BIOL2537 Laboratory in Nutritional Science

Name: Hung Wing Tung

UID: 2009053464
Group: 3
Date: 11/3/2011

Practical 2: Protein Quality and Urea Production in

Saliva
Objective: To demonstrate the quality of protein of a test meal by
determining the concentration of urea in saliva by spectrometry
analysis
Introduction:
After digestion of proteins from diet, amino acids will be released.
These amino acids are either used to synthesize tissue protein or
converted into nitrogenous breakdown products, mainly urea. The
extent of catabolism, and incorporation of dietary amino acids into
tissue protein are determined by several factors, such as the
individual need for growth and repair, energy and protein adequacy
of the diet and the essential amino acid composition of the food
protein (protein quality). The protein quality of a food depends on
whether the essential amino acid composition matches with the
ideal pattern, which can be used with 100% efficiency for tissue
protein synthesis.
For any dietary amino acids not used for protein synthesis will be
available for urea production theoretically. Therefore, the amount of
urea produced after ingestion of a food usually reflects the protein
quality of that food. Since urea is a small molecule that readily
diffuses through all body fluids, in this practical, the urea
concentration in saliva at various time intervals will be used to test
the protein quality of 3 different foods (eggs, cheese and peanuts).
Procedures (Materials and Methods): As shown in lab manual.
Results:

Graph (a) The standard curve of urea concentration

Table (a) Diluted, original and change in urea concentration

in saliva after consuming cheese at different time intervals
Time intervals (hr)
0
0.5
1.0
1.5
Diluted urea
concentration
0.14
0.13
0.15
0.13
(umol/ml)
Original urea
concentration
1.12
1.04
1.20
1.04
(umol/ml)
Change in urea
concentration
0
-0.08
+0.08
-0.08
(umol/ml)

Graph (b) My change in urea concentration against time

0.1
0.08
0.06
0.04
0.02

Change in urea concentration (umol/ml)

0
0
-0.02

0.5

1.5

-0.04
-0.06
-0.08
-0.1

Time (hr)

Table (b) Class result of total and change in average urea

concentration in saliva after consuming 3 test meals at
different time intervals
Time intervals (hr)
0
0.5
1.0
1.5
Average
Peanuts
1.52
2.54
2.45
2.92
urea
1.68
3.07
2.49
2.85
concentra Cheese
tion
Egg
1.61
2.11
2.35
2.29
(umol/ml)
Change
Peanuts
0
+1.02
+0.93
+1.40
in
average
Cheese
0
+1.39
+0.81
+1.17
urea
concentra
Egg
0
+0.50
+0.74
+0.68
tion
(umol/ml)

Graph (c) The class result of change in average urea concentration in saliva after consuming 3 test meals at different time interval
1.5
1.4
1.3
1.2
1.1
1
0.9
0.8

Peanuts

Change in urea concentration (umol/ml) 0.7

Cheese
Egg

0.6
0.5
0.4
0.3
0.2
0.1
0

Time (hr)

Table (c) Change in urea concentration in saliva after

consuming 3 different test meals throughout the
experimental time
Type of food
Peanuts
Cheese
Egg
Change in urea
concentration
throughout the
+1.35
+1.43
+0.81
experimental time (area
under curve) (hrumol/ml)

Graph (d) Change in urea concentration in saliva after consuming 3 different test meals throughout the experimental time
1.6
1.4
1.2
1
0.8

Change in urea concentration over time

0.6
(hr umol/ml)
0.4
0.2
0

Test meals
Discussion:
Change in urea concentration in own saliva sample at different time
intervals
Referring to Graph (b), my individual change in urea concentration
in saliva did not have significant change after the consumption of
cheese. The fasting (0 hr) urea concentration was 1.12 umol/ml.
After consuming the cheese, it decreased to 1.04 umol/ml at 0.5 hr,
and then increased to 1.20 at 1.0 hour and eventually dropped to
1.04 umol/ml again at 1.5 hour. However, all these changes were
too little to reflect the protein quality of the cheese.

