Clinica Chimica Acta 312 Ž2001.

1–11
www.elsevier.comrlocaterclinchim

Review article

Minerals, trace elements and related biological variables in

athletes and during physical activity
Michelle Speich a,) , Alain Pineau b, Françoise Ballereau a
a
´
Departement de Sante´ Publique et de Pharmacie Clinique, Faculte´ de Pharmacie, UniÕersite´ de Nantes, l, rue Gaston Veil, B.P. 53508,
F-44035 Nantes Cedex 1, France
b
` Industrielle, Faculte´ de Pharmacie, UniÕersite´ de Nantes, F-44035 Nantes Cedex 1, France
Laboratoire de Toxicologie et d’Hygiene
Received 10 October 2000; accepted 30 May 2001

Abstract

This review concerns various minerals Žsodium, potassium, calcium, magnesium, phosphorus., trace elements Žzinc,
manganese, selenium, copper, iron, cobalt, iodine, chromium, fluorine, lead, cadmium. and other biological variables Žnitric
oxide, L-carnitine, glutamine, serum transferrin receptor, biopyrrins. in relation to hemorheologic effects, stress, immune
response and infections during physical and sports activities. In athletes, macroelements in the ionized form contribute to
heart and muscle contractions, oxidative phosphorylation and the synthesis and activation of enzymatic systems. Zinc ŽZn.
protects against the effects of increased free reactive oxygen species such as copper ŽCu. and manganese ŽMn. ŽCu–Zn
superoxide dismutases; Mn superoxide dismutase.. Selenium in glutathione peroxidase protects the cardiovascular system
and the muscles, and helps combat allergic and inflammatory diseases. Copper and iron are involved in many aspects of
energy metabolism and are important components in the synthesis of hemoglobin, myoglobin and cytochromes. Fluorine and
Cu protect the ligaments and tendons. Physical activity appears to be beneficial to urban residents who are exposed to metal
pollution Žlead, cadmium.. The data cited in this review are often contradictory and incomplete. It is still unclear in many
cases how minerals are involved in physiological changes, and much work remains. q 2001 Elsevier Science B.V. All rights
reserved.

Keywords: Sports; Physical activity; Minerals; Trace elements; Oxidative stress; Biological variables

1. Introduction constantly occur, involving many enzymatic systems

Human health depends on a delicate balance
among reactions within the organism in which nerve,
muscle, blood, bone, and endocrine and visceral
tissues are continually renewed. Vital exchanges
)
Corresponding author. Tel.: q33-2-4041-2827; fax: q33-2-
4041-2827.
E-mail address: fball@sante.univ-nantes.fr ŽM. Speich..
activated by minerals or trace elements. AMens sana
in corpore sanoB wrote Juvenal ŽSatires X, 356;
60–140 AD. at a time when volcanic ash was con-
sumed and geophagia compensated for mineral defi-
ciencies in the diet.
Sports has long been regarded as an important
cultural value, and the concept of health developed
by the World Health Organization today implies
living a balanced life and neutralizing stress through
physical activity. However, very few publications

0009-8981r01r$ - see front matter q 2001 Elsevier Science B.V. All rights reserved.
PII: S 0 0 0 9 - 8 9 8 1 Ž 0 1 . 0 0 5 9 8 - 8
2 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
have considered the value of minerals and trace
elements in achieving this objective. The present
review investigated most of the literature Žparticu-
larly since 1994. on minerals, trace elements and
some related biological variables in relation to ath-
letes and physical activity.
2. Materials reviewed
Twenty-seven studies published between 1994 and
2000 were analyzed from Medline and ScienceDirect
w1–27x, representing contributions from the USA,
France, Spain, Germany, Finland, Canada ŽOntario.,
China ŽTaiwan., Malaysia, Japan and Sweden. These
studies involved athletes or individuals of both sexes
who performed significant or exceptional physical
activity, in some cases during childhood w6,25x or old
age w19x. Reviews were also included, such as those
of Sobal and Marquart w1x Ž51 studies in 15 different
sports involving 10,274 male and female athletes.
and Fogelholm w2x Ž27 studies concerning 1028 male
and 473 female athletes.. Other studies in the review
relate more particularly to nutritional aspects for
athletes w5,14,22,23,25–27x.
The minerals investigated by these various au-
thors include sodium ŽNa., potassium ŽK., magne-
sium ŽMg., calcium ŽCa., zinc ŽZn., manganese ŽMn.,
selenium ŽSe., copper ŽCu., iron ŽFe., cobalt ŽCo.,
iodine ŽI., chromium ŽCr., fluorine ŽF., lead ŽPb. and
cadmium ŽCd..
3. Findings
All activities of human tissues and organs depend
on minute mineral concentrations that act as enzyme
biocatalysts. For some biological activities, certain
macroelements or toxic trace elements also act as
essential trace elements at concentrations below those
of their constitutive role or with respect to their toxic
nature, such as arsenic ŽAs., vanadium ŽV. and Mn.
