Environmental Toxicology and Pharmacology 22 (2006) 5257

The effect of age, sex, smoking habit and hair color

on the composition of hair
K. Chojnacka

, H. G orecka, H. G orecki
Institute of Inorganic Technology and Mineral Fertilizers, Wrocaw University of Technology, ul. Smoluchowskiego 25, 50-372 Wrocaw, Poland
Received 10 October 2005; accepted 23 November 2005
Available online 7 February 2006
Abstract
Hair samples (N=83) were collected between the years 1996 and 2003 frominhabitants of Wrocaw, a city located in Lower Silesia, south-western
Poland (urbanized and industrialized region). The concentrations of 33 elements: Ca, Mg, Na, K, Cu, Zn, P, Fe, Mn, Cr, Se, B, Co, Mo, Si, V, Ni, Be,
Hg, Cd, Al, Pb, As, Ba, Au, Pt, Ag, Sr, Sn, Ti, W, Sb and Zr in hair were determined by inductively coupled plasma-mass spectrometry (ICP-MS)
and inductively coupled plasma-atomic emission spectrometry (ICP-AES) techniques. The effect of age, sex, smoking habits and hair color on the
elemental composition of hair was investigated. Also, the obtained results were compared with literature data. It was found that the composition of
hair varied with age, sex, hair color and smoking habits. This concerned the majority of elements except Ti, Se, Mn, Ni, Hg, Cd, As, Co (in the case
of effect of age), Al, Cd, Co (sex), Cd, K, Ag, Ca, Co, Be (smoking habit), P, Fe, Al, Mn and Cr (hair color). Therefore, reference values for the level
of the remaining elements should be normalized according to age, sex, smoking habit and hair color. This would make hair analysis more reliable.
2006 Published by Elsevier B.V.
Keywords: Smoking habit; Hair color; Hair analysis
1. Introduction
Human hair has recently been used as a tool that enabled
to assess the environmental exposure to heavy metals (as
biomarker) and metabolic status of an organism concerning
essential trace elements (Bencko, 1995; Ashraf and Jaffar, 1997;
Ashraf et al., 1995). Several advantages of this method were
found, although several limitations were encountered in the
application of hair analysis in, for instance, clinical medicine
(Rodushkin and Axelssom, 2000). The main problems that arose
were related to the interpretation of the results, since the extent to
which elements are accumulated in hair could be a characteristic
of an individual person.
Undoubtedly, hair analysis is a non-invasive method of diag-
nosis. Hair can be easily sampled, collected, stored and trans-
ported. Moreover, it provides a long-terminformation. Also, the
concentration of elements in hair is high when compared with
body tissues or uids (Nowak, 1998).
Release of elements from body tissues and uids to hair is a
method of detoxication of an organism. The excess of a given

Corresponding author. Tel.: +48 71 3203902; fax: +48 71 3203469.

E-mail address: katarzyna.chojnacka@pwr.wroc.pl (K. Chojnacka).
element, either essential or toxic, can be safely deposited in hair.
However, if the content of a given element in hair is elevated, it
does not necessarily mean that a person suffers fromhigh content
of this element. It simply means that this element entered an
organism and was deposited in hair. This process occurs during
hair growth and thus the composition of hair provides a historical
insight of individual exposure and mineral status of an organism.
Difculties with the proper interpretation of results of hair
analysis are due to the absence of well dened reference concen-
tration ranges, problems associated with differentiating between
endogenous and exogenous deposition, inconsistency of hair
concentration anomalies with nutritional status and clinical
symptoms (Rodushkin and Axelssom, 2000). The difculties in
establishing normal or reference ranges are also due to the nat-
ural variance of hair composition as a possible consequence of
age, sex, hair color, ethnical and geographic origin, dietary fac-
tors, etc. (Miekeley et al., 1998; Sharma et al., 2004; Batzevich,
1995).
The aim of the present work was to investigate the composi-
tion of hair of the population group living in an urbanized and
industrialized region of south-western Poland (Wrocaw). The
dependence of hair content on age, sex, tobacco smoking habits
and hair color was examined.
