Biol Trace Elem Res (2010) 134:109–117

DOI 10.1007/s12011-009-8449-z

Toxic and Essential Mineral Elements Content of Black

Tea Leaves and Their Tea Infusions Consumed in Iran

Maryam Salahinejad & Fereydoon Aflaki

Received: 2 June 2009 / Accepted: 25 June 2009 /

Published online: 17 July 2009
# Humana Press Inc. 2009

Abstract The metal contents of eleven black tea samples, four cultivated in Iran and seven
imported, and their tea infusions were determined. Twelve elements consisting toxic metals
(Al, As, Pb, Cr, Cd, and Ni) and essential mineral elements (Fe, Zn, Cu, Mn, Ca, and Mg)
were analyzed using inductively coupled plasma atomic emission spectroscopy (ICP-AES).
Al, Ca, Mg, and Mn ranged in black tea leaves at mg g−1 levels, while Cr, Fe, Ni, Cu, Zn
were at µg g−1 levels. Analysis of variance showed no statistically significant differences
among most elements determined in cultivated and imported black teas in Iran except for Ni
and Cu. The extraction efficiency of each element into tea infusions was evaluated. The
solubility of measured metals in infusion extracts varied widely and ranged from 0 to
59.3%. Among the studied elements, Cr, Pb, and Cd showed the lowest rates of solubility
and Ni had the highest rates of solubility. The amount of toxic metals and essential mineral
elements that one may take up through consumption of black tea infusion was estimated.
The amount of realizing each element into tea infusions and acceptable daily intake, for
safety consumption of black tea, was compared.

Keywords Black tea . Tea infusions . Toxic metals . Essential mineral elements . ICP-AES

Introduction

Tea is one of the most popular beverages in the world, which is prepared from the leaves of a
shrub Camellia sinensis. Tea contains flavonoids, minerals, and trace elements that are
essential to human health. There have been various general surveys of the work on the
medicinal value and beneficial health effects of tea consumption that has taken place in recent
years [1, 2]. Some of the possible beneficial effects of drinking tea are: antioxidative activity
[3], immune system boosting [4], protective effect against a range of cancers including lung,
prostate, and breast cancer [5–7] and reduction of blood cholesterol levels [8].

M. Salahinejad (*) : F. Aflaki

Environmental Laboratory, Nuclear Science Research School,
Nuclear Science & Technology Research Institute, Atomic Energy Organization of Iran (AEOI),
P. O. Box 11365-3486, Tehran, Iran
e-mail: salahinejad@gmail.com
110 Salahinejad and Aflaki

The knowledge of both micronutrients and toxic elements content in beverages is

important, taking into account nutrition requirement and intoxication risk related with their
consumption. It should be stressed that the amount of an element, which can be retained in
the organism, depends on the following parameters: total element content in tea leaves,
fraction of the total content extracted to the infusion, and bioavailability of the element
species present in the infusion.
Due to the importance of minerals in tea, several studies including atomic absorption
spectroscopy (AAS) [9], inductively coupled plasma atomic emission spectrometry (ICP-
AES) [10], inductively coupled plasma mass spectrometry (ICP-MS) [11], capillary
electrophoresis [12], and total reflection X-ray fluorescence spectrometry (TR-XRF) [13]
have been carried out in order to determine their levels in tea leaves and infusions.
Iran is a tea-producing country that relies on substantive imports to supplement its
production to satisfy its considerable consumption needs. Iranians have one of the highest
per capita rates of tea consumption in the world (1.6 kg per capita consumption in the
period 2005–2007).
This article intends to provide information about the content of the essential and toxic
trace elements in this beverage. Therefore, iron, manganese, copper, zinc, calcium,
magnesium (as essential trace elements) aluminum, arsenic, lead, chromium, cadmium, and
nickel (as potentially toxic elements) were determined in the dried leaves and infusions of
black teas available in Iran.

