SERUM CREATININE

Creatinine is formed in the body from the spontaneous degradation of

creatine and creatine phosphate. It serves no useful function in the body and
is excreted in the urine.

Creatine phosphate is also called phosphocreatine. It is the phosphorylated

derivative of creatine found in the muscles. It's a high energy compound that
can reversibly donate a phosphate group to ADP to form ATP. It provides a
small but rapidly mobilized reserve of high energy phosphates that can be
used to maintain intracellular level of ATP during the first few minutes of
intense exercise.

The amount of creatine phosphate is proportional to the muscle mass, varies

little and levels are very stable.

Creatine and creatine phosphate spontaneously cyclize at a slow but

constant rate to form creatinine, which is excreted in urine. The amount of
creatinine is proportional to the total creatine phosphate content of the body
and can be used to estimate the muscle mass.

ANALYTICAL METHOD.
Jaffe's colorimetric two point kinetic method.

PRINCIPLE
Jaffe's reaction is based on the reaction of creatinine with sodium picrate.
Creatinine reacts with alkaline picrate forming a red colored complex called
creatinine picrate. The red color is quantitated by measuring the absorbance
at 492 nm.
REACTION

Creatinine + sodium picrate → creatinine picrate.

SAMPLE
Serum or heparinized plasma.

TEST PROCEDURE
Reagent composition
R-1--- picric acid
R-2--- Sodium hydroxide
Creatinine standard------ 2 mg/dl

Working standard is prepared by mixing equal volumes of R-1 and R-2.

Assay conditions
Wave length : 492nm
Path length : 1 cm
Temperature : 37 ċ
• Adjust the instrument to zero with blank.
• Mark three cuvettes as sample, standard and blank.
• Pipette into cuvette.

Blank Standard Sample

Working reagent (ml) 1.0 1.0 1.0
Standard (µl) --- 100 ---
Sample (µl) --- --- 100

Mix and start the stop watch.

Read the absorbance A1 after 30 seconds and A2 after 90 seconds.

Calculate ∆A = _A2 - A1_______

Time (mins)

CALCULATIONS:

Creatinine in mg/dl = ∆A of sample x concentration of standard.

∆A of standard
REFERENCE VALUES:
Serum or plasma:
Males: 0.7 – 1.4 mg/dl.
Females: 0.6—1.1 mg/dl.

Urine:
Males: 10—20 mg/kg/24 hrs.
Females: 8—18 mg/kg/24 hrs.

CLINICAL SIGNIFICANCE:
Elevated creatinine levels are indicative of renal insufficiency. Values up to
20—30 mg/dl may be seen in later stages of renal failure.
Elevated levels may also occur with:
• Increased dietary intake of roast meats.
• Large muscle mass
• Acromegaly
• Gigantism

Low plasma creatinine concentrations are not clinically important. These

include:
• Small muscle mass particularly in elderly females and infants. They
may have increased plasma creatinine due to renal failure, but have a
level within the reference range.
• Muscular dystrophies.

Merits and demerits of creatinine assay.

Advantages:
• Its concentration is relatively constant from day to day in a given
person.
• It's concentration unlike urea is not affected by endogenous and
exogenous factors like protein intake, dehydration, tissue breakdown
and protein catabolism.
Disadvantages:
• Technical difficulties in its measurement due to the presence of other
substances which react with alkaline picric acid giving high values.
These include hemoglobin and bilirubin.
• Lack of sensitivity of creatinine as plasma creatinine is not raised until
the renal function is substantially impaired.