Open Journal of Endocrine and Metabolic Diseases, 2013, 3, 236-240 Published Online August 2013 (
Evaluation of Serum Trace Elements and Vitamin Levels in
Hashimoto’s Thyroiditis: Single Centre Experience
fr om Turkey
Fatma Dilek Dellal1*, Mutlu Niyazoglu2, Esranur Ademoglu1, Suheyla Gorar1, Zehra Candan1,
Handan Bekdemir1, Ziynet Alphan Uc1, Mehmet Senes3, Aysenur Ozderya4, Yalcin Aral1
1Endocrinology Department, Ankara Training and Research Hospital, Ankara, Turkey
2Endocrinology Department, Cerrahpasa Medical Faculty, Ankara, Turkey
3Biochemistry Department, Ankara Training and Research Hospital, Ankara, Turkey
4Endocrinology Department, Sirnak Government Hospital, Ankara, Turkey
Email: *
Received April 14, 2013; revised May 14, 2013; accepted June 14, 2013
Copyright © 2013 Fatma Dilek Dellal et al. This is an open access article distributed under the Creative Commons Attribution Li-
cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Aim: To determine levels of serum trace elements and vitamins, and to find out possible correlations between these
elements and vitamins with thyroid function tests and thyroid autoantibody levels in patients having Hashimoto’s
thyroiditis (HT). Methods: The study included 51 premenauposal women with untreated HT, aged 18 to 56 years
without any known chronic diseases or chronic medicine usage, and 27 healthy premenauposal women aged 19 to 42
years old. Trace elements (selenium, zinc, copper, iron levels) and vitamins [A, E, B12, 25-OH-D, 1,25(OH)2D and
folic acid levels] were evaluated in patient and control groups. Results: Consequently, serum trace elements and
vitamin B12 levels did not significantly differ in patients with HT and control group. Thyroid functioning tests and
autoantibody levels did not show any correlation with the levels of trace elements, vitamin A, vitamin E and 25-OH
vitamin D. A correlation was detected between vitamin B12 and Anti thyroid peroxidase levels. Conclusion: The
negative correlation between vitamin B12 and Anti thyroid peroxidase levels may demonstrate the necessity to screen
the patients with HT for atrophic gastritis. We believe that more comprehensive studies with larger sample sizes are
needed in which patients are randomized according to their nutritional status.
Keywords: Hashimoto’s Thyroiditis; Trace Element; Vitamin, Vitamin B12, Atrophic Gastritis; Anti Thyroid
Peroxidase Antibody
1. Introduction
Hashimoto’s thyroiditis (HT) is an autoimmune disease
caused by the destruction of thyroid gland in various de-
grees via numerous immune mechanisms. HT is diag-
nosed with the elevated thyroid antibodies in serum. Auto-
antibodies, genetic tendency, intracellular oxidative me-
chanisms and cytokines lead to cellular apoptosis and thus
result in follicular destruction. Hypothyroidism may de-
velop due to the destruction of thyroid gland in HT.
Trace elements are necessary for development, growth
and physiology of the organism. They take part in various
mechanisms in the body, but cannot be synthesized in the
organism. These elements are iron, zinc, copper, selenium,
chloride, florid, iodine, chrome, manganese, bore, cobalt,
molybdenum, vanadium, spelter and silisium. They take
part in functions like immune regulation, nerve conduc-
tion, regulation of membrane potential and maintenance
of mithocondrial activity.
Zinc is an essential element for thyroid hormone func-
tions [1]. Copper is the vital component of numerous oxi-
dative enzymes. [2]. Free copper takes role on cellular
membranes as a pro-oxidant agent. Selenium is essential
for deiodinase activities and thyroid hormone synthesis
and metabolism. Selenium acts as a co-factor in the struc-
ture of glutation peroxidase which has anti-oxidant fea-
tures. Glutation peroxidase takes role in degradation of
hydrogen peroxide to water. After interacting with vita-
min E, selenium protects the cellular membrane against
oxidative damages caused by lipid metabolism [3]. Iron
takes place in structures of many enzymes in the body.
