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Copper in the human organism

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 Trakia Journal of Sciences, Vol. 9, No 1, pp 88-98, 2011
Copyright © 2011 Trakia University
Available online at:
http://www.uni-sz.bg
ISSN 1313-7050 (print)
ISSN 1313-3551 (online)

Mini-Review
COPPER IN THE HUMAN ORGANISM
M. Angelova1, S. Asenova, 1*,V. Nedkova2, R. Koleva-Kolarova3
1
Department of Chemistry and Biochemistry & Physics and Biophysics, University of Medicine –
Pleven, Bulgaria
2
Department of Pediatrics, University of Medicine – Pleven, Bulgaria
3
Department of Medical ethics, healthcare management and information technologies, University of
Medicine – Pleven, Bulgaria

ABSTRACT
Copper is an essential micronutrient for human bodies, whose distribution in different organs and tissues,
metabolism and physiological impact is not thoroughly explained yet. Further issues which need
clarification are the exact composition and constitution of its combinations with other elements, the
physiological impact of copper and copper-enzymes in sickness and health. Biomarkers for identification
of copper status are still being defined. They are necessary in order to determine the prophylactics and
treatment of diseases and conditions, which are related to changes of copper concentrations in human
bodies. Researches continue in an attempt to develop analytical methods for determination of copper in
serum and urine, which are used in clinical laboratory diagnostics. These current developments justify
the purpose of our research: to make a brief review of some issues related to studying the trace element
copper. In the present paper, we have reviewed a part of the essential copper enzymes – cytochrome c
oxidase, lysyl oxidase, feroxidase, monoamine oxygenase, superoxide dismutase, etc., e.g the
physiological functions, which depend on the presence of these essential copper enzymes in the human
organism. The issues of copper metabolism and the interactions of copper with other micronutrients
(zinc, iron, vitamins A, C, E) are briefly discussed. An overview of the influence of copper on several
diseases: Menkes syndrome, Wilson's disease, cardiovascular diseases, oxidative stress, osteoporosis,
diabetes, chronic diarrhea, cystic fibrosis, is performed as well. The results from our study in children,
aged 1 ÷ 3 years, with anemia, cystic fibrosis and chronic diarrhea and a healthy control group, showed
significantly lower levels of copper in blood serum of patients as compared to controls. In conclusion ,
based on our brief review we might state that the results obtained from different researchers in respect
of the influence of copper in different diseases and conditions are conflicting. There is a necessity of
accurate and accessible methods for determination of serum copper in research and clinical biomedical
laboratories. It is obvious that further research work is needed in order to clarify the contradictions and
determine the role of copper in the human organism.

Key words: serum copper and diseases, copper enzymes, copper status.

INTRODUCTION oxidative reduction processes and in disposing

Copper (Cu) is an essential trace element for and removing free radicals from the organism
humans and animals. In the human organism, (1). Although scientists identified copper
copper exists in two forms – the first and compounds to treat diseases in 400 BC (during
second oxidation form, as most of the copper Hippocrates) (2), researchers still discover new
in the human organism is in the second form. information regarding the biochemistry,
The ability of copper to easily attach and physiology, toxicology, many clinical,
accept electrons explains its importance in laboratory and other indicators of the impact of
_________________________________ copper in the organism. In this respect, the
*Correspondence to: Svetla Asenova. Department exact composition and structure of copper
of Chemistry and Biochemistry & Physics and compounds, generated in the organism, are not
Biophysics, University of Medicine - Pleven, St. Kl. yet fully elucidated. The overall and inter-
University 1 St., 5800 Pleven, Bulgaria; Tel. 064 organ distribution of copper is not quite
884 157 e-mail: s.p.asenova @ abv.bg
determined (3-11). The role and the
participation of copper and copper enzymes in

