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Lab Report 3 sbl1023

This laboratory report describes an experiment using spectrophotometry to measure and compare protein levels in rice and porridge samples. Bovine serum albumin standards were prepared and measured to generate a standard curve. Absorbance readings of rice and porridge protein samples were then taken and their protein concentrations determined using the standard curve. The porridge sample had a higher absorbance reading, indicating it contained more protein than the rice sample. The objectives of learning spectrophotometry technique and measuring food protein levels were achieved.

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342 views

Lab Report 3 sbl1023

This laboratory report describes an experiment using spectrophotometry to measure and compare protein levels in rice and porridge samples. Bovine serum albumin standards were prepared and measured to generate a standard curve. Absorbance readings of rice and porridge protein samples were then taken and their protein concentrations determined using the standard curve. The porridge sample had a higher absorbance reading, indicating it contained more protein than the rice sample. The objectives of learning spectrophotometry technique and measuring food protein levels were achieved.

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© © All Rights Reserved
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LABORATORY REPORT

TECHNIQUE IN BIOLOGY AND BIOCHEMISTRY LABORATORY

(SBL1023)

STUDENT NAME ALISYA SYAZRIN ADILA BINTI


MOHD NASIR
REGISTRATION NO. E20161015898
GROUP B
EXPERIMENT EXPERIMENT 3:
SPECTROPHOTOMETER
(protein/enzyme)
LECTURER NAME ASSOCIATE PROF. DR SHAKINAZ
DESA
PRACTICAL 3: SPECTROPHOTOMETRY (PROTEIN/ENZYME
ANALYSIS)

INTRODUCTION
There are many ways to measure protein concentration; one of the common protocol is by
measuring the absorbance of a coloured product formed by the protein and an organic
molecule. Different methods can yield somewhat different results for the same protein as
there is no absolute photometric protein concentration assay. All methods have advantages
and disadvantages and a few aspects will be taken into consideration: specificity, sensitivity,
the measureable range of concentration, the accuracy, the nature of the protein tobe
examined, the presence of materials interfering with the measurement, and the time required
for the measurement.

OBJECTIVES
1. To learn how to use the sperctrophotometry.
2. To find the amount of protein from food sample.

MATERIALS
Stock solution of Bovine Serum Albumin (BSA): 10mg/ml
Deioinized water (dH2O)
Test tubes and stand
Pipette
Biuret reagent
Spectrophotometer
Protein samples

PROCEDURES
(A) Protein preparation
1. 2 sets of test tubes was prepared with the number 1 to 6 and the bovine serum albumin
(BSA) stock solution (10mg/ml) according to the concentration listed below was prepared:

Tube BSA conc. H2O (ml) BSA stock (ml) Total


(mg/ml)
1 0 1.0 0 1
2 1 0.9 0.1 1
3 2 0.8 0.2 1
4 3 0.7 0.3 1
5 4 0.6 0.4 1
6 5 0.5 0.5 1
7 6 0.4 0.6 1

2. Duplicate test tubes was prepared for proteins samples and carefully pipette 1ml of the
protein samples into each tubes.
3. 2ml of biuret reagent was added to every tubes: the 14 tubes for the standard curve and
the duplicate tubes for protein samples.
4. The tubes was covered with parafilm and briefly vortex to ensure that the protein
standards/samples and the biuret reagent are thoroughly mixed.
5. The test tube was stand at 15 minutes.
6. The spectrophotometer was switched on and the wavelength was adjusted to 550nm.

(B) Determine protein concentration


1. 1ml of solution was transferred from tube 1 into a cuvette and gently wipe the cuvette with
a paper towel to remove fingerprints and dust.
2. The absorbance was set into ‘zero’.
3. The absorbance was measured of the other standards and sample proteins using the step
as in (1).
4. The absorbance of each standards and sample was recorded.
5. The graph of standard curve was plot using the absorbance value of protein standards
and interpolate the absorbance values of the protein samples.

RESULTS
Data of results:

TITRATION CONCENTRATION ABSORBANCE


1 0 0
2 1 0.005
3 2 0.039
4 3 0.119
5 4 0.176
6 5 0.313
7 6 0.330
8 Rice (1ml) 0.560
9 Porridge (1ml) 0.880
Absorbance of protein sample
1
0.9
0.8 y-axis = conc.
0.7 x-axis = abso.
ABSORBANCE

0.6
Blue line = rice
0.5
0.4 Green line = porridge
0.3
0.2
0.1
0
1 2 3 4 5 6 7 8 9
CONCENTRATION

Graph 1: Absorbance of protein sample

DISCUSSION
As we can see in Graph 1, the data that we get is not accurate. But we can find the
concentration through the absorbance that we get from the protein sample such as rice and
porridge. When we do this experiment, we can see that the porridge more protein compared
to rice. This is because the porridge less starch than rice. This in turns cause the graph that
we get proven the porridge is more absorbance than rice. The absorbance of porridge is
0.880 but the rice is 0.560. The porridge have higher absorbance compared to rice. From the
experiment that we do, we achieve all objectives that this experiment need.

CONCLUSION
The conclusion that we get from this experiment is the porridge have more protein compared
to rice because the absorbance that we get, the porridge get higher absorbance than rice.
So this experiment prove that porridge is more healthy than rice.
REFERENCES
1. Protein, Nancy Dickmann, Chicago ILL Heinemann Library, 2012.
2. Protein Hydrolystates in Biotechnology, Vijai K Pasupuleti Arnold L Demain, Dordrecht
London Springer, 2010.

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