Kidney Function Tests

Classification of KFTs

Glomerular function tests Tubular function tests

1) Urine examination.
1) Urine concentration test.
2) Estimation of blood
urea & serum 2) Urine dilution test.
creatinine. 3) Plasma electrolyte
3) Urea clearance test. determination.
4) Creatinine clearance
test.
 Glomerular function tests
•The glomerular filtrate is an ultrafiltrate of plasma, and has the
same composition of plasma without most of plasma proteins.

•Plasma is filtered by the glomeruli at a rate of 125 ml/min

(Glomerular filtration rate, GFR).

•GFR: Is the maximum rate that the plasma can be cleared of any
substance.

•GFR can be measured by determination of renal clearance of

some substance either occurring naturally in blood or administered
to the patient.
 1-Clearance

Is the amount of blood which could be completely cleared

a substance per minute.

Clearance (C) [ml/min] = [ U x V ] / P

U = The concentration of the substance in urine (mg/dl).
V = Urine volume per minute ( ml/min).
P = Plasma concentration of the substance (mg/dl).
UV: Is the amount of the substance excreted in urine per
minute.
 The Principle of Clearance

➢ Some substances when filtered & enter the tubules are not
reabsorbed or secreted.
Thus, excreted= GFR

➢ Some substances are filtered, enter tubules, and more of the

substance is secreted into the tubules.
Thus, Clearance>GFR

➢ Some substances are filtered, enter tubules, but are completely

reabsorbed
So, they did not reach the final urine
➢ Clearance is calculated for plasma
constituents which are:
1. Freely filtered at the glomerulus.
2. Not reabsorbed by the tubules.

➢ Example: Urea & creatinine.

 Creatinine clearance test:

➢It is the most sensitive method of assessing renal function (GFR).

➢It needs:
1. 24 hr urine collection.
2. One sample of blood taken.

➢Precautions:
1. The urine should be collected accurately.
2. There are no ketones or heavy proteinuria to interfere with the creatinine
determination.

➢ Normal value of creatinine clearance: 96-126 ml/min.

The decrease in creatinine clearance is a very sensitive indicator of a
decreased GFR as a result of: glomerulonephritis or Reduced blood
flow to the glomeruli.
 2- Estimation of serum levels of certain
substances

➢The kidney is the primary organ involved in the

excretion of urea, creatinine and uric acid.

➢Thus, renal dysfunction is characterized by

measuring the increase in serum levels of these
compounds.
 Determination of serum urea

➢ In human, urea is the major end product of protein (Both dietary

and endogenous tissue protein) and amino acid catabolism.

➢Other species e.g. birds convert protein nitrogen to uric acid

➢fish excrete nitrogen as ammonia.

synthesis
Protein diet Amino acid pool Tissue proteins
catabolism
Xss a.a.

Deamination Urea cycle In Kidney

NH3 Urea Excretion
(In liver) (In liver)
 Normal level of serum urea

❖15-50 mg/dl

❖The conversion factor of mg urea/dl to mmol/l:

mg/dl x 0.166 = mmol/l

 Normal level of serum Urea

It is obvious that this range is wide due to:

➢Dietary effect: Protein rich diet increases serum urea.

➢Gender: Male level is higher than female level.
➢Age: Catabolism increases with age so serum urea
increases.
➢Pregnancy: Anabolism increases with pregnancy so serum
urea decreases.
 Serum Urea
increases decreases

A-Physiologically: In Protein rich diet. A. Physiologically:

B-Pathologically: i. In pregnancy ( ++ Anabolism).
1. Pre-renal disease: ii. In starvation.
i. Decreased blood volume and pressure as in severe iii. In patients whose diet is
vomiting & diarrhea. grossly deficient in proteins.
ii. Increased protein catabolism as in fever and diabetic
coma. B. Pathologically:
2. Renal diseases: In severe liver disease.
i. Acute glomerulonephritis.
ii. Chronic pyelonephritis.

3. Post-renal diseases:
i. Urinary stones
ii. Urinary tumor.
iii. Prostatic enlargement.

4. High doses of highly catabolic drugs e.g. cortisone.

 Principle (Urease-Berthelot method)

Urease hypochlorite phenol

Urea + H2O CO2 + 2NH3 Chloramine Indophenol

Alk. medium

Blue colored complex,

measured colometrically at 550nm
 Procedure
Blank (ml) Standard (ml) Test (ml)
Standard - 0.02 (20 µl)
Sample - - 0.02 (20 µl)
Reagent 2 0.02 (20 µl) 0.02 (20 µl) 0.02 (20 µl)

Mix, incubate for 5 min, at 37 cº

Reagent 3 1.0 1.0 1.0

Reagent 4 1.0 1.0 1.0

•Mix well.
•Incubate for 10 min at 37cº.
•Measure the absorbance of sample (Asample) and the standard
(Astandard) at 550nm (530-570nm).
•The color is stable for 5 hrs.

Urea concentration (mg/dl) = (Asample/ Astandard ) x Standard concentration

 Determination of serum Creatinine

➢ Creatine is synthesized in the liver from glycine and arginine, and

then it passes to the circulation to be taken up by the muscles and
converted to phosphocreatine which serves as a high energy source.
Creatine kinase
Creatine + ATP Creatine~P + ADP

* Creatine phosphate loses phosphoric acid and gives creatine

* Creatine loses water to form creatinine which is released into

plasma. It is removed from circulation by glomerular filtration
and excreted in urine.
Thus, Creatinine is the anhydride of creatine.
The measurement of serum creatinine is a better
indicator of the kidney status than serum urea due to:

❑ Creatinine is excreted at a relatively constant rate

that is proportional to the muscle mass.

❑ Creatinine is not influenced by dietary protein and

protein catabolism.
 Normal value: 0.6 - 1.2 mg/dl.

The conversion factor of mg creatinine/dl to µmol/l:

mg/dl x 88.4 = µmol/l.

Serum creatinine increases in:

i. Increased rate of formation as in gigantism &
acromegaly.
ii. Decreased rate of excretion as in renal failure.
 Principle: Jaffee method (Kinetic method)

Kinetics:
It is the study of the rate of reaction by measuring the increase in product
concentration or the decrease in substrate concentration in a given time interval.

Alk. pH
Creatinine + Picric acid Orange colored complex,
measured Colorimetrically
at 495nm

In kinetic Jaffee method, serum is mixed with alkaline picrate & and the rate
of change in absorbance is measured between 2 time points.
 Why kinetic method?

To increase specificity of the test as:

➢ glucose, ketoacids, ascorbate & uric acid give

false high levels

➢ bilirubin gives false negative results

This is minimized by the use of kinetic method

 Procedure:
Blank (ml) Standard (ml) Test (ml)
Standard - 0.02 (20 µl)
Sample - - 0.02 (20 µl)
1.0 1.0 1.0

(Distilled (Working (Working

water) reagent) reagent)
•Mix well, and after 20 seconds at 20-25cº read the absorbance A1 of the standard
and the sample at 495nm (490-510nm) against blank.

•Exactly 2 min later read the absorbance A2 of the standard and the sample.
•Standard concentration=2 mg/dl.

Calculation:
A2 – A1 = ∆A sample or ∆A standard

∆A sample
Creatinine concentration (mg/dl) = x Standard concentration
∆A standard
Thank You