88 Trakia Journal of Sciences, Vol. 9, No 1, 2011

 ANGELOVA M., et al.
the metabolism, as well as the interaction of 14 years, a positive correlation is found
copper with other micronutrients are not between levels of trace elements in children
clearly specified (1-3, 10, 12-22). Changes in and their parents. It is believed that this is due
copper concentrations in body fluids and to identical dietary habits of children and their
tissues are observed in different diseases and parents (8).
conditions. There are indications of serious
diseases, caused by disorders of the Copper is a trace element which can be found
metabolism of copper in the organism, but the in almost every cell of the human organism.
role of copper in most of them is not The highest concentrations of copper are
completely clarified (11, 19-20, 23-49). discovered in the brain and the liver; the
Investigation of copper functions in human central nervous system and the heart have high
bodies requires accurate, affordable, concentrations of copper as well. About 50%
informative, low-detection-limit methods for of copper content is stored in bones and
determination of trace copper in biological muscles (in skeletal muscle it is about 25%),
samples (7, 51-60). Although a considerable 15% in skin, 15% in bone marrow, 8 to 15% in
number of methods for copper investigation the liver and 8% in the brain (3).
exists, research continues in search of more
Copper is a functional component of several
sophisticated analytical approaches, for its
essential enzymes, known as copper enzymes –
determination in serum and urine, applicable in
cytochrome c oxidase, lysyl oxidase,
clinical laboratory diagnostics. These current
feroxidase, 2-furoate-CoA dehydrogenase,
developments justify the purpose of our
amine oxidase, catechol oxidase, tyrosinase,
research; to make a brief review of some issues
dopamine beta-monooxygenase, D-galaktozo
related to studying the trace element copper.
oxidase, D-hexozo oxidoreductase, indole 2.3-
COPPER STATUS AND dioxygenase, L-ascorbatoxidase, nitratreducta-
PHYSIOLOGICAL IMPACT se, peptidylglycine monooxygenase, flavonol
The human organism contains about 70 ÷ 80 2,4-dioxygenase, superoxide dismutase, PHM
mg of copper (3). There is evidence that its (peptidylglycine monooxygenase hydroxyla-
content varies over the year and depends on tion) and others. (1, 2, 12-18)
gender and age (4-9). With age significant
Some physiological functions, dependent on
differences are observed in the concentrations
the presence these enzymes in the organism,
of ceruloplasmin (the main carrier of copper in
are described below.
the blood) (5). Regarding gender, some studies
Cytochrome oxidase plays an essential role in
have found no statistically significant
cellular energy. As catalyzing the reduction of
differences in the copper content of blood
molecular oxygen (O2) to water (H2O),
serum in healthy children (10), but other
cytochrome c oxidase generates an electrical
studies discovered that the concentration of
gradient, which is used by mitochondria to
serum copper has significantly higher levels in
create vital energy for the organism and stored
women (p <0.05) than in men (7). There can
in molecules of ATP (1).
not be distinguished clear trends in serum
copper of subjects with drinking and smoking Another copper enzyme, lysyl oxidase,
habits. Differences in serum copper are participates in cross-linking of collagen and
reported in people from different regions. elastin, which form the connective tissue. The
According to some authors, the demographic effects of lysyl oxidase helps maintain the
differences are due to variations in copper integrity and elasticity of connective tissue in
content of the soil and / or food habits of the the heart and blood vessels, but also plays a
population. Increased physical activity is also role in bone formation (2).
found to reduce the concentration of serum
copper (p <0.05) (7). A strong positive Two copper-containing enzymes,
correlation is established between serum ceruloplasmin (feroxidase I) and (feroxidase
copper and body mass index (BMI) (R = 0.85, II) have the ability to oxidize iron (Fe2 +) to
p <0.001) in large-scale sample of 2233 iron (Fe3 +), which are connected to the protein
subjects, aged 15-65 years (11). transferrin for transportation to the red blood
cells and blood formation. Although feroxidase
There is evidence for a link between copper activity of these two copper enzymes is still
levels in children and their parents. In a study not thoroughly understood, the physiological
of serum copper in 66 healthy children, aged 3- significance and the involvement of copper in

