Determination of heavy metals in the soils of tea plantations and in fresh and processed tea leaves: an evaluation of six digestion methods

Abstract

Background

The aim of this study was to determine the levels of cadmium (Cd), chromium (Cr), lead (Pb), arsenic (As) and selenium (Se) in (1) fresh tea leaves, (2) processed (black) tea leaves and (3) soils from tea plantations originating from Bangladesh.

Methods

Graphite furnace atomic absorption spectrometry (GF-AAS) was used to evaluate six digestion methods, (1) nitric acid, (2) nitric acid overnight, (3) nitric acid-hydrogen peroxide, (4) nitric-perchloric acid, (5) sulfuric acid, and (6) dry ashing, to determine the most suitable digestion method for the determination of heavy metals in the samples.

Results

The concentration ranges of Cd, Pb, As and Se in fresh tea leaves were from 0.03-0.13, 0.19-2.06 and 0.47-1.31 µg/g, respectively while processed tea contained heavy metals at different concentrations: Cd (0.04-0.16 µg/g), Cr (0.45-10.73 µg/g), Pb (0.07-1.03 µg/g), As (0.89-1.90 µg/g) and Se (0.21-10.79 µg/g). Moreover, the soil samples of tea plantations also showed a wide range of concentrations: Cd (0.11-0.45 µg/g), Pb (2.80-66.54 µg/g), As (0.78-4.49 µg/g), and Se content (0.03-0.99 µg/g). Method no. 2 provided sufficient time to digest the tea matrix and was the most efficient method for recovering Cd, Cr, Pb, As and Se. Methods 1 and 3 were also acceptable and can be relatively inexpensive, easy and fast. The heavy metal transfer factors in the investigated soil/tea samples decreased as follows: Cd > As > Se > Pb.

Conclusion

Overall, the present study gives current insights into the heavy metal levels both in soils and teas commonly consumed in Bangladesh.

Rashid et al. Chemistry Central Journal (2016) 10:7 DOI 10.1186/s13065-016-0154-3

Determination ofheavy metalsinthe soils oftea plantations andin fresh andprocessed tea leaves: an evaluation ofsix digestion methods

Md. Harunur Rashid1, Zeenath Fardous2, M. Alamgir Zaman Chowdhury1*, Md. Khorshed Alam1, Md. Latiful Bari2, Mohammed Moniruzzaman3http://orcid.org/0000-0003-3498-7758

Web End = and Siew Hua Gan4

http://orcid.org/0000-0003-3498-7758

Web End = Background

Tea (Camellia sinensis L.) is one of the most popular nonalcoholic beverages, consumed by over two-thirds of the worlds population for its medicinal, refreshment and mild stimulant eects [1]. Tea leaves contain polyphenols such as epigallocatechin 3gallate, which has

many medicinal properties, including antioxidant [2], cholesterol-lowering [3], hepatoprotective [4] and anti-cancer activities [5]. Moreover, its detoxifying properties are essential in the elimination of alcohol and toxins [5]. However, considering that an estimated 18 billion cups of tea are consumed daily worldwide [6], its economic and social importance is unprecedented. In fact, tea has been reported to be valuable in the treatment and prevention of many diseases [6].

Ideally, tea should be free from contaminants such as heavy metals, which are toxic and harmful to the human

*Correspondence: [email protected]

1 Agrochemical and Environmental Research Division, Instituteof Food and Radiation Biology, Bangladesh Atomic Energy Research Establishment, Savar, Dhaka 1349, BangladeshFull list of author information is available at the end of the article

2016 Rashid et al. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License

(http://creativecommons.org/licenses/by/4.0/

Web End =http://creativecommons.org/licenses/by/4.0/ ), which permits unrestricted use, distribution, and reproduction in any medium,

provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,

and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/

Web End =http://creativecommons.org/

http://creativecommons.org/publicdomain/zero/1.0/

Web End =publicdomain/zero/1.0/ ) applies to the data made available in this article, unless otherwise stated.

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body because of their non-biodegradable nature, long biological half-lives and persistent accumulation in dierent body parts [7]. Tea is consumed in all of Bangladesh throughout the year, and Bangladesh is one of the leading tea producing and exporting countries in the world [8]. In 2006, Bangladesh exported approximately 5 millionkg of tea leaves, and this gure continues to increase even while the total local tea consumption in the country is reported to be 39 millionkg [8].

Tea processing and packaging in Bangladesh is dependent on the type of tea, with a wide variety available in the country that is produced by dierent processing steps. However, the common steps involve the (1) hand plucking of tea leaves by the local farmers, (2) the weighing of tea leaves and (3) transportation to factories. Freshly plucked tea leaves are fragile, and as the rst step in processing, the leaves are laid out to dry for several hours to allow them to wither as their moisture content decreases. The leaves are then rolled and oxidized, which alters their avor and gives the processed tea its nal appearance and color. The above steps are also known as Crush-Tear-Curl (CTC). The next step involves ring (nal drying process), a process that is initiated once the tea leaves have dried. This is followed by visually sorting into various batches of similar sizes and color before being packaged and commercialized both nationally and internationally. For black tea, the leaves are rolled immediately after withering to quickly initiate the oxidation or fermentation processes. The leaves are then completely oxidized before they are dried, which is how they acquire their dark color and rich avor.

Tea safety has piqued great interest because contaminants threaten the life and health of humans, animals and the environment, leading to economic losses [2]. The genetic and epigenetic eects of dietary heavy metals such as cadmium (Cd), chromium (Cr), lead (Pb), arsenic (As) and selenium (Se) in the human body are associated with an increased risk of dierent cancers [9]. Prolonged consumption of heavy metals from food can lead to their accumulation in the kidney and liver, causing disruption of numerous biochemical processes and potentially causing cardiovascular, nervous, kidney and bone diseases [10].

Elemental analysis of a tea sample requires destruction of the organic fraction of the sample, leaving the heavy metals either in solution or in a form that is readily dissolved. Unfortunately, because of a large number of analytes and a variety of sample types, there is no universal sample preparation technique that meets all of the diverse requirements. Among the strategies for sample preparation, dilution, acid digestion and extraction are the most commonly considered [1120]. Microwave digestion, wet digestion and dry ashing are commonly

utilized for the total decomposition of organic matter in samples [11, 21, 22]. Apart from these techniques, ultra-sound-assisted solubilisation/extraction sample preparation procedures were reported to be used for green and black tea samples [23].

