LABORATORY REPORT OF INSTRUMENTAL ANALYSIS OF FOOD (FST 606)

NAME : FATIN NADIAH BINTI SAYPOL ANWAR

MATRIC NUMBER : 2019704597

GROUP : AS246 5A1

LECTURER’S NAME : DR. MARINA ZULKIFLI

EXPERIMENT 2 : DETERMINATION OF MINERAL ELEMENTS IN FOOD

PRODUCTS BY ATOMIC ABSORPTION
SPECTROSCOPY -AAS (ASHING METHOD)
INTRODUCTION

Our health relies on our daily balanced meals. Human body needs diverse kinds of
minerals and vitamins for optimum functional system. Nowadays, a lot of food products are
being fortified as a means to increase the intakes on minerals and vitamins of the populations
without increasing the calorie intake. Example of fortification is iron-fortification in cereal flours
and folic acid fortification in cereal-grain products. (G, V.M.M and Hurrell, 2018)

However, there has been cases of nutrient overdoses where getting too much of vitamin
A could lead to increase in risks of birth defects. “In 2013, a group of physicians from university
schools of public health around the country published a review in the “Annals of Internal
Medicine” asserting that dietary supplements carry few potential benefits and, in some cases, are
more harmful than helpful” according to Schuna (2018).

Therefore, it is important to know the content of our food intake. Determination of a

mineral contents in a food can be done by using an instrument named atomic absorption
spectroscopy (AAS). It is a major role-current database for mineral nutrients and toxicants in
foods. Atomic absorption methods measure the amount of energy (in the form of photons of
light, and thus a change in the wavelength) absorbed by the sample. When the sample is excited
by the radiation, the reading of the spectra produced are recorded. The requirement of using this
instrument is that the food sample has to be in a solubilized solution whether in certain organic
solvents or aqueous form.

Some of the components for atomic absorption spectroscopy are light/radiation source,
atomizer, monochromator detector and readout device. Mineral elements in food that is classified
according to Nutritional Essentiality, potential toxic risk for standard reference is calcium by
contamination of lead toxic, phosphorus by mercury, sodium by cadmium, potassium by nickel
and magnesium by arsenic.
OBJECTIVE
1. To determine the mineral content which is sodium (Na) and calcium (Ca) in canned
mushroom and baby corn.
2. To observe the process of atomic absorption spectroscopy (AAS).

APPARATUS

Muffle furnace Crucibles Ashless filter paper

Bunsen burner Pipettes Distilled water

Water bath Funnels Concentrated hydrochloric

acid

Drying oven Beakers Sodium standard

solution (1000mg/l)

Hot plate Volumetric flasks Magnesium standard

solution (1000mg/l)

Zinc standard Calcium chloride 10% lanthanum

solution (1000mg/l) solution (100mg/L Ca) chloride solution

METHODS

1. Preparation for Sample Ashing

a) 5g of homogenized food sample was weighed into a previously dried, cooled and weighed
crucible.

b) For high moisture foods, dried in an oven at 100°C

c) It was heated gently over a Bunsen burner until food was charred and ceased smoking.

d) The crucible was transferred to a muffle furnace (550°C) and left in the furnace until a white
or light grey ash form was obtained. If the residue is black in colour, moisten with a small
amount of water to dissolve salts, dry in an oven and repeat the ashing process.

e) Cooled in dessicators and weighed the ash obtained.

f) 5mL of concentrated acid was added into the crucible containing the ash and mixture was for 5
minutes on a hot plate in a fume cupboard. Acid was added as to maintain the volume.

g) The contents was transferred by washing the residues in the crucible into a beaker with
deionised water.

h) The volume was adjusted to about 40mL and boiled for another 10 minutes over a Bunsen
burner.

i) It was let cooled and filtered using an ashless filter paper into a 100mL volumetric flask. The
beaker was rinsed with deionised water and make up to a volume. It was mixed well with
repeated inversion of the flask.

j) This ash solution was used for determination of individual mineral elements.
2. Preparation of Sample for Calcium Determination

a) 10mL of ash solution was pipetted into a 500mL volumetric flask

b) 1mL of 10% lanthanum chloride solution was added. It was make up to volume with
deionised water and mixed well with repeated inversion of the flask.

3. Preparation of Sample for Other Mineral Determination

a) 10mL of ash solution was pipetted into a 100mL volumetric flask (for determination of zinc)

b) 10mL of ash solution was pipetted into a 100mL volumetric flask (for determination of
sodium or magnesium)

c) It was make up to volume with deionised water and mixed well with repeated inversion of the
flask.