During strenuous physical activity, the rate of energy
turnover in skeletal muscles may increase several
fold. As exercise augments reactive oxygen species
ŽROS. that can damage cells, the body contains an
elaborate antioxidant defense system that depends
on the endogenous production of antioxidant com-
pounds and the dietary intake of antioxidant vitamins
and minerals w25,27x.
3.1. Minerals in sports and physical actiÕities
3.1.1. Macroelements
The macroelements Ca, Mg, K, Na, and phospho-
rus ŽP. are generally integrated into anatomic struc-
tures Žbone elements, nucleic acids, membranes, pro-
teins, enzymes., although they are also involved in
the ionized active form and regarded as essential
trace elements, as in voltage-gated ionic channels
w28x. In active form, they are of particular importance
for metabolic balance in sports and during physical
exercise w27x. Naq contributes to the maintenance of
osmotic pressure, water regulation, and acid–base
balance. The cations Naq, Kq, Mg 2q and Ca2q are
involved in the propagation of nerve impulses and in
muscle and heart contraction. Ca2q controls vascular
tonicity and coagulation of the blood. Phosphorus
ŽP., in phosphoric ester form ŽATP., results from the
third step of cell respiration. The active cotransport
of Naq and Kq through the plasma membrane in-
volves the energy of ATP hydrolyzed by NaqKq
ATPase and activated by Mg 2q, constituting an es-
sential cell function requiring around 25% of the
energy metabolism of man at rest w29x. The forma-
tion and use of energy-rich bonds require Mg 2q w30x.
After a long-distance run in high heat and humid-
ity, plasma ŽPl. –Na and Pl–K are significantly in-
creased Ž p - 0.01. and Pl–Mg significantly de-
creased Ž p - 0.01. w24x. Magnesium may be lost in
sweat w27x. Lukaski w4x found that Mg movement
from plasma into erythrocytes was greater as exer-
cise became more anaerobic, and that the amount of
Mg lost in urine was related to the degree of exer-
cise-induced anaerobiosis as indexed by post-ex-
ercise oxygen uptake and plasma lactate concentra-
¨ et al. w14x reported that a morning Pl–Mg
tion. Konig
concentration - 0.75 mmolrl is indicative of a Mg
deficit. Marathon runners show significant decreases
of serum ŽSer. –Mg and urine–Mg w18x, whereas
Shephard and Shek w22x found that Pl–Mg increases
and that ultrafiltrable Mg decreases after maximum
exercise. Nuviala et al. w23x observed that serum
ŽSer. –Mg levels for women involved in different
sports were significantly higher in 23 runners than
65 control women, and that urinary excretion of Mg
 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
was significantly greater in 19 basketball players.
Conversely, hypokinesia ŽHK. may induce mineral
changes in bone, cells of many tissues, serum, urine
and feces. In fact, the use of Mg supplements was
not effective in preventing negative Mg balance in
athletes during a prolonged 364-day period of HK
w31x. Concerning Ca, modest levels of exercise stim-
ulate bone mineralization, but very strenuous exer-
cise may promote Ca loss, especially in women w27x.
For all of these macroelements, the data are often
contradictory, and it is unclear in many cases how
concentrations of these minerals translate into phy-
siological changes. Mineral serumrblood levels do
not usually correlate well with body levels, although
Pohl et al. w32x considered that sweat loss and a shift
of Mg into cells without Mg deficiency were respon-
sible for a significant fall in Ser–Mg in football
players after a match.
3.1.2. Trace elements
Essential trace elements are among the most ion-
izable ones. In the metalloenzyme structure, the es-
sential trace element not only serves as an intermedi-
ary for the enzyme and substrate, but also supplies
the energy in electron form needed for the reaction.
Pearson’s principle allows qualitative predictions rel-
ative to the association sites involved in the interac-
tion of a given metallic ion with a multisite receptor.
3.1.2.1. Zinc. The systemic availability of Zn in
tissues is highly influenced by the balance of an-
abolic and catabolic processes regulating the renewal
of soft and skeletal tissues w27x. Most of the body Zn
content is present in muscle Ž60%. and bone Ž30%..
As no hormone specifically and uniquely involved in
Zn metabolism has been identified, Mg- and Ca-reg-
ulating hormones may affect Zn metabolism directly
or indirectly. Zinc is associated with more than 200
enzymatic systems. It is involved in the synthesis of
nucleic acids, protein synthesis, growth inflamma-
tory syndromes, testosterone secretion and cerebral
function. Zinc augments the natural defenses of the
organism and protects against the formation of free
ROS by Cu–Zn superoxide dismutases present in
cell cytoplasm and mitochondrial membrane space as
well as by a Cu–Zn superoxide dismutase that shows
lower activity in plasma and biological fluids w34x.
