Chemosphere 93 (2013) 2230–2238

Contents lists available at ScienceDirect

Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Human health risk from Heavy metal via food crops consumption with
wastewater irrigation practices in Pakistan
Muhammad Usman Khan a, Riffat Naseem Malik a,⇑, Said Muhammad b
a
Environmental Biology and Ecotoxicology Laboratory, Department of Environmental Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, PO 45320, Islamabad, Pakistan
b
Department of Earth Sciences, COMSATS Institute of Information Technology, Abbottabad, Pakistan

h i g h l i g h t s

 Greater Cd, Cr, Ni, Mn, Pb recorded in wastewater irrigated soils and food crops.
 Cd in soil; Pb, Cd, Cr in irrigated water and food crops surpassed permissible limit.
 Wastewater irrigation for food crops is a potential threat to human health.
 Health risk index >1 for Pb, Cd and Mn in food crops cause potential health risk.

a r t i c l e i n f o a b s t r a c t

Article history: The current study was designed to investigate the potential human health risks associated with con-
Received 25 March 2013 sumption of food crops contaminated with toxic heavy metals. Cadmium (Cd) concentration in surface
Received in revised form 20 July 2013 soils; Cd, lead (Pb) and chromium (Cr) in the irrigation water and food crops were above permissible
Accepted 21 July 2013
limits. The accumulation factor (AF) was >1 for manganese (Mn) and Pb in different food crops. The
Available online 26 September 2013
Health Risk Index (HRI) was >1 for Pb in all food crops irrigated with wastewater and tube well water.
HRI >1 was also recorded for Cd in all selected vegetables; and for Mn in Spinacia oleracea irrigated with
Keywords:
wastewater. All wastewater irrigated samples (soil and food crops) exhibited high relative contamination
Wastewater
Heavy metals
level as compared to samples irrigated with tube well water. Our results emphasized the need for pre-
Food crops treatment of wastewater and routine monitoring in order to avoid contamination of food crops from
Health risk the wastewater irrigation system.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction worldwide, which has gained importance nowadays in water scanty

Rapid urbanization and industrialization in 20th century has
dramatically increased the discharge of large amount of wastewater
accompanied with toxic chemicals. Wastewater finds its way to
water channels that are further used for irrigation purposes due to
presence of organic matter and nutrients deposition. In contrast,
wastewater is a rich source of various toxic metals (Ping et al.,
2011). Heavy metals finally become the part of wastewater that
originates from various natural (weathering and erosion of bed rock
material and ore deposits) and anthropogenic (industrial processes,
mining, agricultural practices and waste disposal) activities,
atmospheric deposition and emissions related with traffic trans-
ported with rain/storm water (Karvelas et al., 2003; khan et al.,
2008). Its continuous practice in agricultural lands may cause
contamination and adversely affect the safety of the food crops pro-
duced (Hua et al., 2013). Wastewater irrigation to food crops is the
most familiar practice in the urban, suburban and agricultural areas
⇑ Corresponding author. Tel./fax: +92 5190643017.
E-mail address: r_n_malik2000@yahoo.co.uk (R.N. Malik).
areas. It is posing risk to human via food crops consumption, if it is
used for irrigation without prior treatment.
Food crops composed of carbohydrates, minerals, proteins, vita-
mins and essential metals (macro and micro nutrients) make impor-
tant components of diet (Khan et al., 2008). Food consumption
contaminated with heavy metals is a major contributor pathway
(more than 90%) to human exposure than any other pathways such
as inhalation and dermal contact (Loutfy et al., 2006). Heavy metal
contamination is a known causative of various disorders such as
genomic instability, endocrine disruption, neurotoxicity, carcinoge-
nicity, immunological problems and also impaired psycho-social
behavior (Dyer, 2007). Heavy metals such as Cd and Pb may cause
mutagenesis, teratogenesis and carcinogenesis; high Cd and Pb con-
centrations in food crops were attributed to prevalence of upper gas-
trointestinal cancer (Jarup, 2003; Turkdogan et al., 2003). Besides
these, Pb is also responsible for elevated blood pressure, renal and
tumor infection, improper haemoglobin synthesis and reproductive
system. Manganese is accountable for Parkinson’s disease as a result
of iron oxide deposition (Harmanescu et al., 2011). At high concen-
tration Ni can cause cancer, fatigue, headache, skin rashes, dizziness,

0045-6535/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.chemosphere.2013.07.067
 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238
heart problems and respiratory illness. Dietary Cr intakes studied in
different countries were assumed to be directly correlated with
breast cancer mortalities (Pasha et al., 2010).
The current study area consists of industrial cities of Sialkot and
Wazirabad (Pakistan) known worldwide for their tanneries, phar-
maceuticals, surgical, sports and cutlery industries. Nullah Aik and
Palkhu streams in these areas receive untreated industrial effluents,
urban and domestic sewage, municipal wastes and agricultural run-
off. These streams have been categorized among the most polluted
streams of Pakistan. The aquatic ecosystem is highly stressed due
to release of untreated industrial and urban effluents. It has been
predicted that in near future these streams will lose their aquatic
integrity if no restoration practices are initiated (Qadir et al.,
2008). Agricultural land in a suburb of the aforementioned cities is
irrigated with wastewater from these streams and tube well water.
Information about heavy metal accumulation in food crops and
their dietary intake is necessary for assessing their risk to human
health. Therefore, it is of prime concern to assess the accumulation
of toxic heavy metal in food crops, which are irrigated with water
diverted from polluted streams (Nullah Aik and Palkhu). Informa-
tion is lacking on the heavy metal uptake, bioaccumulation, and
health risks in terms of wastewater irrigation in Sialkot and Waz-
irabad. Therefore, the current study aims to quantify heavy metal
accumulation in water, soil and subsequent contamination in food
crops irrigated with wastewater and tube well water with poten-
tial human health risks.
2. Materials and methods
2.1. Sampling strategy and digestion procedure
The study was conducted in the industrial cities of Sialkot and
Wazirabad, located between 32°240 N–32°370 N latitude and
Fig. 1. Map of the study area showing sampling sites irrigated with wastewater and tube well water.
2231
73°590 E and 75°200 E longitude in the east of Pakistan and
32°270 000 N and 74°70 000 E of Punjab, respectively. The study area is
divided into two main irrigation zones, i.e. wastewater and tube
well water. Nullah Aik and Palkhu are the most polluted streams
used for irrigation purposes of agricultural land in this area. Total
of four sampling sites of wastewater (n = 2 sites from each study
area, Nullah Aik and Palkhu) and four tube well water sites in
the vicinity of the same area (n = 2 sites from each study area) were
selected for collection of soil, water and food crops samples (Fig. 1).
Water and soil samples were collected in triplicate from each
study area. Total of 24 samples of water (n = 12 each for wastewa-
ter and tube well water) and 24 soil samples (12 each for wastewa-
ter and tube well water irrigated soil) were collected randomly
from surface soil (0–20 cm). Water samples were preserved in
pre-rinsed plastic bottles with 10% solution of HNO3 and placed
at 4 °C for further process. Air dried soil samples were grounded
into fine powder, passed through 2 mm mesh sieve and stored in
polythene bags till further analysis.
Twelve food crops species (Coriandrum sativum L. (Coriander),
Allium cepa L. (Onion), Abelmoschus esculentus L. (Okra), Allium sat-
ivum L. (Garlic), Capsicum annum L. (Capsicum), Daucus carota L.
(Carrot), Solanum melongena L. (Brinjal), Spinacia oleracea L. (Spin-
ach), Raphanus sativus L. (Radish), Mentha spicata L. (Mint), Lycoper-
sicum esculentum L. (Tomato) and Triticum aestivum L. (Wheat))
irrigated with wastewater (Nullah Aik and Palkhu) and tube well
water in the same vicinity were collected from agricultural field
in Sialkot and Wazirabad. A total of eight sites (four sites each of
wastewaters and tube well water) were selected for random sam-
pling. All the collected food crops sample were sectioned into edi-
ble and non-edible parts. The edible parts were washed with de-
ionized distilled water to remove all observable dust particles,
and blotted with tissue paper. The edible portion was dried in oven
for 24 h at 70–80 °C, ground into fine powder, sieved through
2232 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238