Dry ashing consists of the ignition of organic compounds by air at atmospheric pressure and at relatively elevated temperatures (450550C) in a muffle furnace. The resulting ash residues are dissolved in an appropriate acid. Wet digestion is used to oxidize the organic portion of samples or to extract elements from inorganic matrices by means of concentrated acids or mixtures there of [24]. Compared to dry ashing, wet digestion may be performed with a wide variety of potential reagents. Although many types of acids, including hydrochloric acid (HCl), nitric acid (HNO3), sulfuric acid (H2SO4), perchloric acid (HClO4), and hydrogen peroxide (H2O2), are used to digest organic samples and soils [11, 25], it remains undetermined which type of acid/acid mixture is the most suitable.

In addition, little is known about the relative recovery of heavy metals from tea leaves, and there are no standard official methods in Bangladesh for the digestion of tea to determine heavy metals. Moreover, to our knowledge, there is limited data on the amount of heavy metals in fresh tea leaves, processed tea or soils from tea plantations in Bangladesh. Therefore, the aims of this study were (1) to determine the concentrations of common heavy metals such as Cd, Cr, Pb, As and Se in tea leaves and soils from tea plantations; (2) to report the degree of contamination and daily intake of toxic heavy metals via tea (3); to measure the interaction of heavy metal concentrations in fresh tea leaves, processed tea and soils from tea plantations by analyzing the transfer factor (TF); and (4) to evaluate six digestion methods using dierent acid combinations and recommend the most appropriate digestion method for determining the levels of ve heavy metals in tea samples.

Experimental

Chemicals andreagents

Heavy metal reference standards for Cd, Cr, Pb, As, and Se were purchased from Kanto Chemical (Tokyo, Japan). Digestion chemicals including HCl, HNO3, H2SO4,

HClO4, and H2O2 were of analytical grade and were purchased from Merck (Darmstadt, Germany).

Description ofstudy area

The samples were collected from two main tea growing areas (Moulvibazar and Sylhet) (Fig. 1). Moulvibazar is also known as the capital of tea production in Bangladesh, with miles and miles of tea gardens that look like green carpets. These areas have over 150 tea gardens,

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including three of the largest tea gardens in the world both in area and production.

Collection andpreservation ofsamples

Fresh tea leaves (n=10) were randomly collected from ve dierent tea gardens in the Sylhet district (n = 5), with the remaining from the Moulvibazar district (n=5)

(Fig.2). Each collection consisted of 500g of tea leaves and was authenticated by a botanist. For black tea, ve processed tea samples were randomly purchased from the local market in Moulvibazar, with another ve from the local market in Sylhet. The samples were supplied by the local tea gardens from the same areas. Purchased tea

sample were processed by plucking, withering, rolling, oxidation and ring. First, the leaves were harvested by hand. After plucking, the leaves were laid out to wilt or wither for several hours to prepare for further processing. During withering, the leaves were gently ued, rotated and monitored to ensure that an even exposure to air. Then, the leaf was put through a rolling machine to mince, twist and break it into even smaller pieces. After rolling, the leaves were laid out to rest for several hours, allowing oxidation (the process in which oxygen in the air interacts with the exposed enzymes in the leaf, turning the sample to a reddish-brown color and changing the chemical composition) to occur. This step also has

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the greatest impact in the creation of the many wonderful and complex avors in tea. The nal step in the production process is to re or heat the leaves quickly to dry them to below 3% moisture content and to stop the oxidation process to ensure that the tea samples were kept well. During rolling and withering step of tea processing, tea may be considered to be contaminated.

Soils from tea plantations (n = 10) were randomly collected from locations similar to where the 10 fresh tea leaf samples were collected (from both Sylhet and Moulvibazar districts, Bangladesh). The soil samples (sandy clay loam) were collected (500g each time) close (110cm perimeter) to the tea plant by digging into the soil (15cm depth). Some of the tea gardens were located near a highway (the closest was within 100 meters), and others were situated very far from the highway.

The collected samples were stored in clean, sterile polyethylene bags and were properly labeled. They were immediately sent to the laboratory of the Agrochemical and Environmental Research Division, Bangladesh Atomic Energy Commission, Dhaka, and were stored at

20C to reduce the risk of hydrolysis or oxidation prior to analysis.

Digestion ofsamples Digestion oftea samples

Before sample digestion, the tea leaves were freeze-dried at 50 C at 100 Pa for 24 h. They were then crushed using a sterile mortar and pestle and sieved (particle size <100m) at room temperature. Finally, 1g of tea leaves was used for digestion (refer to the six digestion methods described below).

Digestion ofsoil samples

Soil samples were oven dried at 60 C for 24 h before being ground into a ne powder using a sterile mortar and pestle. The samples (2.5 g) were transferred into a crucible before being mixed with 10 mL of aqua regia, which consisted of HCl:HNO3 (3:1). The mixture was the digested on a hot plate at 95C for 1h and was allowed to cool to room temperature. The sample was then diluted to 50 mL using deionized distilled water and was left to settle overnight [26]. The supernatant was ltered through Whatman No. 42 lter paper and (<0.45 m) Millipore lter paper, (Merck Millipore, Darmstadt, Germany) prior to analysis by graphite furnace atomic absorption spectrometry (GF-AAS).

Method 1 (HNO3 digestion)

Based on the method previously described by Huang et al. [27] and Narin et al. [28], the sample (1 g) was placed in a 50mL crucible before the addition of 10mL of concentrated HNO3. The sample was heated on a hot

plate until the solution became semi-dry. This was followed by the addition of 10mL of concentrated HNO3.

The solution was kept on a hot plate for 1h to allow the formation of a clear suspension. After the sample was semi-dried, it was cooled and ltered through Whatman No. 42 lter paper. It was then transferred to a 50mL volumetric ask by adding deionized distilled water to the mark [27, 29] before GF-AAS analysis.

Method 2 (HNO3 overnight digestion)

Concentrated HNO3 (10 mL) was added to the sample (1 g) and allowed to stand overnight at room temperature. The sample was then heated on a hot plate until the solution became clear and semi-dried. The solution was then cooled and ltered through Whatman No. 42 lter paper. It was then transferred quantitatively to a 50mL volumetric ask by adding deionized distilled water [30]. Finally, the solution was analyzed using GF-AAS.

Method 3 (HNO3H2O2 digestion)

In this method, the sample (1 g) was weighed into a 50 mL crucible and treated with 10 mL of concentrated HNO3. The solution was placed on a hot plate for 3045min to allow for oxidation. After cooling, 4mL of H2O2 (20%) was added, and the solution was reheated on a hot plate until the digest became clear and semi-dried. After cooling, the suspension was ltered into a 50mL volumetric ask and diluted with deionized distilled water to the mark [30] before GF-AAS analysis.

