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CREATININE CLEARANCE
TEST
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OBJECTIVES
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What’s Cratine and Creatinine…?
1. The creatine is an
amino acid that does
not found in proteins.
2. Creatine is
a nitrogenous organic
acid.
Structure of Creatine
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Structure of Creatine Phosphate
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Formation of Creatinine from
Creatine Phosphate
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What’s the Relationship between
Creatine and Creatinephosphate?
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Creatine Phosphate is a Compound
with High-Energy Phosphate Bond
• Creatine can be charged with energy:
The enzyme creatine kinase (CK) which
transfers the high-energy phosphate
group from ATP to creatine to make
creatine phosphate, a compound with
high-energy phosphate bond.
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The Reversible Equilibrium between
Creatine and Phosphocreatine
Creatine
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Kinase
Formation of Creatinine
Spontaneous, non-
enzymatic cyclization Spontaneous
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Spontaneous One-Way Conversion of PCr and
Creatine
into Creatinine
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The Diagnostic Function of
Creatinine
If the kidneys are damaged or impaired
and cannot work normally
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Serum Creatinine may be Affected
Partly by….
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How is Creatinine Used to Monitor
Renal Function....?....cont.
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The Glomerular Filtration Rate
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Glomerulus Structure
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Structure and Function of Glomerulus
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How is Creatinine Used to Monitor
Renal Function? ……cont.
• In renal failure, the filtering (GFR ) of the
kidney is deficient, blood levels of some
substances will rise and urine levels will
decrease.
• Creatinine Clearance. Therefore, creatinine
levels in blood and urine may be used to
calculate the creatinine clearance (CrCl),
which reflects the GFR.
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How to do Creatinine Clearance
Test ?
The Principle Protocol of Creatinine
Clearance Test :
• A creatinine clearance test is done on both
a blood sample and on a sample of urine
collected over 24 hours (24-hour urine
samples).
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Are there any Substances Used
to Assess GFR other than
Creatinine ?
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Different Clearance Tests Used to
Assess Renal Function
• Kidney function tests (Clearance tests):
is a collective term for a variety of individual
tests and procedures that can be done to
evaluate how well the kidneys are functioning.
There are 3 major clearance tests:
1. Creatinine clearance test.
2. Urea clearance test.
3. Inulin clearance.
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2- Urea Clearance Test
• Urea is a:
1. Waste product of protein; it is created by
protein catabolism
2. Excreted in the urine.
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2- Urea Clearance Test ….cont.
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How to do Urea Clearance
Test ?…Method…
Urea clearance test in order to
determine the amount of urea that is
filtered, or cleared, by the kidneys into the
urine we require the following :
1. A blood sample to measure the amount of
urea in the bloodstream
2. Two urine specimens, collected one hour
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apart.
3- Inulin Clearance Test
What’s Inulin ?
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The Preference of Creatinine
Clearance Test in Evaluating
Kidney Function …..cont.
• Creatinine clearance is preferred because:
1. It is a normal constituent of blood and no
infusion is needed unlike inulin.
2. Moreover it is not reabsorbed by the tubules as
in the case of urea.
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Glomerular Filtration Rate
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Clearance Tests
Clearance Tests
- One of the most important kidney function
tests, it is; the amount of plasma (in ml) that is
cleared from a certain substance by the
kidneys per minute.
Clearance = UV/P
• U = concentration of the substance in urine.
• P = concentration of the same substance in
plasma
• V = volume of urine (ml/min)
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High Levels of Creatine
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Principle
Determination of Creatinine
Creatinine Picrate
Orange color
Measured colorimetrically at 520 nm
Materials
1. Creatinine stock standard: 150 mg creatinine in
100 ml water (1.5 mg/ml)
2. Creatinine working standard for serum
(3mg/dl): dilute 10 ml of stock and increase the
volume up to 500 ml with water.
3. Creatinine working standard for urine (0.75
mg/dl): dilute 50 ml of stock and bring the
volume up to 200 ml with water.
4. Serum and urine samples.
5. NaOH (2.5 M).
6. Picric acid.
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• Method
1. Serum Creatinine :
Standard (A,B) Test (C,D) Blank (B)
Serum - 0.5 ml -
Water 1.5 ml 1.5 ml 2 ml
Standard 0.5 ml - -
for serum
Picric acid 6 ml 6 ml 6 ml
1. Mix well
2. Add 0.4 ml of 2.5 M NaOH
3. Allow to stand for 20 minutes
4. Read the absorbance against the blank at 520
nm
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Method ……cont.
2) Urinary Creatinine
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Method ……cont.
2) Urinary Creatinine
5- mix well
6- Add 0.4 ml of 2.5 M NaOH
7- Allow to stand for 20 minutes
8- Read the absorbance against the blank at
520 nm
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Calculations
• Serum creatinine = Abs of test X concentration of
standard Abs of std
• Urinary creatinine = Abs of test X concentration of
standard Abs of std
• Creatinine clearance = Urinary creatinine X Volume of
urine ml/min/1.73 m2 Serum creatinine
Normal creatinine clearance:
100-130 ml/min/1.73m2.
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