Table 1
Heavy metal concentrations in waste and tube well water (mg L1) and soil (mg kg1) irrigated with wastewater and tube well water.

Heavy metals Wastewater irrigated soil Tube well water irrigated soil P values Wastewater Tube well water P value
Mean ± SD Range Mean ± SD Range Mean ± SD Range Mean ± SD Range
Cr 43.03 ± 11.10 31.65–61.65 24.87 ± 6.009 14.20–37.70 0.0001 4.74 ± 4.43 1.80–18.20 0.85 ± 0.55 0.27–1.67 0.0063
Ni 42.09 ± 11.73 30.05–64.30 30.71 ± 6.15 22.80–43.60 0.001 0.46 ± 0.29 0.21–1.29 0.12 ± 0.04 0.05–0.20 0.0006
Mn 36.47 ± 15.59 18.33–66.78 19.84 ± 5.50 12.72–28.03 0.001 1.99 ± 1.23 0.64–4.88 0.35 ± 0.16 0.12–0.72 0.0001
Pb 36.91 ± 29.37 11.50–90 12.75 ± 9.44 0.00–29.50 0.01 5.03 ± 1.59 3.08–7.32 2.47 ± 0.74 1.32–4.12 0.000
Cd 9.15 ± 1.33 7.13–11.13 2.89 ± 1.00 1.28–4.28 0.001 0.55 ± 0.09 0.27–0.64 0.27 ± 0.08 0.14–0.44 0.000

SD = standard deviation.

2 mm mesh and kept in pre-cleaned polythene bags at ambient
temperature till further analysis.
A 0.5 g of dried samples of food crops and soil were taken. For
digestion, a 5:1:1 acid mixture of (HNO3, H2SO4 and HCLO4) was
added and heated at 80 °C until the solution became transparent
(Allen et al., 1986).
Digestion of water samples was carried out according to the
procedure reported by USEPA (1990). Well digested solutions of
water, soil and food crops samples were poured through Whatman
filter paper No. 42 and volume was made up to 50 mL with de-ion-
ized distilled water.
All samples were analyzed for selected heavy metals (Pb, Cr,
Mn, Cd and Ni) using Flame Atomic Absorption Spectrophotometer
(Varian FAAS-240).
2.2. Quality control analysis
All the reagents used in the experimental procedure were of ana-
lytical grade (Merck Germany). Glass-ware and plastic-ware used
were thoroughly rinsed with 10% HNO3 solution followed by wash-
ing with de-ionized distilled water. For quality data assurance, each
sample was analyzed in triplicate. After every five samples three
standards; one blank and two of 2.5 and 5.0 lg L1 concentration
of respective metal were analyzed for atomic absorption. The repro-
ducibility was found to be at 90 ± 7% confidence level. Therefore, an
average value of three replicates was used for interpretation of re-
sults. Standard solutions (1000 mg L1) of all five metals were pre-
pared by dilution of certified standard (NIST-SRM, 1570a for
spinach leaves, SRM 1643e for water, and SRM 2709 for soil) of cor-
responding metal ions with de-ionized distilled water.
2.3. Statistical analysis
All the data were statistically have been presented in the form
of means, range and standard deviation. Mean values of heavy
metals in soil, water and food crop samples from different sites
were compared using Independent sample t-test. A probability
P < 0.05 was used as a statistical significance level. Statistical anal-
ysis was performed using Statistical package SPSS (version 13).
2.4. Translocation factor calculation
Heavy metals have the capability of translocation from soil to
the edible parts of food crop that can be determined by accumula-
tion factor (AF) (Li et al., 2012). The AF’s for the selected heavy
metals were calculated as in the following equation:
Heavy metal concentration in food crops edible parts
AF ¼
Heavy metal concentration in soil
2.5. Human health risk assessment
The local population of the Sialkot and Wazirabad were inter-
viewed for their age, body weight and daily consumption of each
food crop which was further used for calculation of human health
risk assessment. A total of 65 questionnaires from the local popu-
lations in the age group of 14–50 years (male and female) were
evaluated for health risk assessment.
2.5.1. Daily intake of metals (DIM)
Daily intake of metals was calculated using the following
equation:
C metal  C factor  Dfood intake
DIM ¼ ð2Þ
Baverage weight
where Cmetal, Cfactor, Dfood intake and Baverage weight represent the heavy
metal concentrations in food crops, conversion factor, daily intake
of food crops and average body weight, respectively. The conversion
factor (CF) of 0.085 was used for the conversion of fresh vegetables
to dry weight. The average daily intakes of food crops were
0.527 kg person1 d1 and their average body weight for adult’s
populations were 55.5 kg that were further used for calculation of
HRI.
2.5.2. Health Risk Index (HRI)
HRI refers to the ratio of the daily intake of metals in the food
crops to the Oral reference dose (USEPA, 2002) and was calculated
using the following equation:
HRI ¼ DIM=RfD ð3Þ
An HRI > 1 for any metal in food crops means that the consumer
population faces a health risk.
3. Results
3.1. Heavy metal concentrations in irrigation water, soil and food
crops
In our study, Pb was the leading contaminant followed by
Cr > Mn > Cd > Ni in wastewater and tube well water. In general
the heavy metal concentrations were significantly higher in waste-
water as compared to tube well water (Table 1). Similar trends
were recorded in the soil irrigated with wastewater. In wastewater
concentrations of Cr, Ni, Mn, Pb and Cd ranged between 1.80 and
18.20, 0.21 and 1.29, 0.64 and 4.88, 3.08 and 7.32, 0.27 and
0.64 mg L1, while in tube water it was 0.27–1.67, 0.05–0.20,
0.12–0.72, 1.32–4.12, 0.14–0.44 mg L1 respectively.
Heavy metals were in the order of Cr > Ni > Pb > Mn > Cd with
concentrations ranges from 31.65 to 61.65, 30.05 to 64.3, 18.33
to 66.78, 11.50 to 90, 7.13 to 11.13 mg kg1 in soil irrigated with
wastewater. The concentrations of Ni was highest in tube well
ð1Þ water irrigated soils, followed by Cr, Mn, Pb and Cd ranges between
14.20–37.70, 22.80–43.60, 12.72–28.03, 0.00–29.50 and 1.28–
4.28 mg kg1 respectively.
In general the wastewater irrigated food crops have higher me-
tal concentrations as compared to tube well water irrigated coun-
terparts (Fig. 2). The level of Cr was significantly higher in
wastewater irrigated food crops (1.46–6.08 mg kg1) as compared
 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238 2233

