FST 606

INSTRUMENTAL ANALYSIS OF FOOD

PRACTICAL 2: DETERMINATION OF MINERAL

ELEMENTS IN FOOD PRODUCTS BY
AAS(ASHING METHOD).

STUDENT NAME: NUR MUSFIRAH BT AHMAD FAZILAH

STUDENT ID: 2019583853
GROUP: AS2465B
LECTURER’S NAME: Dr RASEETHA VANI SIVA
MANIKAM
DATE OF EXPERIMENT: 8/5/2020
DATE OF SUBMISSION: 15/5/2020
INTRODUCTION
Atomic absorption spectrometer(AAS) is a widely used technique in the
determination of trace levels(ppm, ppb, ppt) of mineral and metal elements. Next, AAS is
based on the fact where the atoms of most metals will absorb energy when the radiation
containing their characteristic excitation wavelengths is passed through an atomised sample
of a solution containing the particular element .Then, when the intensity of the radiation is
reduced, it will be proportional to the concentration of the element present (Aishah&et.al,
2015). In the simpler word, AAS is based on the absorption of UV light or visible radiation by
free atoms in the gaseous state(Serna-Saldivar, 2012). Besides, the analyte atoms need to
be in the gas phase or atomic vapour in order to accomplish this technique. Therefore, the
sample to be analysed can be converted into an atomic vapour by using flame
atomiser(Aishah&et.al, 2015).
Then, the sample to be analyzed is normally ashed and followed by dissolving it
in an aqueous solution where this solution will be heated to vaporize and atomize the
minerals. Next, a beam of radiation is moves through the atomised sample and the
absorption of radiation is measured at specific wavelength corresponding to the mineral of
interest(Anonymous,n.d). In addition, the mineral composition in food is determined in wet or
dry ash samples that are diluted with acid for the analysis in AAS(Serna-Saldivar, 2012).
Moreover, inorganic elements constitute the major part of dry ash that remains after ignition
of the organic matter and it shows that the dry ashing technique are still the primary way to
determine the total minerals in the food(Poitevin, 2016). Furthermore, the type of minerals
present in the food are determined by measuring the position of the peaks in the emission or
absorption spectra. In addition, the concentration of the mineral components is determined
by measuring the intensity of a spectral line known to correspond to the particular element of
interest(Anonymous,n.d). In this experiment, dry ashing was conducted to determine the
present of minerals such as calcium and sodium in the baby corn and mushroom.

OBJECTIVE
To determine the mineral elements in food products by using AAS(ashing method).
EQUIPMENT AND REAGENTS
AAS instrument Volumetric flasks
Muffle furnace Measuring cylinder
Bunsen burner Ashless filter paper
Water bath Distilled water
Drying oven Concentrated hydrochloric acid
Hot plate Sodium standard solution(1000mg/L)
Blender Magnesium standard solution (1000mg/L)
Crucibles Zinc standard solution(1000mg/L)
Pipettes Calcium standard solution
Funnels 10% Lanthanum chloride solution
Beakers

PROCEDURE
1. Preparation for sample ashing.
a) 5g of homogenised food sample was weighed into a previously dried, cooled and
the crucible was weighed.
b) For high moisture foods, it was dried in an oven at 100oC.
c) The sample was heated gently over a bunsen burner until the food is charred and
ceased smoking.
d) The crucible was transferred to a muffle furnace and it was leaved until a white or
light grey ash form is obtained. If the residue is black in colour, it was moisted
with a small amount of water to dissolve salts and the sample was dried in an
oven and the ashing process is repeated.
e) The sample was cooled in desiccators and the ash obtained was dried.
f) 5 ml of concentrated hydrochloric acid was added into the crucible containing ash
and the mixture was boiled for 5 minutes on a hot plate in a fume cupboard. Acid
was added as necessary to maintain the volume.
g) The contents were transferred by washing the residue in the crucible into a
beaker with deionised water.
h) The volume was adjusted to about 40ml and it was boiled for another 10 minutes
over a bunsen burner.
i) It was cooled and the sample was filtered using an ashless filter paper into 100
ml volumetric flask. The beaker was rinsed with deionised water and make up to
volume. The sample was mixed well with repeated inversion of the flask.
j) This ash solution was used for the determination of individual mineral elements.
2. Preparation of sample for calcium determination.
a) 10 ml of the ash solution was pipetted into a 500 ml volumetric flask.
b) 1ml of 10% lanthanum chloride solution was added. The volume was make up
with deionised water and it was mixed well with repeated inversion of the flask.

3. Preparation of sample for other mineral determination.

a) 10ml of the ash solution was pipetted into a 100 ml volumetric flask(for
determination of zinc).
b) 10ml of the ash solution was pipetted into a 100 ml volumetric flask(for
determination of sodium and magnesium).
c) The sample was make up to volume with deionised water and it was mixed well
with repeated inversion of the flask.