Experimental

Materials

ICP-AES standard stock solutions of Al, Mg, Ca, Mn, Zn, Fe, Pb, Cd, Cr, Ni, Cu, and As at
a concentration of 1,000 mg L−1 were obtained from Merck (Darmstadt, Germany), and
working solutions were prepared by dilution of stock solutions in 5% (v/v) HNO3
immediately before use. Purified water obtained from a Milli-Q System (Millipore, Milford,
MA, USA) was used for standard dilutions and other necessary preparations. All chemicals
and reagents employed were of analytical grade.

Instrumentation

A Perkin Elmer model Optima 2100DV ICP-AES (Norwalk, CT, USA) was used for
determination of selected elements in black tea leaves and their infusions. The operation
conditions are listed in Table 1. These conditions were established after optimization of the
instrumental parameters.

Sample Preparation

Eleven commercial black teas were purchased at the local markets in Tehran, Iran. The
collected black tea brands were Nushineh, Bahar lahijan, Langar shomal, and Golestan
cultivated in Iran (northern part of the country, Gilan province), Jahan, Golabi, and
Darjeeling imported from India and Shahrzad, Chakoshe sabz, Tala, and Silan Golestan
imported from Ceylon.
For each sample, about 1 g of ground tea sample was accurately weighed in a glass
beaker, after adding 5 ml concentrated nitric acid, the samples were kept standing overnight
Toxic and Essential Mineral Elements Content of Black Tea Leaves 111

Table 1 Operation Parameters for ICP-AES

Parameters

RF Power (watt) 1,300

Nebulizer gas flow rate (L min−1) 0.8
Auxiliary gas flow rate (L min−1) 0.2
Plasma gas flow rate (L min−1) 15
Sample gas flow rate (L min−1) 1.5
Plasma viewing Axial
Processing mode Area
Delay time (sec) 30

at room temperature for predigestion. The samples were heated on a hot plate (80°C,
30 min), cooled down, and 1 ml concentrated nitric acid and 4 ml concentrated perchloric
acid mixture were added. The digestion was carried out for about 1 h at 130°C until the
solutions became transparent. After cooling to room temperature, the solutions were filtered
into 25-mL volumetric flask and diluted. The solutions were subsequently analyzed for
selected metals using ICP-AES.
Tea infusions were prepared by adding 45 ml of hot distilled water to 1 g black tea
particles (or tea leaves from a tea bag). After five minutes of infusion, the solution was
filtered (or the tea bag was taken out). When the solution reached room temperature, 1 mL
concentrated nitric acid and appropriate amount of distilled water was added in a volumetric
flask to make up a 50-mL volume.

Statistical Analysis

In order to verify differences among the mineral content of the varieties of studied tea
samples, one factor analysis of variance was applied with the level of significance set at
0.05. The SPSS statistical program (SPSS Inc, IL, USA) was used to perform all statistical
calculations.

Method Validation

For all elements, calibration was performed using aqueous standard solutions. To evaluate
the analytical characteristics of the method for each element, the detection limits were
calculated according to IUPAC rules [14].
The precision of method was evaluated in ten replicate determinations on each of five
different randomly chosen samples. The reliability of the method was confirmed by using a
certified reference material (National Bureau of Standard SRM 1571, Orchard Leaves).

Result and Discussion

The precision (as relative standard deviation, RSD), detection limits (µg g−1) and analytical
wavelength (nm) are presented in Table 2. The results showed a good precision under the
analytical conditions used since the relative standard deviations are lower than 10%. In
order to verify the accuracy and analytical performance of the method, a certified reference
material was used to determination of selected elements and the results are given in Table 3.
112 Salahinejad and Aflaki

Table 2 Analytical Characterization of Measured Elements

Element Wavelength (nm) Precision (RSD %) Detection limit (µg g−1)a

Ca 317.933 5.6 0.01

Mg 285.213 3.1 0.002
Mn 257.61 0.8 0.01
Cu 327.393 1.2 0.009
Zn 206.200 2.3 0.006
Fe 238.204 1.6 0.005
Al 396.153 4.8 0.03
Ni 231.604 3.5 0.015
Pb 220.353 8.2 0.04
Cr 267.716 0.9 0.007
Cd 228.802 1.5 0.003
As 193.696 7.5 0.05
a
Calculated according to 3 times the SD of the blank (n=10)