Fig. 2. Heavy metal concentrations in the edible parts of food crops grown with wastewater and tube well irrigation for (a) Cr, (b) Ni, (c) Mn, (d) Pb and (e) Cd. Asterisks
represent significance level among wastewater and tube well water irrigated food crops for heavy metals concentrations at P < 0.05(), P < 0.01() and P < 0.001().

to tube well water irrigated food crops (0.48–2.08 mg kg1) i.e. A.
esculentus, S. oleracea and S. melongena (Fig. 2a). Similar trend
was recorded in the case of Ni (3.36–6.40 and 1.34–2.72 mg kg1)
in all the food crops irrigated with waste and tube well water ex-
cept A. esculentus grown on both sites (Fig. 2b). Mn ranged from
14.89 to 201.26 mg kg1 in waste and 8.49 to 52.58 mg kg1 in
tube well water irrigated crops. A. cepa, A. sativum, C. annum, S.
oleracea, S. melongena, L. esculentum and T. aestivum showed higher
pattern of accumulation in wastewater irrigation system (Fig. 2c).
The food crops irrigated with wastewater and tube well water
had Pb contents between 22.41–40.85 and 15.28–27.02 mg kg1
and Cd ranged between 1.18–3.81 and 0.41–1.20 mg kg1 in waste
and tube well water irrigated food crops respectively. Pb concen-
trations in A. cepa, A. esculentus, A. sativum, D. carota, R. sativus, S.
melongena, L. esculentum and T. aestivum and Cd concentration in
all food crops except C. annum showed significantly higher
accumulation under wastewater irrigation system as compared
to tube well irrigation (Fig. 2d and e). As in case of accumulation
perspective maximum concentrations of Cr, Mn, and Cd were ob-
served in leafy vegetables, while S. melongena and A. cepa had accu-
mulated Ni and Pb in higher concentrations respectively (Fig. 2).
3.2. Heavy metals transfer from soil to plant
The AF values for food crops irrigated with wastewater and tube
well water are shown in Tables 2 and 3 respectively. Highest AF
was observed in wastewater ranging between 4.94–6.07 for Mn,
and 0.91–1.43 for Pb in S. oleracea and A. cepa respectively. In tube
well water irrigated food crops, AF values for Pb (2.66–3.96) and
Mn (2.20–3.43) was high. AF values of wastewater irrigated food
crops were in the order of Mn > Pb > Cd > Ni > Cr, while trends in
tube well water irrigated food crops were as;
Pb > Mn > Cd > Ni > Cr. The AF of wastewater irrigated food crops
for Cr, Ni, Mn, Pb and Cd ranged between (0.03–0.14),
2234 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238

Table 2
Accumulation factor (AF) (on dry weight basis) for food crops grown in wastewater.

Species Statistics Cr Ni Mn Pb Cd
Coriandrum sativum Mean ± SD 0.10 ± 0.09 0.11 ± 0.03 2.09 ± 1.37 0.93 ± 0.36 0.41 ± 0.05
Range 0.03–0.22 0.07–0.14 0.45–3.51 0.57–1.44 0.37–0.48
Allium cepa Mean ± SD 0.07 ± 0.06 0.12 ± 0.04 1.23 ± 0.57 1.11 ± 0.24 0.41 ± 0.02
Range 0.01–0.13 0.09–0.17 0.66–1.93 0.91–1.43 0.38–0.43
Abelmoschus esculentus Mean ± SD 0.09 ± 0.05 0.10 ± 0.04 1.38 ± 0.84 0.96 ± 0.26 0.23 ± 0.03
Range 0.03–0.13 0.06–0.14 0.33–2.37 0.65–1.29 0.19–0.26
Allium sativum Mean ± SD 0.06 ± 0.01 0.12 ± 0.03 0.67 ± 0.30 0.84 ± 0.16 0.34 ± 0.06
Range 0.05–0.07 0.08–0.16 0.23–0.92 0.66–1.04 0.25–0.38
Capsicum annum Mean ± SD 0.06 ± 0.05 0.11 ± 0.02 1.07 ± 0.49 0.76 ± 0.20 0.20 ± 0.07
Range 0.01–0.12 0.09–0.13 0.35–1.48 0.57–0.95 0.12–0.27
Daucus carota Mean ± SD 0.08 ± 0.06 0.10 ± 0.04 1.42 ± 1.12 0.73 ± 0.08 0.29 ± 0.03
Range 0.02–0.13 0.07–0.15 0.42–2.63 0.65–0.84 0.25–0.31
Spinacia oleracea Mean ± SD 0.14 ± 0.04 0.10 ± 0.01 5.52 ± 0.55 0.61 ± 0.07 0.36 ± 0.13
Range 0.10–0.18 0.09–0.12 4.94–6.07 0.52–0.68 0.20–0.49
Raphanus sativus Mean ± SD 0.06 ± 0.03 0.13 ± 0.05 0.60 ± 0.49 0.92 ± 0.10 0.28 ± 0.03
Range 0.02–0.08 0.07–0.17 0.16–1.31 0.77–0.99 0.27–0.33
Mentha spicata Mean ± SD 0.07 ± 0.04 0.09 ± 0.02 1.51 ± 0.24 1.01 ± 0.52 0.42 ± 0.06
Range 0.04–0.12 0.08–0.11 1.20–1.78 0.55–1.50 0.34–0.48
Solanum melongena Mean ± SD 0.06 ± 0.01 0.15 ± 0.04 0.41 ± 0.03 0.86 ± 0.06 0.15 ± 0.03
Range 0.05–0.07 0.12–0.19 0.38–0.45 0.81–0.92 0.12–0.19
Lycopersicum esculentum Mean ± SD 0.03 ± 0.02 0.11 ± 0.04 0.65 ± 0.06 0.88 ± 0.06 0.26 ± 0.04
Range 0.02–0.05 0.08–0.15 0.58–0.71 0.81–0.93 0.20–0.30
Triticum aestivum Mean ± SD 0.06 ± 0.02 0.08 ± 0.01 1.57 ± 0.10 0.87 ± 0.17 0.13 ± 0.03
Range 0.04–0.09 0.05–0.09 1.50–1.73 0.61–1.02 0.11–0.1 8

Table 3
Accumulation factor (AF) (on dry weight basis) for food crops grown in tube well water.