1382-6689/$ see front matter 2006 Published by Elsevier B.V.
doi:10.1016/j.etap.2005.11.006
K. Chojnacka et al. / Environmental Toxicology and Pharmacology 22 (2006) 5257 53
2. Materials and methods
2.1. Hair samples
The studied material was sampled in the years 19962003 from 83 subjects
living in an urban population group in south-western Poland (Wrocaw). Hair
(directly after washing with shampoo and drying) was sampled from nape of the
neck. Samples were washed using acetone and distilled water (Nowak, 1998).
Hair was digested and afterwards underwent multielemental analysis with ICP-
MS and ICP-AES.
2.2. Digestion of samples
Hair samples were digested in Microwave Digestion System CEM 2000
(USA) in closed Teon bombs. Hair (ca. 1.3 g) was mineralized with concen-
trated nitric acid (7.5 ml). The reagent and digestion conditions were chosen in
Fig. 1. (ah) Concentrations of elements in human hair for 015 years (N=8), 1525 years (N=21), 2545 years (N=29) and 4565 years (N=25) of age (mg/kg).
54 K. Chojnacka et al. / Environmental Toxicology and Pharmacology 22 (2006) 5257
order to achieve complete mineralization and decomposition of solid phase into
liquid phase. All the used reagents were of trace pure grade (from Merck). After
digestion, the solutions were lled up to 75 ml with deionized water.
2.3. Analytical methods
The concentrations of 33 elements: Ca, Mg, Na, U, Cu, Zn, P, Fe, Mn, Cr,
Se, B, Co, Mo, Si, V, Ni, Be, Hg, Cd, Al, Pb, As, Ba, Au, Pt, Ag, Sr, Sn,
Ti, W, Sb and Zr in hair were determined. Multielemental analysis of digested
hair samples was performed with inductively coupled plasma mass spectrometry
(ICP-MS; VarianUltraMass 700, Australia) for the elements, except alkali metals
and alkaline earth metals, that were analyzed with inductively coupled plasma-
atomic emission spectrometry (ICP-AES; PU 7000 from Philips, Cambridge,
UK) connected with ultrasonic nebulizer CETAC(USA). The analytical process
was controlled by the use of matrix standard NCS Reference MaterialHuman
Hair NCS ZC81002 from China National Analysis Center. The samples were
Fig. 2. (ah) Concentrations of elements in female (F, N=41) and male (M, N=42) hair as well as in hair of non-smokers (N-S, N=63) and smokers (S, N=20)
(mg/kg).
K. Chojnacka et al. / Environmental Toxicology and Pharmacology 22 (2006) 5257 55
Fig. 2. (Continued).
analyzed in three repeats (the relative standard deviation of the measurement
was as accepted for Certied Reference Material). Mercury was analyzed with
an atomic absorption mercury analyzer (AMA 254, Czech Republic).
3. Results and discussion
Hair analysis of 83 individuals living in south-western Poland
was carried out. The statistical analysis, including multi-variant
analysis, was shown previously (Chojnacka et al., 2005). Table 1
presents mean values and standard deviations of the concen-
tration of 33 elements in hair (Chojnacka et al., 2005). When
comparing with reference ranges elaborated for an unexposed
population (Rodushkin and Axelssom, 2000), the concentrations
of Ag, Al, Ca and Na elements were higher in the examined sub-
jects.
The studied population was divided into four age groups:
015 years (N=8), 1525 years (N=21), 2545 years (N=29)
and 4565 years (N=25). Fig. 1 shows mean concentration of
elements for each studied group. It was found that the level of
the majority of elements differed signicantly between the age
groups (when comparing the average levels). Moreover, tenden-
cies of the variation in the content of elements were observed.
For this reason, the elements could be classied into the follow-
ing categories (years):
- (Ca) High level at
1545
Low level at <15, >45
- (Na, K) High level at <15,
>45
Low level at 1545
- (P, Al, B, Pb, Fe, Cr,
Au, Pt, Sb, Be, W)
High level at <15 Low level at >15
- (Cu, V, Sn) High level at >45 Low level at <45
- (Zn, Si, Mg, Sr) High level at
1525
Low level at <15, >25
- (Ag, Zr) High level at
2545
Low level at <25, >45
- (Ba) High level at >25 Low level at <25
- (Ti, Se, Mn, Ni, Hg,
Cd, As, Co)
The level does not depend on age
The concentration of elements from the group (Ti, Se, Mn,
Ni, Hg, Cd, As, Co) was rather related to either environmen-
tal exposure or entered an organism from food and drinking
water.