The paired t test showed good agreement (p>0.05) between the certified values and the
obtained values.
The contents of toxic and essential elements of the black tea leaves and their infusions
that determined in this study are summarized in Tables 4 and 5, respectively.
Aluminum, calcium, magnesium, and manganese are present in tea leaves at mg g−1
level, while chromium, iron, nickel, copper, and zinc are present at µg g−1 level [15]. In this
work, Ca (3,454 to 5,891µg g−1), Mg (1,540 to 2,716µg g−1), and Al (584 to 1,546µg g−1)

Table 3 Analytical Method Validation Against a Certified Reference Material (Orchard Leaves SRM 1571
NBS)

Element Content (µg/g) Accuracy (%) Precision (RSD %)

a
Certified Measured

Ca 2.09±0.05b 2.07±0.1b 99 5.2

b
Mg 0.62±0.02 0.61±0.05b 98.4 4.5
Mn 91±4 90±6 98.9 3.2
Cu 12±1 11.5±2 95.8 6.8
Zn 25±3 24±5 96 6.5
Fe 300±20 298±27 99.3 4.7
Al c
– – – –
Ni 1.3±0.2 1.2±0.5 92.3 8.2
Pb 45±3 42±6 93.3 6.3
Cr 2.6±0.3 2.5±0.5 96.2 8.4
Cd 0.11±0.01 0.1±0.03 91 12.2
As 10±2 9.4±3 94 8.7
a
Analytical values are based on the “dry weight” material, mean±SD (n=6)
b
Percentage value
c
Non-certified
Toxic and Essential Mineral Elements Content of Black Tea Leaves 113

Table 4 Toxic Metals Contents (µg g−1) in Total and Infusion Black Tea Samples

Sample Al Ni Cd Pb Cr

Total Infusion Total Infusion Total Infusion Total Infusion Total Infusion

Iranian teas
Nushineh 1,546 426.4 10.48 4.54 n.d n.d 2.92 n.d 1.79 n.d
Bahar lahijan 954 252.4 12.40 6.19 0.76 n.d 1.67 n.d 1.94 n.d
Langar shomal 843 361.3 9.71 7.65 n.d n.d 2.13 n.d 0.89 n.d
Golestan 1,229 757 7.54 4.92 n.d n.d 0.92 n.d n.d n.d
Mean 1,143 449.3 10.03 5.82 <0.76 – 1.91 – <1.54 –
Indian teas
Jahan 616.5 314 4.50 2.0 n.d n.d 1.22 n.d n.d n.d
Golabi 759.9 366 5.21 2.38 0.77 n.d 1.83 n.d 1.28 n.d
Darjeeling 1,297 702 4.94 2.65 0.59 n.d 0.98 n.d 1.84 n.d
Mean 891.2 460.67 4.88 2.34 <0.68 – 1.34 – <1.56
Ceylon teas
Shahrzad 874.2 340.5 4.37 2.08 0.65 n.d 2.55 n.d 0.95 n.d
Chakoshe sabz 584.5 289.2 4.6 2.99 0.45 n.d 0.94 n.d 0.97 n.d
Tala 1,076 353.9 4.13 1.86 n.d n.d 1.44 n.d n.d n.d
Cylan Golestan 1,338 492.5 7.28 3.36 n.d n.d 1.93 n.d 1.71 n.d
Mean 968.2 369.1 5.09 2.57 <0.55 – 1.71 – <1.21 –

n.d not detected (below quantification limit)