Species Statistics Cr Ni Mn Pb Cd
Coriandrum sativum Mean ± SD 0.06 ± 0.04 0.06 ± 0.01 1.43 ± 1.07 2.12 ± 0.11 0.34 ± 0.14
Range 0.01–0.15 0.05–0.07 0.19–3.71 2.00–2.26 0.16–0.46
Allium cepa Mean ± SD 0.02 ± 0.01 0.08 ± 0.03 0.74 ± 0.49 1.81 ± 0.75 0.41 ± 0.14
Range 0.00–0.03 0.05–0.11 0.25–1.42 1.29–2.90 0.21–0.53
Abelmoschus esculentus Mean ± SD 0.06 ± 0.02 0.09 ± 0.04 1.12 ± 0.73 1.56 ± 0.13 0.23 ± 0.09
Range 0.03–0.08 0.06–0.14 0.39–1.98 1.40–1.69 0.11–0.33
Allium sativum Mean ± SD 0.05 ± 0.03 0.07 ± 0.03 0.48 ± 0.16 1.49 ± 0.26 0.40 ± 0.18
Range 0.01–0.10 0.03–0.10 0.32–0.69 1.22–1.85 0.19–0.62
Capsicum annum Mean ± SD 0.04 ± 0.02 0.08 ± 0.02 0.60 ± 0.20 1.20 ± 0.60 0.37 ± 0.23
Range 0.00–0.07 0.05–0.09 0.38–0.85 0.74–2.07 0.12–0.67
Daucus carota Mean ± SD 0.06 ± 0.04 0.06 ± 0.02 0.57 ± 0.39 1.33 ± 0.11 0.22 ± 0.13
Range 0.02–0.11 0.04–0.09 0.26–1.14 1.18–1.45 0.04–0.33
Spinacia oleracea Mean ± SD 0.02 ± 0.01 0.04 ± 0.01 1.4 ± 0.15 1.38 ± 0.39 0.23 ± 0.05
Range 0.01–0.03 0.04–0.05 1.23 – 1.58 0.91–1.80 0.17–0.29
Raphanus sativus Mean ± SD 0.06 ± 0.04 0.05 ± 0.03 0.58 ± 0.39 1.70 ± 0.30 0.29 ± 0.09
Range 0.02–0.10 0.00–0.08 0.20–1.30 1.45–2.12 0.19–0.40
Mentha spicata Mean ± SD 0.08 ± 0.05 0.09 ± 0.02 2.94 ± 0.52 3.29 ± 0.63 0.28 ± 0.16
Range 0.02–0.18 0.06–0.10 2.20–3.43 2.66–3.96 0.12–0.50
Solanum melongena Mean ± SD 0.06 ± 0.01 0.06 ± 0.00 0.43 ± 0.08 1.52 ± 0.24 0.19 ± 0.07
Range 0.05–0.06 0.06–0.06 0.32–0.52 1.23–1.80 0.09–0.26
Lycopersicum esculentum Mean ± SD 0.04 ± 0.02 0.05 ± 0.02 0.63 ± 0.05 1.41 ± 0.19 0.28 ± 0.05
Range 0.01–0.06 0.03–0.06 0.58–0.70 1.17–1.60 0.22–0.34
Triticum aestivum Mean ± SD 0.06 ± 0.04 0.05 ± 0.03 1.18 ± 1.01 1.48 ± 0.42 0.14 ± 0.08
Range 0.03–0.11 0.01–0.07 0.18–2.57 1.13–2.07 0.06–0.24

(0.08–0.15), (0.41–5.52), (0.61–1.11), (0.13–0.42) respectively and
for tube well water irrigated food crops it ranged from 0.02 to
0.08 for Cr, 0.04 to 0.09 for Ni, 0.43 to 2.94 Mn, 1.20 to 3.29 Pb
and 0.14 to 0.41 for Cd.
3.3. Daily intake of metals and human health risk assessment
Results for the evaluation of DIM and HRI from heavy metals
contaminated food crops are presented in Tables 4 and 5 for tube well
water and wastewater irrigated crops. Results showed that DIM and
HRI values were high in food crops irrigated with wastewater as
compared to tube well irrigated food crops. DIM of tube well water
irrigated food crops ranged from 4.0E04 to 1.7E03, 1.1E03 to
2.2E03, 6.9E03 to 4.2E02, 1.2E02 to 2.2E02 and 3.0E04 to
1.0E03 mg kg1 person1 d1 for Cr, Ni, Mn, Pb and Cd, respectively
(Table 4). Similarly, for wastewater irrigated food crops DIM values
were 1.2E03 to 4.9E03, 2.7E03 to 5.2E03, 1.2E02 to
1.6E01, 1.8E02 to 3.3E02 and 1.0E03 to 3.1E03 mg kg1 per-
son1 d1 for Cr, Ni, Mn, Pb and Cd, respectively (Table 5). The HRI
values for Cr, Ni, Mn, Pb and Cd for tube well water irrigated food
crops were in the range of 3.0E04 to 1.1E03, 5.4E02 to
1.1E01, 4.8E02 to 3.0E01, 3.5E + 00 to 6.2E + 00 and 3.2E01
to 9.7E01, respectively (Table 4). Similarly, in wastewater irrigated
food crops HRI values for Cr, Ni, Mn, Pb and Cd ranged from 8.0E04
to 3.3E03, 1.4E01 to 2.6E01, 8.6E02 to 1.2E + 00, 5.2E + 00 to
9.4E + 00 and 9.5E01 to 3.1E + 00, respectively (Table 5).
 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238 2235

Table 4
Daily intakes of metals (DIM) (mg kg1 person1 d1) and Health Risk Index (HRI) for individual heavy metals in food crops irrigated with tube well water.