Method 4 (HNO3HClO4 digestion)

Approximately 1g of sample was placed in a 50mL crucible before the addition of 10mL of concentrated HNO3.

The mixture was placed on a hot plate for 3045min to allow for oxidation. After cooling, 5mL of HClO4 (70%)

was added, and the mixture was reheated on a hot plate until the digest became clear and semi-dried. Then, the sample was cooled and ltered through Whatman No. 42 lter paper before being quantitatively transferred to a 50 mL volumetric ask by adding deionized distilled water [29, 30]. Finally, the solution was analyzed using GF-AAS.

Method 5 (H2SO4 digestion)

The sample (1 g) was placed in a 50 mL crucible followed by the addition of 7 mL of concentrated H2SO4.

The mixture was allowed to stand for 30 min at room temperature. Approximately 7 mL of H2O2 (30 %) was added to the crucible, and the sample was reheated on the hot plate for 40min. Thereafter, 1mL of H2O2 (30%)

was added until the digest appeared clear upon cooling. Then, deionized distilled water was added to bring the nal sample volume to 50mL. The solution was ltered

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through Whatman No. 42 lter paper [29] and then analyzed using GF-AAS.

Method 6 (dry ashing)

Initially, 1g of sample was placed in a crucible on a hot plate at 100150C for 1h. It was transferred to a muffle furnace set at 480C. After 4h, the sample was removed from the furnace and cooled. Then, 2mL of 5M HNO3

was added, and the sample was evaporated to dryness on a hot plate. The sample was placed in a cool furnace and reheated to 400C for 15min before being removed, cooled and moistened with four drops of deionized distilled water. Then, 2mL of concentrated HCl was added, and the sample was evaporated to dryness before the addition of 2M HCl (2 mL). The solution was ltered through Whatman No. 42 lter paper and <0.45m Millipore lter paper and then quantitatively transferred to a 25 mL volumetric ask by adding deionized distilled water [29, 30].

GFAAS analysis

An atomic absorption spectrophotometer (model AA-6300, Shimadzu, Kyoto, Japan) equipped with a Shimadzu model GFA-EX7i graphite furnace atomizer was used to determine the heavy metals. Pyrolytic graphite tube was used for detection of As, Cr and Se while in case of Pb and Cd, high-density graphite tube was used. The absorption wavelength for the determination of each heavy metal type and other operating parameters and temperature programming of GF-AAS for the working

elements are given in Tables1, 2 and each analysis was performed in triplicate.

Calibration curves

Calibration curves for Cd, Cr, Pb, As and Se were prepared at seven dierent concentrations (0.0, 0.1, 1.0, 5.0, 10.0, 20.0 and 40.0g/L).

Recovery analysis

To calculate the percent recovery, the samples were spiked with known amounts of the analytical standards of Cd, Cr, Pb, As and Se. The mean percent recoveries for the various metals were calculated using the following equation:

where CE is the experimental concentration determined from the calibration curve, and CM is the spiked concentration.

Determination ofthe transfer factor (TF)

The transfer factor or transfer coefficient was calculated by dividing the concentration of the heavy metal in present in the tea by that of the total heavy metal concentration in the soil [31]:

Results anddiscussion

Heavy metal contents infresh tea leaves

Analysis of heavy metals such as As, Cr, Cd, Pb and Se in fresh tea leaves is important because they are toxic

Percent recovery = (CE/CM) 100

TF = Concentration in tea leaves/Concentration in soil.

Table 1 Operating parameters forthe GF-AAS analysis ofheavy metals

Elements Wavelength (nm) Detection limit (g/g) Slit width (nm) Lamp current (mA) Gas ow (L/min)

Cd 228.8 0.00005 0.7 8 1

Cr 357.9 0.00002 0.7 10 1

Pb 283.3 0.00005 0.7 10 1

As 193.7 0.00010 0.7 12 1

Se 196.0 0.00005 0.7 23 1

Table 2 Temperature programming ofGF-AAS forthe analysis ofCd, Cr, Pb, As andSe intea leaves andsoil samples

Stages Cd temperature C, hold time (s)

Cr temperature C, hold time (s)

Pb temperature C, hold time (s)

As temperature C, hold time (s)

Se temperature C, hold time (s)

Stage-1 150, 20 150, 20 150, 20 150, 20 150, 20

Stage-2 250, 10 250, 10 250, 10 250, 10 250, 10

Stage-3 500, 10 800, 10 800, 10 600, 10 600, 10

Stage-4 500, 10 800, 10 800, 10 600, 10 600, 10

Stage-5 500, 3 800, 3 800, 3 600, 3 600, 3

Stage-6 2200, 2 2300, 2 2400, 2 2200, 2 2200, 2

Stage-7 2400, 2 2500, 2 2500, 2 2500, 2 2400, 2

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and can be transported into humans and animals via the food chain. The concentration ranges of Cd, Pb, As and Se in fresh tea leaves were (0.030.13), (0.051.14), (BDL to 2.06) and (0.471.31 g/g), respectively (Table 3). Several studies have previously reported on the presence of trace elements in tea leaves and soil of tea gardens in Bangladesh [3235]. The mean Cd concentration in fresh tea leaves was 0.09 0.03 g/g (Fig. 3), which was lower than the World Health Organization (WHO)

recommended limit of 0.10g/g [36]. The Cd concentration was also lower than that reported for fresh tea leaves from India (0.430.01g/g), China (0.770.02g/g),

Japan (0.150.01g/g), and Italy (0.090.01g/g) [37] (Table 4). Moreover, our result was also lower than Cd content of tea samples from Turkey (0.50 0.10 g/g)

[28]. The variations in heavy metal contents of dierent samples may be due to dierences in geographical location, environmental conditions, seasonal changes, physiochemical characteristics of the growing regions and matrix-to-matrix transfer.

In comparison, the levels of Cr were low (below the detection limit) (Fig. 4), indicating that these fresh tea leaves were free from Cr contamination. The WHO-recommended limit for Cr is 0.05g/mL [36], and contamination by this heavy metal has been reported in Japanese, Chinese, Iranian and Thai green teas at 0.024, 0.14, 0.05 and 0.06g/g, respectively [38, 39]. Cr has been reported to cause cancer in humans, especially bronchial and lung cancers [40].

