its a small slide preparation on copper

2. Presented by :-
Jijo George
4th MLT
COPPER

3. COPPER
• It is a relatively soft yet tough metal with
excellent electrical and heat conducting
properties
• Copper is widely distributed in nature both in
element form and in compounds
• It forms alloys with Zinc, Tin, nickel
• In body contains about 100 mg copper
distributed in different organs.
( muscle, Liver, Bone marrow, Brain, Kidney,
Heart and in Hair )

4. DIETARY REQUIREMENTS
Adults :- 2-3 mg/day
Infants and children's : 0.5 -2 mg/day

5. SOURCES
 Liver
 Kidney
 Meat
 Green leafy vegetables
 Egg yolk
 Cereals
 Nuts
 Milk is a poor source

6. FUNCTIONS
• It is necessary for iron absorption and
incorporation of iron into hemoglobin.
• It is necessary for tyrosine activity
• It is a co-factor for vit.c requiring hydroxylation
• It increases HDL and so protects the heart
• Copper is necessary for the synthesis of
melanin and phospholipids

7. ABSORPTION
About 10% of dietary copper is absorbed,
mainly in the duodenum
Metallothionein is a transport protein that
facilitates copper absorption
Phytate, Zinc and Molybdenum decreases
copper up take.

8. EXCRETION
Only 0 to 32 mg of copper is excreted in normal
urine in 24 hours
The daily biliary excretion of copper is 0.5 to 1.3
mg and 0. to 0.3 mg is excreted across the
intestinal mucosa into the bowel lumen

9. BLOOD COPPER
The normal conc. Of copper in serum is 75-160 mg/100ml
Both red blood cells and serum contain copper
In 80% of red cell copper is present as superoxide
dismutase (erythrocuprein )
The copper in plasma occur in firmly bound and loosely
bound forms.
The firmly bound copper consists of ceruloplasmin.
The loosely bound coper is known as direct reacting
copper and is loosely bound to serum albumin
The plasma copper levels increase in pregnancy because
of their estrogen content
Oral contraceptives have a similar effect.

10. DISEASE STATES
Copper deficiency :- severe deficiency of copper
cause
Demineralization of bones
Demyelination of neutral tissue
Anemia
Fragility of arteries
Myocardial fibrosis
Hypopigmentation of skin
Greying of hair

11. 1. WILSON'S DISEASE
 Also known as hepatolenticular degeneration
 It is a rare disorder of abnormal copper metabolism and
is characterized by the following manifestations
• Copper is deposited in abnormal amounts in liver and
lenticular nucleus of brain. This may lead to hepatic
cirrhosis and brain necrosis
• Low levels of copper and ceruloplasmin in plasma with
increased excretion of copper in urine.
• Copper deposition in kidney causes renal damage. This
leads to increased excretion of amino acids, glucose,
peptides, and hemoglobin in urine.
• Intestinal absorption of copper is very high, about 4-6
times higher than normal.

12. 2. COPPER DEFICIENCY ANEMIA
Copper is essential for the formation of Hb
Copper containing ceruloplasmin helps in iron
transport
Copper is an integral part of ALA synthase, which
is the key enzyme in heme synthesis
Copper helps the uptake of iron by normoblasts
Copper deficiency is manifested as anemia
In this case RBC count is reduced, cell size is
small, but Hb conc. Is more or less normal

13. 3. CARDIO VASCULAR DISEASES
 Copper is a constituent of lysyl oxidase
 It oxidize four lysine residues together to form
desmosine which makes cross linkages in
elastin
 In copper deficiency, elastin becomes
abnormal, leading to weakening of walls of
major blood vessels.
 This favors aneurysm and fetal rupture of the
wall of aorta
 Fibrosis of myocardium leading to cardiac
failure.

14. 4. MENKE’S KINKY HAIR SYNDROME
 It is an X-linked disorder of intestinal copper
absorption.
 The absorption of copper and its uptake in the
mucosal cells and its intracellular transport with
in the mucosal cells are normal in patients with
this disease
 But the transport of copper across the serosal
aspect of the mucosal cell membrane is defective
 Symptoms :- mental retardation, temperature
instability, abnormal bone formation and
susceptibility to infection are not to be cured
 The clinical features show that unless therapy is
started promptly at birth.

15. TREATMENT
Wilson’s disease :- Administration of pencil
amine, a naturally occurring copper chelating
agent
Menke’s disease :- No effective treatment

16. DETERMINATION OF SERUM COPPER

17. SODIUM DIETHYL DITHIO CARBAMATE
METHOD
 Principle :-
The copper is released from its linkage
to protein by means of hydrochloric acids, the
proteins precipitated by trichloro acetic acid, and
the copper extracted from protein free fluid
obtained, into an amyl alcohol – ether mixture as a
golden yellow – coloured complex with sodium
diethyl dithio carbamate. The amount of this
formed is read colorimetrically

18. REAGENTS
HCl – 0.1 N
Hydrochloric acid – 6N
Trichloro acetic acid 20% aqous solution
Sodium pyrophosphate, 6% solution
Ammonia
Sodium diethyl dithio carbomate – 0.4%
solution
Amyl alcohol – ether mixture
Powdered anhydrous sodium sulphate

19. Stock standard solution :- Dissolve 0.398 g
of copper sulphate in H2O, add 0. ml of conc.
Sulphuric acid and make up to 1 litter
Standard for use :- Dilute the above 1 to
100 to obtain a solution containing 1
micro gram/ ml

20. TECHNIQUE
 To 3 ml of serum add 1 ml of 0.1 N HCl and
warm in boiling water, stirring continuously,
until the mixture begins to cloud
 Cool & then add 1.5 ml of 6 N hydrochloric acid
& stand for 10’
 Add 3 ml of trichloro acetic acid, mix & stand
for few minute and centrifuge.
 Remove the supernatant and wash the
precipitate with 3 ml of 5% trichloro acetic acid
 Centrifuge again and combine the supernatant
fluids
 Add 1 ml of 0.4% sodium di ethyl dithio amyl
alcohol - ether mixture to extract the copper
.

21.  Remove the amyl alcohol layer and dry by
shaking with a little powdered anhydrous
sodium sulphate
 Read using a violet filter or 440 milli microns.
 Treat 5 ml of standard in the same way as the
serum, and carry through a complete blank,
against which both standard and unknown are
read

22. CALCULATION
Micrograms copper/100 ml serum = Reading of unknown
×5×100/3
Reading of std
= Reading of unknown
×167
Reading of std

23. REFERENCE

24. REFERENCE
Text book of biochemistry for medical students
– D.M Vasudevan & Sreekumari.s, Kannan
vaidyanathan
Fundamentals of biochemistry - AC Deb
Biochemistry – satyanarayana
Clinical chemistry , techniques, principles,
correlations - Michael L. bishop, Edward .P.
fody, Larry .E. schoeff
Practical clinical biochemistry – Haroid Varley

25. Prepared by
Jijo George
4th MLT