had the highest concentration in black tea leaves, respectively. As shown by other authors
[16], the most abundant heavy metal in tea leaves was Mn which, in the present study,
ranged from 362 to 674µg g−1. The total contents of other heavy metals determined in tea
leaves arranged in the following order: Fe (105 to 407µg g−1), Cu (19.84 to 58.48µg g−1),
and Zn (21.2 to 33.12µg g−1). It should be noted that the concentrations of some toxic
heavy metal such as As and Cd in black teas and their infusions were too low to be detected
using the available analytical technique.
Many factors may be contributing to the metals accumulation in the tea leaves, such as
soil [17], its organic matter contents [18], manufacturing process [10] and environmental
pollutions. It was confirmed that the content of metals might be an adequate discriminator
of tea varieties and their geographical origin [19].
Figure 1 shows the mean concentration in milligrams per gram (Fig.1a) and micrograms
per gram (Fig.1b) of studied metals in black tea samples imported and cultivated in Iran.
One factor analysis of variance (ANOVA) of the data showed no statistically significant
differences in mean contents of most determined element among the different black tea
samples (p>0.05). By performing the t test, the mean content of nickel and copper in
cultivated black teas is significantly higher than imported black teas (p<0.05).
The factor affecting the metal contents in tea leaves influence subsequently the metal
concentrations in tea infusions. It has been recognized that the amount of trace elements
extracted into the tea infusions depends principally on whether the compound is strongly
bound to the matrix or more soluble in the solution employed [20]. Therefore, the amount
of minerals in tea infusion depends on extraction efficiencies and total concentration of
metal in tea leaves. According to extraction efficiencies from black tea leaves, Natesan and
Ranganathan [21], classified analyte elements into three groups: highly extractable element
 114

Table 5 Essential Metals Contents (µg g−1) in Total and Infusion Black Tea Samples

Sample Ca Mg Mn Cu Zn Fe

Total Infusion Total Infusion Total Infusion Total Infusion Total Infusion Total Infusion

Iranian teas
Nushineh 3,655 260 1,782 418.6 674.3 136 46.78 3.48 26.88 7.83 307.2 5.41
Bahar lahijan 3,644 291.2 1,724 556.9 651.2 164 52.14 3.9 24.64 10.38 199.9 5.71
Langar shomal 3,682 238 2,021 405 668 136.9 40.19 5.26 22.20 11.39 105.55 6.41
Golestan 3,454 271 1,918 526.8 439.9 163.3 58.48 9.0 22.69 5.86 139.6 5.59
Mean 3,609 265 1,861 476.8 608.3 150 49.39 5.41 24.10 8.86 188.1 5.78
Indian teas
Jahan 3,457 233.5 1,979 738.2 436.2 125.8 23.16 2.38 27.48 18.03 111.5 5.44
Golabi 3,939 203.5 1,799 728.5 508.7 162.9 26.64 3.72 33.12 16.05 117.2 3.34
Darjeeling 4,182 202.5 2,182 399.2 549.9 153.4 19.84 3.31 21.20 6.24 212.2 5.48
Mean 3,859 216.2 1,987 621.9 496.3 147.4 23.21 3.13 27.26 13.44 146.9 4.75
Ceylon teas
Shahrzad 3,929 210.5 1,540 610.6 362 88.7 39.57 4.35 32.34 6.23 94.93 5.34
Chakoshe sabz 3,804 210.5 1,824 663.5 333.9 93.2 37.98 4.39 23.37 6.48 133.55 5.98
Tala 5,891 272.2 2,716 608.6 655.4 249.4 35.79 7.15 32.03 12.63 310.1 4.36
Cylan Golestan 3,488 279.7 2,036 703.8 604 136.2 39.32 8.63 25.53 12.54 133.98 2.27
Mean 4,278 243.2 2,029 646.6 488.8 141.8 38.16 6.13 28.31 4.97 168.14 4.48
Salahinejad and Aflaki
Toxic and Essential Mineral Elements Content of Black Tea Leaves 115

Fig. 1 Mean concentration of

studied metals in black teas 4.5 Iran
imported and cultivated in Iran 4 India
[a: mg g−1 and b: µg g−1 units]
Ceylon
3.5
3