Comparison of mychange in urea concentration with class results

Graph (e) Comparison of the my change in urea concenration in saliva aftering consuming cheese with the class results
1.6
1.4
1.2
1

Change in urea concentration (umol/ml)

0.8
0.6

My results
Class results

0.4
0.2
0

1.5
-0.2-0.5

Time (hr)
My results were deviated from class results that my urea
concentration did not have significant change throughout the
experimental time, while the class results had a fluctuated but
general and significant increase in urea concentration. Besides, my
directions of changes were totally opposite to class results. First, the
urea concentration in class saliva samples increased after
consuming the cheese for 0.5 hr, but my results was a reduction in
urea concentration. When the class results showed a significant drop
in urea concentration at 1.0 hr, my urea concentration increased a
bit at same time interval. At 1.5 hr, when the class urea
concentration increased, my urea concentration dropped a little.
The difference between mine and class results can be attributed to
personal factors. For instance, although I ate all assigned cheese
according to my weight and energy needs, I may have greater
protein needs for tissue growth and repair than the others and thus
more amino acids from the cheese protein were absorbed and
incorporated for the synthesis of tissue protein. Therefore, less urea
were produced and diffused to the saliva. Besides, the difference
might be due to the presence of drinking water in my mouth cavity
during collection of saliva samples, which greatly diluted the urea
concentration in my saliva.
Difference in the change of average urea concentrations in different
test meals
From the class results, the change in average urea concentrations in
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saliva samples for all test meals increased. This was because part of
the dietary amino acids wasused for urea production, causing more
urea diffused to the saliva.
The consumption of egg caused the lowest increase in average urea
concentration, followed by peanuts, and lastly cheese. This reflects
that a higher portion of amino acids from egg protein are
incorporated for tissue protein synthesis than cheese and peanuts.
Thus,egg has the highest protein quality among the 3 test meals. In
fact, egg is regarded as the food having excellent protein quality (a
complete protein) since it is rich in leucine, lysine, glutamate, and
all other essential amino acids. These essential amino acids cannot
be synthesized in the body and are required to taken in from diet.
Therefore, when food containing these essential amino acids is
consumed, the essential amino acids will be readily absorbed by the
body as building blocks of many cellular compounds but not
converted to nitrogenous compounds for urea production. This is the
reason why less urea was present in the saliva after taking in egg
than taking in other 2 test meals.
Although the experimental results showed that consumption of
cheese caused a higher increase in average urea concentration than
peanuts, theoretically, the urea production owing to intake of
cheese should be less than that of peanuts. This is because cheese
is a dairy product, which contains allessential amino acids derived
from milk protein (also a complete protein), while peanuts only
contain legume protein with low concentration of lysine and
methionine. More urea should be produced from the non-essential
amino acids present in peanuts. The inconsistency between the
experimental results and theory might be attributed to some
experimental errors, or the fact that the cheese might have been
undergone some food processing procedures, and some of these
procedureshave caused great milk protein loss.
Possible experimental errors which affect the overall class results
There were several errors that might affect the class results. First,
some subjects might have eaten meals within the 3 hours before the
consumption of test meals. This caused the measurement of urea
concentration in the saliva less reliable since the urea produced
might be derived from the food other than the test meals. Second, if
the subjects did not consume the test meals according to their
weight and energy needs, for example, eating less than the amount
assigned, more food protein from test meals might be oxidized for
ATP synthesis. This might overestimate the protein quality of the
test meals. Third, for subjects who had difficulties in producing
saliva, they might rely on drinking lots of water to assist the saliva
production. This might lead to the presence of water in the mouth
cavity which diluted the urea concentration of saliva, causing
overestimation of protein quality of the test meals. Fourth, since the
7

subjects were all unique individuals, they might have different body
metabolism and hence different protein utilization. This caused the
results of some subjects much deviated from the average. Fifth, the
standard curves derived by different groups might be different, so
the urea concentrations in saliva determined by the standard curve
for one group might not fit with that determined by another
standard curve. Sixth, the poor experimental skills of experimenters
in preparing the sample solutions might also affect the results.
5 common foods in Hong Kong that are considered as High Quality
Protein diet
The recommended dietary intake of all essential amino acids is
illustrated in the following table:
Recommended dietary
Amino acids
allowances for adults
(mg/kg/day)
Histidine
14
Isoleucine
19
Leucine
42
Lysine
38
Methionine + cysteine
19
Phenylalanine + tyrosine
33
Threonine
20
Tryptophan
5
Valine
24
A High Quality Protein must be a complete protein which contains all
these essential amino acids in the approximate amounts needed by
humans.Most of the high quality proteins come from an animal
origin. In Hong Kong, the 5 common foods which belong to High
Quality Protein diet are namely egg, milk, cheese, chicken and beef.
Conclusion:
The protein quality of egg was the highest, followed by peanutsand
then cheese.
Reference:
S. Gropper, J. Smith, J. Groff. 2009. Advanced Nutrition And Human
Metabolism (5th Edition).