*Corresponding author.
opyright © 2013 SciRes. OJEMD
Vitamins are the essential elements which are necessary
for occurrence of metabolic events and maintenance of
health status, while they cannot be synthesized in the body
or synthesized inadequately, and need to be intaken.
Non-enzymatic anti-oxidants, like vitamin E and A,
contribute to decrease the oxidative damage caused by
oxygen radicals by taking their high-energy electrons [4].
Another function of vitamin E is to increase the absorp-
tion of vitamin A from the intestines and its level in the
tissues. Concomitance of hypothyroidism and pernicious
anemia is very frequent, and vitamin B12 deficiency is
observed in pernicious anemia. Due to its antiinflam-
matory and immunomodulatory features and potential
effects on cytokine levels, decreased levels of vitamin D
is associated with the increased risks of many disorders,
particularly autoimmune diseases [5,6]. Folic acid, which
is actually a pro-vitamin, is changed to dihydrofolat by
dehydrofolat reductase enzyme after being absorbed, and
then it is converted to tetrahydrofolate. Using single car-
bon units, the nascent tetrahydrofolate transfers single
carbon to some endogenous substances via various oxi-
dating mechanisms.
In this study, we aimed to determine the levels of serum
trace elements like selenium, zinc, copper and iron, and
vitamins like A, E, B12, 25-OH-D, 1,25(OH)2D, folic acid
in patients with HT and evaluate the association between
thyroid antibodies and these elements and vitamins.
2. Material and Methods
This prospective study included 51 premenopausal wo-
men aged between 18 to 56 years and 27 healthy pre-
menopausal women aged between 19 to 42 years, who
had applied to our clinic. Only female participants are
involved in order to create a homogeneous group. Pa-
tients were newly diagnosed and untreated with L-thy-
roxine. Patients with any known diseases (diabetes mel-
litus, hypertension, hyperlipidemia, coronary artery dis-
ease, chronic liver or kidney diseases, gastrointestinal
absorption problems, collagen tissue disease, bone me-
tabolism disease, thyroid disease or malignancy) or chro-
nic medicine users were excluded. Demographic charac-
teristics, information on current smoking and alcohol
consumption and personal and familial histories were re-
corded, and detailed physical examinations were per-
formed. Informed consent forms were obtained from all
patients. Approval of Local Ethics Committee was taken.
The study was performed in accordance with Helsinki
Declaration and Good Clinical Practice.
HT was diagnosed by elevated anti thyroid peroxidase
(anti-TPO), anti thyroglobulin (anti-TG) levels and thy-
roid ultrasonography evaluation which revealed hetero-
geneity and fibrotic bands in thyroid glands [7]. Subjects
with normal thyroid antibodies were considered to be
Weight, waist circumference, hip circumference and
height were measured in fasting status and with daily
clothes by the same person. The waist circumference was
accepted as the narrowest diameter between the arcus
costarum and spina iliaca anterior superior, and the hip
circumference was considered as the largest diameter over
the gluteus maximus posteriorly and symphysis pubis
anteriorly. Body mass index (BMI) was the ratio of the
weight to the square of height (weight/heigh2- kg/m2).
Blood samples were collected following 12 hours of
fasting. In order to study selenium levels (N: 46 - 143
µg/L), the collected blood samples were centrifuged at
5000 rpm/min after coagulation and stored at 80˚C until
testing. The test was performed manually by AAS Hydro
System Management. Blood samples were collected from
antecubital veins to evaluate iron, copper, zinc, albumin,
thyroid stimulating hormone (TSH), Anti-TG, Anti-TPO,
TSH receptor antibody (TRAB), vitamin A, vitamin E,
vitamin B12, 25-OH vitamin D, 1.25-(OH)2 vitamin D
and folic acid. Iron (N: 70 - 180 µg/dL), copper (N: 12.6 -
24 µmol/L) and zinc (N: 10.4 - 22.9 µmol/L) were de-
tected by Olympus AU 2700 equipment using the original
kits. 25-OH vitamin D (N: 10 - 80 ng/mL), 1.25-(OH)2
vitamin D (N: 10 - 60 pg/mL) and TRAB (N: 0 - 14 U/L)
levels were detected by radioimmunaassay (RIA) method
using Dia Source kit. fT3 (N: 2.3 - 4.2 pg/mL), sT4 (N:
0.7 - 1.76 ng/dL), TSH (N: µIU/mL), anti-TPO (N: 0 - 60
U/mL), anti-TG (N: 0 - 60 U/mL), vitamin B12 (N: 190 -
911 pg/mL), folic acid (N: 5.38 - 20 ng/mL) levels were
detected using Advia Centaur System (Siemens) and its
original kits by chemiluscent method. Vitamin A (N: 1.05
- 2.8 µmol/L) and vitamin E (N: 11.6 - 46.4 µmol/L) were
tested by HPLC method using Agilent 1200 equipment
and Chromosystems kits.