3
In inflammatory processes and in stress and post-
traumatic pathologies in athletes, Pl–Zn is decreased
when Zn is redistributed for the repair of damaged
tissues. Zn is taken up in the liver, kidney and spleen
by induction of metallothionein synthesis, but is
decreased in erythrocytes, bone, muscle and urine
w4,8,14,20,22,27x. Rodriguez Tuya et al. w9x reported
a higher Pl–Zn level in anaerobic Žjudo and fencing.
than aerobic Žcycling and endurance. athletes due to
lower Zn mobilization resulting from decreased en-
ergy metabolism and antioxidizing functions. In a
study of women athletes w23x, 16 karate participants
and 23 middle- and long-distance runners had signif-
icantly higher levels of Ser–Zn, whereas 19 basket-
ball players showed significantly lower values. How-
ever, no differences in Zn urinary excretion were
found among these groups. On the whole, these
results indicate that data are very contradictory for
Zn.
3.1.2.2. Manganese. Very little has been reported
about Mn concentrations and their changes during
sports and physical activities, although a superoxide
dismutase found in the mitochondrial matrix func-
tions with Mn w5x. With respect to exercise stress and
antioxidant status, a decrease in Mn superoxide dis-
mutase subsequent to nutritional intervention with
enzymatic yeast cells and antioxidants can be inter-
preted as a sign of reduced free radical stress w35x.
The metalloenzymes of Mn include arginase, pyru-
vate carboxylase and glutamine synthetase w32x. Glu-
tamine is the most abundant amino acid in the
organism, especially in muscles, and glutamic acid is
one of the three constitutive amino acids of glu-
tathione, an important antioxidant w25,36,37x. Man-
ganese, which acts on skeletal growth, the synthesis
of nucleic acids, proteins and hemoglobin, and the
metabolism of lipids and carbohydrates, also has
anti-allergic properties. It acts on the nervous system
as a calcium antagonist and is several times as
efficient as Mg 2q in inhibiting the release of neuro-
mediators into synaptic space, probably because of
competition with Ca2q at the level of calcium chan-
nels.
3.1.2.3. Selenium. During physical activity, the ox-
idative stress due to excessive oxygen consumption
is compensated by higher levels of free radical scav-
4 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
engers and by an increase in the activities of antioxi-
dant enzymes to preserve the structure and functions
of lipids, DNA and proteins w25–27,33,36–39x. For
example, free radicals may interact with albumin,
which contains several disulfide bonds, and cause
variations in its three-dimensional configuration and
binding capacities with metals. Glutathione peroxi-
dase Žan enzyme containing Se. and other antioxi-
dant enzymes act to preserve the cell and its mem-
brane w15,22,27x. In association with vitamin E and
glutathione, Se is anticarcinogenic and delays aging
and degenerative neurological diseases. It protects
the muscles, heart and arteries and helps combat
inflammatory and allergic diseases w22,33x.
Few results are available concerning Se concen-
tration during or following physical activity, and the
levels of glutathione peroxidase reported in athletes
are divergent w15,37x. Endurance training may induce
heterogeneous effects on oxidative and antioxidant
adaptation independently of Se supplementation w15x.
However, athletes are generally not affected by Se
deficiencies w27x.
3.1.2.4. Copper. Although Cu plays an important
role during physical activity, the results for this
mineral are divergent in reports concerning athletes
w2,4,40x. About 23.3% of an adult’s total body Cu is
found in muscle w23x. Copper is a critical nutrient
involved in many aspects of energy metabolism and
an important component for the synthesis of hemo-
globin, myoglobin, cytochromes and some peptide
hormones w27x. It is needed for proper utilization of
Fe and protection against oxidative damage to cells
w4,40x. Copper was mentioned above with respect to
Cu–Zn superoxide dismutases. It is also the cofactor
of proteins involved in redox processes, such as
mitochondrial cytochrome oxidase. Plasma cerulo-
plasmin, a Cu-transport protein, is also an oxido-
reductase. Copper and iron are interdependent both
in the intestine, where Cu allows assimilation of Fe
and in ceruloplasmin, where it catalyzes the transfor-
mation of ferrous into ferric ions. Any Cu deficit can
lead to iron deficiency ŽID. anemia. High Zn intake
impairs Cu retention and decreases SOD activity w4x.
Copper contributes to the synthesis of elastin and
collagen and to the restoration of cartilage in arthro-
sis and to bone structure. It possesses antiviral and
antibacterial properties and also facilitates the heal-
ing of wounds, burns and inflammation. Copper
improves some cases of fatigue and ID anemia,
participates in energy metabolism and intervenes in
neuroactive compounds Žcatecholamine, endorphin.
w35x.