Conc. (mg/g)
2.5
2
1.5
1
0.5
0
Al Ca Mg Mn Fe
Elements
(a)
60
Iran
50 India
Ceylon
40
Conc. (ug/g)

30

20

10

0
Ni Zn Cu Pb
Elements
(b)

(>55%), e.g., Ni, moderately extractable elements (22–55%), e.g., Mn, Mg, Al, Zn, and Cu
and poorly extractable elements (<20%), e.g., Fe and Ca.
The extraction efficiency or realizing of each element was estimated as the ratio of its
concentration in tea infusions to total concentration in black tea leaves. Table 6 presented
mean percentage of realized minerals in black tea infusions and average daily intake of each
element for black tea samples. The results showed that the concentration values of Mn for
all samples were the highest concentration values and the concentration levels of Cu and Fe
were the lowest concentration values among the essential heavy metals in tea infusions.
Toxic heavy metals such as Cd and Pb are poorly extractable elements, therefore, the
concentrations of these elements in tea infusions were below the detection limit of the
proposed method.
With 1.6 kg per capita consumption of black tea leaves annually, Iranian daily
consumption averages 4.3835 g of black tea leaves per person. Average daily intake of
116 Salahinejad and Aflaki

Table 6 Mean Percentage of Realized Minerals in Tea Infusions and Their Average Daily Intake

Element Iranian tea samples Indian tea samples Ceylon tea samples

Realizing (%) Intake(mg/day) Realizing (%) Intake(mg/day) Realizing (%) Intake(mg/day)

Ca 7.35 1.163 5.58 0.944 5.88 1.105

Mg 25.82 2.106 32.03 2.79 33.25 2.957
Mn 25.74 0.686 29.58 0.644 28.25 0.605
Cu 10.85 0.023 13.64 0.014 16.12 0.027
Zn 37.1 0.039 47.83 0.057 33.88 0.042
Fe 3.56 0.029 3.43 0.022 3.3 0.024
Al 37.69 1.888 51.07 1.995 39.53 1.678
Ni 59.32 0.026 47.92 0.01 50.94 0.011

elements for black tea infusions consumption for Iranian was estimated (Table 6), for both
cultivated and imported black teas.
Table 7 represents the allowable or acceptable daily intake (ADI) of toxic metals and
reference or recommended daily intake (RDI) of essential minerals [22]. Average daily
intake of Al and Ni by consumption of black tea infusions was low rather than ADI.
Content of other potentially toxic elements such as: Pb, As, Cd, and Cr were below the
detection limits of our instrument (ICP-AES), therefore, a more sensitive instrument, e.g.,
ICP-MS or preconcentration methods must be used for determination of these elements in
tea infusions.

Conclusion

Twelve elements consisting potentially toxic metals (Al, As, Pb, Cr, Cd, and Ni) and
essential mineral elements (Fe, Zn, Cu, Mn, Ca, and Mg) were analyzed in eleven black tea
samples using ICP-AES. The reported levels for toxic and essential mineral elements in the

Table 7 Acceptable Daily Intake (ADI) of Toxic Metals and Recommended Daily Intake (RDI) of Essential
Minerals for Adults

Element ADI (mg/day) RDI (mg/day)

Ca – 1,000–1,300
Mg – 240–420
Mn – 1.6–2.3
Cu – 0.7–0.9
Zn – 8–11
Fe – 8–18
Al <50 –
Ni <1 –
Pb <0.2 –
Cr 0.05–0.2 0.02–0.035
Cd <0.1 –
As <0.13 –
Toxic and Essential Mineral Elements Content of Black Tea Leaves 117

black tea samples in present study were in agree with other studies. Tea is potentially a rich
source of some dietary metals such as manganese. With widespread consumption of tea in
the world, for many people, tea drinking may be a major source of dietary of essential
elements.
With respect to acceptable daily intake of Al and Ni as toxic elements in daily dietary
and safety standards, the infusion of black tea samples analyzed in the present study were
found to be safe for human consumption. However, for other toxic metals, because of their
low content, judgment about safety needs equipment that are more sensitive.

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