FACULTY OF APPLIED SCIENCES

UNIVERSITI TEKNOLOGI MARA (UiTM)

CAMPUS SHAH ALAM, SELANGOR

FST606: INSTRUMENTAL ANALYSIS OF FOOD

EXPERIMENT 4: DETERMINATION OF MINERALS ELEMENTS IN CANNED

FOOD PRODUCTS BY AAS (NON-ASHING METHOD)

NAME: NURUL AYU NATASHA BINTI MAT RANI (AS246-5C)

STUDENT ID: 2022961819

NAME OF GROUP MEMBERS STUDENT ID GROUP

NUR AMIRA AISYA BINTI AMIR SHARIFUDIN 2022912935 AS246-5C
MUHAMMAD OMAR BIN ZAKARIA 2022844848 AS246-5B
MUHAMMAD AMIR IZZUDDIN BIN MAT NAWI 2022815956 AS246-5B

DATE OF EXPERIMENT: 10th NOVEMBER 2023

NAME OF LECTURER: DR. AISHAH BINTI BUJANG

INTRODUCTION

Minerals are among the six vital elements needed for human bodybuilding, including
the growth of bones, teeth, and soft tissue structure. According to Geren (2020), minerals are
inorganic solid materials that crystallise and are categorised based on our relative needs. They
are frequently divided into two groups which are macro minerals and micro minerals. Macro
minerals are needed in greater quantities in the food or are found in greater concentratins in
the bodies of animals. Some macro minerals include calcium, sodium, magnesium, and
potassium. Meanwhile, micro minerals also known as trace minerals are needed in lesser
amounts by animals or are present in the body at low concentrations. Iron, manganese, zinc,
and iodine are examples of micro minerals (Central Ag Supply, 2018). Among the mineral
elements that humans require are calcium, magnesium, sodium, and potassium.

One method for determining the amounts of chemical elements in environmental

samples is atomic absorption spectroscopy (AAS), which measures the radiation that the
chemical element of interest absorbs. The fundamental idea behind an atomic absorption
spectrometer (AAS) is that most metal atoms absorb energy when light with specific
excitation wavelengths passes through an atomized sample of the element's solution. To put it
simply, measuring the radiation absorbed by the target element is how elements are analysed.
Because AAS is relatively interference-free and provides sufficient sensitivity for a wide
range of applications, it has emerged as one of the most widely used analytical methods for
elemental analysis (Hill & Fisher, 2017). Additionally, this approach may be used to evaluate
complex materials, such food or other biological and environmental samples, in order to
identify substances present.

Modern AAS systems consist of a radiation source, an atomizer, a monochromator, a

detecting system, and a computer. The monochromator isolates a line that uniquely identifies
an element while scattering light waves that do not. This will then detect UV and visible light
since the isolation from the monochromator has eliminated background light interference. It
requires the employment of radiation sources whose emission spectra are more restricted than
the absorption line width of the trace elements under study. In atomic absorption
spectroscopy, two fundamental atom cells are used: flame and electrothermal heating of a
sample cell (Cámara & Moreno, 2016). It is widely accepted that the flame approach, which
has the added benefits of being quick and extremely easy to use, should be used to evaluate if
there is sufficient analyte present in the sample.
 Food samples are treated with procedures like acid treatment of the ash or wet
oxidation to get a solution of the metal to be calculated. The organic matter is removed by
either wet digestion or dry ashing, which uses powerful acids under high pressure in closed
vessels (Hosry et al., 2023). After that, this solution is exposed to a hot flame to undergo
nebulization. Atomization process then occurs by which the flame releases the sample solvent
and disintegrates metal complexes into free metal atoms or free radicals. These unbound
atoms absorb some of the radiation from the hollow cathode lamp as it travels through them.
The concentration of the metal in solution affects the degree of absorption.