Copper homeostasis is essential in physical activ-
ity and sports w40x. During intense physical exercise,
SOD activity is increased, favoring the adaptation of
Cu metabolism w4,9,25x. Significantly, lower serum
ceruloplasmin biological activity was found in trained
professional soccer players than in a control group,
but with no difference in Ser–Cu w40x. Anuradha and
Balakrishnan w38x reported lower total proteins and
albumin and higher ceruloplasmin in trained male
athletes after long-distance running. Elevated cerulo-
plasmin may be related to its ferroxidase action and
to endogenous response to an acute phase reaction.
No significant differences in Ser–Cu and cerulo-
plasmin were found by Nuviala et al. w23x between
four groups of women athletes and control women.
In fact, Cu measurement in urine was of limited
value because of the slight urinary excretion of this
metal Ž1–2% of that absorbed.. Only the group of
karate women had significantly lower Cu excretion
w23x.
Copper and iron are more abundant in the sweat
of athletes after exercise, particularly on limbs. This
ultrafiltrable Cu appears to be bound to small
molecules capable of stimulating membrane lipid
peroxidation, thereby providing a cytotoxic effect in
controlling skin bacterial flora. As in the case of
Pl–Zn, Pl–Cu was increased in athletes vs. controls,
and Pl–Cu was lower in an aerobic than an anaero-
bic group w9x. Conversely, Ser–Cu was lower Ž p -
0.02. in 49 male ice hockey players Ž12 " 0.5 years.
than in 35 control males Ž12.4 " 0.5 years. w6x. In
fact, age and sex are important in Cu metabolism
w32,41x. Copper concentration in the blood may be
higher, unvaried or lower in athletes than in control
subjects due to varying concentrations of cerulo-
plasmin and Cu-dependent enzymes w40x. The contra-
dictory results reported are inexplicable, but it is
noteworthy that ceruloplasmin works not only by
binding Cu ions, but also Žand mainly. by means of
its ferroxidase activity.
3.1.2.5. Iron. Iron, a component of oxygen-binding
molecules Žhemoglobin and myoglobin. of cy-
 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
tochromes and of many enzyme cofactors Žheme and
Fe–S clusters., can accept and donate electrons,
interconverting between ferric and ferrous forms
w29,42x. During athletic activity, Fe may damage
tissues by catalyzing the conversion of hydrogen
peroxide to free radical ions. However, proteins se-
quester Fe to reduce this threat. Iron ions circulate
bound to plasma transferrin and accumulate within
cells in the form of ferritin. Both ID and Fe overload
are deleterious. During periods of Fe insufficiency or
impaired Fe utilization, Zn becomes an alternative
metal substrate for ferrochelatase, leading to in-
creased Zn protoporphyrin ŽPP. formation w43x. Iron
interacts positively in metabolism with Cu, Cr and F,
and negatively with Zn, Mn, Co and Se w5,23x.
Serum ferritin concentration is lower in women
than men, and the prevalence of low serum ferritin
concentration is higher in female athletes than in
untrained female controls w2x, which is also the case
for male child athletes vs. controls w6x. Males have
greater Fe requirements in early puberty, but large
postpubertal Fe stores may be a source of potentially
harmful effects w6,10,26x. Recently, Constantini et al.
w44x reported that adolescent athletes of both genders
Žgymnasts particularly. are prone to nonanemic ID,
which might compromise their health and athletic
performance Žlow transferrin saturation, low ferritin
concentration.. The organism is particularly Fe-spar-
ing w27,42x. The Fe pool, consisting of about 4 g in
the adult male Žcirculating hemoglobin: 1800 mg,
liver parenchyma: 1000 mg, reticuloendothelial ma-
crophages: 600 mg, bone marrow and muscle myo-
globin: 300 mg each, plasma transferrin: 3 mg. and
2.5 g in the adult female, is continually renewed. In
the balanced state, 1–2 mg of Fe enters and leaves
the body each day. Iron is released from cells when
tissues are damaged and Ser–Fe concentrations in-
crease w18x. Various recent studies have dealt with
the influence of ID on health, even before marked
anemia appears, i.e. when alterations are apparent in
numerous enzymatic systems. Any ID leads directly
to a hemoglobin deficit, resulting in poor cell oxy-
genation affecting the heart and the entire muscula-
ture Žcramps. and in decreased exercise performance
w27x. Heart rate accelerates in order to increase irriga-
tion and compensate for oxygen deficiency in the
blood. Iron needs to be administered to augment
physical capacity during exercise, while relieving
5
heart strain w45x. In addition to causing anemia, ID
may affect physical exercise by altering glycolysis
through production of excess lactate, which gener-
ates highly damaging free radicals w25x. This could
lead to muscular disorders responsible for apathy,
somnolence, irritability and loss of attention, with
resulting risk of accident and incapacity to concen-
trate during sports events w45x. Iron also contributes
to regulating body temperature after physical exer-
cise and to controlling immune defenses w22,33x. The
signs of abnormal encephalin and endorphin meta-
bolism in individuals with Cu deficiency are similar
to those associated with the early stages of ID.
