Lab-3

Diabetic Profile
Measurement of Blood Glucose

Diabetes mellitus is a group of

metabolic disorders that requires
continuing medical care and patient-self
management education to prevent
acute complications and to reduce the
risk of long term complications.
- It is characterized by hyperglycemia
and abnormal protein, fat and
carbohydrate metabolism due to
defects in insulin secretion, i.e.,
inadequate and deficient insulin action
on target tissues.

It is classified into 4 clinical classes:

Type I diabetes mellitus (TIDM)
Type 2 diabetes mellitus (TIIDM)
Gestational diabetes mellitus (GDM)
Other specific types due to other causes e.g. Drugs or chemical induced.

Diabetic Profile Tests:

Group of tests that are used to diagnose diabetes and to measure

response to treatment. They include:

1-C-peptide…..Differentiates between type I and type II.

2-Blood Glucose

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 • FBG (= FBS= PG=FPG)….Fasting blood glucose.
• PP glucose = Post prandial.
• GTT= Glucose tolerance test, OGGT= Oral glucose tolerance test
3-HbA1c = Glycosylated hemoglobin.
4-Ketones.
5-Microalbuminurea.
6-Insulin.
7-ICA = islet cell antibodies.

1- C-peptide:

Measurement of C-peptide exhibits a number of advantages over insulin

measurement.
- It is better indicator of B-cell function than peripheral insulin.
- It doesn’t measure the exogenous insulin.
- A C-peptide test can be done when diabetes has just been found and it
is not clear whether type 1 diabetes or type 2 diabetes is present. A
person whose pancreas does not make any insulin (type 1 diabetes) has a
low level of insulin and C-peptide. A person with type 2 diabetes has a
normal or high level of C-peptide.
Normal value: Fasting …..0.51-2.72ng/ml
Blood glucose:
- It is a vital component of diabetes management.
- In most cases, significantly elevated fasting glucose levels (>140mg/dl
or > 7.77 mmol/l) are, in themselves, usually diagnostic of diabetes.
However mild or borderline cases may present with normal FBS values. If
diabetes is suspected, GTT can confirm the diagnosis.
- Occasionally, other diseases may produce elevated plasma glucose
levels, therefore, a comprehensive history, physical examination and
other tests should be carried out to confirm the diagnosis

Types of blood glucose tests:

(1)Fasting blood sugar (FBS) measures blood glucose after fasting for
at least 8 hours. It often is the first test done to check for diabetes.

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(2)2-hour postprandial blood sugar (2-hour PP) measures blood
glucose exactly 2 hours after eating a meal.
(3)Random blood sugar (RBS) measures blood glucose regardless of
when the person last ate. Several random measurements may be taken
throughout the day. Random testing is useful because glucose levels in
healthy people do not vary widely throughout the day. Blood glucose
levels that vary widely may indicate a problem. This test is also called a
casual blood glucose test.
(4)Oral glucose tolerance test (OGTT) measures the body's ability to
use glucose. It is used mainly to diagnose prediabetes and diabetes. An
oral glucose tolerance test is a series of blood glucose measurements
taken after you drink a sweet liquid that contains glucose. This test is
commonly used to diagnose diabetes that occurs during pregnancy
(gestational diabetes). This test is not commonly used to diagnose
diabetes in a person.
How to Prepare?
Oral Glucose tolerance test (OGTT)
• On the day of testing, the following steps will be done:

• A blood sample will be collected when the subject arrives. This is

the fasting blood glucose value. It provides a baseline for comparing
other glucose values.
• Then a sweet liquid containing a measured amount of glucose will
be given to the subject. It is best to drink the liquid quickly. For the
standard glucose tolerance test, the subject will drink 75 g to 100
g; pregnant women drink 100 g of glucose.
• Blood samples will be collected at timed intervals of 1, 2, and 3
hours after drinking the glucose.

