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Objectives :
Background :
Glucose, a monosaccharide (or simple sugar), is the most important carbohydrate in biology.
Transported via the blood stream, it is the primary source of energy for the body’s cell.
Glucose levels are tightly regulated in the human body. Failure to maintain blood glucose in
the normal range leads to conditions of persistently high (hyperglycemia) or low
(hypoglycemia) blood sugar. Diabetes mellitus, characterized by persistent hyperglycemia, is
the most prominent disease related to improper blood sugar regulation.
Where A = absorbance
ε = molar absorptivity of the substance
C = concentration (for clinical measurements, the concentration often used is
mg/dl)
L = pathlength (the length the light travels through the cuvette)
We assume that the y-intercept is zero. In other words we are assuming that the
absorbance of a solution is zero when none of the substance we are measuring is present. We
do this by using a blank. A blank solution contains all the substances present in the solution
except the substance to be measured. Before making measurements with the Spec 20, the
blank is measured, and its absorbance value is set to zero. By using a blank, if water or sugar
absorbs light, that absorption will be set to zero, and thus the y-intercept on our linear Beer’s
Law equation would indeed be zero.
RESULT AND CALCULATIONS
We pour three identical solutions that we have prepared into three cuvettes to measure the
absorbance and take the average. From there we got 0.0833 absorbance of unknown solution
(average). From the given standard curve equation y = 3.5329 x, we divide the absorbance by
the slope of the standard curve. We calculated our concentration (x) to be 0.0235 mg/mL.
From here, we know that on average, only 0.0235 mg/mL of glucose present in the
0.0833 mg/mL of our unknown solution. In other words, glucose appears to be 28.2% of the
actual unknown concentration. Our result might differ from other groups based on our
pipetting process. If our pipetting has been accurate and precise, everyone should have the
same result.
Heating is necessary to accelerate the chemical reaction. We heat our mixture of DNS
solution and unknown solution until it develops to red-brown color. It is aware that the
heating process should only be between 5 to 15 minutes. Overheating will cause the chemical
reaction to stop. Therefore, we use a timer to make sure we didn’t stop the reaction.
Some safety precautions that we need to observe includes using test tube holders to
handle hot tubes to avoid hand injuries. Since we are dealing with DNS solution which is
toxic, we need to wear goggles at all times to avoid any contact with the skin. If any is spilled,
wash thoroughly with soap and water. At the end of the session, we need to clean up before
we go for others convenience.
CONCLUSION
REFERENCES
Dr. Mohamed E. S. Mirghani, Lab Manual for Biotechnology Engineering Lab 1 BTE
2221
http://www.bio.davidson.edu/courses/bio111/bio111labman/preface%20d.html
http://www.jenway.com/adminimages/A09_008A_Determination_of_glucose_in_drin
ks.pdf
http://employees.csbsju.edu/mcampos/bio114/labmaterials/lab.2.writeup.03.pdf
http://biochem.ncsu.edu/faculty/brown/Lab%20Exercise%200ne%202008.ppt
http://bric.postech.ac.kr/myboard/down.php?
Board=exp_qna&filename=Determination%20of%20sugar%20as
%20glucose.pdf&id=42071&fidx=1
http://www.kau.edu.sa/GetFile.aspx?id=156654&fn=bioc_211(lab1).doc