On the basis of the above results we concluded that when
elaborating the reference ranges for hair analysis it is necessary
to take into consideration the age of the studied person. Hair of
Table 1
Descriptive statistics for element concentration in hair froman urban population
group living in south-western Poland (Wrocaw) (83 subjects) (Chojnacka et al.,
2005)
Element Mean (mg/kg) S.D.
Ag 0.395 0.78
Al 14.938 29.40
As 0.044 0.11
Au 0.049 0.14
B 2.041 2.09
Ba 2.025 2.53
Be 0.055 0.05
Ca 1087.8 970
Cd 0.114 0.14
Co 0.034 0.04
Cr 0.568 1.04
Cu 12.352 12.05
Fe 15.000 16.07
Hg 0.500 0.39
K 209.68 407.2
Mg 66.991 65.14
Mn 0.601 0.59
Mo 0.017 0.02
Na 217.330 268.78
Ni 0.838 1.13
P 132.031 271.84
Pb 1.046 1.39
Pt 0.0004 0.0003
Sb 0.455 0.32
Se 0.679 0.94
Si 57.51 145.9
Sn 1.205 0.92
Sr 2.882 3.69
Ti 1.491 2.28
V 0.092 0.12
W 0.002 0.00
Zn 156.48 74.5
Zr 0.568 1.77
56 K. Chojnacka et al. / Environmental Toxicology and Pharmacology 22 (2006) 5257
children (<15 years) has a tendency to contain more Na, K, P,
Al, B, Pb, Fe, Cr, Au, Pt, Sb, Be and W and less Ca, Cu, V,
Sn, Be, Ag and Zr than hair of older people. Hair of individuals
in the age 1525 years (young people) has higher levels of Ca,
Zn, Si, Mg and Sr and lower levels of Na and K than people
from other age groups. Hair of adults in the age 2545 years
has higher level of Ca, Ag and Zr and lower level of Na and K.
People from the oldest group 4565 years have higher level of
Na, K, Cu, V and Sn and lower level of Ca, Zn, Si, Mg, Sr, Ag
and Zr in their hair.
Also, a group of males and females was distinguished from
the studied population (Fig. 2). In females hair, the level of the
following elements was higher when comparing with males hair
(the average concentrations were compared): Ag (2000%), W
(1700%), Sr (350%), Ca (280%), Mg (270%), Si (240%), Ba
(190%), Cr (170%), P (150%) and Mn (150%). In males hair the
level of the following elements was higher than in females hair:
Sn (3400%), V (1100%), Au (780%), Sb (340%), Pb (280%),
Zr (280%), K (230%), B (210%), As (210%), Na (180%), Mo
(170%) and Hg (150%). The content of Al, Cd and Co was
similar and thus, we concluded that it did not depend on sex.
Also, a group of tobacco smokers and non-smokers was dis-
tinguished (Fig. 2). Hair of non-smokers generally contained
more V (3000%), Se (420%), P (350%), Sr (240%), Si (240%),
Ba (190%), Na (180%) and Cr (170%) than hair of smokers.
Hair of smokers contained more Zr (550%), Mo (270%) and As
(150%) than hair of non-smokers. The level of Cd, K, Ag, Ca,
Co and Be was similar. Surprisingly, the content of the majority
of elements in hair of non-smokers was higher than in the hair of
smokers. Similar results were obtained (Ellis et al., 1981; Frery
et al., 1993; Hoffmann et al., 2000). They found that smokers
have elevated blood cadmium levels when compared to non-
smokers. These studies showed conicting results with respect
to hair cadmiumconcentrations in smokers versus non-smokers.
However, they found little correlation with cadmium in hair and
active cigarette smoking, although it was the major reason for
elevated blood and urine cadmium concentrations.