Trakia Journal of Sciences, Vol. 9, No 1, 2011 89

 ANGELOVA M., et al.
iron metabolism has been clearly demonstrated cell storage of copper, can be included here as
(2, 12). well (1).
Many enzymatic reactions, which are essential There are two main forms of copper in serum,
for the proper functioning of the brain and the one form is tightly associated with the plasma
nervous system, are catalyzed by copper protein ceruloplasmin, and the other is bound
enzymes. Dopamine-beta-mono oxidase reversibly to serum albumin. Serum also
catalyses the conversion of the contains copper enzymes – cytochrome c
neurotransmitter dopamine into norepinephrine oxidase and monoamine oxidase. (3)
(12). Monoamine oxidase (MAO) plays a role
in the metabolism of the neurotransmitters There is still no consensus on the best
norepinephrine, epinephrine and dopamine. biomarker for determination of copper status.
MAO functions in the breakdown of the Serum copper or serum ceruloplasmin are most
neurotransmitter serotonin, which justifies the commonly used in order to investigate copper
use of MAO inhibitors as antidepressants (13). levels. According to some authors, these
Myelin sheath is made of phospholipids whose markers should be combined with more
synthesis depends on the activity of the sensitive, specific and functional markers such
cytochrome c oxidase copper enzyme (2). as cytochrome c oxidase, platelets and
Copper enzyme tyrosinase is required for the superoxide dismutase in erythrocytes. In the
formation of melanin pigment. Melanin is future it is possible to use as functional
produced in cells, called melanocytes, and indicators the antioxidant status, the changes in
plays a role in the pigmentation of hair, skin immune function, in combination with other
and eyes (2). biochemical markers. (3)

Superoxide dismutase (SOD) functions as an METABOLISM AND INTERACTIONS

antioxidant, which catalyses the conversion of WITH MICRONUTRIENTS
superoxide radicals (free radicals) in hydrogen Common daily intake of copper is 2 ÷ 5 mg,
peroxide, that can subsequently be reduced to which exceeds the required amount. Copper is
water by other antioxidant enzymes (14). Two carried by the blood protein ceruloplasmin.
forms of SOD contain copper: 1) copper / zinc Once absorbed from the digestive system, it is
SOD is found in most cells of the organism, transported to the liver. Copper is absorbed in
including red blood cells, and 2) extracellular the duodenum and upper sections of the small
SOD is a copper-containing enzyme, located in intestine. It is believed that it is mainly
large quantities in the lungs and in low levels – absorbed in the small intestine, although there
in plasma (2). is evidence for its absorption in the stomach. In
circulation, copper ions are connected to
Ceruloplasmin can function as an antioxidant proteins: ceruloplasmin by 95% and the
in two different ways. Copper and iron ions are remainder – to albumin and amino acids. Small
powerful catalysts in neutralizing of free quantities of copper are excreted in the urine.
radicals. Ceruloplasmin facilitates the The main part of the copper intake in the
catalyzation oxidative processes of the disposal organism is excreted in bile juice. The proper
of free radicals. In feroxidase, the activity of amount of copper which an adult person
ceruloplasmin facilitates iron binding and the requires per day is about 0.9 mg. There is no
degradation of the transport protein transferrin, evidence that high dietary intake of copper is a
and may also prevent free ions (Fe2+) from problem for human health. In the USA, the
participating in the generation of harmful free recommended intake of copper is: for adults ≥
radicals (15). 19.10 mg/d.; for pregnant and lactating women
and adolescents – 8 mg/d (2001). (3)
Copper-dependent enzymes are involved in the
regulation of gene expression. Cellular copper Copper is necessary in human nutrition for
levels may affect the synthesis of proteins in normal iron metabolism and the formation of
the organism by enhancing or inhibiting the red blood cells. Anemia is a clinical sign of
transcription of specific genes. Enzymes, deficiency of both iron and copper. Infants,
which are involved in the regulation of gene who receive food with high iron content,
expression, are copper / zinc superoxide absorb less copper than babies who take food
dismutase (Cu/Zn SOD), catalase (another with low iron content. This suggests that high
antioxidant enzyme); proteins, associated with doses of iron can interfere with copper
absorption in infants (15). There are