4. Preparation of the standard solution. Prepare stock solution of each mineral

element(100mg/L).
a) 10 ml of each(Mg, Na or Zn) standard solution(1000mg/L) were pipetted into a
100ml volumetric flask and then the volume was make up with deionised water. It
was mixed well.
b) A series of standard solutions of each element to be analysed were prepared by
dilution of the mineral stock solution using deionised water in a 100 ml volumetric
flask. Use equation: C1V1 = C2V2 to prepare the standard solutions.
c) In the preparation of calcium standard solution: 1ml of 10% lanthanum chloride
was added to each flask before making up to volume. This is to minimise the
interference effects of phosphate. Each solution was mixed well.

5. Absorbance measurements
a) The AAS instrument was set up for the element to be analysed.
b) The absorbance of each of the standard solutions prepared was measured.
c) The absorbance of the sample ash solution was measured. If the absorbance of
this ash solution is too high, a known volume was diluted with deionised water
and the measurement was repeated.
RESULTS
1. Absorbance Data for Samples:
Mineral measured: Na
Absorbance reading at λ: 589.0nm

ABSORBANCE
SAMPLE
1 2 3 Average ± S.D.

MUSHROOM 0.613 0.612 0.611 0.612 ± 0.001

BABY CORN 0.757 0.758 0.756 0.757 ± 0.001

Mineral measured: Ca
Absorbance reading at λ: 422.7nm

ABSORBANCE
SAMPLE
1 2 3 Average ± S.D.

MUSHROOM 0.035 0.035 0.035 0.035 ± 0

BABY CORN 0.036 0.036 0.036 0.036 ± 0

2. Absorbance Data for Standard Solution

 Mineral measured: Na
Absorbance reading at λ: 589.0nm

STANDARD ABSORBANCE
CONCENTRATION Average ± S.D
(mg/l) 1 2 3

1 0.267 0.266 0.269 0.267 ± 0.002

2 0.523 0.526 0.529 0.526 ± 0.003
3 0.714 0.712 0.710 0.712 ± 0.002
4 0.859 0.853 0.855 0.856 ± 0.003
5 0.971 0.970 0.968 0.970 ± 0.002

Average Absorbance VS Standard Concentration

1.2

0.8
Axis Title

0.6

0.4

0.2

0
0 1 2 3 4 5 6
Axis Title

Mineral measured: Ca
Absorbance reading at λ: 422.7nm

STANDARD ABSORBANCE
CONCENTRATION Average ± S.D
(MG/L) 1 2 3
1 0.082 0.082 0.082 0.082 ± 0
2 0.134 0.134 0.133 0.134 ± 0.0006
3 0.190 0.190 0.190 0.190 ± 0
4 0.242 0.241 0.242 0.242 ± 0.0006
5 0.295 0.295 0.296 0.295 ± 0.0006

Average Absorbance VS Standard Concentration

0.35

0.3
Average Absorbance

0.25

0.2

0.15

0.1

0.05

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

CALCULATION:
 A. Determination of sodium in canned food products

Take equation from curve,

y=0.2133x

SODIUM CONTENT IN MUSHROOM,

y, average absorbance of sodium in mushroom = 0.612
y = 0.2133x
0.612 = 0.2133x
x=
=2.87 mg/L
% Sodium in mushroom:
% Sodium in mushroom =
Where,
M ppm : Concentration of mineral in sample solution.
Wg : Weight of food used.
V ml : Volume of ash solution diluted to 100 ml.

% Sodium in mushroom = 2.87 mg/L

%Sodium in mushroom = 0.014 %

SODIUM CONTENT IN BABY CORN:

y, average absorbance of sodium in baby corn = 0.757
y = 0.2133x
0.757 = 0.2133x
 x=
=3.55 mg/L
% Sodium in baby corn:
% Sodium in baby corn =
Where,
M ppm : Concentration of mineral in sample solution.
Wg : Weight of food used.
V ml : Volume of ash solution diluted to 100 ml.

% Sodium in baby corn = 3.55 mg/L

%Sodium in baby corn = 0.018%

B. Determination of calcium in canned food products

Take equation from curve,

y=0.0611x

Calcium content in mushroom

y, average absorbance of calcium in mushroom = 0.035
y = 0.0611x
0.035 = 0.0611x
x=
=0.573mg/L
% Calcium in mushroom:
% Calcium in mushroom =
Where,
M ppm : Concentration of mineral in sample solution.
Wg : Weight of food used.
V ml : Volume of ash solution diluted to 100 ml.

% Sodium in mushroom = 0.573 mg/L

%Sodium in mushroom = 2.865 × 10-3 %

Calcium content in baby corn
y, average absorbance of calcium in baby corn = 0.036
y = 0.0611x
0.036 = 0.0611x
x=
=0.589 mg/L
% Calcium in baby corn:
% Calcium in baby corn =
Where,
M ppm : Concentration of mineral in sample solution.
Wg : Weight of food used.
V ml : Volume of ash solution diluted to 100 ml.