The differences in the composition of hair of people having
various hair colors were also studied (Fig. 3). Naturally colored
hair were also compared with those that were articially col-
ored. It was found that colored hair contained more Sr (400%),
Ba (270%), Ca (270%), Mg (260%), W (250%), Mo (180%),
Ag (150%) and Mn (140%) and less V (900%), Zr (800%), Sb
(380%), Pb (250%), As (200%), Si (190%), K (180%) and Hg
(170%) than naturally colored hair. Generally, colored hair con-
tained 1800% more of all the elements studied when compared
with other groups. Among naturally colored hair, the highest
level of a given element was denoted for Si (1600%, dark blond),
Ni (360%, dark hair), Co (340%, blond), Au (330%, dark hair),
Cd (230%, dark hair), Sb (230%, dark hair), V (200%, auburn
air), Pt (200%, auburn hair), Ag (180%, dark hair), Sr (170%,
blond hair), Mo (170%, dark hair), Ca (170%, blond) and Mg
(170%, blond) and the lowest level was denoted for: V (2400%,
dark hair), Mo (150%, blond hair), Si (360%, dark hair), Se
(350%, blond), Al (340%, dark blond), Cd (330%, blond), Ba
(340%, dark blond), K (300%, dark hair) and Mn (300%, dark
blond).
Fig. 3. (ac) The concentration of elements in hair of different color (mg/kg).
Few literature studies report that the elemental composition
of human hair depends on age, sex and hair color. Nowak (1998)
examined the subjects living in Poland in a non-industrialized
region (Silesian Beskid). The author examined the effect of age,
sex and hair color on elemental composition of hair. Nowak
(1998) divided the studied population into two age groups (030
and 3180 years). In the present paper, four age groups are dis-
cussed. Therefore, it was difcult to compare the effect of age.
When comparing the effect of sex, similar trends of elevated
concentration of Pb in males hair and Ca in females hair, and
no effect of sex on Co content, were detected. More similarities
were found when comparing the effect of hair color. In the paper
of Nowak (1998) and also in the present paper the level of Ca in
fair hair was signicantly higher than in dark hair. The level of
K. Chojnacka et al. / Environmental Toxicology and Pharmacology 22 (2006) 5257 57
the following elements was higher in dark hair: Pb, Ni, Cd, Co
and Cu. The level of the following elements was not the function
of hair color: Mn, Fe and Na.
The results were also compared with data reported by other
authors. The following similarities were observed:
- Higher level of Ni in females, higher level of Cu and Pb in
males; the level of Cd did not depend on sex (Sharma et al.,
2004).
- Higher level of Zn in smokers, higher level of Fe and Hg in
non-smokers; the level of Cd did not depend on smoking habits
(Sharma et al., 2004).
- Higher level of Se and Sb in non-smokers (Hosseini et al.,
1996).
- Higher level of Pb in children hair, lower level of Zn in hair
of older people; the level of Cd was not the function of age
(Rivai, 2001).
- Ashraf and Jaffar (1997) examined the effect of age on element
contents in hair. Similar tendencies in metal accumulation in
hair with age were obtained for Cu, Ca, Mg, Fe and K.
4. Conclusions
Hair analysis might soon become a powerful diagnostic tool
in monitoring of environmental exposure and in studying the
nutritional status of individuals. However, it is necessary to
precisely elaborate standard ranges of hair composition for dif-
ferent groups of people, with consideration of age, sex, hair
color and living habits. The results of the present study might
be useful in elaboration of reference values for the concen-
tration of elements in hair with the consideration of age, sex,
smoking habit and hair color. It is necessary to perform fur-
ther, wider examination of people living in different geographic
regions.
In the present paper it was found that the composition of
hair varied with age, sex, hair color and smoking habits. This
concerned essential elements as well as heavy metals with few
exceptions. The level of the following elements was not the
function of age: Ti, Se, Mn, Ni, Hg, Cd, As and Co; sex: Al,
Cd and Co; smoking habit: Cd, K, Ag, Ca, Co and Be; hair
color: P, Fe, Al, Mn and Cr. The obtained results were com-
pared with the ndings of other authorssome tendencies were
conrmed.
Therefore, in order to evaluate global reference values for hair
analysis it is necessary to study the effect of age, sex, smoking
habit and hair color on large group from various populations
living in different regions. In this way, hair analysis would be
more reliable.
Acknowledgment
This research was nancially supported by Polish State Com-
mittee for Scientic Research (No. PBZ KBN 060/T09/2001).
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