4. Preparation of Standard solution. Preparation stock solution of each mineral element

(100mg/L)

a) 10mL of each (Mg, Na or Zn) standard solution (1000mg/L) was pipetted into a 100mL
volumetric flask and then make up to volume with deionised water. It was mixed well.

b) A series of standard solution of each element to be analysed by dilution of the mineral stock
solution using deionised water in a 100mL volumetric flask was prepared. Use equation

C1V1 = C2V2 to prepare the standard solution. [Note: The concenration used should be in the
linear range of the instrument and appropriate for the amount of the element likely to be present
in the food extract. Typically, these woukd be the order of 1,2,3,4 abd 5mg/L. except for zinc
prepare a standard series of 0.2, 0.4, 0.6, 0.8 and 1.0 mg/L].
c) In the preparation of calcium standard solutions: 1mL if 10% lanthanum chloride should be
added to each flask before making up to volume. This is to minimise the interfernce 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 abosorbance was measured for each of the standard solutions prepared

c) In a similar manner, the absorbance was measured for the sample ash solution. If the
absorbance of this ash solution is too high, a known volume is diluted with deionised water
measurement is 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

3. Absorbance Data for Standard Solution

Mineral measured: Na
Absorbance reading at λ: 589.0nm

STANDARD ABSORBANCE
CONCENTRATIO Average ± S.D
N (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

Graph of Average Absorbance against standard concentration for Na

AVERAGE ABSORBANCE VS STANDARD CONCENTRATION OF Na

1.2

f(x) = 0.21 x
1 R² = 0.99 0.97

0.86
0.8
Average Absorbance

0.71

0.6
0.53

0.4

0.27
0.2

0
1mg/L 2mg/L 3mg/L 4mg/L 5mg/L

Standard Concentration mg/L

Mineral measured: Ca
Absorbance reading at λ: 422.7nm

STANDARD ABSORBANCE
CONCENTRATIO Average ± S.D
N (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
3 0.190 0.190 0.190 0.190 ± 0
4 0.242 0.241 0.242 0.242 ± 0.001
5 0.295 0.295 0.296 0.295 ± 0.001

Graph of Average Absorbance against standard concentration for Ca

AVERAGE ABSORBANCE AGAINST STANDARD CONCENTRATION OF Ca

0.35
f(x) = 0.06 x
R² = 1
0.3 0.3

0.25
0.24
Average Absorbance

0.2
0.19

0.15
0.13

0.1
0.08

0.05

0
1 mg/L 2 mg/L 3 mg/L 4 mg/L 5 mg/L

Standard Concentration (mg/L)

CALCULATION
A. Determination of sodium (Na) in canned food products

FOR MUSHROOM

Equation from curve,

y=0.2133 x

Average absorbance of sodium in mushroom,

y = 0.612

Thus,
0.612
x = 0.2133
mg
2.87
= L

SODIUM CONTENT IN CANNED MUSHROOM,

M
% mineral in sample = W x Z
M = Concentration mineral in food
mg sample
2.87
L W (g) = Weigh of food used
100mg  1L 
(5 g x ) 40ml x  V (ml) = Volume ash solution diluted in
= 1g  1000ml 
100ml

= 0.014%
FOR BABY CORN

Equation from curve,

y=0.2133 x

Average absorbance of sodium in baby corn,

y = 0.757

Thus,
0.757
x=
0.2133

= 3.56 ppm

SODIUM CONTENT IN BABY CORN,

M
% mineral in sample = W x Z

M = Concentration mineral in food

mg sample
3.56
L
100mg  1L  W (g) = Weigh of food used
(5 g x ) 40ml x 
= 1g  1000 ml  V (ml) = Volume ash solution diluted in
100ml
= 0.018%
B. Determination of calcium (Ca) in canned food products

FOR MUSHROOM

Equation from curve,

y = 0.0611

Average absorbance of sodium in mushroom,

y = 0.035

Thus,
0.035
x = 0.0611
mg
0.57
= L

CALCIUM CONTENT IN MUSHROOM,

M
% mineral in sample = W x Z
M = Concentration mineral in food
sample

mg W (g) = Weigh of food used

0.57
L V (ml) = Volume ash solution diluted in
100mg  1L  100ml
(5 g x ) 40ml x 
= 1g  1000ml 

= 2.936 x 10-3 %
FOR BABY CORN

Equation from curve,

y = 0.0611

Average absorbance of sodium in mushroom,

y = 0.036

Thus,
0.036
x = 0.0611
mg
0.59
= L

CALCIUM CONTENT IN BABY CORN,

M
% mineral in sample = W x Z

M = Concentration mineral in food

mg
0.59 sample
L
100mg  1L  W (g) = Weigh of food used
(5 g x ) 40ml x 
= 1g  1000 ml  V (ml) = Volume ash solution diluted in
100ml

= 2.585 x 10-3 %
DISCUSSION

Atomic absorption spectroscopy (AAS) is used for tracing any contents of metals in a
variety of sample. The analysis is done by a implementing technique of measuring quantities of
chemical elements present in wanted samples by measuring the absorbed radiation by the
chemical element of interest. This method can be used to trace metals in drinking water,
beverages, food, pharmaceuticals and even cosmetics. The process of atomic absorption
spectroscopy revolves around 2 steps. Atomization of the sample and absorption of radiation
form a light source by the free atoms. The calculation of concentration is based on BEER-
Lambert Law. The absorbance of an absorbing analyte is proportional to its concentration.