% Calcium in baby corn = 0.589 mg/L

%Calcium in baby corn = 2.945 × 10-3 %

DISCUSSION
The aim of this experiment is to determine the mineral elements in the food
products by using atomic absorption spectrometer(AAS) technique. In this experiment, there
are 2 type of samples to be analyzed such as mushroom and baby corn. These samples are
prepared for the calcium and sodium determination. Next, AAS is an analytical technique
that is used for the mineral analysis where it is based on the absorption of UV light or visible
radiation by free atoms in the gaseous state(Serna-Saldivar, 2012). Hence, in order to
accomplish this technique, these samples need to be converted into the gas phase or atomic
vapour by using flame atomiser(Aishah&et al., 2015). This can be done by exposing the
samples to high temperature but not too high as the atoms may become ionized which it is
undesirable.Then, the absorption of radiation can be measured as the beam of radiation is
passed through the atomized sample at specific wavelengths which corresponding to the
mineral of interest. Besides, the concentration of minerals in the samples can be measured
by measuring the reduction in amplitude of the beam of radiation that has passed through
the samples, rather than the radiation emitted by the excited sample. This can be done by
using a chopper(Anonymous,n.d).
Moreover, a calibration curve for atomic absorption spectrometer(AAS) was
plotted by aspirating into the flame and the absorption of each samples that contains known
concentration of calcium and sodium were measured and then plotting a graph of average
absorption against the standard concentration. Besides, the samples need to be diluted as it
can cause the absorbance values to be too high and would not fit into the range of the
calcium and sodium standard solutions(Bissessur, Goburdhun, & Ruggoo, 1999). Based on
the results obtained, it shows that mushroom has an average absorbance sodium content of
0.612 with standard deviation of 0.001 whereas for baby corn, the average absorbance is
0.757 with standard deviation of 0.001. Next, for calcium determination, the average
absorbance of calcium content in mushroom is 0.035 and the standard deviation is zero
meanwhile for baby corn, the average absorbance is 0.036 with standard deviation of zero.
Therefore, the higher the absorbance value, the higher the concentration of mineral
elements present in the food products. Next, the linear regression(R2) for both of the mineral
measured in the standard solutions are less than 0.98 in which it is required to repeat the
preparation of the standard solution. This is because of the error might be occurred or the
used range of the graph may not be linear(Anonymous,2020). Furthermore, calcium and
sodium contents in the original food had been calculated by using this formula as shown
below:
% Mineral in food =
Therefore, the percentage of sodium present in the mushroom is 0.014 % meanwhile in the
baby corn is 0.018%. From this calculation, it indicates that the baby corn is contained more
sodium than the mushroom. Hence, the higher the percentage of mineral component, the
higher the concentration of the mineral present in food. Next, for the calcium determination,
the percentage of calcium present in the mushroom is 2.865 × 10-3 % whereas in the baby
corn is 2.945 × 10-3 %. This shows that the baby corn is contained more calcium than the
mushroom. In the overall, there is high concentration of calcium and sodium in the baby corn
than in mushroom. There might be sources of error that occurred during the experiment such
as, low ashing temperature, short duration and large sample size might lead to the
incomplete combustion which it will results in a higher ash content than the true value(Liu,
2019).
.
CONCLUSION
In the conclusion, we can conclude that the objective for this experiment is achieved where
we have determine the mineral element in the food products which are mushroom and baby
corn by using AAS(ashing method). Next, dry ashing was conducted to determine the
present of minerals such as calcium and sodium in these samples. In overall, baby corn is
contained more calcium and sodium than in mushroom.

REFERENCES
1. Aishah, B., Fadhilah, J., Hariyah, H., Norahiza, M. S. (2015). Instrumental
Analysis of Food: Practical Manual for Undergraduates. Shah Alam: UiTM Press
2. 7 quality of analytical procedures(2020). Retrieved from
http://www.fao.org/3/w7295e/w7295e09.htm

3. Bissessur, V., Goburdhun, D., & Ruggoo, A. (1999). 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.
4. Liu, K. (2019). Effects of sample size, dry ashing temperature and duration on
determination of ash content in algae and other biomass. Algal Research, 40,
101486. doi: https://doi.org/10.1016/j.algal.2019.101486
5. (n.d). Retrieved from https://people.umass.edu/~mcclemen/581Ash&Minerals.html
6. Poitevin, E. (2016). Official methods for the determination of minerals and trace
elements in infant formula and milk products: a review. Journal of AOAC
International, 99(1), 42-52.
7. Serna-Saldivar, S. O. (2012). Cereal Grains: Laboratory Reference and
Procedures Manual: CRC Press.