The Beer-Lambert law is used to calculate the concentration. Under the current set of
conditions, absorbance is directly proportional to the concentration of the analyte absorbed.
Typically, a calibration curve obtained using standards of known concentration is used to
determine the concentration. To remove the organic materials in this experiment, the wet
digestion method has been used. In order to reduce sample contamination from compounds in
the air, surrounding environment, and vessel walls, Twyman (2005) states that the different
acid and flux treatments are carried out at high temperatures in specially built containers. In
this experiment, the mineral elements calcium and magnesium found in canned green peas
and mushrooms were determined. The following formula was used to determine the amounts
of calcium and magnesium in each sample:

𝑀×50
Mineral in food = 𝑊

OBJECTIVES

• To determine the mineral elements, present in canned mushroom and green peas by
AAS.
• To determine the amount of Calcium and Magnesium present in canned mushroom
and canned green peas.

APPARATUS AND REAGENT

AAS instrument, Analytical balance, concentrated hydrochloric acid, Calcium chloride
solution (100mg/L Ca), 10% Lanthanum chloride solution, Magnesium standard solution
(1000mg/L), Deionised water, Pipettes, Volumetric flasks, Hot plate, Beakers, Measuring
cylinder, Funnels, Can opener, Ashless filter paper and Glass wool.

Sample: canned mushroom and canned green peas.

PROCEDURE

A. Preparation of Sample
1. Firstly, the can of the sample food is opened and poured into a separate container
without scrapping the can.
2. 10g of the drained sample was weighed accurately and homogenised using a
mortar and a pestle.
3. After that, the homogenised sample was transferred into a beaker and added 20mL
of deionised water followed by 5mL of concentrated acid. (Conducted in fume
hood).
4. The beaker was then placed on a hot plate and bring to the boil for 1 minute.
Then, the beaker was removed from the hot plate and left cool. (Conducted in
fume hood)
5. After cool, the content was filtered using glass wool into a 50mL volumetric flask,
washed with deionised water and make up the volume.
6. Lastly, the solution was well mixed and filtered again through an ashless filter
paper to provide sufficient solution for analysis (10 to 12mL).
B. Preparation of Sample for Calcium Determination
1. Firstly, 1 mL of the filtered solution from (f) was pipetted into a 100mL
volumetric flask.
2. Next, 1mL of lanthanum chloride was added and make up to volume with
deionised water.
3. Lastly, the solution was mixed well with repeated inversion of the flask.
C. Preparation of Sample for other Mineral Determination (Magnesium)
1. 1mL of the filtered solution from (f) was pipetted into a 100mL volumetric flask.
2. The solution was made up to volume with deionised water and mixed well with
repeated.
D. Preparation of Standard Solutions
1. Firstly, a mineral stock solution was prepared (100 mg/L).
2. 10mL of each (Mg) standard solution (1000 mg/L) into a 100mL volumetric flask
and then make up to volume with deionised water. The solution was mixed well
with repeatable inversion of flask.
3. After that, a range of standard solutions of each element was prepared to be
analysed by dilution of the mineral stock solution using deionised water in a
100mL volumetric flask.
4. C1V1 = C2V2 equation was used to prepare the standards.
5. Lastly, in the preparation of calcium standard solution: 1mL of 10% lanthanum
chloride per 100mL was added to each flask before making up to volume. This is
to minimise the interference effects of phosphate. Each of the solution were mixed
well.
E. Absorbance Measurement
1. The AAS instrument was set up for the element to be analysed.
2. The absorbance of each of the standard solutions prepared was measured.
3. In a similar manner, the absorbance of the sample solutions prepared from the
canned food was measured. If the absorbance of the sample solution is high, dilute
a known volume with deionised water and repeat the measurement.
RESULTS
Table 1: Weight of Sample and Absorbance Data Obtained
Sample Weight (g)
Green Peas 10.5403
Mushroom 10.0292

Table 2: Absorbance Data for mineral elements (Ca and Mg) measured
Mineral Element: Calcium (Ca)
Absorbance reading at 𝝀 = 422.67 nm
Standard/sample Abs 1 Abs 2 Abs 3 Abs average ± SD
concentration
Blank 0.000 0.001 0.001 0.001 ± 0.0003
1 mg/L 0.064 0.065 0.065 0.064 ± 0.0006
2 mg/L 0.094 0.093 0.093 0.093 ± 0.0003
3 mg/L 0.141 0.146 0.144 0.144 ± 0.0023
4 mg/L 0.206 0.208 0.208 0.207 ± 0.0010
5 mg/L 0.286 0.280 0.282 0.282 ± 0.0009
Green peas 0.032 0.033 0.033 0.033 ± 0.0004
Mushrooms 0.019 0.019 0.019 0.019 ± 0.0001

