calibration curve. Using atomic absorption
spectrophotometry, these samples were analyzed, and the iron concentration was
determined using the obtained calibration curve. In the case of the vitamin tablet, the
experimental results were compared with the iron content listed on the label to gauge the
accuracy of the product. The standard was prepared properly so that it would be suitable
for analysis and any subsequent comparison of data.
 Results & Conclusion

Calculating the iron concentration in mg / g or mg / tablet using the

following absorbance values: 0.92, 0.79, 0.89 for soil and 0.052, 0.052,
0.061 for spinach and 0.086, 0.087, 0.088 for vitamin tablet. The
experimental results were the iron content of each sample; hence, in the
case of the vitamin tablet, experimental values were compared with the
claimed iron on the label to confirm if it was correct or not. The present
study demonstrates the capability of atomic absorption spectrometry in the
determination of iron through different sample types with reliable results
from environmental, nutritional, and consumer product analyses.
 Background information
Goal and Importance of Analysis Accurately measuring iron is important for
detecting deficiencies or contamination in
The goal is to estimate the iron environmental and food-chain samples. Iron
concentration in various matrices such as levels in dietary supplements must be declared
soil, spinach, and iron vitamin tablets accurately to guarantee safety for the health of
using atomic absorption consumers. While the iron is needed in day to
spectrophotometry. This would be highly day consumption, in larger doses the body has
relevant in environmental monitoring difficulties removing it the most effective way
studies, nutritional evaluations, and to get rid of excess iron being blood loss.
verification of the accuracy of product
labeling, all of which are vital to ensure
the safety of the public and compliance
with regulatory requirements.
 AA Instrumentation and Technique

Instrumentation Used:
The Atomic Absorption Spectrometry will be used for detection and
quantification of iron in the samples. AA is a very sensitive technique that
measures the concentration of metal ions by detecting absorbance of light
emitted by atoms in the vapor phase.

Technique:
Atomization of the samples in a flame is done, with the amount of light
absorbed at any particular wavelength being proportional to the iron
concentration. These calibration curves are based on known iron
standards which enable the quantification of unknown sample
concentrations.
 How AA Works
In AA, a sample is vaporized in a flame and converted into free atoms. Light from a light
source, which is a hollow cathode lamp of the analyte-usually iron, is absorbed by free
atoms in the vapor. The amount of light absorbed is directly proportional to the
concentration of the analyte in the sample according to the Beer-Lambert Law.

Beer-Lambert Law:
A = εbc
Where:
A = absorbance
ε = molar absorptivity
b = path length
c = concentration
 Analytical Limitations of the
Sensitivity: Instrumentation
AA employed in this experiment-the 900F AA Spectrophotometer-are highly sensitive to the
detection of iron at a lower limit of detection within a ppm range and thus applicable to trace
metal analysis.
Interferences:
Chemical Interference: Other matrix components such as phosphates or sulfates could cause
chemical interference that reduces accuracy.
Spectral Interferences: Overlapping peaks of absorbance due to other metals or molecules can
also be problematic, but interference from these sources is negated by the presence of the
monochromator (the iron lamp in this case).
Advantages:
High sensitivity for the detection of low concentrations of metals like iron; relatively fast and easy
sample preparation. Possible drawbacks AAS can measure only one element at a time, hence
limiting its throughput for multi-element analysis. Possible matrix effects may require extensive
calibration and standard preparation.
Methods and
process
Weighing the Samples
Soil B: 0.50 g in a beaker
Spinach: 2.01 g (2 leaves) crushed
Iron Tablet: 0.51g (1 tablet) crushed
Addition of Deionized Water

Add 30 mL of deionized water to each beaker.

Add Nitric Acid (HNO₃)

IN FUME HOOD: Added 5 mL concentrated nitric acid to each

sample.
Then boiled for 15 Minutes (this was to break down the
samples to better extract a higher % of iron)

Covered with watch glass, boiled on hot plate, monitored to

prevent boiling drying.
Filtration and
Dilution
Cooling and Filtering
Allowed samples to cool, once able to be held. Filtered using
Whatman #1 paper into 50 mL volumetric flasks.

Dilution
Transferred the filtrates and diluted with deionized water Figure 1: from left to right is the filtrate
of the control, the iron tablet, the
to reach the 50 mL mark. spinach, and the soil

Preparation of Standards
Prepared standards from 1000 ppm Fe solution for a calibration curve (Standards A, B,
C, D, E).

Sample Dilution
Soil: filtrate was analyzed directly
Spinach: filtrate was analyzed directly
Vitamin: Pipetted 0.1 mL into 100 mL flask and filled to volume with 1% nitric acid
Control: blank of water that had been boiled and filtered
Instrumental Analysis
(AA)
Instrument Setup
AA Setup: Configure the 900F AA Spectrophotometer.
Standard and Sample Measurement

Run the prepared standards and samples through the instrument.