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• The oral glucose tolerance test is not commonly used to diagnose
diabetes in people who are not pregnant. Many experts recommend
using an oral glucose tolerance test if the result of a fasting blood
glucose test is between 100 mg/dL (5.5 mmol/L) and 126 mg/dL
(7.0 mmol/L).
• Glucose tolerance test screening by age 30 is recommend for all
women who have polycystic ovary syndrome.

Glycosylated hemoglobin HbA1c:

• Glycosylated hemoglobin is an indicator of the blood glucose
concentration over a longer period of time than either a single blood
glucose measurement (which reflects the glucose concentration at
the time of blood collection)
• A glycohemoglobin test indicates how well diabetes has been
controlled in the 2 to 3 months before the test.
• The A1C level is directly related to complications from diabetes:
(The lower the A1C level, the lower the risk for complications)
• Normal values vary from lab to lab, depending on the test method
used.
Normal Values:
Glycohemoglobin A1c:4.5%-5.7%
Total glycohemoglobin:5.3%-7.5%
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 Sample collection:

When whole blood is drawn, allowed to clot

and kept centrifuged at room temp, the
average decrease in serum glucose is
approximately 7% in 1 hour. It is due to
glycolysis by RBCs. The rate of glycolysis
after 2 hours increases considerably, so the
decrease in serum glucose may be about
50% or more.
Serum: Remove from clot within 30 minutes of collection in
order to prevent glycolysis.
Plasma: An anticoagulant containing fluoride is recommended,
but any of the common anticoagulants may be used

- In separated non-hemolyzd serum, the

glucose conc. is generally stable for 8 hours
at 25C (or up to 72 hours at 4C)
- Glycolysis can be prevented by collecting
blood in fluoride tube.
-For HBA1c EDTA tube (whole blood) is used

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Lab Practices:
-Collect blood for FBS in an appropriate tube.
-Follow the method in the leaflet.
-Compare the results to the normal value.

Principal:
-Glucose oxidase (GOD) catalyses the oxidation of glucose to gluconic
acid.

-The formed hydrogen peroxide (H2O2), react under catalysis of

peroxidase (POD) with phenol and 4-aminoantipyrine to a red-violet
quinoneimine dye as indicator.
The intensity of the color formed is proportional to the glucose
concentration in the sample.
Procedure:
1. Assay conditions:
Wavelength: . . . . . . . . . . ..... . 500 nm
Cuvette: . . . . . . . . . . . . . . . . . . . . .. 1 cm light path
Temperature. . . . . . . . . . . . . . . . . . . 37ºC / 20-25ºC

2. Measurement against reagent blank. Only one reagent blank per series
is required.

3. Pipette into a cuvette

Blank Standard Sample

WR µl 1000 1000 1000
Standard µl ----- 10 -----
Sample µl ----- ----- 10

4. Mix and incubate for 5 min at 37ºC or 10 min at room temperature

(20-25ºC).

5. Read the absorbance (A) of the samples and standard, against the
Blank. The colour is stable for at least 60 minutes.

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Calculations:

ΔA sample
Conc. = 100 ∗ 𝑚𝑔/𝑑𝐿
ΔA STD

Conversion factor: mg/dL x 0.0555= mmol/L.

Normal values:

Serum or plasma ------70-115 mg/dl (3.33-6.10mmol/L)

Linearity:
From detection limit of 0.04 mg/dL to linearity limit of 500 mg/dL.
If the results obtained were greater than linearity limit, dilute the sample
1/2 with NaCl 9 g/L and multiply the result by 2.
References:

1. Kaplan L.A. Glucose. Kaplan A et al. Clin Chem The C.V. Mosby Co. St
Louis. Toronto. Princeton 1984; 1032-1036.
2. Trinder P. Ann Clin Biochem 1969; 6: 24-33.
3. www.webmed.com.

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 Lab Report

Palestine Polytechnic University

College of Medicine
Clinical Biochemistry Lab

Patient Name: Age/Sex:

Patient ID: Sample Date:
Doctor Name: Test Date:

BIOCHEMISTRY REPORT

DESCRIPTION RESULT UNIT REFERNCE RANGE

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EXAMINED BY:
DATE/ TIME:

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