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Experiment 1

Title: Atomic Absorption Spectroscopy

Introduction

Atomic-absorption spectroscopy (AAS) is a technique used for analysis of major,

minor, and trace elements in foodstuffs (Chemicool.com, 2016). A liquid sample containing

the metal analyte is aspirated into an air-acetylene flame, causing evaporation of the solvent

and vaporization of the free metal atoms (Chemicool.com, 2016). This process is called as

atomization. The common emission source that emits radiation characteristic of a particular

metal is a hollow cathode lamp. A monochromator is used to select the specific wavelength

of light. In this experiment, flame atomization will be used to determine iron in the solution.

Spectroscopic measurements are easily performed with liquid samples (Chemicool.com, 2016).

Flame atomic absorption spectroscopy (FAAS) can only analyze ions or atoms in solution

(Chemicool.com, 2016). Therefore, the preparation of sample either in a known or unknown

sample should be done in solution form in order to allow FAAS to analyze the sample

accurately and precisely (Chemicool.com, 2016).

The objective of the experiment is to determine the amount of iron present in a given

food sample. The known iron sample is prepare with different concentration from 2.5ppm to

10ppm, they are used to design the calibration such that the unknown sample concentration

locates somewhere in the middle of set range.

Materials and apparatus

The materials used in the experiment were 100 ppm Fe standard, reagent-grade water, 2 %

(v/v) nitric acid, unknown solid sample, 0.1 % (w/w) 1, 10-phenanthroline monohydrate, 10 %

(w/w) hydroxylamine hydrochloride, 0.01 M, pH 4 acetate buffer and 50 % (w/w) sodium

hydroxide (NaOH). The apparatus and instruments that used are hotplate, beakers, plastic tubes,

0.45m filter paper, measuring cylinder, volumetric flask, watch glass, atomic absorption

spectrometer, pestle and mortar, pipette.


Methodology

A) Preparation of Unknown

A solid unknown would be provided by the instructor. The sample was digested by

dissolving approximately 0.1 g in 10 mL of nitric acid (conc.), to which 30 mL of

hydrochloric acid (conc.) was added in a beaker. A graduated cylinder was used to deliver

the correct acid volumes. These steps were done in the fume hood. The beaker was warmed

on a hotplate with a watch glass on top. The reaction was watched carefully, the heat was

turned down when the reaction became violent. There should be no solid at the bottom of

the beaker once the digestion was completed. The mixture was let cool and then it was added

slowly to a 100 mL volumetric flask containing approximately 30 mL of reagent-grade

water. The beaker was rinsed with reagent-grade water several times to ensure that all

materials were transferred. It was then diluted to the mark with reagent-grade water and

mixed well. The unknown solution was transferred to a plastic bottle and saved.

B) Investigation of Instrumental Parameters and Calibration Design

First, 5 ppm iron standard was prepared. 5 mL of the provided 100 ppm iron standard was

pipetted into a 100 mL volumetric flask and diluted to the mark with 2 % nitric acid. Second,

test solution of unknown was prepared. 0.1 mL of the unknown solution prepared in Part A

was pipetted into a 100 mL volumetric flask and diluted to the mark with 2 % nitric acid.

Then, the absorbance of the two prepared solutions was recorded using 2 % nitric acid as

the blank.

C) Final Calibration Design

The recommended linear range of response for atomic spectroscopic measurements of iron

was 2.5 to 10 ppm. A set of four standards that span this range was designed with even

concentration spacing. Aliquot volume of the 100 ppm iron standard was rounded to fit the

available pipettes. The results from the 5 ppm iron standard were used to predict the

absorbance range corresponding to 2.5 to 10 ppm iron. This information was used together
with the measured absorbance of the unknown solution based on a 10 mL aliquot volume

that would produce an absorbance that you would predict to fall in the middle of the 2.5 to

10 ppm calibration range.

