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The document discusses an experiment that aims to compare how quickly the enzyme amylase can break down different types of flour by measuring the time it takes for a purple solution to turn clear. It will test kamut flour, white refined flour, and potato flour.

The experiment aims to see how fast the enzyme amylase can break down starch into glucose in each type of flour by measuring the time it takes for a purple solution to turn clear after the addition of amylase to each flour-water mixture. This will indicate the relative digestion speeds between the flours.

The independent variable is the type of flour, and the dependent variable is the time taken for the purple solution to turn clear. The temperature, amounts of flour and amylase, and pH will be controlled.

INTERNAL ASSESMENT BIOLOGY

Purpose:
My interest in finding out more on how different types of food that we eat have a
different effects on our digestive system developed as I decided to follow a diet.
Before, I used to eat carbohydrates made just by white refined flour which I was later
told to be the type of flour that lacks the most of good nutrients. For this reason Ive
cached the opportunity of the internal assessment to investigate and see practically
how and why different types of flours that contain different quantities of the same
nutrients are processed and digested differently by our digestive system, more
specifically by amylase. Ive chosen Alpha-amylase, which is the enzyme found in
saliva, as it is one of the fist processes needed for digestion. Moreover, its optimum pH
is quite neutral and this would help me to set the experiment more easily.
Research question:
How and why does the action of amylase occur differently on different types of flour
(white refined, kamut and potato flour) which nutritional values differ significantly on
the amounts of proteins, fibre and carbohydrates?
Background:
Nutritional values for 100g:
Type of flour

Protein (g)

Fat (g)

kamut
Potato
Whiterefined
[Table1]

14.70
6.90
10.33

2.20
0.34
0.98

Total carbs
(g)
70.38
83.10
76.31

Fibre (g)
9.1
5.9
2.7

Net carbs
(g)
61.28
77.20
73.61

Alpha-Amylase is a digestive enzyme whose function is to catalyse the 1, 4--Dglucosidic linkages in starch, polysaccharide, into maltose, disaccharide, which contain
several 1,4--linked D-glucose units, monosaccharide (Figure 1). Glucose (Figure 2)
is one of the most important molecules for the life of human beings as it is the starting
point of both aerobic and anaerobic respiration.
Since flour is mainly made of starch, as it is clearly visible in the table above thanks to
the big values of Net carbs, this experiment aims to see how fast the action of
amylase to break down starch into glucose is, in white- refined flour, kamut flour and
potato flour. I chose these three types of flour because I also want to see if the amount
of proteins and fibre have a significant effect on amylase action. As a matter of fact
Ive chosen white-refined flour because it has a high amount of net carbs and proteins
but a very little amount of fibre while kamut flour has a higher amount of fibre and
proteins compared to white refined flour but a significantly lower amount of net carbs.
Instead, Ive chosen potato flour because it has quite peculiar nutritional values as it
has very little amount of proteins and fibre but a very big amount of net carbs.
The experiment will be displaced in four different backers containing an
heterogeneous mixture made of water and one type of flour (one in each backer).
Then I would add to the heterogeneous mixture another mixture made of amylase,
potassium permanganate and sulphuric acid. These last two compounds give a purple

solution that would get later oxidised by the presence of glucose. This process cause a
change in colour of the solution, going from purple to transparent. This change allows
the gathering of quantitative data expressed in the time taken for the solution to
become transparent.

Hypothesis:
From my experiment I expect kamut flour to take more time to be digested as the hull
of the grain is still partially present while in white refined flour the hulls has been
removed during the refinetion process. This difference should merge significantly in
this experiment as starch is closed in the grains hulls which are mostly made of fibres.
As a consequence amylase would act more laboriously and less glucose would be
present in the solution at the end of the experiment. Following this logic, as potato
flour also has a quite high amount of fibre, the action of amylase on it should be lower
compared to white refined but a little bit higher compared to kamut. However, as
potato flour doesnt come from grain but from dried potatoes, this logic could also be
wrong.
I would rely on the experiment made by Janelle Tang(1) in which she states as the
concentration of glucose solution increased, the amount of time it took for the solution
to decolourise decreased exponentially. Basing my experiment on such statement I
would be able to observe on which of the three final solutions, that contain the three
different types of flours, amylase would have a bigger action over the same period of
time. My result would be expressed in time as the solution that takes more time to
decolorize would be the one with a smaller concentration of glucose, so the one on
which the action of amylase occurred the less.
http://www.academia.edu/3772012/Unknown_Glucose_Concentration
(Figure 1)

(Figure 2)

Variables:
- Independent, type of flour
- Dependent, time taken for the pink colour of potassium permanganate solution
to disappear (demonstrative of glucose concentration)
- Controlled, temperature (room temp), amount of flour and amount of amylase,
pH of the solution
Materials:
- Alpha amylase
2mg (=60U) diluted in 1,2ml of water 50mM solution of maltose
6mg (=180U) diluted in 3,6ml of water 50mM solution of maltose
333mg(=999U) diluted in one 1ml of water 1M solution of maltose
1g(=3000U) diluted in 3ml of water 1M solution
-

2ml of diluted Potassium permanganate (KMnO4, 0.025M), dissolved in 1:4


water solution
- 5ml Sulphuric acid (H2SO4) 1M
- 2,5 g Kamut flour
- 2,5 g Refined white flour type 00
- 2,5 g Potato flour
- 100 ml Water
- 3 backers
- Cylinders
- Balance
- Spoon
- Stopwatch
Procedure
-

Heat water at 40C


Put flour in hot water
Mix the solution for one minute

Filter the solution


Take the filtered solution and add to it ?ml of amylase diluted in physiological
solution
- Wait for 5 minutes
- Transfer 10ml of the solution in another backer
- Pour 5ml of sulphuric acid and 2ml of potassium permanganate in the solution
- Turn the stopwatch on
- Turn it off as soon as the solution turns clear (transparent) again
- Do it at least 3 times for each flour
http://www.immuneweb.org/lowcarb/food/grains.html

Unit Definition
1 U corresponds to the amount of enzyme which liberates 1 mol maltose per minute at
pH 6.0 and 25C (starch acc. to Zulkowsky, Cat. No. 85642, as substrate).
http://www.sigmaaldrich.com/catalog/product/sigma/10065?lang=it&region=IT

http://www.biolog.de/technical-info/umol-um-conversion/

SOURCES
(Figure 1)
Pujari, Saritha. "3 Major Classifications of Carbohydrate and Its
Significance." YourArticleLibrarycom The Next Generation Library. Your Article Library,
03 Jan. 2014. Web. 11 May 2016.
(Figure 2)

Ophardt, Charles E. "Carbohydrates - Glucose." Virtual Chembook. El Mhurst College,


2003. Web. 11 May 2016.
[Table 1]
[Taken from: Norwitz, Cyndi. "Nutritional Values of Grains & Flours." Nutritional Values
of Grains & Flours. N.p., 11 Nov. 2007. Web. 10 Apr. 2016 ]
"Carbohydrate Analysis." Enzyme Explorer. SIGMA-ALDRICH, n.d. Web. 24 Apr. 2016.

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