Statistical analyzes were performed by SPSS 16 pro-
gram. The values were presented as mean ± standard de-
viation. Mann Whitney U test was used to compare the
means between the two groups. P < 0.05 was accepted to
be significant.
3. Results
Features of Hashimoto and control groups are presented
on Table 1.
While there was a significant difference between Has-
himoto and control groups regarding TSH, fT4, Anti-
TPO, Anti-TG and TRAB, there was no significant dif-
ference in terms of FT3 (Table 2).
The levels of serum selenium, zinc, copper, iron, vita-
min E, vitamin A, vitamin B12, folic acid, 25-OH vita-
min D and 1.25-(OH)2 vitamin D did not differ between
Hashimoto and the control groups (Table 3).
A correlation was detected between vitamin B12 and
Anti-TPO levels (r = 0.226, p = 0.04, Figure 1).
Copyright © 2013 SciRes. OJEMD
Table 1. Features of patient and control group.
Control Patient
Age (year) 33.14 ± 6.87 35.37 ± 8.000.2
BMI (kg/m²) 26.76 ± 7.08 27.48 ± 5.170.64
Waist circumference
(cm) 87.08 ± 14.14 87.14 ± 11.680.98
Hip circumference
(cm) 105.57 ± 12.23 106.89 ± 9.490.63
Table 2. Thyroid function tests and thyroid autoantibody
levels of patient and control group.
Control Patient p
TSH (µIU/mL) 1.99 ± 1.22 5.64 ± 6,32 0.0001
fT3 (pg/mL) 3.37 ± 0.33 3.26 ± 0.27 0.132
fT4 (ng/dL) 1.04 ± 0.15 0.91 ± 0.16 0.002
(U/mL) 41.56 ± 13.72 236.74 ± 266.95 0.0001
(U/mL) 31.53 ± 12.48 145.03 ± 118.98 0.0001
TRAB (U/L) 7.4 ± 4.16 4.92 ± 3.32 0.019
Normal ranges: TSH: µIU/Ml, fT3: 2.3 - 4.2 pg/mL, fT4: 0.7 - 1.76 ng/dL,
anti-TPO: 0 - 60 U/mL, anti-TG: 0 - 60 U/mL, TRAB: 0 - 14 U/L.
Table 3. Trace elements and vitamin levels of patient and
control group.