Metals Index Cr Ni Mn Pb Cd
Coriandrum sativum DIM 1.30E03 1.40E03 2.28E02 2.18E02 8.00E04
HRI 9.00E04 6.89E02 1.63E01 6.23E + 00 8.01E01
Allium cepa DIM 4.00E04 1.90E03 1.19E02 1.86E02 1.00E03
HRI 3.00E04 9.68E02 8.48E02 5.32E + 00 9.69E01
Abelmoschus esculentus DIM 1.10E03 2.20E03 1.80E02 1.61E02 5.00E04
HRI 8.00E04 1.09E01 1.28E01 4.60E + 00 5.43E01
Allium sativum DIM 1.10E03 1.80E03 7.70E03 1.54E02 9.00E04
HRI 7.00E04 9.04E02 5.50E02 4.39E + 00 9.32E01
Capsicum annum DIM 8.00E04 1.90E03 9.70E03 1.23E02 9.00E04
HRI 5.00E04 9.75E02 6.92E02 3.52E + 00 8.62E01
Daucus carota DIM 1.20E03 1.60E03 9.00E03 1.37E02 5.00E04
HRI 8.00E04 7.83E02 6.46E02 3.90E + 00 5.19E01
Spinacia oleracea DIM 4.00E04 1.10E03 2.25E02 1.42E02 5.00E04
HRI 3.00E04 5.42E02 1.61E01 4.07E + 00 5.35E01
Raphanus sativus DIM 1.20E03 1.20E03 9.30E03 1.75E02 7.00E04
HRI 8.00E04 6.12E02 6.64E02 5.00E + 00 6.84E01
Mentha spicata DIM 1.70E03 2.20E03 4.24E02 2.16E02 6.00E04
HRI 1.10E03 1.10E01 3.03E01 6.18E + 00 6.50E01
Solanum melongena DIM 1.10E03 1.50E03 6.90E03 1.56E02 5.00E04
HRI 8.00E04 7.30E02 4.89E02 4.46E + 00 4.53E01
Lycopersicum esculentum DIM 8.00E04 1.30E03 1.01E02 1.46E02 7.00E04
HRI 5.00E04 6.33E02 7.22E02 4.16E + 00 6.64E01
Triticum aestivum DIM 1.20E03 1.30E03 1.89E02 1.52E02 3.00E04
HRI 8.00E04 6.56E02 1.35E01 4.34E + 00 3.27E01

Table 5
Daily intakes of metals (mg kg1 person1 d1) and HRI for individual heavy metals in food crops irrigated with wastewater.

Metals Index Cr Ni Mn Pb Cd
Coriandrum sativum DIM 3.40E03 3.60E03 6.16E02 2.78E02 3.00E03
HRI 2.30E03 1.79E01 4.40E01 7.95E + 00 3.02E + 00
Allium cepa DIM 2.50E03 4.00E03 3.61E02 3.30E02 3.10E03
HRI 1.60E03 2.01E01 2.58E01 9.42E + 00 3.06E + 00
Abelmoschus esculentus DIM 3.30E03 3.20E03 4.06E02 2.85E02 1.70E03
HRI 2.20E03 1.62E01 2.90E01 8.14E + 00 1.67E + 00
Allium sativum DIM 2.10E03 4.20E03 1.97E02 2.50E02 2.50E03
HRI 1.40E03 2.11E01 1.41E01 7.15E + 00 2.48E + 00
Capsicum annum DIM 2.10E03 3.70E03 3.14E02 2.26E02 1.50E03
HRI 1.40E03 1.85E01 2.25E01 6.46E + 00 1.49E + 00
Daucus carota DIM 2.80E03 3.50E03 4.18E02 2.17E02 2.10E03
HRI 1.90E03 1.73E01 2.98E01 6.19E + 00 2.11E + 00
Spinacia oleracea DIM 4.90E03 3.40E03 1.62E01 1.81E02 2.60E03
HRI 3.30E03 1.68E01 1.16E + 00 5.17E + 00 2.63E + 00
Raphanus sativus DIM 2.20E03 4.30E03 1.77E02 2.75E02 2.10E03
HRI 1.50E03 2.14E01 1.26E01 7.86E + 00 2.10E + 00
Mentha spicata DIM 2.60E03 3.10E03 4.43E02 3.01E02 3.10E03
HRI 1.70E03 1.57E01 3.17E01 8.60E + 00 3.07E + 00
Solanum melongena DIM 2.20E03 5.20E03 1.20E02 2.57E02 1.10E03
HRI 1.50E03 2.58E01 8.58E02 7.34E + 00 1.13E + 00
Lycopersicum esculentum DIM 1.20E03 3.80E03 1.91E02 2.62E02 1.90E03
HRI 8.00E04 1.91E01 1.36E01 7.47E + 00 1.89E + 00
Triticum aestivum DIM 1.90E03 2.70E03 4.61E02 2.59E02 1.00E03
HRI 1.30E03 1.36E01 3.30E01 7.41E + 00 9.52E01

