Creatine is a nitrogenous organic acid that occurs

naturally in vertebrates.

Creatine was identified in 1832 when Michel

Eugène Chevreul isolated it from the basified
water-extract of skeletal muscle.

Its main role is to facilitate recycling of adenosine

triphosphate (ATP), the energy currency of the
cell, primarily in muscle and brain tissue, which
we’ll have a word about in the next slides.
 Creatine is synthesized in liver, kidneys
and pancreas by two enzymatically
mediated reactions:
 Transamidination of arginine and glycine to
give guanidoacetic acid.
 Methylation of guanidoacetic acid in
presence of S- adenosyl methionine as
methyl donner to give Creatine.
Non-phosphorylated creatine is released into
the blood by the liver where it travels mainly to
the muscle cells (95% of the body's creatine is in
muscles), and to a lesser extent the brain, heart,
and pancreas.

Once inside the cells it is transformed into

phosphocreatine by the enzyme
complex creatine kinase, which makes it able
to donate its phosphate group to
convert adenosine diphosphate (ADP)
into adenosine triphosphate (ATP).
This process is an important component of all
vertebrates' bioenergetic systems.
Creatine phosphate is broken
down into creatinine, which is
then excreted in the urine.

A 70-kg man has about120g of creatine,

with 40% being the unphosphorylated form
and 60% as creatine phosphate.
Of that amount, 1–2% is broken down and
excreted each day as creatinine.
Reference values:

Gender Creatinine Uric acid Urea

Men 71-115 mmol/L 180-400 mmol/L 1,7-8,3

Women 53-106 mmol/L 90-350 mmol/L mmol/L
Principle:
In the method of Jaffe (used first in 1886)
which is often applied for determination
of creatinine, creatinine reacts with
picric acid in alkaline solution to give red
colour complex.
The absorbtion of this complex is
proportional to creatinine concentration
I the serum sample.
Material:
Reagent 1 – solution of picric acid –
8,73mmol/L
Reagent 2 – solution of NaOH – 312,5
mmol/L and 12,5 mmol/L of sodium
phosphate
Before it, the working solution is made by
mixing reagent 1 and reagent 2 in a ratio
4:1. Standard solution of creatinine is 132
mmol/L
WR(Working Reagent): 6mL R1 + 1,5 mL R2
Biological material is serum or plasma
Method:
- colour reaction, 3 test tubes
Material Sample Standard Blank

WR 2,0 mL 2,0 mL 2,0 mL

Stand. - 100 mL -
solut. of
creatinine
serum 100 mL - -

water - - 100 mL
Read the absorbance of standard at
sample after 1 min at d=490nm
After 5 min, read again the absorbance at
490nm using blank as reference

Concetration of creatinine

A2-A1 sample
132 mmol/L X
A2-A1 standard