Mineral Element: Magnesium (Mg)

Absorbance reading at 𝝀 = 285.21 nm
Standard/sample Abs 1 Abs 2 Abs 3 Abs average ± SD
concentration
Blank 0.001 0.001 0.001 0.001 ± 0.0004
1 mg/L 0.426 0.420 0.418 0.421 ± 0.0043
2 mg/L 0.777 0.769 0.766 0.771 ± 0.0057
3 mg/L 1.029 1.025 1.028 1.027 ± 0.0023
4 mg/L 1.200 1.215 1.212 1.209 ± 0.0084
5 mg/L 1.319 1.326 1.322 1.322 ± 0.0036
Green peas 0.075 0.073 0.071 0.073 ± 0.0021
Mushrooms 0.038 0.036 0.036 0.037 ± 0.0013
GRAPH

Concentration (mg/L) vs Absorbance for Calcium

0.3

0.25

0.2
Absorbance

y = 0.0531x
R² = 0.9947
0.15

0.1

0.05

0
0 1 2 3 4 5 6
Concentration (mg/L)

Figure 1: Linear graph for absorbance against standard concentration (mg/L) of Calcium (Ca)
at 422.67nm

Concentration (mg/L) Vs Absorbance for Magnesium

1.6

1.4

1.2
y = 0.2998x
1
Absorbance

R² = 0.9818
0.8

0.6

0.4

0.2

0
0 1 2 3 4 5 6
Concentration (mg/L)

Graph 2: Linear graph for absorbance against standard concentration (mg/L) of Magnesium
(Mg) at 285.21nm
CALCULATIONS

1. Calculation for preparing a desired concentration of standard solution of mineral

element from the stock solution given.

𝐶1 = 100 𝑚𝑔/𝑚𝑙 𝐶2 = 1 𝑚𝑔/𝑚𝑙 𝐶1 = 100 𝑚𝑔/𝑚𝑙 𝐶2 = 2 𝑚𝑔/𝑚𝑙

𝑪 𝟏 𝑪 𝟐 = 𝑽𝟏 𝑽𝟐 𝑪 𝟏 𝑪 𝟐 = 𝑽𝟏 𝑽𝟐

(100 mg/ml) (𝑉1 ) = (1 mg/ml) (100 ml) (100 mg/ml) (𝑉1 ) = (2 mg/ml) (100 ml)

𝑉1 = 1 ml 𝑉1 = 2 ml

𝐶1 = 100 𝑚𝑔/𝑚𝑙 𝐶2 = 3 𝑚𝑔/𝑚𝑙 𝐶1 = 100 𝑚𝑔/𝑚𝑙 𝐶2 = 4 𝑚𝑔/𝑚𝑙

𝑪 𝟏 𝑪 𝟐 = 𝑽𝟏 𝑽𝟐 𝑪 𝟏 𝑪 𝟐 = 𝑽𝟏 𝑽𝟐

(100 mg/ml) (𝑉1 ) = (3 mg/ml) (100 ml) (100 mg/ml) (𝑉1 ) = (4 mg/ml) (100 ml)

𝑉1 = 3 ml 𝑉1 = 4 ml

𝐶1 = 100 𝑚𝑔/𝑚𝑙 𝐶2 = 5 𝑚𝑔/𝑚𝑙

𝑪 𝟏 𝑪 𝟐 = 𝑽𝟏 𝑽𝟐

(100 mg/ml) (𝑉1 ) = (5 mg/ml) (100 ml)

𝑉1 = 5 ml
 2. Determination of mineral elements in canned mushrooms

Determination of Calcium (Ca) in canned Determination of Magnesium (Mg) in canned

mushroom mushroom
Average absorbance reading = y = 0.019 Average absorbance reading = y = 0.037
Equation Equation
y = 0.0531x y = 0.2998x
y = 0.0531x y = 0.2998x
0.019 = 0.0531x 0.037 = 0.2998x
X = 0.3578 mg/L X = 0.1234 mg/L