The mean Pb concentration in all of the fresh tea leaves investigated was 0.270.35g/g (Fig.3), which is lower than the WHO-recommended limit of 0.30g/g [36]. This is also lower than the Pb content of tea leaves

from Turkey (17.907.10g/g) [22] as well as tea leaves from India (1.860.04g/g), China (1.490.03g/g)

and Japan (1.550.03g/g), but is slightly higher than that from Italy (0.230.01g/g) [37]. Pb is a cumulative toxin that can primarily aect the blood, nervous system and kidneys. If present in high concentrations, Pb inhibits red blood cell formation, which can result in anemia [36].

The mean As concentration in fresh tea leaves was 1.21 0.74 g/g (Fig. 4), which is higher than the WHO-recommended limit (0.10g/g) [36] and higher than that of green tea from China (0.28 g/g) [41], Thailand (0.013 g/g) [38], Canada (0.04 g/g) [42] and Japan (0.00 g/g). A potential source of As is the high amount of As present in the soils of the studied tea plantations. As is toxic to humans, especially in its methylated forms produced by glutathione s-transferase (GST), As III methyltransferase (AS3MT) and S-adenosyl methionine (SAM). These enzymes can compete with DNA methyltransferase (DNMT) for DNA methylation, hence indirectly inhibiting DNA methyltransferase and inducing the reactivation of silenced tumor suppressor genes (Mishra et al. 2009). Chronic toxicity from high exposure to inorganic As is associated with arsenicosis, melanosis, keratoses of the skin and cancer [36].

The Se content of all investigated fresh tea leaves was 0.64 0.50 g/g (Fig. 4), and the WHO-recommended limit and contents of Japanese sencha green tea, Japanese jasmine tea, Chinese pai mu tan tea and Chinese gunpowder tea were 0.125, 0.092, 0.089, 0.075 and 0.070 g/g, respectively [39]. Se can lead to selenosis if taken in doses exceeding 400g per day [43]. Symptoms

Table 3 Heavy metal contents infresh tea leaves (FTL)

Sample ID MeanSD (g/g)

Cd Pb As Se

FTL-1 0.07 0.0001 0.05 0.0005 1.84 0.0001 BDLFTL-2 0.03 0.0008 0.31 0.0001 1.66 0.0062 0.90 0.0024

FTL-3 0.08 0.0064 0.40 0.0003 BDL BDLFTL-4 0.13 0.0038 1.14 0.0005 2.06 0.0056 1.28 0.0004

FTL-5 0.07 0.0040 0.05 0.0003 1.17 0.0013 0.78 0.0015 FTL-6 0.09 0.0015 0.09 0.0001 1.45 0.0054 1.31 0.0098

FTL-7 0.11 0.0056 0.22 0.0006 1.48 0.0237 BDLFTL-8 0.06 0.0025 0.46 0.0004 1.81 0.0064 0.47 0.0004

FTL-9 0.13 0.0041 BDL 0.44 0.0022 0.77 0.0002 FTL-10 0.12 0.0036 BDL 0.19 0.0001 0.85 0.0075

Mean 0.089 0.272 1.210 0.636

The limit of detection were 0.0052, 0.0026, 0.0046, 0.01 and 0.0084g/g for Cd, Cr, Pb, As and Se, respectively. The data (g/g) shown in Table is reported on dry weight basisn=3 (n no. of analyses), SD standard deviation, BDL below detection limit

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of selenosis include a garlic odor of the breath, gastrointestinal disorders, hair loss, sloughing of nails, fatigue, irritability and neurological damage. Extreme cases of selenosis can result in cirrhosis of the liver, pulmonary edema and death [43].

Heavy metal contents inblack tea

In the present study, heavy metal contents were also analyzed in the black tea produced from Bangladesh. The concentration ranges of Cd, Cr, Pb, As and Se were 0.040.16, 0.4510.73, 0.071.03, 0.891.90 and 0.7610.79g/g, respectively using HNO3 overnight digestion procedure (Table5).

The mean concentration of Cd in black tea (0.08 0.04 g/g) (Fig. 3) was lower than the World Health Organization (WHO)-recommended limit of 0.10g/g [36], but higher than that reported in black tea from Canada (0.026 g/g) [42], Thailand (0.0071 g/g) [41] and Turkey (0.0100 g/g) [44]. However, its level was lower than that reported in India (0.8900g/g) [3], Nigeria (0.1200 g/g) and Saudi Arabia (0.9890 g/g) [41]. Moreover, our result was also lower than Cd content of black teas from Turkey (2.300.40g/g) [22]. In a previous study, the concentration of Cd was 0.03g/g [34] which is slightly lower than that of our ndings. In another study, the presence of some trace elements (Cu,

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Table 4 Level ofCd, Pb, As andSe (g/g) intea leaves fromvarious countries

Country Cd Cr Pb As Se

Bangladesh 0.02 [34] 32.87 [34] 0.34 [34] NA ND [34]

India 0.43 [37]0.590.77 [50]0.010.03 [51]

0.090.37 [39]1.281.84 [50]0.431.14 [51]

1.86 [37]0.981.83 [50]0.100.51 [51]

NA 0.050.07 [39]2.122.47 [51]

0.28 [41] 0.050.09 [39]2.553.97 [51]

Japan 0.15 [37] 0.110.24 [39] 1.55 [37] NA 0.050.09 [39]

Italy 0.090.17[37]0.04 [51]

1.31 [51] 0.190.52 [37]0.55 [51]

NA 2.65 [51]

China 0.77 [37]0.043 [52]0.040.08 [51]

0.070.37 [39]1.232.20 [51]

1.49 [37]0.86 [52]0.601.08 [51]

Turkey 0.70.9 [22, 28]

3.13.5 [22, 28]

3.13.7 [22, 28]

NA

Thailand 0.0010.086

[38]

0.0403.294 [38]

0.10822.245 [38]

0.000.01 [38]0.0140.508 [53]

Sri Lanka 0.030.24 0.050.11 [39] 0.59 [34] 0.050.09 [39]

ND [34]

Iran 0.76 [50]

134.5 [54]

0.891.79 [50]8.2 [54]

0.922.92 [50] 209.5 [54]

0.013 [38]

0.0100.238 [53]

0.280.56 [41] NA

NA not available data, ND not detected

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Table 5 Heavy metal contents inprocessed tea leaves (PTL, black tea)

Sample ID MeanSD (g/g)

Cd Cr Pb As Se

PTL-1 0.16 0.0013 9.31 0.0493 0.27 0.0008 1.90 0.0006 1.44 0.0038 PTL-2 0.10 0.0031 7.03 0.0156 0.70 0.0004 BDL BDL

PTL-3 0.12 0.0023 10.73 0.0348 0.40 0.0009 1.17 0.0153 0.80 0.0002 PTL-4 0.04 0.0030 2.10 0.0004 0.07 0.0021 1.40 0.0036 0.76 0.0023