4. Discussion manufactories, surgical instruments and garages wastes are also

4.1. Heavy metals in irrigation water and their subsequent deposition
in agricultural soil
In Pakistan a well-known source of irrigation water is surface
water of streams and rivers which is either diverted or pumped
into nearby agricultural fields (Qadir et al., 2008). Among other
possible sources of such contamination are excessive wastewater
irrigation practices (Singh et al., 2010). The industrial, domestic
and urban wastewaters discharge into the surface water without
prior treatment is prominent. Moreover, various other wastewater
sources including oil and ghee industries, sports industries,
tanneries and cutlery industries wastes, while electroplating, col-
oring, metal surface treatment, fabric printing, batteries and paints
found to be the major contributors. Same wastewater bodies are
used for the purpose of irrigation. Wastewater of Nullah Palkhu
and Aik is being practiced for the irrigation of crops from last
few decades. There has been found a linear relationship of heavy
metals accumulation in soil with irrigation time (Lucho-Contantino
et al., 2005). As a matter of fact the wastewater irrigation of agri-
cultural soils for longer period may be one the factor responsible
for contamination of heavy metals (Chary et al., 2008). Agrochem-
icals and atmospheric deposition are other possible factors that
cause soil contamination in the study area. According to ETPI
(1998) the wastewater discharge into Nullah Aik and Palkhu by
the Sialkot city is approximately 547–814 m3 d1 of effluents with
no effluent treatment plant for toxic chemicals. A total of 19
million m3 per annum of wastewater is discharged directly or
2236 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238
indirectly by the Sialkot city into Nullah Aik and Palkhu (WWF,
2007). However the wastewater discharge capacity of Wazirabad
is still unknown. Agricultural practices in urban and sub-urban
areas and geological sources (Himalayas of Jammu and Kashmir)
are also among the contributors of heavy metals in water bodies
of currently studied areas. It is therefore obvious that with long
term wastewater irrigation, agricultural soil seems to be deposited
with different toxic metals, and this fact is clearly reflected from
our results. The accumulation of heavy metals in the agricultural
field is also already reported (Yu et al., 2006). In our study, we re-
corded high concentrations of Cd, Pb and Cr in the wastewater of
Nullah Aik and Palkhu which exceeded the permissible limits for
heavy metals in irrigation water, i.e. 0.01, 5.0, 2.0, 2.0 and
0.10 lg mL1 for Cd, Pb, Mn, Ni and Cr respectively (FAO, 1985).
Similarly Cd and Cr concentrations in tube well water also surpass
the permissible limits. Higher concentration of Cr in the earlier
study was recorded in the ground water of Sialkot (Ullah et al.,
2009). In our findings the higher heavy metal concentrations in
tube well water are however attributable to the presence of tan-
neries and leather industry in Sialkot city. Our results of certain
metals are above the previously detected values from the neigh-
boring countries, e.g. Singh et al. (2010) reported lower values
for Cd (0.02), Pb (0.09), Ni (0.06) and Cr (0.05) lg mL1 in wastewa-
ter of Varanasi, India. Similarly, Sharma et al. (2006) have also re-
ported concentration of Cd (0.03), Pb (0.26), Ni (0.070) and Cr
(0.09) in the wastewater collected from Dinapur, India. However,
Gupta et al. (2008) have recorded higher concentrations of Pb, Cu
and Ni in treated and untreated wastewater samples, India.
The permissible limits for metals in soil are 3.0 (Cd), 300 (Pb),
75 (Ni), 150 (Cr) lg g1 and 2000 (Mn) mg kg1 as given by Euro-
pean Union Standards (2002) and Pendias and Pendias (1992).
Among studied metals, Cd concentration ranged from 7.13 to
11.13 mg kg1 which exceeded permissible limits set by European
Union (2002). Cd has greater exchangeable capacity, easily become
available and soluble in soils and become bio-available and accu-
mulates in edible parts of the plants (Luo et al., 2011). As compared
to the current study, higher level of Cd (22.2–51.0 mg kg1) were
found in wastewater irrigated soil samples collected from Titagarh,
India, while lower concentrations (0.41–1.71 mg kg1) in the
wastewater irrigated soils from Beijing, China (Gupta et al.,
2008). In contrary, Mapanda et al. (2005) reported lower level of
Cd (3.4 lg g1) in the wastewater irrigated soil, Zimbabwe.
4.2. Heavy metals in edible part of food crops
The present study indicated that all the heavy metals concen-
trations in the wastewater irrigated food crops were higher than
tube well water irrigated food crops. Our results showed that Cd,
Pb exceeded the permissible limits in all the wastewater and tube
well water irrigated food crops. Cr was also beyond the permissible
limits in wastewater irrigated food crops except L. esculentum.
These findings are a clear reflection of wastewater irrigation and
subsequent accumulation of heavy metals in the vegetables.
The permissible limits for food crops are 500, 67, 0.2, 5.0,
2.3 mg kg1 for Mn, Ni, Cd, Pb and Cr respectively (Codex Alimen-
tarius Commission, 2001; WHO/FAO, 2007). Apart from aforemen-
tioned sources of heavy metal in the selected food crops
contamination, the use of phosphate fertilizers is a prominent
practice, and is the most important cause of Cd accumulation in
the agricultural field. Cd is known to be present as impurity in
phosphate rocks (Zhou et al., 1994) and gets transferred to plants
(edible parts) due to high mobility. Leafy vegetables (S. oleracea,
M. spicata, C. sativum) had maximum metal concentrations (except
Ni and Pb) among food crops studied that is consequence of their
large surface area of leaf, higher transpiration and faster growth
rate that enhances the metals translocation in leafy vegetables
(Itanna, 2002; Muchuweti et al., 2006). The expanded leaf of leafy
vegetables makes them sensitive as a recipient of dust and
splashing of rainwater (Luo et al., 2011).
Low metal concentrations in some species such as T. aestivum, A.
sativum and S. melongena and some variant concentrations of heavy
metals in food crops could be ascribed to the variability in metals
absorption in plant and physical and chemical nature of soil (Zurera
et al., 1989). Morphological and physiological changes play a vital
role in metal translocation, elimination, and accumulation (Itanna,
2002). The non-essential heavy metals become toxic via consump-
tion of food crops. These results indicated that accumulation of cer-
tain metal beyond permissible limits is a serious health risk factor
for the human population of the two selected cities of Pakistan.
Our findings show that continuous wastewater irrigation accumu-
lates heavy metal in food crops that was beyond the safe limits for
most of the food crops. No doubt, the local population is barely
