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CHEMISTRY I
MKEB 2404
Title:
Effects of specimen handling on analytes
Objectives:
To determine the effect of several preanalytical factors on the level of
analytes
Principle:
Results from the laboratory test are used for detection and diagnose
of disease. Its also can be use to monitor the progress or its response to
treatment and to screen for disease in otherwise healthy individuals. If the
result is abnormal, it shows a disease state in patient
However, many factors also can affect the composition of body
fluids. These factors may be analytical or preanalytical. Thus, wherever
possible, both preanalytical and analytical variability should be controlled
so that correct interpretation of the result can be made In this experiment
we are going to see some effect of preanalytical factors on the level of
glucose and potassium in serum
Procedure:
1. 10 ml blood was drawn from each student
2. Blood was divided into 5 tubes
(a) Room temperature. The blood was centrifuge at 3000rpm for 15
minutes. The plasma was taken for the result of glucose and
potassium concentration
(b) Lysed, room temperature. The blood was centrifuge at 3000rpm for
15 minutes. The plasma was taken for the result of glucose and
potassium concentration
(c) At room temperature and was incubated for over night. The blood
was centrifuge at 3000rpm for 15 minutes. The plasma was taken
for the result of glucose and potassium concentration
(d) 40C and incubate for over night. The blood was centrifuge at
3000rpm for 15 minutes. The plasma was taken for the result of
glucose and potassium concentration
(e) -200C and incubate for over night. The blood was centrifuge at
3000rpm for 15 minutes. The plasma was taken for the result of
glucose and potassium concentration
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3. The serum was separated on the specified day
4. The glucose and potassium level in tube (a) and (b) was measured using a
reflotron analyzer
5. The analyte level in tube (c), (d), and (e) was measured on the next day
6. The result and observe of any changes in the reading was recorded.
Result:
Reference of glucose and potassium level:
Glucose: 4.22 – 6.11 mmol/L
Potassium: 3.5 – 5.1 mmol/L
Tubes Glucose level Potassium level
(a) 5.29 mmol/L 4.23 mmol/L
(b) 5.87 mmol/L 4.40 mmol/L
(c) <0.555 mmol/L 4.45 mmol/L
(d) 3.15 mmol/L 9.19 mmol/L
(e) 8.08 mmol/L >12.0 mmol/L
Discussion:
Calculations of K+ concentration in blood
The principle of the reaction involve the diffusion of potassium from the
aqueous phase into the organic phase where it forms a complex with
valinomycin
In order to balance the charge, a pH indicator was dissolved in the organic
phase. It gives off a proton and forms a colored anion.
An optimum change of the reflectance was achieved by addition of a
strong acid, which forms a colorless anion that compete with the indicator.
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However the glucose and potassium concentration in this tube is still
within the range of the normal potassium and glucose level.
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Conclusion:
Usually normal human body contains 50mmol/L potassium per kg body
weight.
It is important to maintain the concentration gradient between
extracellular and intracellular in order to maintain the membrane
potential and excitability.
Glucose plays an important role as body energy source. It is stored as
glycogen and was broken down by glycolysis process.
The normal glucose level in body should be 4.22 – 6.11 mmol/L
The normal potassium level in body should be 3.5 – 5.1 mmol/L
The result can be affected by various ways such as an error in blood intake
(vein puncture) and other methods.