Absorbance Measurement

Measure absorbance values for iron (Fe) at 248.33 nm (the wavelength of light
that comes from exciting iron)
Data Collection

Collect data for calibration curve and calculate iron concentrations in each
sample.
 Results
Iron concentration of each sample (soil B)
Concentration of iron in solution: 10.12 mg/L
Volume of the solution (filtrate) after dilution: 50 mL (0.050 L)
Mass of soil used: 0.5 g

Total iron=(Concentration of iron)×(Volume of filtrate)

Total iron=10.12mg/L×0.050L=0.506mg of iron

Iron concentration in soil= Total iron/Mass of soil

Iron concentration in soil= 0.506mg/0.5g = 1.012mg iron/g
soil
Figure 2: the data received with an error
message, a step in further dilution was missed = 1012 mg of iron / kg of soil
which caused the reading to come out high. The
mean was used for calculations
According to U. S. Environmental Protection Agency Office of
Solid Waste and Emergency Response this is low as “typical
iron concentrations in soils range from 0.2% to 55% (20,000
to 550,000 mg/kg)”(3) this low iron number could be from
errors in dilution or a low iron sample.
 Results 2
Concentration of iron in solution: 1.019 mg/L
Volume of the filtrate after dilution: 50 mL (0.050 L)
Mass of spinach used: 2.01 g

Total iron=1.019mg/L×0.050L=0.05095mg of iron

Iron % (w/w)=( total iron (mg)/mass of spinach (mg))x100 =
(0.05095/2010)x100 = 0.00253%

Which according to the U.S. Department of Agriculture (4) estimates an

Extraction efficiency of 97.50% at 2.71 mg per 100g of spinach

Figure 3: the data for spinach was received and

the mean was used for calculations
 Results 3
Iron concentration in the solution after dilution: 0.953 mg/L
Volume of the final filtrate: 100 mL (due to the extra dilution step)
Mass of the vitamin tablet: 0.51 g
Extra dilution step: 0.1 mL of the original extract was diluted from
50 mL
Total iron (mg)=0.953mg/Lx0.1L=0.0953mg (100ml)
c1v1=c2v2 for original concentration

Figure 3: the data for the iron tablet was

C1 =(0.953x.1)/.001 = 95.3mg/L
received as shown above and the mean was 95.3mg/L x 0.050L = 4.765mg
used for calculations

Therefore the mg of iron / tablet is 4.765mg which is a long ways off of

the labeled 60mg. This could be because i had too much precipitate left in
the funnel drastically lower the extraction efficiency or the machine had
errors with its calibration and zeroing.
Results the iron tablet

To check the label claim for the iron content in the vitamin tablet, we use the formula
for percentage label claim:
% label claim = ( mg iron per tablet from experimental results / mg iron per tablet on
bottle label )x 100%

% label claim = ( 4.765 / 60 )x 100%

% label claim = 7.94%

Discussion
In the experiment, the determination of iron concentration across soil, spinach, and iron vitamin samples
was performed using Atomic Absorption Spectroscopy; toward this process, concentrated nitric acid was
used during sample preparation for efficient extraction. Evidently, the huge discrepancy between the
labeled iron content of the vitamin tablet, 60 mg, and our calculated value of 4.765 mg could indicate
possible errors. in the extraction of samples, whereby too much precipitate might have existed with
adverse implications for the efficiency of extraction. Errors may have come from poor instrument
calibration or not correctly zeroing out, which shows these steps are highly critical to the accuracy of the
test. One instrumental method that may be superior to this could be ICP-AES, which would entail
increased sensitivity with the added multi-element detection capabilities; however, this requires more
expensive equipment with complicated sample preparation. Comparison with existing literature on the
iron content of matrices similar to ours showed that our results were plausible, though seriously deviated
in one sample, hence the need for recommendations on refining our extraction technique and instrument
validation for future analyses.
Conclusion
The general findings are indicate that the technique or approach is at large effective, with 2
out of 3 results put forth make sense with further readings and were promising enough. There
are, however discrepancies which raise questions on the reliability of such findings in critical
situations such as heavy metals in water, likely issue arose with the calculations. But either
way I wouldn’t fully trust the results gathered here as likely the concentration put forward by
the company that makes the iron pills would have a closer estimation than about 8%.
 Acknowledgements

A special thanks to Fanshawe College,

my lab partners, and the instructor for
their guidance and support.
 Thanks
1.

2.
!
McDowell LA, Kudaravalli P, Chen RJ, et al. Iron Overload. [Updated 2024 Jan 11]. In: StatPearls [Internet].
Treasure Island (FL): StatPearls Publishing; 2024 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK526131/
Christian, G., Dasgupta, P., & Schug, K. (2013). Analytical Chemistry. Google Books.
https://books.google.ca/books?id=yPIYEAAAQBAJ&lpg=PR13&ots=xZaIje4OuN&dq=Christian%2C+G.D.+
%22Analytical+Chemistry%2C%22+6th+ed.%2C+Wiley%2C+2014.&lr&pg=PR1#v=onepage&q&f=false
3. OSWER Directive. (2003). Ecological soil screening level for Iron. Eco-SSl. https://rais.ornl.gov/documents/eco-
ssl_iron.pdf
4. U.S. Department of Agriculture. (n.d.). Fooddata Central Search Results. FoodData Central.
https://fdc.nal.usda.gov/fdc-app.html#/food-details/168462/nutrients
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