90 Trakia Journal of Sciences, Vol. 9, No 1, 2011

 ANGELOVA M., et al.
contradictive data on the copper content of impaired intestinal resorption, parenteral
blood serum in anemia. The results of some nutrition, protein loss (nephrosis syndrome,
studies in children with IDA (19, 20) showed exudative enteropathy and others.
increased serum concentrations of copper. The
authors concluded that high levels of copper Increased concentration of copper is observed
reduced the absorption of iron and adversely in: pregnancy, cholestasis, increased
affected haematological indices. ceruloplasmin – inflammation, tumors,
lymphomas, liver cirrhosis, myeloid leukosis;
High additional intake of zinc - 50 mg/d or Wilson’s disease. Hypercupremia is considered
more for an extended period of time can lead to be related to several acute and chronic
to copper deficiency. Zinc supplemented diet infections and malignancies – leukemia,
increases the intestinal synthesis of cellular Hodgkin's disease, severe anemia
proteins, called metallothioneins. They bind hemohromatosys, myocardial infarction,
metals and do not allow their absorption by hyperthyroidism, etc. Serum levels of copper
intestinal cells. Metallothioneins have a are higher in patients who use contraceptives
stronger affinity for copper than for zinc, so or estrogens.
high levels of metallothioneins due to
increased zinc can cause reduced absorption of Two genetic diseases, which are caused by
copper. On the other hand, it is found that high impaired metabolism of copper, are well
doses of copper affect zinc nutritional status (2, studied.
15).
Menkes syndrome is an acquired condition and
The effect of supplements of vitamin C and a recessive disease which leads to copper
copper on nutritional status of humans is not deficiency (23). Intake and transport of copper
clear. Two studies in healthy men showed that is changed, whereby mineral substances in
the activity of ceruloplasmin oxidase may be cells and organs are allocated abnormally.
impaired by relatively high doses of Symptoms include sparse and coarse hair,
supplementary vitamin C (21, 22). Adverse weak muscle tone (hypotonia), sagging facial
effects of vitamin C supplements on copper features, seizures, mental retardation. Menkes
nutritional status are not reported in any of disease is characterized as a recessive disorder
these studies. with retardation of growth, brittle hair and
focal cerebral and cerebellar degeneration (24).
In another study, the correlation between The neurodegenerative processes change the
serum content of copper, zinc, iron and fat grey matter of the brain – impaired twisted
soluble vitamin A and E in healthy pre-school cerebral arteries (25). This can lead to rupture
children is examined. A strong correlation or blockage of arteries. Weakened bones
between serum zinc and serum copper and iron (osteoporosis) may result in fractures.
is discovered. Serum levels of vitamin A are Symptoms appear during infancy and are
found to be significantly correlated to serum largely due to abnormal intestinal absorption of
zinc and vitamin E as well. The authors believe copper with an average deficit in mitochondrial
that in order to investigate thoroughly the trace copper-dependent enzymes. Reduced supply of
elements and fat-soluble vitamins, they should copper decreases the activity of copper
include in the study nutritional surveys, enzymes (e.g. lysine oxidase), which are
metabolic balance and correlations between necessary for structuring and functioning of
micronutrient levels and anthropometric bones, skin, hair, blood vessels and nervous
variables (stature, weight, body mass index) system. (26, 27). In rare cases, symptoms
(10). appear later in childhood and are less severe.
Early treatment with subcutaneous or
CHANGES IN SERUM COPPER IN intravenous injection of copper supplements
VARIOUS DISEASES AND CONDITIONS (in the form of acetate salts) may be applied in
Deficiency or excess of copper in the organism the treatment of this disease. (28) Wilson's
is observed in metabolic disturbances and in disease is an autosomal recessive disorder,
various diseases and conditions. which causes copper overload and toxicity in
the organism. This impairment of copper
In several studies, the authors concluded that
metabolism causes copper accumulation and
medical conditions in which low
toxic damage to cells primarily in the liver and
concentrations (abnormal) of copper are found
the brain tissue, but also in the kidneys, eyes,
are: Menkes syndrome, Parkinson's disease,