Result

Table 1.0 Absorbance data of 0.1g wheat flour


Concentration Absorbance Mean Standard
of Standards Deviation
(ppm) Replicate 1 Replicate 2 Replicate 3
Blank 0.001 -0.000 0.002 0.001 0.0011
2.5 0.095 0.093 0.089 0.092 0.0027
5.0 0.199 0.200 0.199 0.199 0.0004
7.5 0.277 0.280 0.279 0.279 0.0016
10.0 0.359 0.364 0.363 0.362 0.0027
Unknown 0.058 0.023 0.015 0.032 0.0226
Sample

Table 1.1 (0.1g wheat flour)


Standard Concentration (mg/L) Absorbance
Blank 0.0 0.001
1 2.5 0.092
2 5.0 0.199
3 7.5 0.279
4 10.0 0.362

Graph 1.0 The concentration versus absorbance of standard curve


Calculation
y = absorbance, x = concentration, c = 0 Absorbance readings of sample:
y = mx + c 0.058, 0.023, 0.015
y = 0.0364x + 0 Mean absorbance of sample: 0.032
Intercept: 0.0 Standard deviation of sample: 0.0226
Slope: 0.0364
Correlation Coefficient: 0.9973

Concentration of sample: M1V1 = M2V2


y = 0.0364x M1 0.1ml = 0.8791ppm 100ml
Absorbance, y = 0.032 M1 = 879.1ppm =879.1mg/kg =0.8791mg/g
0.032 = 0.0364x Every 1g of the sample contains 0.8791mg
x = 0.8791ppm of iron.

Table 1.2 (0.5g wheat flour)


Standard Concentration (mg/L) Absorbance
Blank 0.0 0.001
1 2.5 0.087
2 5.0 0.204
3 7.5 0.300
4 10.0 0.304

Calculation
y = absorbance, x = concentration, c = 0 Absorbance readings of sample:
y = mx + c 0.006, 0.000, 0.001
y = 0.0328x + 0 Mean absorbance of sample: 0.003
Intercept : 0.0 Standard deviation of sample: 0.0031
Slope : 0.0328
Correlation Coefficient : 0.9442
Concentration of sample : M1V1 = M2V2
y = 0.0328x M1 0.1ml = 0.0914ppm 100ml
Absorbance, y = 0.003 M1 = 91.4ppm = 91.4mg/kg = 0.0914mg/g
0.003 = 0.0328x Every 1g of the sample contains
0.0914mg of iron.
x = 0.0914ppm
Table 1.3 (Cream cracker)
Standard Concentration (mg/L) Absorbance
Blank 0.0 -0.000
1 2.5 0.096
2 5.0 0.189
3 7.5 0.265
4 10.0 0.359

Calculation
y = absorbance, x = concentration, c = 0 Absorbance readings of sample:
y = mx + c 0.032, 0.029, 0.026
y = 0.0355x + 0 Mean absorbance of sample: 0.029
Intercept : 0.0 Standard deviation of sample: 0.003
Slope : 0.0355
Correlation Coefficient : 0.9986
Concentration of sample : M1V1 = M2V2
y = 0.0355x M1 0.1ml = 0.8169ppm 100ml
Absorbance, y = 0.029 M1 = 816.9ppm =816.9mg/kg =0.8169mg/g
0.029 = 0.0355x Every 1g of the sample contains 0.8169mg
x = 0.8169ppm of iron.

Table 1.4 (Coco powder)


Standard Concentration (mg/L) Absorbance
Blank 0.0 0.001
1 2.5 0.107
2 5.0 0.214
3 7.5 0.301
4 10.0 0.362

Calculation

y = absorbance, x = concentration, c = 0 Absorbance readings of sample:


y = mx + c 0.012, 0.013, 0.008
y = 0.0366x + 0 Mean absorbance of sample: 0.011
Intercept : 0.0 Standard deviation of sample: 0.0028
Slope : 0.0366
Correlation Coefficient : 0.9889
Concentration of sample : M1V1 = M2V2
y = 0.0366x M1 0.1ml = 0.301ppm 100ml
Absorbance, y = 0.011 M1 = 301ppm = 301mg/kg = 0.301mg/g
0.011 = 0.0366x Every 1g of the sample contains 0.301mg
of iron.
x = 0.301ppm

Table 1.5 (Milk powder)


Standard Concentration (mg/L) Absorbance
Blank 0.0 0.001
1 2.5 0.101
2 5.0 0.205
3 7.5 0.296
4 10.0 0.380