Control Patient p
Selenium (µg/L) 70.03 ± 11.43 67.32 ± 10.29 0.34
Zinc (µmol/L) 13.26 ± 1.77 12.85 ± 2.58 0.42
Copper (µmol/L) 15.75 ± 3.19 16.57 ± 3.57 0.31
Iron (µg/dL) 69.51 ± 40.23 68.68 ± 36.94 0.92
Vitamin E
(µmol/L) 27.64 ± 6.99 29.20 ± 7.22 0.36
Vitamin A
(µmol/L) 1.50 ± 0.35 1.61 ± 0.49 0.25
Vitamin B12
(pg/mL) 298.33 ± 88.17 279.09 ± 98.160.38
Folic acid (ng/mL) 9.64 ± 3.13 9.76 ± 4.37 0.86
25-OH vitamin D
(ng/mL) 17.3 ± 8.0 21.8 ± 15.2 0.18
1.25-(OH)2 v i tamin
D(pg/mL) 23.76 ± 16.75 17.18 ± 11.64 0.09
Normal ranges: selenium: 46 - 143 µg/L, zinc: 10.4 - 22.9 µmol/L, iron: 70 -
180 µg/dL, copper: 12.6 - 24 µmol/L, vitamin E: 11.6 - 46.4 µmol/L, vita-
min A: 1.05 - 2.8 µmol/L, vitamin B12: 190 - 911 pg/mL, folic acid: 5.38 -
20 ng/mL, 25-OH vitamin D: 10 - 80 ng/mL, 1.25-(OH)2 vitamin D: 10 - 60
4. Discussion
Levels of serum trace elements and vitamins did not dif-
fer between HT and control groups in our study. We also
did not determine a correlation between the levels of
trace elements and thyroid antibodies. We determined a
negative correlation only between vitamin B12 and Anti-
TPO levels.
In the recent studies, oxidative mechanisms are con-
sidered to play a role in thyroid autoimmunity. In this
case, thyroid autoantibodies could be expected to be as-
sociated with selenium, vitamin E and vitamin A, which
are called antioxidants, as well as zinc and copper, which
are called antioxidant enzyme co-factors. However, we
could not detect such an association in our study. There
was also no association between vitamin D, folic acid
and iron with thyroid autoantibodies. We did not find any
reports demonstrating correlations between thyroid auto-
antibodies and vitamin E, vitamin A, vitamin, D folic
acid, copper and zinc. While some reports showed an
association between the levels of selenium [8] and auto-
antibodies, some did not [9-11]. Different outcomes in
these studies might be due to nutrition alterations among
the individuals, genetic tendency, different stages of thy-
roid diseases, differences in numbers of patients and their
distributions and/or alterations in laboratory techniques.
There are a few reports regarding the association be-
tween thyroid and vitamin B12. It is known that HT may
accompany other autoimmune diseases. Pernicious ane-
mia is one of them. Of the patients with hypothyroidism,
7% to 12% have evident pernicious anemia and 10%
have latent pernicious anemia [12]. Vitamin B12 defi-
ciency is observed in pernicious anemia [13]. Ness-
Abramof et al. suggested that patients with autoimmune
diseases should be screened for pernicious anemia by
screening vitamin B12 levels in every 3 or 5 years [14].
We did not determine any vitamin B12 deficiency among
patients in our study, but detected a correlation between
vitamin B12 and Anti-TPO levels (r = 0.226). This rela-
tionship might be indicative of an underlying similar
autoimmune pathology. By these results, necessities ap-
pear for the investigation of the level of gastric parietal
cell antibody and for the determination of the co-mor-
bidity of atrophic gastritis, which also an autoimmune
disease, in patients with high levels of Anti-TPO.
Our study had some limitations. Selenoprotein levels
for selenium status, seruloplasmin for copper status, and
zinc levels in urine, erythrocyte for zinc status and parie-
tal cell antibody could not be detected. Dietary habits of
patients were not asked.
Consequently, serum trace elements and vitamin levels
did not significantly differ in patients with HT and the
control group. Thyroid function tests (TSH, fT3, fT4) did
not show any correlation between the levels of trace ele-
Copyright © 2013 SciRes. OJEMD
Copyright © 2013 SciRes. OJEMD
Figure 1. Vitamin B12 and Anti-tpo levels in patient and control groups.
ments and vitamins. Thyroid autoantibody levels did not
show any correlation between the levels of trace ele-
ments and vitamin A, vitamin E and 25-OH vitamin D. A
correlation was detected between vitamin B12 and Anti-
TPO levels. This result may show that patients with HT
should be screened for atrophic gastritis. We believe that
more comprehensive studies with larger sample sizes are
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Anti-TG: Anti thyroglobulin;
Anti-TPO: Anti thyroid peroxidase;
BMI: Body mass index;
HT: Hashimoto’s thyroiditis;
TSH: Thyroid stimulating hormone;
TRAB: Thyroid stimulating hormone receptor antibody.
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