Anemia is often diagnosed erroneously in athletes
because endurance training leads to an expansion of
blood volume and thus to low hemoglobin concentra-
tion, despite normal total hemoglobin content
w22,27,46x. Latent ID may be observed in the en-
durance competitor because of the involvement of
transferrin in antioxidant reactions and a restriction
of intestinal Fe absorption associated with the adop-
tion of a high-proteinrfat diet w22x. Conversely, sig-
nificant increases in hemoglobin, hematocrit and
plasma osmolality, as well as a decrease in mean
plasma volume, were observed immediately after a
race in high heat and humidity w24x. Zinc protopor-
phyrin quantification is useful as a sensitive and
specific tool for evaluating Fe nutrition and meta-
bolism w43x.
3.1.2.6. Cobalt. The human organism is incapable of
transforming cobalt ions into vitamin B12 Žcyanoco-
balamin.. In the human diet, cobalt must be supplied
entirely in its active physiologic form. Vitamin B12
deficiency is the cause of megaloblastic anemia ŽBi-
ermer’s disease.. Cobalt is a regulatory agent of the
sympathetic nervous system, limiting arterial blood
pressure. It is used as a supplement by athletes,
although there is no evidence for increased require-
ment or for any beneficial effect of specific supple-
mentation on performance w27x.
3.1.2.7. Iodine. The only known role of iodine in
humans is to provide an element essential to the
composition of hormones secreted by the thyroid
gland, namely tetraiodothyronine Žthyroxine or T4.
and triiodothyronine ŽT3., which are involved in the
metabolism of all cells of the organism during the
6 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
growth process and in the development of most
organs, particularly the brain. Iodine stimulates the
cardiovascular system and the metabolic combustion
of sugars, lipids and proteins, making its role essen-
tial in athletes w5,27x. Generally, intake is excessive,
and there is no evidence of a greater requirement in
physically active individuals w27x.
3.1.2.8. Chromium. The input of Cr in the diet is
below the optimal value because of the consumption
of refined foods. Chromium is involved in a large
number of enzymatic processes. It increases toler-
ance to sugars through the glucose tolerance factor
ŽGTF., a complex of unknown structure, which po-
tentiates insulin activity. Chromium increases the
number of insulin receptors and can improve matu-
rity-onset diabetes. Chromium andror GTF also seem
important in lipid metabolism through their role as
cofactors of insulin, a hormone involved in both lipid
and carbohydrate metabolism, and by increasing the
HDL cholesterol concentration. Chromium con-
tributes to gene expression and influences protein
metabolism w35x. Chromium metabolism is altered
during stress.
Chromium metabolic variations during sports ac-
tivity have rarely been studied, notably because of
the lack of a good indicator of Cr status or of an
appropriate reference database with sensitive and
reliable assay methods w47x. Endurance activity may
influence Cr status negatively and enhance urinary
Cr excretion due to a complexing of Cr with lactate
and an increase in plasma cortisol w12x. However,
similar Pl–Cr concentrations have been reported in
athletes and untrained controls w2x. Chromium sup-
plementation of young men and women did not
promote muscle accretion, fat loss, or gains in
strength w47x. Chromium picolinate supplementation
of football players was ineffective in bringing about
changes in body composition or strength during a
program of intensive training in weightlifting w48x.
3.1.2.9. Fluorine. Fluorine, cited once by the refer-
enced authors, is the most electronegative nonmetal
w5x. It combines directly with all elements except
oxygen and nitrogen and reacts strongly with most
organic compounds. More than 98% of F in the
organism is found in the skeleton. It constitutes one
of the major elements of bone and tooth structures,
activates certain enzymes, notably those that elabo-
rate collagen during the initial stage of fracture
restoration, and has a positive effect in primary
osteoporosis. In athletes, F may help strengthen the
resistance of interosseous ligaments or muscle ten-
dons during dislocations and sprains and prevent
tendinitis. It binds Ca to bone, thereby avoiding
deposits within vascular walls, and may thus prevent
arteriosclerosis.
3.1.2.10. Cadmium. Cadmium enters the organism
through the lungs Žindustrial discharges, cigarette
smoke., in food, or in drinking water from pipes
with galvanized welds. It has been suggested that Cd
dissolved from galvanized iron pipes might be the
Awater factorB causing hypertension w49x. Its toxic
effects are apparent in erythrocytes, kidney and liver
and by blockage of the biocatalytic action of Cu, Zn
and Co. The manifestations experienced are bother-
some for the athlete since they can produce joint
pain, bone demineralization and a progressive rise in
blood pressure. Athletes, as compared to controls,
showed a significant decrease of Pl–Cd Ž p - 0.005.,
and a similar trend was apparent after athletes were
classified according to aerobic and anaerobic activi-
ties w9x.