Trakia Journal of Sciences, Vol. 9, No 1, 2011 91

 ANGELOVA M., et al.
joints and other organs. In some cases, large rheumatic heart disease (35), atherosclerosis
quantities of copper can destruct many red (35) , myocardial infarction (36). It is believed
blood cells which results in severe anemia. that these results are due to a lack of a reliable
(29) Symptoms of Wilson's disease usually biomarker of copper status and it is not clear
appear at the age of 5-6 years. Kayser- how copper is associated with cardiovascular
Fleischer ring is a significant diagnostic diseases.
indicator, because of the accumulation of
copper in the eye. Neurological symptoms of Contradictory results were obtained in
the disease include behavioral abnormalities, experimental studies on oxidative stress. When
tremor of the hands, unclear speech, mask the dietary intake of copper was low, adverse
expression on the face. Once the symptoms changes had been observed in blood
emerge, they usually quickly progress. cholesterol, including increased total and LDL-
cholesterol and decreased HDL-cholesterol
Disease diagnosis is confirmed by low levels (37). Results from other studies did not
of the protein ceruloplasmin, increased confirm these findings (38). High copper
excretion of copper in the urine, high levels of supplements for four to six weeks did not lead
copper in the liver or through confirmation of to clinically significant changes in cholesterol
the genetic defect. levels (33), e.g. it was not proved that
increased intake of copper lead to increase of
A retrospective study was performed in oxidative stress.
Bulgaria, which analyzed the clinical and
laboratory profile of 13 patients with Wilson’s It is known that copper plays an important role
disease for a period of 12 years. The onset of in the development and maintenance of
clinical symptoms includes predominantly immune system function, whose exact
neurological manifestations (61.5%). In 54% mechanism is not revealed yet. Neutropenia
of the patients there are clinical and laboratory (abnormally low number of white blood cells –
evidence of liver cirrhosis. The ring of K-F is neutrophils) is a clinical sign of copper
observed in 69% of the cases. Serum deficiency in the human organism. Adverse
concentration of ceruloplasmin was reduced in effects of copper deficiency on immune
92% of the patients, urine excretion of copper function is most pronounced in infants. Infants
in 24-hour urine was increased in 92% and with Menkes disease, which causes severe
copper in serum was increased in all patients copper deficiency, suffer from frequent and
(30). It was discovered that the level of copper severe infections (39). In a study of 11
in the liver in Wilson's disease is about 25 malnourished children with copper deficiency,
times higher than in healthy subjects. A the ability of white blood cells to absorb
content of 89.8 µg/g copper in the liver is pathogens has increased significantly one
reported in a 22-year-old woman who was in month after administration of copper
the subclinical stage of the disease and supplementations (40).
suddenly died. (31)
Osteoporosis is a socially significant disease of
Wilson's disease has a specific treatment. If the the bones. Copper-containing enzyme lysyl
treatment is not implemented on time, it can oxidase is required for the development (cross-
lead to death after the age of 30. linking) of collagen, which is a key element in
the organic matrix of bone. Osteoporosis
Cardiovascular diseases are widely spread and occurs in children and adults with severe
their diagnostics and treatment are essential for copper deficiency, but it is not clear whether
human life and health. Studies on copper deficiency contributes to the
atherosclerosis have shown contradictory manifestation and development of the disease.
results. Some scientists suggest that elevated Serum levels of copper in elderly patients with
levels of copper can increase the risk of hip fractures were found to be significantly
atherosclerosis (32) and others that copper lower than these of controls (41). Studies in
deficiency rather than copper excess holds an healthy adult men and women showed that
increasing risk of cardiovascular disease (33). copper supplements significantly increased
Contradictory results were discovered in bone density (42). However, in another study,
epidemiological studies on serum copper and the addition of copper did not have effect on
copper enzymes in cardiovascular diseases – biochemical markers of bone resorption and
coronary heart disease (34), chronic heart
failure and coronary artery disease (34),