The objective of this experiment is to determine the mineral content of Sodium (Na) and
Calcium (Ca) in canned mushroom and baby corn. A few procedures had been done starting
from the preparation for sample ashing, Sodium and Calcium determination, other mineral
determination, standard solution and absorbance measurements. Sample ashing was done for
decomposition of large sample sizes by heating under muffle furnace. Heating under high
temperature turns the sample into ashes. For preparation of sample, it can be seen that in calcium
preparation sample, 1mL of 10% lanthanum chloride solution was added. According to Mostyn,
Newland and Hearn (1970), “lanthanum is particularly effective as a releasing agent in the
determination of calcium and act as as a chemical interference suppressor ”.
 Picture above shows the components for atomic absorption spectroscopy (AAS). It works
on the basis of atomization. A sample will enter by changing its form into aerosol form by
nebuliser. Hollow cathode lamp consists of a hollow tube filled with argon/neon, an anode (made
from tungsten) and cathode (metallic form of element being measured). The lamp gives out the
exact wavelength required for the analysis. The atoms of the metal tested are present in the lamp.
Therefore, when the lamp is turned on, these atoms are supplied with energy, causing them to
elevate to the excited levels. While the monochromator works by making sure only the desired
wavelength reaches the detector. Lastly, before getting the full data, detector will converts the
radiant energy into electrical signal which will produce analog or digital readout.

From the graph of average absorbance against standard concentration of Na, it can be
seen that there is a deviation. Deviation occurs when the concentration of metallic analytes
increases. It might happen due to un-absorbed radiation, stray light or disproportionate
decomposition of molecules at high concentrations. This can be avoided by preparing a blank
and sufficient samples to fit standard curve appropriately.

In canned mushroom, the amount of sodium (Na) present is 0.014% while for baby corn,
it consists of 0.018% sodium (Na) content. Whereas, the amount for calcium (Ca) in canned
mushroom is 2.936 x 10-3% while for baby corn it is 2.585 x 10-3% . Baby corn has a higher
content in both sodium (Na) while canned mushroom has high content in calcium (Ca). The
linear regression (R2) of calcium and sodium were 0.9118 and 0.9742 respectively. This indicates
that the preparation of standard solution were successful as it is above 0.9. The value of x for

mg mg
calcium (Ca) and sodium (Na) in mushroom is 0.57 L and 2.87 L respectively. For sodium
 mg mg
(Na), in baby corn, it is 3.55 L and for calcium (Ca) in mushroom is 0.59 L respectively.
According to Intech (2012), fresh mushroom consists of 1mg/100g of calcium. As can be seen
from the result, canned mushroom has half of the nutrient.

A few things should be considered while using this atomic absorption spectoroscopy
(AAA) is that samples and instrument should not be prepared under direct AC. Instead, sample
digestion should be done under fume hood. Gloves should also be use to avoid contamination of
sample.

CONCLUSION

As a conclusion, both objectives have been successfully achieved. The analytical

technique which Atomic Absorption Spectroscopy (AAS) is used in order to determine the
concentration of the calcium and sodium content in canned mushroom and baby corn has been
learnt. This techniques uses sample that has turnt into ash. The concentration of the calcium and
sodium content in the unknown solution can be determined by calibration curve. For sodium
(Na) content, it seems that, baby corn has a higher content than canned mushroom with 0.018%
and for calcium (Ca), canned mushroom has a higher content of 2.936 x 10 -3%. From the
standard solution which was prepared, the linear regression (R2) of calcium and sodium were
0.9118 and 0.9742 respectively.
REFERENCES

G. , V. M. M., & Hurrell, R. (2018). Food fortification in a globalized world. Amsterdam:

Academic Press.
INTECH. (2012). Atomic Absorption Spectroscopy.
Mostyn, R., Newland, B., & Hearn, W. (1970). Lanthanum as a releasing agent in atomic
absorption spectroscopy in organic solvents. Analytica Chimica Acta, 51(3), 527–
528. doi: 10.1016/s0003-2670(01)95752-6
Schuna, C. (2018, November 19). What Are the Dangers of Fortified Foods and Supplements?
Retrieved from https://healthyeating.sfgate.com/dangers-fortified-foods-
supplements- 11621.html