aware of the nutritious status of vegetables which they consume
daily. In addition, there is no alternative source of vegetable sup-
plies. Therefore, the population in this area is at a serious risk.
4.3. Heavy metal transfer in edible parts of food crops
Accumulation factor (AF) for Mn and Pb > 1 in most of the stud-
ied food crops indicates greater accumulation. Those metals which
have high transfer factor make their way to edible part of plant
easier than those into low transfer factor (Luo et al., 2011). That
is why these metals reflect their high accumulation in various food
crops in such high ratio. AF values varied for heavy metal in various
food crops and also with Wastewater and Tube well water irrigated
sites (Cui et al., 2004). High AF values was observed for Pb and Cd
in tube well irrigated food crops and that is considered to be one of
the possible reasons to cause health risk in human via their con-
sumption. Low AF value for heavy metals (Pb and Cd, Cr and Mn
in various food crops) was indicated in wastewater irrigated food
crops than those of tube well water irrigated food crops, while
for other studied metals (Ni and Cr, Mn, Cd in different food crops)
a reverse trend was observed (Tables 2 and 3). This indicated that
the uptake of heavy metals by food crops did not increase linearly
with the increased metal concentration in soil. Our study is in
agreement with previous findings of Hooda et al. (1997) and Rattan
et al. (2005). This phenomenon is very significant in term of long-
term wastewater irrigation that the same proportions would not
be the part of food chain.
4.4. Daily intake of metals and human health risk assessment
Human risk assessment quantification from the pathway of
food chain is of prime importance in countries like Pakistan, where
wastewater irrigation practice is still unchecked. There are several
exposure pathways which mainly depend on contaminated
sources of air, water, soil, food and consuming population (Caussy
et al., 2003), but the routes of exposure via food chain is one of the
key pathways of heavy metals exposure to human (Muchuweti
et al., 2006). Pakistani populations are mostly vegetarian and also
rely on cereal crop (T. aestivum L.) as a staple food. Food crops pro-
duced in the present study are consumed directly by the local
inhabitants or sold to the market for dispersed population con-
sumption. Studied food crops are consumed daily in their daily diet
(meals) by the local population as a Salad and/or as in cooked form
in higher proportion. Some of the leafy vegetables (C. sativum, M.
spicata), C. annum, L. esculentum, D. carota, R. sativus, and A. cepa
are traditionally used as salad with almost 1–2 times in their meal
on daily basis and also some used in sandwich, burger, ketchup,
jelly, etc., and posing greater health risk as they get contaminated
with toxic metals. Oral reference dose (RfD) is the daily exposure of
individuals to toxins or pollutants which can pose no appreciable
 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238
hazardous in their lifetime. The RfDs for toxic metals like Cd, Cr, Ni,
Pb, and Mn are 1E03, 1.5E0, 2E2, 3.5E3, 1.4E1 mg kg1 d1
respectively (USEPA IRIS, 2006). In our study the heavy metals ex-
cept Cr and Ni have HRI > 1 indicates a possible future human
health risk via food crops intake. Cd showed HRI > 1 in all the
wastewater irrigated food crops except T. aestivum, while tube well
water irrigated food crops have HRI < 1. The highest HRI values > 1
was observed in the case of Pb in all the food crops irrigated with
wastewater and tube well water, while Mn revealed HRI > 1 only in
S. oleracea irrigated with wastewater. However Cd and Pb are con-
sidered as the non-essential metals contributing to the health haz-
ards even at extreme low concentrations (Ikeda et al., 2000).
Zhuang et al. (2009) have also reported HRI for Cd and Pb above
the permissible limits in vegetables and cereals. The present study
is very important in-term of health perspectives indicating the
health risk to the human population from heavy metals consump-
tion in food crops.
5. Conclusion
Significant accumulation of toxic heavy metals in soils and food
crop samples is due to the wastewater irrigation in Sialkot and
Wazirabad cities of Pakistan. As far as we know, no data is available
from Pakistan on thorough hazards assessment from the intake of
contaminated food and their outcomes on human health. The cur-
rent study indicates that wastewater irrigation is the main source
of metal accumulation in food crops, which on human consump-
tion may lead to adverse health outcomes on large scale. Results
reveal that heavy metal (Pb, Cd and Mn) showed high AF. Further-
more, HRI values >1 were also observed for Pb, Cd and Mn con-
sumption in food crops. This study gives a brief insight into the
current scenario of food crops contamination and possible future
health risk estimates. There is a dire need to monitor strictly the
wastewater irrigation system and also tube well irrigated system
of the study area and develop different strategies to prevent heavy
metals accumulation in food crops that may ultimately minimize
chronically health risk to the exposed population.
Acknowledgements
The authors pay thanks to Dr. Abdul Qadir for his tedious and
exhaustive work during sample collection and also Department
of Biochemistry in regard of heavy metal analysis.
References
Allen, S.E., Grimshaw, H.M., Rowland, A.P., 1986. Chemical analysis. In: Moore, P.D.,
Chapman, S.B. (Eds.), Methods in Plant Ecology. Blackwell, Scientific Publication,
Oxford, London, pp. 285–344.
Caussy, D., Gochfeld, M., Gurzan, E., 2003. Lessons from case studies of metals
investigating exposure, bioavailability and risk. Ecotoxicol. Environ. Saf. 56, 45–
51.
Chary, N.S., Kamala, C.T., Raj, D.S.S., 2008. Assessing risk of heavy metals from
consuming food grown on sewage irrigated soils and food chain transfer.
Ecotoxicol. Environ. Saf. 69, 513–524.
Codex Alimentarius Commission (FAO/WHO), Food Additives and Contaminants.
Joint FAO/WHO Food Standards Program 2001, ALINORM 01/12A:1-289.
Cui, Y.J., Zhu, Y.G., Zhai, R.H., Chen, D.Y., Huang, Y.Z., Qiu, Y., Ling, J.Z., 2004. Transfer
of metals from soil to vegetables in an area near a smelter in Nanning, China.
Environ. Int. 30, 785–791.
Dyer, C.A., 2007. Heavy metals as endocrine disrupting chemicals. In: Gore, A.C.
(Ed.), Endocrine-Disrupting Chemicals: From Basic Research to Clinical Practice.
Humana Press, Totowa, pp. 111–133.
ETPI, 1998. The Leather Sector: Environmental Report, Environmental Technology
Programme for Industry (ETPI), Federation of Pakistan Chamber of Pakistan.
http://www.cpp.org.pk/etpirpt/LeatherSectorReport.Pdf (accessed 04.11.11).
European Union, 2002. Heavy Metals in Wastes, European Commission
on Environment. <http://ec.europa.eu/environment/waste/studies/pdf/heavy
metalsreport.Pdf> (accessed 12.06.12).