92 Trakia Journal of Sciences, Vol. 9, No 1, 2011

 ANGELOVA M., et al.
bone formation in healthy adult men and copper sulphate and copper amino acid
women (43). chelates (50). Often they are combined with
vitamins and other micronutrients. These
The role of copper in glucose homeostasis in supplements are produced by renowned
diabetic patients has not been thoroughly pharmaceutical companies and could be found
determined. In a study of plasma selenium, under different brand names – Suppravit,
zinc and copper in patients with diabetes Type Doppelherz aktiv products, etc.
1and healthy controls, there were not found
any significant differences in plasma copper In patients with Wilson's disease penicillamine
concentrations of males and females – controls is used to bind with copper and enhance its
and patients with diabetes. The reduction in elimination from the organism. Since
plasma copper concentration in patients with penicillamine dramatically increases urine
poor metabolic control is less strong in women excretion of copper, the dosing should be very
than in men with diabetes. (44) precise (2).
Concentrations of copper in blood serum in It is known that antacids can interfere with
type 2 diabetes were significantly higher than copper absorption when applied in very large
those in controls. In recent years, many quantities (2).
experiments with animals are performed in
order to discover a treatment for impaired CLINICAL LABORATORY METHODS
copper status in diabetes. A copper chelating AND BENCHMARKS (REFFERENCE
agent was used in these experiments. As a VALUES) FOR DETERMINATION OF
result, serum copper was reduced to levels COPPER
consistent with those of controls, treatment The analytical methods which are applied most
with copper-chelating agent decreased insulin often for determination of copper are: atomic
resistance and improved glucose intolerance in absorption spectroscopy – flame and
diabetic patients. In addition, the treatment electrotermal (FAAS and ETAAS) atomic
reduced triglyceride levels in blood serum. In emission spectroscopy (AES) and some
conclusion, the results show that copper was photometric kinetic methods. Reference values
related to the manifestation of type 2 diabetes for copper concentrations in human body fluids
and should be applied in the treatment of and tissues of healthy people are listed in
diabetic patients. (45) Table 1.

A research on patients with obesity and Rükgauer et al. (3) have published reference
hypertension showed statistically significant values for serum copper in healthy children
higher serum levels of copper as compared to aged 1 to 18 years who had not received any
healthy subjects (p <0.001). (11) vitamin and mineral supplements – Table 2.

Studies in children with chronic diarrhea Copper content in serum and urine are often
investigated zinc and copper status (46). The used to determine Cu status in clinical
level of both trace elements in serum was laboratory tests. In some of these methods,
reduced. The authors have found deficit of preliminary preparation of samples for analysis
serum copper in chronic diarrhea (47). – such as extraction, salinity and others, is
required, in order to obtain lower limits of
Best K et al, 2004 (48) found moderate copper detection and / or to improve their accuracy
deficiency in cystic fibrosis patients. Other and precision.
authors (49) found that patients with cystic
fibrosis were at risk of nutritional deficiency Recommended analytical methods, applied in
due to the malabsorption syndrome, associated clinical and chemical determination of trace
with endocrine pancreatic insufficiency. elements (Al, Cu, Zn, As, Cd, Co, Cr, Hg, Mn,
According to them, serum levels of copper and Ni, Pb, Se, etc.), are flame or electrothermal
zinc demonstrate deficiency of these atomic absorption spectrophotometry. They
micronutrients in cystic fibrosis patients. have been adopted by the International
Federation of Clinical Chemistry and
Trace elements alone or in combination can be Laboratory Medicine (IFCC), the Institute of
used as additives to the treatment of different Clinical and Laboratory Standards (CLSI) and
diseases. Copper supplements are used in the the International Committee for
form of copper oxide, copper gluconate, Standardization and Haematology (ICSH).

Trakia Journal of Sciences, Vol. 9, No 1, 2011 93

 ANGELOVA M., et al.
Table 1. Current reference values for copper concentrations in human body fluids and tissues of
healthy people
Sample Units Values, Analytical Reference list
X ± SD or values method
range from – to**
Blood serum mg/l 1.23 ± 0.16 FAAS 51
mg/l 1.16 ± 0.52 AAS 7
µg/dl (15-19 years)* from 64 to160 AAS 52
µg/dl (18–60 years)* 77.11 ± 17.67 AAS 53
Urine µg/l 26.2 ± 10.2 AAS 54
µg/24 h < 51 AAS 31
Liver µg/g 3.58 ± 1.71 FAAS 31
Total blood µg/ml 0.88 AAS 31
count mg/l 0.989 ± 0.254 AAS 54
Hair µg/g from 22.9 to 46.6 AAS 54
µg/g (9-10 год.)* 12.0 ± 10.3 ICP-AES 56
mg/kg (3-15 год.)* 22.1 ICP-AES 6
µg/g from 3.2 to 32.8 FAAS 55
Nails µg/g 2.6 ± 1.4 AAS 54
*Age of investigated patients
** X – average value; SD – standard deviation