PTL-5 0.06 0.0031 1.71 0.0032 0.31 0.0008 0.89 0.0017 BDLPTL-6 0.11 0.0030 0.45 0.0026 0.22 0.0006 1.78 0.0066 10.79 0.0065

PTL-7 0.05 0.0001 2.75 0.0086 0.66 0.0002 1.02 0.0030 0.44 0.0003 PTL-8 0.07 0.0004 1.19 0.0084 1.03 0.0011 1.16 0.0006 BDL

PTL-9 0.05 0.0035 BDL 0.72 0.0006 1.00 0.0060 1.89 0.0101 PTL-10 0.07 0.0020 0.54 0.0049 BDL 1.30 0.0004 0.21 0.0016

Mean 0.083 3.581 0.438 1.162 1.633

The limit of detection were 0.0045, 0.003, 0.0028, 0.0032 and 0.0064g/g for Cd, Cr, Pb, As and Se, respectively. The data (g/g) shown in Table is reported on dry weight basisn=3 (nno. of analyses), SD standard deviation, BDL below detection limit

Ni, Mn and Zn) in three commercially available tea from Bangladesh were analyzed [32]. Nevertheless, they were dierent from that of the current investigation.

Cr was detected in rather high amounts in black tea (3.581 3.941 g/g), but it was not detected in fresh tea leaves or tea plantation soils (Fig. 3). Its level is higher than the recommended limit for Cr by the WHO of 0.05g/mL [36]. Moreover, Cr concentrations in black tea from India, China, Sri Lanka and Turkey were reported at 0.371, 0.155, 0.050 and 3.000g/g [39, 44], respectively. It is plausible that Cr contamination occurred during the fermentation process, which is one of the important processing steps of black tea in Bangladesh. In particular, it may occur during the CTC rolling steps involved in the production of black tea. However, this nding is lower than the previously reported Cr concentration (32.87g/g) in some tea samples from Bangladesh [34] which may be contributed to the dierent types of tea samples used as well as variance in the type of soil in the tea garden.

The Pb concentration in black tea was 0.438 0.328 g/g (Fig. 3), which is higher than the WHO recommended limit of 0.30 g/g [36]. Moreover, our ndings are also similar to the previously reported concentration of Pb (0.34 g/g) [34] in tea samples from Bangladesh but is higher than those reported for Nigeria (0.330 g/g) [6], Egypt (0.395 g/g) and Thailand (0.0237 g/g) [41], but lower than that in Turkey (2.500 g/g) [44], Iran (2.915 g/g), Saudi Arabia (1.250 g/g), China (3.270 g/g), Pakistan (2.500 g/g) and India (0.810g/g) [41].

The mean As concentration in black tea was 1.162 0.524 g/g (Fig. 4, which was higher than the

WHO-recommended limit (0.10 g/g) [36], as well as higher than in Thailand (0.00084 g/g) and China (0.280g/g) [41]. However, it was lower than that reported in Nigeria (2.220 g/g) [6]. The Se content in black tea from Bangladesh was higher (1.6333.280g/g) (Fig.4)

than that reported in black tea from Nigeria [6], India, China and Sri Lanka [39], which were 0.520, 0.070, 0.087 and 0.050g/g, respectively.

Heavy metal contents insoils fromtea plantations

In this part of the study, the heavy metal contents in the soils from tea plantations in Bangladesh have been reported. This analysis is important because of the metals potential toxicity and transportation through the root system into the buds and tea leaves. The concentration ranges of Cd, Pb, As and Se in tea plantation soils were 0.110.45, 2.8066.54, 0.784.49 and 0.030.99 g/g, respectively (Table6).

Similar to the ndings for fresh tea leaves, Cr was not detected in the tea garden soil samples (Fig.4). However, Cr has been reported in agricultural soils in the United States (48.5 g/g) [45], India (1.23 g/g) [46] and Kunshan, China (87.73g/g) [47]. Low concentrations of Cd (mean 0.222 0.103 g/g) were observed in all investigated soils from the tea plantations samples (Fig. 3).

These levels were lower than that previously reported in U.S. agricultural soils (13.5 g/g) [45], but higher than in Indian agricultural soils (0.05g/g) [46] and soil from Kunshan in China (0.20g/g) [47].

Because of the toxicological importance of Pb, many studies have investigated the levels of this element in soil from several countries. Among all of the soil samples investigated, STP-1 had the highest Pb

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Table 6 Heavy metal contents insoils fromtea plantations (STP)

Sample ID MeanSD (g/g)Cd Cr Pb As Se

STP-1 0.16 0.0013 BDL 66.54 0.5200 BDL BDL STP-2 0.15 0.0042 BDL 10.65 0.0120 2.75 0.0013 BDL

STP-3 0.34 0.0019 BDL 63.63 4.2400 1.13 0.0041 BDL STP-4 0.24 0.0016 BDL 8.65 0.0420 4.49 0.0128 BDL

STP-5 0.22 0.0021 BDL 11.86 0.0660 1.79 0.0017 BDL STP-6 0.45 0.0015 BDL 6.90 0.2000 2.32 0.0022 BDL

STP-7 0.16 0.0038 BDL 2.80 0.0100 1.08 0.0117 0.03 0.0003 STP-8 0.23 0.0015 BDL 9.48 0.0260 3.03 0.0004 BDL

STP-9 0.16 0.0011 BDL 3.60 0.0350 BDL 0.99 0.0000 STP-10 0.11 0.0059 BDL 10.22 0.4400 0.78 0.0157 0.81 0.0032

Mean 0.222 19.433 1.737 0.183

The limit of detection were 0.036, 0.0018, 0.0093, 0.0051 and 0.0012g/g for Cd, Cr, Pb, As and Se, respectively. The data (g/g) shown in Table is reported on dry weight basisn=3 (n no. of analyses), SD standard deviation, BDL below detection limit

Table 7 Transfer factors ofheavy metals fromtea plantation soils oftea leaves

Sample ID Cd Pb As Se

FTL-1 0.4375 0.0008

FTL-2 0.2000 0.0291 0.6036

FTL-3 0.2353 0.0062

FTL-4 0.5417 0.1318 0.4588

FTL-5 0.3182 0.0042 0.6536

FTL-6 0.2000 0.0130 0.6250

FTL-7 0.6875 0.0786 1.3704

FTL-8 0.2609 0.0485 0.5974

FTL-9 0.8125 0.0000 0.0000 0.7778

FTL-10 1.0909 0.0000 0.2436 1.0494

Mean 0.4784 0.0312 0.4552 0.1827

concentration (66.54 0.520 g/g) potentially because of its location, which was adjacent to a highway. Overall, the mean level of Pb in the tea plantation soil samples was 19.4324.25g/g (Fig.3). This is higher than that reported for agricultural soils in India (2.82g/g) [46] but lower than agricultural soils in the U.S. (55.00g/g) [45] and Kunshan, China (30.48g/g) [47].