𝑀×50 𝑀×50
Mineral in food = Mineral in food =
𝑊 𝑊

Where; Where;
M = concentration of mineral in sample M = concentration of mineral in sample
solution solution
W = weight of the sample used W = weight of the sample used
Hence, Hence,
X=M X=M
Concentration of mineral in sample Concentration of mineral in sample
𝑀×50 𝑀×50
= =
𝑊 𝑊
𝑚𝑔 𝑚𝑔
0.3578 ×50 𝑚𝑙 0.1234 ×50 𝑚𝑙
𝐿 𝐿
= =
1000 𝑚𝑙 1000 𝑚𝑙

= 0.0179 mg in 10 g mushroom = 0.0062 mg in 10 g mushroom

Concentration in mg/kg Concentration in mg/kg
0.01789 𝑚𝑔 0.00617 𝑚𝑔
= 1 𝑘𝑔 = 1 𝑘𝑔
10 𝑔 × 10 𝑔 ×
1000 𝑔 1000 𝑔

= 1.79 mg/kg of Ca in canned mushroom = 0.62 mg/kg of Ca in canned mushroom

 3. Determination of mineral elements in canned peas

Determination of Calcium (Ca) in canned Determination of Magnesium (Mg) in canned

peas peas
Average absorbance reading = y = 0.033 Average absorbance reading = y = 0.073
Equation Equation
y = 0.0531x y = 0.2998x
y = 0.0531x y = 0.2998x
0.033 = 0.0531x 0.073 = 0.2998x
X = 0.6215 mg/L X = 0.2435 mg/L

𝑀×50 𝑀×50
Mineral in food = Mineral in food =
𝑊 𝑊

Where; Where;
M = concentration of mineral in sample M = concentration of mineral in sample
solution solution
W = weight of the sample used W = weight of the sample used
Hence, Hence,
X=M X=M
Concentration of mineral in sample Concentration of mineral in sample
𝑀×50 𝑀×50
= =
𝑊 𝑊
𝑚𝑔 𝑚𝑔
0.6215 ×50 𝑚𝑙 0.2435 ×50 𝑚𝑙
𝐿 𝐿
= =
1000 𝑚𝑙 1000 𝑚𝑙

= 0.0311 mg in 10 g mushroom = 0.0122 mg in 10 g mushroom

Concentration in mg/kg Concentration in mg/kg
0.0311 𝑚𝑔 0.0122 𝑚𝑔
= 1 𝑘𝑔 = 1 𝑘𝑔
10 𝑔 × 10 𝑔 ×
1000 𝑔 1000 𝑔

= 3.11 mg/kg of Ca in canned mushroom = 1.22 mg/kg of Ca in canned mushroom

DISCUSSION

The objective of this experiment is to determine the amount of Calcium and

Magnesium present in canned food by AAS. Magnesium and Calcium are used as a standard
solution in order to determine the concentration of mineral elements present in canned green
peas and canned mushrooms. In this experiment, the mineral content of the canned food
sample is measured using the non-ashing method of an atomic absorption spectrophotometer.
The quickest method of determination is AAS non-ashing as it requires less sample
preparation and omits the ashing step. The atom cell in AAS utilised in this experiment was a
flame. Samples are converted by the flame into excitable free ground state atoms. When light
from a lamp that emits light at a wavelength that is particular to the atoms passes through the
flame, the atoms' electrons are stimulated and light energy is absorbed (Agilent, 2023). It is a
very useful method in determining how much of a certain mineral is present in a food sample.
Chanda et al. (2014) states that this apparatus will measure the intensity change, which will
then be translated by computer-based software into an absorbance correlated with the sample
concentration.

The Beer-Lambert equation was used to determine the concentration in AAS. The
concentration of the analyte absorbed under the current set of circumstances is directly
proportional to the absorbance. 10% lanthanum chloride solution was also utilised in calcium
analysis to serve as a releasing agent whilst the amount of calcium was being measured. As a
result, the interfering impact of the phosphate in the samples can be reduced. According to
Sommer et al. (2010), in the calcium analysis, lanthanum chloride was added to avoid the
production of calcium pyrophosphate, which causes an underestimation of calcium. Large
molecules like calcium pyrophosphate have the potential to precipitate out and escape the
flame. It might not be easy to atomize even if it reached the flame. Canned green peas and
mushrooms were utilised as samples in this investigation.