3.1.2.11. Lead. Lead enters the organism by the
lungs and in food, drinking water and wine. Its toxic
effects are produced in the kidneys, liver, heart,
arteries and bones, and it inhibits enzymatic reac-
tions essential to hemoglobin synthesis w49x. These
disturbances may be harmful for athletes, leading to
fatigue, headaches, dizziness, intestinal disorders,
muscular pain and cardiovascular disease. Plasma–Pb
concentrations were significantly lower in athletes
than in a control group Ž p - 0.05., which was also
true after the athletes were classified according to
aerobic and anaerobic activities w9x.
Physical activity could provide a positive protec-
tive effect against diseases related to the action of
toxic metals, especially for individuals in large cities
who are more exposed to pollution w9x.
3.1.3. Conclusions and research prospects
The data cited in this review concerning minerals
in relation to sports and physical exercise are often
contradictory and incomplete. In fact, it is still un-
 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
clear in many cases how minerals are actually in-
volved in physiological changes. Reports often fail to
include various essential matters: a description of the
methods for assaying minerals and for their quality
control, the type of samples studied Žwhether total
blood, platelets, erythrocytes, serum, plasma, plasma
fractions, saliva, urine, sweat, feces, or tissues., sam-
pling time, the subject’s condition Žfasting or not.
and age, the sports or type of physical activity
involved Žduration, intensity, phase., nutritional
habits, and possible mineral, vitamin or supplement
intakes. In addition, healthy athletes are not inclined
to provide blood or tissue samples, which makes it
difficult to perform follow-up analyses.
A good mineral and essential trace element status
is fundamental to sports activities, particularly as
physical exercise often creates additional needs, de-
pending on the nature, intensity and duration of
exercise, training periods and pre-competition, the
opportunities for recuperation, and the age and sex of
the individual. The objective should be a varied,
well-balanced diet, although there is certainly no
AwonderB nutrient available w22,44,46,50x. Physically
active subjects should be advised to select and con-
sume food with high nutrient densities rather than to
rely on nutritional supplements w47x. In fact, much
work needs to be done in this area.
3.2. The effects of sports and physical actiÕities on
other markers
3.2.1. Nitric oxide (NO)
Disturbances in the metabolism of nitric oxide
ŽNO. can be related to many pathophysiological
events, in particular, the oxidative stress of sports
activity w28,43,51–53x. Jay-Gerin and Ferradini w53x
have postulated that NO-reductase or NO-dismutase
regulates the high local NO concentrations released
within NO-generating cells in response to oxidative
stress. The active site of these enzymes results from
the formation of Cu–nitrosyl complexes by a mecha-
nism involving oxidation–reduction of Cu, as in
superoxide dismutation. Generally, acute exercise in
healthy subjects causes only mild oxidative stress,
which may be related to an increase in antioxidant
activity and down-regulation of NO formation w52x.
However, NO has a high affinity for Fe and, as an
endothelium-derived relaxing factor ŽEDRF., is a
7
vasodilator that acts on guanyl cyclase, a hemin
enzyme w28,53x. Moreover, possible protective ef-
fects of exercise training against the risk of cardio-
vascular diseases may relate to NO-mediated platelet
functional changes Ždecreased platelet adhesiveness.,
together with changes in Mg and Ca platelet concen-
trations w16,30,53x. In fact, the trained group in the
study of Wang et al. w16x had significantly lower
resting heart rates and blood pressures compared to
the control group Ž p - 0.05. and to pretraining val-
ues and showed improved exercise performance. In-
creased Zn protoporphirin formation after Fe deple-
tion may affect nerve cell metabolism and EDRF
directly through inhibition of NO synthase and solu-
ble guanyl cyclase enzyme activity w28,43x.
3.2.2. Hemorheologic effects
Zinc status affects exercise performance by he-
morheologic effects. Zinc cations protect red cells
against calcium-induced binding of the erythrocyte
cytoskeleton to the membrane w13x. Hyperviscosity is
observed in soccer players with low Ser–Zn, which
can be attributed to increased erythrocyte rigidity,
low blood glucose and higher blood lactate. Hyper-
viscosity leads to changes of erythrocyte distribution
in muscle microvessels.
3.2.3. Physical stress and oxidatiÕe stress
An increase in muscular stress is indicated by a
rise in creatine kinase, myoglobin, protein turnover
and N-methylhistidine and by changes in K, Mg, Zn,
Cu and Fe concentrations w4,14,32,34,54x. During
and following exercise, the course of cytokines and
acute phase proteins, as well as of cortisol levels, is
an indirect indicator of exercise intensity and indi-
vidual ability to cope with physical stress w8,14,20,
34,54x. Athletes with increased interleukin ŽIL.-6
have decreased Pl–Zn and increased Zn sweat loss
w14,20,22x. Several authors have suggested that Zn
and Mg redistribution and modifications in the con-
centrations of Zn, Cu, Fe, Cr, K, Mg, Na and Ca
occur in sweat and urine during and after physical
activity w4,8,20,24,27,32x. Corticoids, catecholamines
and IL-6 induce metallothionein synthesis, resulting
in the sequestering of circulating Zn, Pb and Cd by
the liver and other tissues w8,20,49x.