The concentrations of As ranged from 0.78 to 4.49g/g. The highest As level was 4.49g/g in STG-4, but As was not detected in STP-1 or STP-9. The mean concentration of As was 1.741.429g/g (Fig.4), which is lower than that reported in Kunshan, China (8.15 g/g) [47].

Among all of the investigated soil samples, the mean Se concentrations in STP-1, STP-2, STP-3, STP-4, STP-5, STP-6 and STP-8 were below the detection limit. Low Se contents (mean 0.180.398g/g) (Fig.4) have also been reported in soils from garlic (0.026 g/g), radish (0.028 g/g), carrot (0.011 g/g) and orchard grass (0.069 g/g) plantations [48]. In comparison, higher Se concentrations were detected in the soils of oilseed rape (0.316 g/g), white clover (0.211 g/g), red clover (0.223g/g) and English plantain (0.277g/g) plantations [48]. These higher Se concentrations may be attributed to fertilizer (sodium selenite) use in tea plantations. High levels of heavy metals such as Se and As can potentially be easily transported to the tea leaves through the roots of the plant from contaminated soils. In addition, the acidic nature of tea garden soils can increase the extraction of As and hence the detected As concentration.

Heavy metal transfer fromsoils totea leaves inBangladesh

Soil-to-plant transfer is one of the key components of human exposure to metals through the food chain. The

transfer factor (TF) describes the transfer of heavy metals from soils to the plant body. In the present study, the TFs for Cd, Pb, As and Se were 0.47845, 0.03122, 0.45524 and 0.18272, respectively (Table 7). The transfer factors for heavy metals in the investigated tea samples decreased as follows: Cd>As>Se>Pb. In general, the TFs increased with decreasing metal concentrations in soils. Thereby, lower TFs in tea plants could be explained by uptake saturation [49]. In another study, the TFs of lettuce, spinach, radish and carrot followed a trend of Mn>Zn>Cd>Pb (Intawongse and Dean, 2006). To our knowledge, our study is the rst to report TFs in tea.

Method validation

The analytical results for the recovery of spiked metals in tea using the six digestion methods and LODs for

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Table 8 Recovery analysis (n=2) ofheavy metals andLODs ofinvestigated methods formethod validation

Method Percentage, LOD (g/g)

Cd Cr Pb As Se

Method 1 (HNO3) 96.80, 0.0076 95.20, 0.094 70.70, 0.012 80.00, 0.152 72.50, 0.0049 Method 2 (HNO3 overnight)

99.50, 0.0052 97.30, 0.0026 100.00, 0.0047 89.30, 0.014 100.03, 0.0084

Method 3 (HNO3 and

H2O2)

94.80, 0.016 74.90, 0.062 75.80, 0.186 95.60, 0.021 72.00, 0.0124

Method 4 (HNO3 HClO4) 76.20, 0.002 93.50, 0.068 74.90, 0.0052 90.20, 0.065 86.10, 0.0325 Method 5 (H2SO4) 80.60, 0.018 56.00, 0.128 65.00, 0.194 58.80, 0.176 71.20, 0.0982

Method 6 (Dry ashing) 113.60, 0.0142 87.40, 0.0014 84.70, 0.024 93.20, 0.052 60.20, 0.0018

The uncertainty of results was less than 1%. The data (g/g) shown in Table is reported on dry weight basis

each method are presented in Table8. Method 2 (overnight digestion with HNO3) was the most efficient for recovering Cd, Cr, Pb, As and Se with mean percent recoveries of 99.50, 97.30, 100.00, 89.30 and 100.03 %, respectively. For this reason, all tea samples were subsequently digested using this method, which is recommended as the best method for the destruction of tea. The method likely provided sufficient time for HNO3

to digest the tea matrix. On the other hand, Method 5 (H2SO4) yielded the lowest recoveries, possibly due to the incomplete digestion of tea samples or losses of elements through volatilization. Recoveries of Cd, Cr, Pb, As and Se were 80.60, 56.00, 65.00, 58.80 and 71.20%, respectively, all of which were below the acceptable limits (75125 %), except for Cd (80.60 %). Thus, the digestion method using H2SO4 is not recommended for tea samples. However, in a previous study, tea samples digested with three dierent acids at similar ratio [HNO3/H2SO4/H2O2(2: 2: 2)] showed shorter digestion time with better recovery and precision than other acid mixtures [28].

Method 1 (destruction with HNO3) and Method 3 (digestion using HNO3 and H2O2) yielded acceptable recoveries of Cd, Cr, Pb, As and Se. However, only 70.70 % of Pb was recovered by Method 1, which is below the acceptable limit. Therefore, Methods 1 and 3 could also be used as relatively inexpensive, simple and rapid substitutes. Method 6 (Dry ashing) is not recommended because of the high cost incurred due to the requirement of a muffle furnace. Method 4 (HNO3

HClO4 procedure) is also not recommended because HClO4 is potentially hazardous during digestion. This method also yielded poor recoveries. For all procedures, recovery of Cd was signicantly higher, while recovery of Pb was relatively lower. The likely reason for the lower recovery of Pb is the eect of the acidic pH used during sample digestion, which does not favor sample extraction.

Conclusions

Six digestion methods followed by GF-AAS have been successfully optimized in the present study. An overnight digestion with nitric acid (method no. 2) oered adequate time to digest the tea matrix and was the most efficient method for recovering Cd, Cr, Pb, As and Se. Moreover, Methods 1 and 3 were also satisfactory, relatively cheap, simple and fast. Method no. 5 is not recommended for the digestion of tea samples while method no. 6 was expensive. Cd, Pb, As and Se were detected in fresh tea leaves, but Cr was not detected. The concentrations of As were high in both fresh and black tea, while the concentration of Pb and Cr in black tea was higher than the recommended level set by the WHO. The soil from tea plantations was contaminated with As and Se, levels of which were at times higher than the WHO recommendation. High levels of heavy metals can easily be transported to tea leaves through the roots of tea plants. However, Cr was not detected in the soil samples. The trend in heavy metal TFs in the investigated tea samples was Cd>As>Se>Pb. An overnight digestion with HNO3 was the most efficient digestion method for recovering heavy metals.