FAO, 1985. Water Quality for Agriculture. Paper No. 29 (Rev. 1) UNESCO,
Publication, Rome. Ge, K.Y. 1992. The Status of Nutrient and Meal of Chinese
in the 1990s. Beijing People’s Hygiene Press.
2237
Gupta, N., Khan, D.K., Sandra, S.C., 2008. An assessment of heavy metal
contamination in vegetables grown in wastewater-irrigated areas of Titagarh,
West Bengal, India. Bull. Environ. Cont. Toxicol. 80, 115–118.
Harmanescu, M., Alda, L.M., Bordean, D.M., Gogoasa, I., Gergen, I., 2011. Heavy
metals health risk assessment for population via consumption of vegetables
grown in old mining area; a case study: Banat County, Romania. Chem. Cent. J.
5, 64.
Hooda, P.S., McNulty, D., Alloway, B.J., 1997. Plant availability of heavy metals in
soils previously amended with heavy applications of sewage sludge. J. Sci. Food
Agric. 73, 446–454.
Hua, J., Wua, F., Wua, S., Cao, Z., Lin, X., Wong, M.H., 2013. Bioaccessibility, dietary
exposure and human risk assessment of heavy metals from market vegetables
in Hong Kong revealed with an in vitro gastrointestinal model. Chemosphere
91, 455–461.
Ikeda, M., Zhang, Z.W., Shimbo, S., Watanabe, T., Nakatsuka, H., Moon, C.S., Matsuda-
nInoguchi, N., Higashikawa, K., 2000. Urban population exposure to lead and
cadmium in east and south-east Asia. Sci. Total Environ. 249, 373–384.
Itanna, F., 2002. Metals in leafy vegetables grown in Addis Ababa and toxicological
implications. Ethiopian J. Health Dev. 6, 295–302.
Jarup, L., 2003. Hazards of heavy metal contamination. Br. Med. Bull. 68, 167–182.
Karvelas, M., Katsoyiannis, A., Samara, C., 2003. Occurrence and fate of heavy metals
in the wastewater treatment process. Chemosphere 53, 1201–1210.
Khan, S., Cao, Q., Zheng, Y.M., Huang, Y.Z., Zhu, Y.G., 2008. Health risks of heavy
metals in contaminated soils and food crops irrigated with wastewater in
Beijing, China. Environ. Pollut. 152, 686–692.
Li, Q., Chen, Y., Fu, H., Cui, Z., Shi, L., Wang, L., Liu, Z., 2012. Health risk of heavy
metals in food crops grown on reclaimed tidal flat soil in the Pearl River Estuary,
China. J. Hazard. Mater. 227–228, 148–154.
Loutfy, N., Fuerhacker, M., Tundo, P., Raccanelli, S., Dien, A.G., Ahmed, M.T., 2006.
Dietary intake of dioxins and dioxin-like PCBs, due to the consumption of dairy
products, fish/seafood and meat from Ismailia city, Egypt. Sci. Total Environ.
370, 1–8.
Lucho-Contantino, C.A., Alvarez-Suárez, M., Beltrán-Hernádez, R.I., Prieto- Garcia, F.,
Poggi-Varaldo, H.M., 2005. A multivariate analysis of the accumulation and
fractionation of major and trace elements in agricultural soils in Hidalgo State,
Mexico irrigated with wastewater. Environ. Int. 31, 313–323.
Luo, C., Liu, C., Wang, Y., Liu, X., Li, F., Zhang, G.G., Li, X., 2011. Heavy metal
contamination in soils and vegetables near an e-waste processing site, south
China. J. Hazard. Mater. 186, 481–490.
Mapanda, F., Mangwayana, E.N., Nyamangara, J., Giller, K.E., 2005. The effect of long-
term irrigation using wastewater on heavy metal contents of soils under
vegetables in Harare, Zimbabwe. Agric. Ecosyst. Environ. 107,
151–165.
Muchuweti, M., Birkett, J.W., Chinyanga, E., Zvauya, R., Scrimshaw, M.D., Lester, J.N.,
2006. Heavy metal content of vegetables irrigated with mixtures of wastewater
and sewage sludge in Zimbabwe: implications for human health. Agri. Ecosyst.
Environ. 112 (1), 41–48.
Pasha, Q., Malik, S.A., Shaheen, N., Shah, M.H., 2010. Investigation of trace metals in
the blood plasma and scalp hair of gastrointestinal cancer patients in
comparison with controls. Clin. Chim. Acta 411 (7–8), 531–539.
Pendias, A.K., Pendias, H., 1992. Elements of group VIII. In: Trace Elements in Soils
and Plants. CRC Press, Boca Raton, pp. 271–276.
Ping, L., Hai-jun, Z., Li-li, W., Zhao-hui, L., Lin, J.W., Yan-qin, W., Li-hua, J., Liang, D.,
Yu-feng, Z., 2011. Analysis of heavy metal sources for vegetable soils from
Shandong Province, China. Agri. Sci. China 10, 109–119.
Qadir, A., Malik, R.N., Husain, S.Z., 2008. Spatio-temporal variations in water quality
of Nullah Aik-tributary of the river Chenab, Pakistan. Environ. Monit. Assess.
140, 43–59.
Rattan, R.K., Datta, S.P., Chhonkar, P.K., Suribabu, K., Singh, A.K., 2005. Long-term
impact of irrigation with sewage effluents on heavy metal content in soils, crops
and groundwater—a case study. Agric. Ecosyst. Environ. 109,
310–322.
Sharma, R.K., Agrawal, M., Marshall, F.M., 2006. Heavy metals contamination in
vegetables grown in wastewater irrigated areas of Varanasi, India. Bull. Environ.
Cont. Toxicol. 77, 311–318.
Singh, A., Rajesh, K.S., Madhoolika, A., Fiona, M.M., 2010. Health risk assessment of
heavy metals via dietary intake of foodstuffs from the wastewater irrigated site
of a dry tropical area of India. Food Chem. Toxicol. 48, 611–619.
Turkdogan, M.K., Fevzi, K., Kazim, K., Ilyas, T., Ismail, U., 2003. Heavy metals in soil,
vegetables and fruits in the endemic upper gastrointestinal cancer region of
Turkey. Environ. Toxicol. Pharmacol. 13, 175–179.
Ullah, R., Malik, R.N., Qadir, A., 2009. Assessment of groundwater contamination in
an industrial city, Sialkot, Pakistan. Afr. J. Environ. Sci. Technol. 3, 429–446.
United State, Environmental Protection Agency, 2002. Region 9, Preliminary
remediation goals. <http://www.epa.gov/region09/waste/sfind/prg> (accessed
07.08.12).
US EPA-IRIS, 2006. United States, Environmental Protection Agency, Integrated Risk
Information System. <http://www.epa.gov/iris/subst> (accessed 03.12.11).
USEPA, 1990. Recommended protocols for measuring conventional water quality
variables and metals in fresh water of the Puget Sound, 1200 6th Avenue
Seattle, WA 98101. <http://www.psat.a.gov/Publications/protocols/protocol_
pdfs/freshwater.Pdf> (accessed 6.10.12).
WHO/FAO, 2007. Joint FAO/WHO Food Standard Programme Codex Alimentarius
Commission 13th Session. Report of the Thirty Eight Session of the Codex
Committee on Food Hygiene. Houston, United States of America, ALINORM 07/
30/13.
2238 M.U. Khan et al. / Chemosphere 93 (2013) 2230–2238

WWF, 2007. Pakistan’s water at risk: and health related issues in Pakistan and key
recommendation (special report), fresh water and toxic programme. WWF,
Pakistan.
Yu, L., Yan-bin, W., Xi, G., Yi-bin, S., Gang, W., 2006. Risk assessment of heavy metals
in soils and vegetables around non-ferrous metals mining and smelting sites,
Baiyin, China. J. Environ. Sci. 18 (6), 1124–1134.
Zhou, Q., Wu, Y., Xiong, X., 1994. Compound pollution of Cd and Zn and its
ecological effect on rice plant. Chin. J. Appl. Ecol. 5, 428–441.
Zhuang, P., Murray, B., McBride, X.H., Lia, N., Li, Z., 2009. Health risk from heavy
metals via consumption of food crops in the vicinity of Dabaoshan mine, South
China. Sci. Total Environ. 407, 1551–1561.
Zurera, G., Moreno, R., Salmeron, J., Pozo, R., 1989. Heavy metal uptake from
greenhouse border soils for edible vegetables. J. Sci. Food Agr. 49, 307–314.