Table 2. Plasma copper concentrations in children according to age (X ± SD)

Age of healthy subjects (years) n Copper concentrations in blood serum (µmol/l)
1-2 15 21.1 ± 4.5
2-4 23 21.5 ± 3.9
4-6 19 19.4 ± 5.3
6-10 25 23.4 ± 2.5
10-14 21 21.1 ± 3.7
14-18 17 20.5 ± 4.4

FAAS method are applied in linear correlation laboratories utilise the spectrophotometric
analysis to examine the correlation between methods. The manual application of tests
serum copper, amniotic copper, lysyl oxidase usually requires using one or two reagent as
and collagen III (57). For toxicological some of the methods are direct – without prior
diagnostic purposes, atomic emission protein separation; they only require the
spectrometry with inductively coupled plasma application of masking agents, which are often
(ICP-AES) can be used as a method for included in the composition of reagents. These
determining Cu in the urine, because of its low methods are automated and do not require
detection limit (below 10 µg/l) (58). expensive and technologically sophisticated
equipment. Modern large and small clinical
Detection limit of the spectrophotometric laboratories have at their disposal
methods may be used for determining copper spectrophotometers and autoanalysers with
in biological matrices. The method should be spectrophotometric indications.
applied after all interfering or masking
components are pre-concentrated and separated ORIGINAL ANALYTICAL RESEARCH
from the samples. Copper in serum could be OF SERUM COPPER
determined spectrophotometrically through In our study, we investigated copper in serum
different reagents: 4-(2-quinolylazo)phenol of children with anemia, cystic fibrosis and
(59), disodium salt batokuproin disulfonat chronic diarrhea. Spectrophotometric method –
(BCDS) (60), etc. tests AUDIT diagnostics (Ireland) was applied.
The reagent, used in this method, is 4-(3,5-
The development of tools and methodology dibromo-2pyridylazo)-N-ethyl-N-sulfopropy-
has significantly facilitated the application of lanine, which forms a chelated copper complex
spectrophotometric copper tests for biological and measures the absorption at a wavelength of
fluids and tissues. Therefore, many clinical 580 nm. The absorption is proportional to the

94 Trakia Journal of Sciences, Vol. 9, No 1, 2011

 ANGELOVA M., et al.
concentration of copper in the sample. We supplementation and the use of appropriate
conducted a study of the copper status of supplements can lead to rapid relief of clinical
children aged 1 ÷ 3 years – patients with symptoms. Application of current findings on
anemia, cystic fibrosis and chronic diarrhea, the synergistic correlation between clinical
and a group of healthy controls. indicators and micronutrient levels can reduce
the burden of disease and mortality rates.
The obtained results show significantly lower Further research is needed in order to clarify
copper content – 12.80 ± 2.30 µmol/l, in serum the metabolism of trace elements in sickness
of patients with cystic fibrosis and chronic and health, and enlarge the scope of knowledge
diarrhea (n = 11) and low content of copper – about micronutrient status. Copper is an
12.8 ± 2.1 µmol/l, in serum of 25 children with essential micronutrient for different
anemia. biochemical and physiological processes in
health and disease conditions. Copper can be
CONCLUSION
used in diagnosis, therapy and prevention of
High serum levels of copper are associated
many diseases and conditions. The availability
with increased cardiovascular risk, but the
of accurate and accessible methods for
significance of these findings is still unclear
determination of serum copper in biomedical
because of the effect of ceruloplasmin and
research and clinical laboratories is very
inflammatory conditions on copper serum.
important, because it provides possibilities for
Further research is required in order to clarify
performing studies that will clarify the role and
the relationship between copper nutritional
participation of copper in metabolism of
status, ceruloplasmin levels and cardiovascular
healthy people and patients.
diseases.
ACKNOWLEDGEMENT
While unbound copper and ceruloplasmin can
This study was performed with financial
lead to LDL oxidation in a tube, there is little
support from the University of Medicine –
evidence that the increase of copper in the
Pleven, Bulgaria (project № 12/2008).
dietary intake could increase the oxidative
stress in the human organism. The results from REFERENCES
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