Compared to age- and sex-matched control sub-
jects not engaged in training, female and male ath-
8 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
letes showed significant increases in superoxide
dismutase ŽSOD. activity in erythrocytes during
physical training w4,9,34,36x. There was a significant
correlation between SOD and the degree of training.
A decrease in Mn–SOD concentration might be in-
terpreted as a sign of reduced free radical stress w34x.
Although these studies suggest that training enhances
antioxidant capacity, the results are not as clear as
for NO. In particular, the studies used different
markers of antioxidant status and involved different
training levels for subjects w15,25x. Moreover, in-
creased or decreased blood concentrations of anti-
oxidants or of oxidative metabolites do not reflect
concurrent changes at the tissue level and may have
minimal physiological implications. The high degree
of variability in resting concentrations of some of
these indicators, and in their response to exercise, is
indicative of the complexity of the relationships
among minerals, exercise, oxidative stress and the
generation of free radicals w25x.
3.2.4. Immune response and infections
Intense physical exercise has been found to be
associated with immunosuppression and an increased
rate of infection. Some athletes can withstand intense
training periods without health problems, while oth-
ers are prone to infections. Many factors may inter-
fere with immunoregulation: nutritional imbalance,
fatigue, stress, and the fragility of the organism
w21,22x. In particular, it has been shown that trace
elements are related to cell-mediated and humoral
immunity w21,33,42x. Deficiencies in Se, Zn, Fe, Mg
and Ca may have effects on exercise tolerance and
immune function w21,22,30x. Iron deficiency causes
overall atrophy of immune tissues, and major Zn
deficiency leads to lymphoid atrophy w22x. Supple-
ments may be required after heavy sweating, but
excessive intake of Fe facilitates bacterial growth
w22x. It is important in dietary recommendations to
athletes to recognize that immune response can be
jeopardized not only by deficiencies, but also by
excessive intake of certain nutrients w42x.
3.3. Related biological Õariables
Finally, certain other currently assayable biologi-
cal variables should not be overlooked since they are
related to mineral metabolism in athletes.
3.3.1. L-Carnitine and glutamine
L-Carnitine, a quaternary amine related to L-lysine
and L-methionine, is a physiological compound that
exists in all cells of the body and is particularly
abundant in muscle and myocardium Žtissues rich in
K, Mg, and Zn., where paradoxically, it is not
synthesized. L-carnitine plays a fundamental role in
lipid utilization and is also involved in intermediary
metabolism. L-Carnitine is provided by the exoge-
nous food supply Žred meat, milk. as well as by
endogeneous synthesis Žmainly in liver and kidney..
Carnitine deficiencies occur when the diet is inade-
quate or in conditions causing abnormal renal loss or
overconsumption notably during physical activity
Žincreased amino acid catabolism, and loss of body
proteins and minerals. w55x. In elite athletes, good
L-carnitine balance appears to be quite useful during
competition. In particular, L-carnitine is involved in
transporting long-chain fatty acids, energy meta-
bolism, prevention of hemolysis and protection of
membrane fluidity and mineral permeability w32,55x.
During sports activity, increased secretion of stress
hormones and cytokines, increased excretion of N-
methylhistidine, K, Mg and Zn, and intracellular
glutamine depletion are potential mediators of al-
tered protein metabolism w22,30,32x. Biolo et al. w55x
found that both carnitine and alanyl-glutamine lim-
ited whole-body nitrogen loss in severely trauma-
tized patients.
3.3.2. Serum transferrin receptor
Red-cell production is not disturbed until Fe stores
are depleted, i.e. until serum ferritin levels become
low w11,42,46x. When the stores have been used up,
Fe saturation of transferrin decreases and subjects
begin to show evidence of iron-deficient erythro-
poiesis. The levels of free protoporphirin ŽPP. and
Zn PP increase in erythrocytes, whereas those of
soluble transferrin receptor ŽsTfR., a plasma protein-
cleavage product, increase when ID limits the pro-
duction of new red cells. Most circulating receptors
come from erythroid marrow precursors w56x. The
synthesis of ferritin and transferrin receptors, which
are present on all human cells, but more frequent on
erythroblasts, is regulated in an opposite manner:
regulation occurs at the mRNA level and depends on
intracellular Fe concentration w42,56,57x.