Calculation
y = absorbance, x = concentration, c = 0 Absorbance readings of sample:
y = mx + c 0.003, 0.004, 0.001
y = 0.0381x + 0 Mean absorbance of sample: 0.003
Intercept : 0.0 Standard deviation of sample: 0.0013
Slope : 0.0381
Correlation Coefficient : 0.9982

Concentration of sample : M1V1 = M2V2


y = 0.0381x M1 0.1ml = 0.341ppm 100ml
Absorbance, y = 0.0013 M1 = 341ppm = 341mg/kg = 0.341mg/g
0.0013 = 0.0381x Every 1g of the sample contains 0.341mg
of iron.
x = 0.0341ppm
Discussion

Atomic Absorption Spectroscopy is a type of spectrometry that is used to determine the


concentration of a specific element in a sample, using a combination of a known absorption spectra
and measured absorbance to determine analyte concentration, using the Beer-Lambert Law to
correlate absorption and concentration (Melville, 2014). This works because each element has
characteristic absorption spectra, relating to the specific, quantized transitions of electrons to
excited states (Melville, 2014).

The Beer-Lambert law (or Beer's law) is the linear relationship between absorbance and
concentration of an absorbing species (Hplc.chem.shu.edu, 2016). The Beer-Lambert law is
necessary in order to compare its absorbance with that of the sample solution of unknown
concentration. The relationship between the absorbance of a solution and the concentration of the
absorbing species is known as Beers law, which written as A=abc. If absorption measurements
are conducted by same spectrometer, the cells should have the same pathlengths (b), the
absorptivity(a) and the cell thickness(b) (Food chemicals codex, 1981). The concentration of the
sample solution is directly proportional to the absorbance of the sample solution.

Based on the graph, the calibration curve is more accurate to the plotted data points. The
graph had generated a regression of R = 0.996932. The R2 value obtained was higher than the
acceptable R2 value which is 0.9973. This mean that the regression of R model has a good fit. The
y-intercept of the trendline is near-zero. The trendline followed trivially from the Beer-Lambert
Law, which stated that A = kbC. If C = 0, A must obviously also equal to 0 (Melville, J. 2014).
Among all the sample products, wheat flour contained a highest concentration of iron in every 1g
of the sample, which is 0.8791mg of iron per gram.

The advantages of AAS is high sample throughput, easy to use and has a high precision.
The concentration of sample is calculated by using the equation y = mx + c from the plotted graph.
FAAS calculation had showed a slight higher value by comparing with two concentration of
unknown sample.
Conclusion

The objective of this experiment which is to determine the concentration of iron ions, Fe2+
in the iron pill achieved. The concentration of unknown iron sample is fall between the ranges of
standard calibration from 2.5ppm to 10ppm. The trendlines produced is a direct correlation
between concentration and absorption, hence it means that the linear fit is obey to Beer-Lambert
law.The high values of the correlation coefficients (0.9973) obtained demonstrate good linear
correlation of the absorbance with trace element concentrations. The higher the concentration of
iron pill, the higher the absorbance. The absorbance of the unknown sample is 0.8791 mg/L. Every
1g of the sample contains 0.8791 mg of iron.

Reference

1. Hplc.chem.shu.edu. (2016). Beer-Lambert Law. [online] Available at:


http://hplc.chem.shu.edu/NEW/Undergrad/Molec_Spectr/Lambert.html [Accessed 8 July
2017].

2. Food chemicals codex. (1981). Washington, D.C.: National Academy Press [Accessed 8
July 2017]

3. Chemicool.com. (2016). Definition of atomic_absorption_spectroscopy_aas - Chemistry


Dictionary. [online] Available at:
http://www.chemicool.com/definition/atomic_absorption_spectroscopy_aas.html
[Accessed 8 July 2017].

4. Melville, J. (2014). Atomic Absorption Spectroscopy of Metal Alloys. [online] California:


UC Berkeley College of Chemistry. Available at:
https://www.ocf.berkeley.edu/~jmlvll/lab-reports/AASalloy/AASalloy.pdf [Accessed 8
July 2017].