Authors contributions

HR and ZF conducted the experiments, analyzed the data, and wrote the manuscript. AZC, KA and LB designed the experiments and supervised the work. MM analyzed the data and wrote the manuscript while SHG critically revised the manuscript. All authors read and approved the nal manuscript.

Author details

1 Agrochemical and Environmental Research Division, Institute of Foodand Radiation Biology, Bangladesh Atomic Energy Research Establishment, Savar, Dhaka 1349, Bangladesh. 2 Food Analysis and Research Laboratory, Center for Advanced Research in Sciences, University of Dhaka, Dhaka, Bangladesh. 3 Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150 Kota Bharu, Kelantan, Malaysia. 4 Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150 Kota Bharu, Kelantan, Malaysia.

Acknowledgements

The authors would like to acknowledge IAEA for nancial assistance under IAEA Research Contact no. 15052/R2 and Universiti Sains Malaysia Research University Team (RUT) Grant (1001/PPSP/853005). We would also like to thank

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Bangladesh Atomic Energy Commission for providing laboratory facilities to carry out the study and Abdullah-Al-Masud Mazumder, Botanist of Bangladesh Forest Research Institute, for authenticating the tea leaf samples.

Competing interests

The authors declare that they have no competing interests.

Received: 27 August 2015 Accepted: 5 February 2016

References

1. Karak T, Bhagat R (2010) Trace elements in tea leaves, made tea and tea infusion: a review. Food Res Int 43(9):22342252

2. Li X, Zhang Z, Li P, Zhang Q, Zhang W, Ding X (2013) Determination for major chemical contaminants in tea (Camellia sinensis) matrices: a review. Food Res Int 53(2):649658

3. Srividhya B, Subramanian R, Raj V (2011) Determination of lead, manganese, copper, zinc, cadmium, nickel and chromium in tea leaves. Int J Pharm Pharm Sci 13:257258

4. Issabeagloo E, Ahmadpoor F, Kermanizadeh P, Taghizadieh M (2012) Hepatoprotective eect of green tea on hepatic injury due to leunomide in rat. Asian J Exp Biol Sci 3:136141

5. Dufresne C, Farnworth E (2000) Tea, Kombucha, and health: a review. Food Res Int 33(6):409421

6. Achudume AC, Owoeye D (2010) Quantitative assessment of heavy metals in some tea marketed in Nigeria-Bioaccumulation of heavy metals in tea. Health 2(09):10971100

7. Sharma RK, Agrawal M, Marshall F (2007) Heavy metal contamination of soil and vegetables in suburban areas of Varanasi, India. Ecotoxicol Environ Saf 66(2):258266

8. Nasir T, Shamsuddoha M (2012) Tea production, consumption and exports in global and Bangladeshi perspective. USV Ann Econ Public Adm 11(2):7480

9. Bower JJ, Leonard SS, Shi X (2005) Conference overview: molecular mechanisms of metal toxicity and carcinogenesis. Mol Cell Biochem 279(12):315

10. Jrup L (2003) Hazards of heavy metal contamination. Br Med Bull 68(1):167182

11. Welna M, Szymczycha-Madeja A, Pohl P: Quality of the trace element analysis: sample preparation steps: INTECH Open Access Publisher; 2011

12. Salarian M, Ghanbarpour A, Behbahani M, Bagheri S, Bagheri A (2014) A metal-organic framework sustained by a nanosized Ag12 cuboctahedral node for solid-phase extraction of ultra traces of lead (II) ions. Microchim Acta 181(910):9991007

13. Behbahani M, Akbari AA, Amini MM, Bagheri A (2014) Synthesis and characterization of pyridine-functionalized magnetic mesoporous silica and its application for preconcentration and trace detection of lead and copper ions in fuel products. Anal Methods 6(21):87858792

14. Behbahani M, Abandansari HS, Salarian M, Babapour M, Bagheri A, Nabid MR (2014) Synthesis and application of a thermosensitive tri-block copolymer as an efficient sample treatment technique for preconcentration and ultra-trace detection of lead ions. Microchim Acta 181(12):129137

15. Nabid MR, Sedghi R, Behbahani M, Arvan B, Heravi MM, Oskooie HA (2014) Application of Poly 1, 8-diaminonaphthalene/multiwalled carbon nanotubes-COOH hybrid material as an efficient sorbent for trace determination of cadmium and lead ions in water samples. J Mol Recognit 27(7):421428

16. Fouladian HR, Behbahani M (2014) Solid phase extraction of Pb(II) and Cd (II) in food, soil, and water samples based on 1-(2-pyridylazo)-2-naphtholfunctionalized organicinorganic mesoporous material with the aid of experimental design methodology. Food Anal Methods 8(4):98299317. Ebrahimzadeh H, Behbahani M (2013) A novel lead imprinted polymer as the selective solid phase for extraction and trace detection of lead ions by ame atomic absorption spectrophotometry: synthesis, characterization and analytical application. Arabian J Chem. doi:http://dx.doi.org/10.1016/j.arabjc.2013.09.017

Web End =10.1016/j. http://dx.doi.org/10.1016/j.arabjc.2013.09.017

Web End =arabjc.2013.09.017

18. Behbahani M, Hassanlou PG, Amini MM, Omidi F, Esrali A, Farzadkia M, Bagheri A (2015) Application of solvent-assisted dispersive solid phase

extraction as a new, fast, simple and reliable preconcentration and trace detection of lead and cadmium ions in fruit and water samples. Food Chem 187:828819. Ghorbani-Kalhor E, Behbahani M, Abolhasani J, Khanmiri RH (2014) Synthesis and characterization of modied multiwall carbon nanotubes with poly (N-phenylethanolamine) and their application for removal and trace detection of lead ions in food and environmental samples. Food Anal Methods 8(5):13261334

20. Behbahani M, Abolhasani J, Amini MM, Sadeghi O, Omidi F, Bagheri A, Salarian M (2015) Application of mercapto ordered carbohydrate-derived porous carbons for trace detection of cadmium and copper ions in agricultural products. Food Chem 173:12071212

21. Soylak M, Tuzen M, Souza AS (2007) Korn MdGA, Ferreira SLC: optimization of microwave assisted digestion procedure for the determination of zinc, copper and nickel in tea samples employing ame atomic absorption spectrometry. J Hazard Mater 149(2):264268

