CLINICAL CHEMISTRY 1 LABORATORY

EXERCISE NO. 3- BSMT 3S

GLUCOSE MEASUREMENT

Pre-Lab Activities:
Collect fasting blood sample for serum glucose measurement (10-12 hours fasting). Prepare all
the materials and equipment needed in the activity.
Materials:
Serum
Phlebotomy kit (SST, Gold top)
Gluco- Toluidine reagent and standard solution
Control sera
Pipettes
Test tubes
Nescofilm
Spectrophotometer
Facial tissue

METHOD: Ortho-Toluidine, Dubowski Method (Chemical Condensation Method)

PRINCIPLE:
Glucose condenses with O-toluidine in glacial acetic acid when heated at 100-C forming
N- glucosylamine which is blue- green in color. The absorbance is measured at 600-640.
Glacial HAC
Glucose + Aromatic Amines Glucosylamine + schiff’s base (blue-green)
heat
Heat
Procedure:
1. Label all tubes accordingly as blank, standard, normal control, abnormal control and
patient sample. Heat
2. To each tube, transfer the desired volume.

REAGENT Standard Sample/Control

Glucose Standard 50 ul -
Test Sera/Controls - 50 ul
Gluco-Toluidine Reagent 1500 ul 1500 ul

3. Mix well and place in boiling water AT THE SAME TIME for THREE MINUTES.
4. Immerse test tube in room temperature water or tap water for 1-2 minutes to cool.
5. Read the absorbance for each tube against the water blank at 600 nm wavelength.
6. Record the results and compute the serum glucose concentration.

Calculation:
Absorbance of Sample
Glucose (mg/dL) = Absorbance of Standard x 100 mg/dL (Value of Standard )

CONVERSION TO SI UNIT: mg/dl x 0.055 = mmol/l

NORMAL RANGE: 65-110 mg/dl (3.58-6.05 mmol/l)
 CLINICAL CHEMISTRY 1 LABORATORY
EXERCISE NO. 3- BSMT 3S
GLUCOSE MEASUREMENT

Name: Date:
POST- LAB ACTIVITY
A. Data and Results (10 pts)
Absorbance of Standard:
Absorbance of Normal Control:
Absorbance of Abnormal Control:
Absorbance of Unknown:

1. Draw and label the results. (5pts)

2. Compute the serum blood glucose: (Show your computation. Express the
values in mg/dL and mmol/L) (5pts)

a. Normal Control

b. Abnormal Control

c. Unknown Sample
B. Questions (Maximum of 3-4 sentences only):

1. Differentiate diabetes mellitus from diabetes insipidus. (5pts)

2. Define the following terms: (5pts)

a. Glycogenesis-
b. Glycogenolysis-
c. Gluconeogenesis-
d. Glycolysis-

3. Why do we need to separate the serum intended for glucose measurement from
the packed red cells immediately after centrifugation? (5pts)

4. Define Hemoglobin A1C and its relation to glucose measurement. (5pts)