RESULTS:

Part A – Determination of Glucose in Urine

Figure 1: Showing the results obtain for Benedict test with urine sample.

Urine samples Observation

Before taking oral glucose dose (left) No changes

Oral glucose dose taken after 2 hours (right) Blue-green solution formed
Table 1: Showing the tabulated results of Benedict test.

Part B – Determination of blood glucose

Sample Absorbance (505 nm)

Blank 0.000
Standard 1.146

T0 0.278

T1 0.370

T2 0.453

T3 0.485
Table 2: Showing the tabulated results of the blood glucose determination.
 Graph of Absorbance vs. Concentration of Glucose Standard
1.4

1.2
Absorbance (at 505 nm)

0.8

0.6

0.4
y = 0.041x
0.2

0
0 5 10 15 20 25 30
Concentration of glucose standard (mmol/L)

Figure 2: Showing the graph of absorbance at 505nm against the concentration of glucose
standard, mmol/L.

From the graph equation, we can calculate the concentration of blood glucose in each sample

y = 0.041ϰ

For urine samples of T0,

y = 0.041ϰ

0.278 = 0.041ϰ

ϰ = 6.78 mmol/L

Therefore, the concentration of blood glucose in urine samples of T0 is 6.78 mmol/L.

For urine samples of T1,

y = 0.041ϰ

0.370 = 0.041ϰ
ϰ = 9.02 mmol/L

Therefore, the concentration of blood glucose in urine samples of T1 is 9.02 mmol/L.

For urine samples of T2,

y = 0.041ϰ

0.453 = 0.041ϰ

ϰ = 11.04 mmol/L

Therefore, the concentration of blood glucose in urine samples of T2 is 11.04 mmol/L.

For urine samples of T3,

y = 0.041ϰ

0.485 = 0.041ϰ

ϰ = 11.83 mmol/L

Therefore, the concentration of blood glucose in urine samples of T3 is 11.83 mmol/L.

Concentration of blood
Urine samples Time (hours)
glucose (mmol/L)

T0 0 6.78

T1 1 9.02

T2 2 11.04

T3 2.5 11.83

Table 3: Showing the tabulation of the calculation, hours and the respective concentration of
blood glucose mmol/L.
 Graph of Concentration of Blood Glucose vs Time
14

12

10
Time (hrs)

4
y = 2.0349x + 6.8695
2

0
0 0.5 1 1.5 2 2.5 3
Concentration of blood glucose (mmol/L)

Figure 3: Showing the graph of time, hours, against the concentration of blood glucose,
mmol/L.

From the graph obtained, the concentration of blood glucose increases when the fasting
period gets longer after taking an oral glucose dose.

DISCUSSIONS:

The purpose of this experiment is to study the presence of glucose in the urine sample and
blood sample provided. The glucose tolerance test is a test whereby the patient will be asked
to ingest glucose and blood sample is taken afterward to determine how whether it has been
cleared from the blood. Firstly, Benedict test, a simple reducing sugar test were done. Benedict
test is a very simple test as the basis of this test under alkaline condition is able to reduce cupric
ions 𝐶𝑢2+ to cuprous ions 𝐶𝑢+ , which is responsible for the change in color of the reaction
mixture. (Refer Figure 1) The color changes is proportional to the amount of reducing sugars
present in the sample. The normal amount of glucose in urine is 0 to 0.8 mmol/L. A higher
measurement could be a sign of a health problem. The presence of glucose in urine is called
glucosuria and is usually an indication of diabetes mellitus. In some cases, a high amount of
glucose in urine can be due to pregnancy. Pregnant women tend to have higher urine glucose
levels than women who aren’t pregnant. Anyhow, pregnant lady should be careful as elevated
glucose level in their urine should be carefully screened for gestational diabetes. Comparing
the result obtained, blue-green solution (Figure 1) and the Figure 2 below, it can be said that
there is a traces of glucose in the urine. Overall, it is important to carried out this benedict test
two hours after sugar intake, as in the initial stages of the disease, a diabetic patient does not
lose sugar in his urine, when on empty stomach. Hence if the Benedict's test is performed in
the fasting state, it is possible to miss the diagnosis of the disease.
Even if sugar is detected in the urine by Benedict's test, the diagnosis of diabetes should be
confirmed by blood-analysis.

FIGURE 4: Showing the color coordination that may formed when the Benedict reagent react
with urine sample.

Second test is using blood plasma sample to estimate the amount of glucose present. Time
interval observation were done after the patient ingested the oral glucose. The reference value
of glucose at 1 hour GTT (Glucose Tolerance Test) glucose level below 10 mmol/L is
considered normal. While for 2 hour GTT (Glucose Tolerance Test), a glucose level below
7.8 mmol/L considered normal. Blood plasma glucose between (7.8 – 11.1) mmol/L indicate
impaired glucose tolerance and 11.1 mmol/L above is considered as diabetic patient.
Comparing the results obtained through a simple calculation, the glucose level at 1 hour and 2
hour are (9.02 and 11.04) mmol/L respectively. This shows that the patient is suffering from
slightly high level of blood glucose. The reason why time interval test were done as in human
digestive system, it take approximately two too two and a half hours to digest sugars completely
before the body revert back to its normal fasting state. Therefore, if there is still sugar retain
the blood, it may be marker of hyperglycemia, which is the beginning of diabetes mellitus.
Among many of the precautions steps that were taken during the experiments is that gloves
were worn throughout the experiment to prevent any contamination. Besides, that all the
mixture in Part B, were discarded in separate prepared waste discarding bottle as it contains
harmful reagents.

REFERENCES:

1. DiabetesMellitus-information.com. (n.d.). Diabetes Diagnosis. [Online]. Available at:

http://www.diabetesmellitus-information.com/diabetes-diagnosis.htm [Accessed on 13
March 2018].

2. LaboratoryInfo.com. (2016). Benedict Test: Principle, Reagent, Preparation, Procedure

and Interpretation. [Online]. Available at: https://laboratoryinfo.com/benedicts-test-
principle-reagent-preparation-procedure-interpretation/ [Accessed on 13 March
2018].

3. Amrita.olabs.edu.in,. (2014). Detection of Sugar in Urine. [Online]. Available at:

http://amrita.olabs.edu.in/?sub=79&brch=17&sim=207&cnt=1 [Accessed on 13
March 2018].