22. Narin I, Colak H, Turkoglu O, Soylak M, Dogan M (2004) Heavy metals in black tea samples produced in Turkey. Bull Environ Contam Toxicol 72(4):844849

23. Szymczycha-Madeja A, Welna M, Pohl P (2015) Determination of essential and non-essential elements in green and black teas by FAAS and ICP OES simpliedmultivariate classication of dierent tea products. Microchem J 121:122129

24. Sneddon J, Hardaway C, Bobbadi KK, Reddy AK (2006) Sample preparation of solid samples for metal determination by atomic spectroscopy-An overview and selected recent applications. Appl Spectrosc Rev 41(1):114

25. Welna M, Szymczycha-Madeja A, Pohl P (2013) A comparison of samples preparation strategies in the multi-elemental analysis of tea by spectro-metric methods. Food Res Int 53(2):922930

26. Mwegoha W, Kihampa C (2010) Heavy metal contamination in agricultural soils and water in Dar es Salaam city, Tanzania. African Journal of Environmental Science and Technology 4(11):763769

27. Huang L, Bell R, Dell B, Woodward J (2004) Rapid nitric acid digestion of plant material with an open-vessel microwave system. Commun Soil Sci Plant Anal 35(34):427440

28. Narin I, Tuzen M, Soylak M (2004) Comparison of sample preparation procedures for the determination of trace heavy metals in house dust, tobacco and tea samples by atomic absorption spectrometry. Anal Chim 94(11):867873

29. Hseu Z-Y (2004) Evaluating heavy metal contents in nine composts using four digestion methods. Bioresour Technol 95(1):5359

30. Du Laing G, Tack FM, Verloo MG (2003) Performance of selected destruction methods for the determination of heavy metals in reed plants (Phragmites australis). Anal Chim Acta 497(1):191198

31. Kachenko AG, Singh B (2006) Heavy metals contamination in vegetables grown in urban and metal smelter contaminated sites in Australia. Water Air Soil Pollut 169(14):101123

32. Tarafdar S, Khan A, Rahman M (1985) Determination of Trace Elements in Bangladesh Tea Leaves. Agric Biol Chem 49(1):201202

33. Karim Z, Rahman A (1980) Cationic environment in some selected tea soils of Bangladesh. Tea Quarterly 49(1):5357

34. Marcos Ana, Fisher Andrew, Rea Gerry (1998) Hill SJ: preliminary study using trace element concentrations and a chemometrics approach to determine the geographical origin of tea. J Anal At Spectrom 13(6):521525

35. Moreda-Pineiro A, Fisher A, Hill SJ (2003) The classication of tea according to region of origin using pattern recognition techniques and trace metal data. J Food Compos Anal 16(2):195211

36. Organization JFWECoFAWH: Safety evaluation of certain mycotoxins in food, vol. 74. Food and Agriculture Org. 2001

37. Melucci D, Locatelli M, Locatelli C (2013) Trace level voltammetric determination of heavy metals and total mercury in tea matrices (Camellia sinensis). Food Chem Toxicol 62:901907

38. Limmatvapirat C, Charoenteeraboon J, Phaechamud T (2012) Determination of Heavy Metals in Herbal Teas by Inductively Coupled Plasma Mass Spectrometry. Res J Pharm, Biol Chem Sci 3(3):952958

39. Cabrera C, Gimnez R, Lpez MC (2003) Determination of tea components with antioxidant activity. J Agric Food Chem 51(15):44274435

40. Kim K-C, Park Y-B, Lee M-J, Kim J-B, Huh J-W, Kim D-H, Lee J-B, Kim J-C (2008) Levels of heavy metals in candy packages and candies likely to be consumed by small children. Food Res Int 41(4):411418

Rashid et al. Chemistry Central Journal (2016) 10:7

Page 13 of 13

41. Shekoohiyan S, Ghoochani M, Mohagheghian A, Mahvi AH, Yunesian M, Nazmara S (2012) Determination of lead, cadmium and arsenic in infusion tea cultivated in north of Iran. Iranian J Environl Health Sci Eng 9(1):16

42. Schwalfenberg G, Genuis SJ, Rodushkin I (2013) The benets and risks of consuming brewed tea: beware of toxic element contamination. J Toxicol 2013:370460

43. Selenium PHS: Agency for Toxic Substances and Disease Registry. Retrieved 01 May 2009, 2009

44. Soylak M, Aydin A (2011) Determination of some heavy metals in food and environmental samples by ame atomic absorption spectrometry after coprecipitation. Food Chem Toxicol 49(6):12421248

45. Jean-Philippe SR, Labbe N, Franklin JA, Johnson A (2012) Detection of mercury and other metals in mercury contaminated soils using mid-infrared spectroscopy. Proc Intern Acad Ecol Environ Sci 2(3):139149

46. Prajapati SK, Meravi N (2014) Heavy metal speciation of soil and Calotropis procera from thermal power plant area. Proc Intern Acad Ecol Environ Sci 4(2):6871

47. Chen F (2007) Pu L-j: relationship between heavy metals and basic properties of agricultural soils in Kunshan County. Soils 39:291296

48. Metka S, Walter G. Accumulation of selenium in natural plants and selenium supplemented vegetable and selenium speciation by HPLC-ICPMS. Chem Speciation Bioavailability. 2005, 17(2)

49. Karak T, Paul RK, Sonar I, Sanyal S, Ahmed KZ, Boruah RK, Das DK, Dutta AK (2014) Chromium in soil and tea (Camellia sinensis L.) infusion: does soil amendment with municipal solid waste compost make sense? Food Res Int 64:114124

50. Salahinejad M, Aaki F (2010) Toxic and essential mineral elements content of black tea leaves and their tea infusions consumed in Iran. Biol Trace Elem Res 134(1):109117

51. Barone G, Giacominelli-Stuffler R, Storelli MM (2016) Evaluation of trace metal and polychlorinated biphenyl levels in tea brands of dierent origin commercialized in Italy. Food Chem Toxicol 87:113119

52. Ning P, Gong C, Zhang Y, Guo K, Bai J (2011) Lead, cadmium, arsenic, mercury and copper levels in Chinese Yunnan Puer tea. Food Addit Contam 4(1):2833

53. Nookabkaew S, Rangkadilok N, Satayavivad J (2006) Determination of trace elements in herbal tea products and their infusions consumed in Thailand. J Agric Food Chem 54(18):69396944

54. Falahi E, Hedaiati R (2013) Heavy metal content of black teas consumed in Iran. Food Additives & Contaminants: Part B 6(2):123126