The Mighty Egg By Nazathul Syakirah Ang (D20101037476)

Introduction Egg is an excellent source of protein. We can get the protein throughout the entire egg. Protein monomers are made up of amino acids. Therefore, by eating egg we can get all the essential amino acids that are needed by our body. Not only that egg provided us with protein, it is also a valuable source of vitamins and minerals. Before this, I never thought that the concentration of protein in egg would vary in different type of eggs. Furthermore, it never crossed my mind that we can actually determine the protein concentration of an egg in a school laboratory only by using a simple instrument which is the spectrophotometer. Spectrophotometer is a very powerful tool yet the operations are simple, a beam of light is shine, filtered to a specific wavelength, through a sample and onto a light meter. Some basic properties of the sample can be determined by the wavelengths and amount of light absorbed by the sample. Using spectrophotometer, we can determine the absorbance of protein, hence determining its concentration.

Spectrophotometer Protein Assay In experiment that I had conducted with my friends on protein assay last week, we are using Biuret assay and Lowry assay to determine the concentration of protein in different egg types. We put the solution in a sample holder called cuvette and place it in spectrophotometer. Cuvette comes in many types which are plastic, glass or fused quartz. When using Biuret assay, the absorbance are measured at 540 nm, while for Lowry assay at 750 nm.

Before determining the concentration of protein of the test samples, we must plot a standard curve. To be able to do so, the absorbance of the different concentration of gelatine is measured using the spectrophotometer. Solution of gelatine is prepared at 1, 2, 3, 4, 5, and 6 mg/mL for Biuret assay and 0.1, 0.2, 0.3, 0.4, 0.5, and 0.6 mg/L for Lowry method. From the absorbance value that we get for each solution, a standard curve for Biuret assay and Lowry assay is plotted. For both graph 1 and graph 2, I used Microsoft Excel. This is the first time that I used this Microsoft application and I am very pleased with it as this Microsoft Excel is easy to use. It took only few minutes to learn the basic steps in plotting graph and getting the best fit line for both graphs.

Graph of Absorbance Vs Concentration of Gelatin for Biuret Assay

0.350 y = 0.0438x 0.300 0.250 Absorbance 0.200 0.150 0.100 0.158 0.250 0.296

0.199
0.190

0.222

0.050
0.000 1 2 3 4 mg/mL, Concentration 5 6 7

Graph 1

Graph of Absorbance Vs Concentration of Gelatin for Lowry Assay

0.400 0.38 0.350

0.300
Absorbance 0.250 0.200 0.150 0.100 0.050 0.000 1 2 3 4 mg/mL, Concentration 5 6 0.040 0.062 0.25

0.32

y = 0.041x

0.111

Graph 2 From graph 1, the equation of the standard curve is y = 0.043x, while the equation for the standard curve in graph 2 is y = 0.041x. It is important to get the best standard curve because the mathematical formula describing the fitted curve will be used to calculate the concentration of the test samples. Comparing the equation of the standard curve for both Biuret assay and Lowry assay, we can see that there is not much difference in their value. Test samples Five different eggs are tested using both Biuret and Lowry assay to determine which eggs contain the highest protein. The five samples are: Sample 1 2 3 4 5 Type of Egg Ducks egg Chickens egg Grade C egg Omega 3 egg Quail egg

Protein samples are processed in the same manner by mixing them with assay reagent and using a spectrophotometer to measure the absorbance. Absorbance values of the five samples are then interpolated onto the formula for the standard curve to determine their concentration. Below are the bar chart showing which samples contained the highest protein.

Concentration of Protein for Test Sample (Biuret Assay)

20.000 17.605 14.047 17.419

concentration

15.000 10.000 5.000 0.000 1 2 3

12.907
10.791

sample Figure 1

Concentration of Protein for Test Sample (Lowry Assay)

69.390 70.000 56.537 62.439 60.000 50.000 40.000 30.000 20.000 10.000 0.000 1 2 3 Sample 4 5

Concentration

51.732

55.610

Figure 2 From both bar chart in figure 1 and figure 2, the protein concentration inside test samples in order are duck egg > grade C egg > Chicken egg > quail egg > omega 3 egg. The egg contained the highest protein for both Biuret assay and Lowry assay is the same

which is the duck egg. However, the concentration is different. In Biuret assay, the concentration of protein in duck egg is 17.605 mg/mL, while in Lowry assay the concentration is 60.390 mg/L. Both Biuret assay and Lowry assay shown that omega 3 eggs give the lowest concentration of protein. Omega 3 eggs are from chickens that have been fed by a diet supplemented with a source of omega-3 fatty acids. Even though this results shows that omega-3 eggs contained the lowest protein, we cannot say that this type of eggs will not give benefits to us when we consumed it. Omega 3 fatty acids in the egg are also essential for health, immunity and brain development. Omega 3 fatty acids also possess anti-inflammatory qualities. Even though eggs are high in protein and vitamins, it is low in calories and carbohydrates. Furthermore, eggs are excellent source of choline and a good source of riboflavin. The protein in eggs helps adults build and preserve muscle strength and allow them to stay energized. Throughout the lab session, Ive learn a lot about protein assay, how to use the spectrophotometer and how to use Microsoft Excel. Before starting the experiment, we were questioned on which types of egg contained the highest protein. I answered that the chicken egg should contain the most protein as we normally consume chicken egg compared to the other sample. The final result is different from what I imagine it would be. I have a lot of fun conducting this experiment. Conclusion From five samples that are tested, duck egg contained high concentration of protein followed by grade C egg, chicken egg, quail egg and lastly omega 3 eggs. From this experiment, the duck egg is the mighty egg compared to the others if we referred to its protein concentration. References Shereen J. (2012). How Much Protein Is In an Egg? Retrieved November 27, 2012 from http://nutrition.about.com/od/askyournutritionist/f/eggs_protein.htm Donald A. (2011). Omega 3 in Eggs. Retrieved November 27, 2012 from

http://www.livestrong.com/article/283853-omega-3-in-eggs/