Based on the results obtained in table 1, the average absorbance of the calcium
standard solution in 1mg/mL, 2mg/mL, 3mg/mL, 4mg/mL, 5mg/mL is 0.064, 0.093, 0.144,
0.207, and 0.282, respectively. Meanwhile, the average absorbance reading for magnesium
standard of 1mg/mL, 2mg/mL, 3mg/mL, 4mg/mL, 5mg/mL are 0.421, 0.771, 1.027, 1.209,
and 1.322 respectively. This shows higher concentrations of standard solution yield a higher
absorbance reading. This indicates that a greater absorbance measurement is produced by a
higher standard solution concentration. One element that influences a sample's absorbance is
its concentration. More radiation should be absorbed as the concentration rises, raising the
absorbance. As a result, there is a direct proportionality between the concentration and
absorbance. According to Beer-Lambert Law, the absorbance is directly correlated with the
route length through the sample and the absorbing species' concentration. Additionally,
absorbance is closely correlated with path length, which is the measure of how far light goes
through a material. As the light travels through the solution over a longer distance, it interacts
with more molecules due to its longer path length. Consequently, the absorbance rises.

Then, a standard calibration curve was plotted in figure 1 and figure 2. By using a
standard calibration curve, the concentrations of calcium and magnesium were determined. Y
= 0.0531x is the equation obtained from the standard curve for calcium, while y = 0.2998x is
the equation obtained from the standard curve for magnesium. The average absorbance value
for Mg and Ca in the mushroom and green peas was substituted with y in the linear standard
curve equation using the resulting equation. As a result, the x that was obtained shows the
concentration of each mineral element (Mg and Ca). Green peas and mushrooms had Ca
concentrations of 0.6215 mg/ml and 0.3578 mg/ml, respectively. The content of magnesium
in green peas and mushrooms was found to be 0.2435 mg/mL and 0.1234 mg/mL,
respectively. The following formula was then used to determine the mineral content:

𝑀×50
Mineral in food = 𝑊

Calculations show that canned mushrooms have 1.79 mg of calcium and 0.62 mg of
magnesium per kilogramme, respectively. In canned green peas, the amounts of calcium and
magnesium are 3.11 mg/kg and 1.22 mg/kg, respectively. These results indicate that the
concentration of calcium and magnesium in green peas is higher than the concentration of
calcium and magnesium in canned mushrooms. The US Department of Agriculture (2019)
states in their article that mushrooms have a calcium level of 3 mg per 100 g and a
magnesium content of 9 mg per 100 g. This shows that mushrooms contain 30 mg/kg of
calcium and 90 mg/kg of magnesium. It can be seen that there are a huge amount of
differences compared to the result that has been mentioned before which was 1.79 mg/g of
calcium content in mushrooms and 0.62 mg/g of magnesium content in mushrooms. In
addition, Kumari et al. (2021) state that green peas contain 9.5 mg/g of calcium and 5.5 mg/g
of magnesium. This indicates that there are 95 mg/kg of calcium and 55 mg/kg of magnesium
in green peas. This demonstrates that the result obtained deviates significantly from the
theoretical value as well. It is clear that there are significant variations from the previously
reported results, which showed that green peas contained 1.22 mg/g of magnesium and 3.11
mg/g of calcium in mushrooms. This can be the result of the erroneous approach taken.
According to Bissessur (2007), the AAS-Non-Ashing approach for analysis proved to be the
least accurate when compared to the AAS-Ashing method and titrimety (potassium
permanganate titration). On the other hand, the latter approach requires the least amount of
sample preparation.