 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
During physical and sports activities, the mea-
surement of sTfR can be a powerful tool for the
diagnosis of ID in a variety of physiological and
clinical situations. It is a sensitive means of detecting
early-developing ID erythropoiesis, and it retains its
specificity to changes in Fe status, irrespective of
concurrent inflammatory or infectious status or of
cytolysis w56,57x. Contrary to the situation for fer-
ritin, there is no sex-related difference for sTfR in
the adult and no difference between pre- and post-
menopausal women. Its intraindividual biological
variability is markedly lower than that of ferritin,
iron and the saturation coefficient of transferrin w57x.
sTfR determination is particularly indicated in groups
of subjects with low physiological stores that limit
the value of assaying ferritin, i.e. young children,
adolescents, pregnant women and trained athletes.
An intensive training session Ž90 min. in athletes did
not modify basal sTfR concentration, whereas a 50%
increase occurred under recombinant human erythro-
poietin Žr-Hu EPO., a forbidden doping substance.
Thus, the sTfR assay might be one means of detect-
ing the illegal use of this substance as part of the
battle against doping and associated problems within
the Public Health sector w46,56x.
3.3.3. Bilirubin and biopyrrins
A last group of substances consists of bilirubin
and biopyrrins w58–60x. Bilirubin has been shown to
scavenge ROS produced by oxidative stress in vivo,
in which case the oxidative metabolites of bilirubin
Žbiopyrrins. are excreted into urine. Heme oxygenase
isoenzyme 1, an inducible enzyme of bilirubin bio-
synthesis, is enhanced by oxidative stress, heavy
metals or hormones w58x. Shimoharada et al. w60x
have confirmed that biopyrrins in urine may prove
useful as a new practical test for estimating oxidative
stress in vivo. It would be useful to determine these
indicators in athletes whose ROS concentrations in-
crease because the high reactivity of ROS, together
with their very short life span, complicates the direct
measurement of these species in human subjects
w60,61x.
4. Conclusion
It would seem useful in research concerning ath-
letes to relate NO metabolites, hemorheologic ef-
9
fects, immune response, infectious and allergic dis-
eases, L-carnitine, glutamine, sTfR andror biopy-
rrins to the minerals involved in energy metabolism
and oxidative stress. In this respect, minerals have
many functions: transportation of electrons in redox
reactions ŽFe.; components of mitochondrial cy-
tochrome oxidase ŽCu., superoxide dismutases ŽCu,
Zn, Mn., glutathione peroxidase ŽSe. and thyroid
hormone ŽI.; cofactors of dehydrogenases and car-
bonic anhydrase ŽZn.; catalysts in protein, lipid and
carbohydrate metabolism; or activators of adenyl
cyclase and membrane ATPases ŽMg.. The cellular
role of macroelements, in the form of highly mobile
ions ŽNa, K, Ca, Mg. in voltage-gated ionic chan-
nels, and of P is also an important factor for cardiac
and muscular contractions, the Krebs cycle, oxida-
tive stress and apoptosis w28x. Trace elements and
macroelements are constantly involved within the
human body, playing a key role in cellular metabo-
lism from the lowest level of intracellular life to the
functional activity of the largest organs. This role
differs depending on age, sex, physiological or
pathological state and the physical activities per-
formed. For athletes, Se, Zn, Fe, Mg and Ca increase
tolerance and the immune function; K, Mg, Ca and
Fe favor muscle and heart contraction and relaxation;
Mg, Zn and I improve endurance; Cr, Fe and Mn
activate the utilization of stored sugars; Mg, Cu and
Mn heighten reflexes; Zn, Cu, Mn and Se eliminate
toxins and free radicals; and F, Ca, Cu and Mn
prevent tendinitis or torn ligaments. Moreover, nickel,
Co and Mn may stimulate the production of EPO
w62x. Physical activity can provide a positive protec-
tive effect against diseases related to the action of
toxic metals ŽPb, Cd., especially for individuals liv-
ing in large urban areas and exposed to various
forms of pollution. However, the need for these
elements, supplied essentially by a balanced diet, is
not the same for everyone. Certain questions remain
unanswered: Is it necessary to provide mineral sup-
plements for athletes according to their age, sex, or
type and duration of sports activity? If so, at what
period of their sports career? Which elements are to
be recommended, and which may be considered as
useless or even harmful? Which elements should be
analyzed with sufficient sensitivity to detect and
correct deleterious abnormalities? Mineral variations
during exercise or during the hours and days of
10 M. Speich et al.r Clinica Chimica Acta 312 (2001) 1–11
training or recuperation often appear to be more
physiological than pathological in populations that
are generally well-nourished and show normal tran-
sient variations in blood volume and homeostasis.
Further research needs to be performed. For many
minerals, there are few ŽMn, Se, Co, I, Cr, F. or
contradictory ŽMg, Zn, Cu. data about the concentra-
tions found in athletes and during physical activity.
Moreover, the relationships between the serumr
blood concentrations of a mineral and its tissue
concentrations are still largely unclear.
Acknowledgements
The authors are grateful to Prof. James Gray for
native linguistic assistance in the preparation of this
article.
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