Lanthanum chloride serves as a releasing agent for the production of the calcium
standard. In order to avoid any phosphate precipitation in the flame and stop it from
interacting with the analyte, LaCl is utilised in the AAS investigation. However, a few
mistakes might be made throughout the experiment, producing an erroneous outcome. In this
experiment, glass must not be utilised; instead, plastic equipment must be used. It is crucial
because the glass equipment could absorb the minerals in the samples and provide a result
that is not accurate. In addition, the residue in the volumetric flask has to be cleaned using
deionized water after that in order to process the sample. This is so because deionized water
is completely contaminant-free and clean. As a result, it can stop any chemical process and
hinder the identification of minerals.
CONCLUSION

In conclusion, the experiment's goal has been accomplished. The graph of the
absorbance against the standard concentration of calcium and magnesium are directly
proportional as the absorbance reading increase with the concentration. Concentration of
calcium and magnesium in canned mushrooms and canned green peas are obtained from the
equation derived in a linear standard curve. From the calculation, canned mushrooms have
1.79 mg of calcium and 0.62 mg of magnesium per kilogramme, respectively. Meanwhile, in
canned green peas, the amounts of calcium and magnesium are 3.11 mg/kg and 1.22 mg/kg,
respectively. These results indicate that green peas have a greater calcium and magnesium
concentration than mushrooms in cans. However, according to the theory, the amount of
calcium content in canned mushroom is higher than canned green peas while magnesium
content in green peas is higher than in mushroom. These results have a big difference with the
expected result. Therefore, there may have been an error occurred during conducting this
experiment.
REFERENCES

Agilent. (2023). Atomic Absorption spectroscopy how does AAS work, AAS FAQs.

https://www.agilent.com/en/support/atomic-spectroscopy/atomic-absorption/flame-

atomic-absorption-instruments/how-does-aas-work-aas-

faqs#:~:text=The%20flame%20converts%20samples%20into,elevated%20to%20an%

20excited%20state

Bissessur, V., Goburdhun, D., & Ruggoo, A. (2007). Evaluation of atomic absorption

Spectrophotometry (ashing, non-ashing) and titrimetry for calcium determination in

selected foods. University of Mauritius Research Journal, 3(1), 115–128.

https://www.ajol.info/index.php/umrj/article/download/130881/120459

Cámara-Martos, F., & Moreno-Rojas, R. (2016). Cobalt: Properties and Determination. In

Elsevier eBooks. https://doi.org/10.1016/b978-0-12-384947-2.00175-6

Central Ag Supply. (2018). The difference between macro and micro minerals – Central AG

Supply Services. https://www.centralagsupply.com/the-difference-between-macro-

and-micro-minerals/

Chanda, S., Das, S., Paul, B. N., & Giri, S. S. (2014). Mineral assay in atomic absorption

spectroscopy. ResearchGate.

https://www.researchgate.net/publication/279195165_Mineral_Assay_in_Atomic_Ab

sorption_Spectroscopy

Green, S. (2020, August 23). Essential nutrients. Pressbooks.

https://open.maricopa.edu/nutritionessentials/chapter/essential-nutrients/

Hill, S. J., & Fisher, A. (2017). Atomic absorption, methods and instrumentation. In Elsevier

eBooks (pp. 37–43). https://doi.org/10.1016/b978-0-12-803224-4.00099-6

Hosry, L. E., Sok, N., Richa, R., Mashtoub, L. A., Cayot, P., & Bou-Maroun, E. (2023).

Sample preparation and analytical techniques in the determination of trace elements in

food: a review. Foods, 12(4), 895. https://doi.org/10.3390/foods12040895

Kumari, T., & Deka, S. C. (2021). Potential health benefits of garden pea seeds and pods: A

review. Legume Science, 3(2). https://doi.org/10.1002/leg3.82

Sommer, M., Rutman, M. G., Rotter, E. W., Wagoner, H., & Fritsche, E. T. (2010).

Determination of Calcium in Serum Samples by AAS Using a Fuel Lean Flame.

Agilent, 1. https://www.agilent.com/Library/applications/aa117.pdf

Twyman, R. M. (2005). Sample Dissolution for Elemental Analysis | Wet digestion. In

Elsevier eBooks (pp. 146–153). https://doi.org/10.1016/b0-12-369397-7/00539-2

U.S Department of Algiculture. (2023). FoodData Central. Algiculture Research Service.

https://fdc.nal.usda.gov/fdc-app.html#/food-details/169251/nutrients

Zhong, J., & Wang, X. (2019). An introduction to evaluation technologies for food quality. In

Elsevier eBooks (pp. 1–3). https://doi.org/10.1016/b978-0-12-814217-2.00001-9