This research article studied the presence of pesticide and phthalate residues in raw tea, branded tea, herbal tea, and tea infusions packaged in different containers. The Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method was used to extract and analyze samples. Some pesticide residues were detected in raw and branded teas, while only two pesticides were found in herbal tea. Phthalates were detected in tea packaged in plastic bags and disposable cups. The study also examined how pesticide residues are transferred from raw tea to infusion over different steeping times and how phthalates leach from plastic containers into tea over time. Pesticide and phthal

1. RESEARCH ARTICLE
Determination of pesticide and phthalate residues in tea
by QuEChERS method and their fate in processing
Sapna Yadav1
& Satyajeet Rai1
& Ashutosh K. Srivastava2
& Smita Panchal3
& D.K. Patel3
&
V.P. Sharma4
& Sudha Jain5
& L.P. Srivastava1
Received: 26 May 2016 /Accepted: 9 September 2016
# Springer-Verlag Berlin Heidelberg 2016
Abstract In this study, the quick, easy, cheap, effective, rug-
ged, and safe (QuEChERS) method was applied for the anal-
ysis of the multiclass pesticide residues of 12 organochlorines
(OCs), 9 organophosphates (OPs), 11 synthetic pyrethroids
(SPs), 4 herbicides, 6 phthalates in raw tea (loose tea, branded
tea and herbal tea), and tea infusion in 4 different containers
(glass cup, earthen cup, plastic bag and disposal cup). In loose
tea and branded tea residues, malathion (0.257 and
0.118 mg kg−1
), cypermethrin (0.065 and 0.030 mg kg−1
),
and fenvalerate (0.032 and 0.030 mg kg−1
) were detected,
respectively. In herbal tea, residues of only cypermethrin
(0.053 mg kg−1
) and fenvalerate (0.045 mg kg−1
) were detect-
ed. Tea infusion samples contained in a plastic bag were found
to be contaminated with only dibutyl phthalate (DBP)
(0.038 mg kg−1
). Disposable cup was found to be
contaminated with DBP (0.026 mg kg−1
) and diethyl phthalate
(DEP) (0.004 mg kg−1
). Further, to know the processing be-
havior of pesticides, the spiked raw tea was subjected to tea
infusion at different brewing times (2, 5, 10 min). The analysis
demonstrated that dimethoate, dichlorvos, and malathion had
shown more than 10 % of translocation at 5 min of brewing
time. Further brewing for 10 min revealed the reduction in
concentration of pesticides. Leaching of phthalate residues
from different plastic containers was also studied at 10, 30,
and 60 min. DBP, benzyl butyl phthalate (BzBP), and di-
2-(ethylhexyl) phthalate (DEHP) were leached in the tea infu-
sion samples packed in plastic bags. On the other hand, in
disposable cups, leaching of DBP, DEP, and dimethyl phthal-
ate were found. The concentration of phthalate residues in-
creased with retention time. Pesticide and phthalate contami-
nants were recorded at low quantities in few samples only.
Keywords QuEChERS . Pesticides . Phthalates . Leaching .
Tea
Abbreviations
OCs Organochlorines
OPs Organophosphates
SPs Synthetic Pyrethroids
QuEChERS Quick, easy, cheap, effective, rugged, safe
HPLC High performance liquid chromatography
PSA Primary secondary amine
HCH Hexachlorocyclohexane
p,p’-DDE para,para’-dichlorodiphenyldichloroethylene
p,p’-DDD para,para’-dichlorodiphenyldichloroethane
p,p’-DDT para,para’-dichlorodiphenyltrichloroethane
o,p’-DDE ortho,para’-dichlorodiphenyldichloroethylene
o,p’-DDT otrho,para’-dichlorodiphenyltrichloroethane
DMP Dimethyl phthalate
Responsible Editor: Ester Heath
* L.P. Srivastava
laxmanprasad13@gmail.com
1
Pesticide Toxicology Laboratory, Regulatory Toxicology Group,
CSIR-Indian Institute of Toxicology Research (CSIR-IITR), MG
Marg, Lucknow, Uttar Pradesh 226001, India
2
Indian Council of Medical Research, Department of Health
Research,Ministry of Health & Family Welfare, National Aids
Research Institute, Plot No.73, G Block, MIDC, Pune, Bhosari 411
026, India
3
Analytical Chemistry Laboratory, Regulatory Toxicology Group,
CSIR-Indian Institute of Toxicology Research (CSIR-IITR), MG
Marg, Lucknow, Uttar Pradesh 226001, India
4
Developmental Toxicology Laboratory, Regulatory Toxicology
Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR),
MG Marg, Lucknow, Uttar Pradesh 226001, India
5
Department of Chemistry, University of Lucknow, Lucknow, Uttar
Pradesh 226007, India
Environ Sci Pollut Res
DOI 10.1007/s11356-016-7673-2

2. DEP Diethyl phthalate
DBP Dibutyl phthalate
DEHP Diethylhexyl phthalate
BzBP Benzyl butyl phthalate
DOC Dioctyl phthalate
GC Gas chromatography
GC-MS/
MS
Gas chromatography tandem mass
spectrometry
LC-MS/MS Liquid chromatography tandem mass
spectrometry
ECD Electron capture detector
PDA Photo diode array
SRM Selected reaction monitoring
UPLC Ultra performance liquid chromatography
ESI Electrospray ionization
MRM Multiple reaction monitoring
IS Ion source
LOD Limit of detection
LOQ Limit of quantification
%RSD Percentage relative standard deviation
BDL Below detection limit
MRL Maximum residue limit
SML Specific migration limit
Introduction
Tea (Camellia sp.) justifies the title of Bqueen of beverages^
for its salubrious properties, pleasant aroma, and flavor.
Globally, tea is the most consumed beverage due to its intrin-
sic properties and acceptance of quality by consumers. India
ranked second after China in the production of tea and it ac-
counts for 23.34 % of global tea production; hence, it can be
considered as one of the largest global producers and ex-
porters of tea (Tea board of India). It is one of the popular
industries where India has entrenched itself completely in
the global market.
Tea crop is subjected to various biotic stresses during their
life cycle. This Bhealth beverage^ has to go through several
challenges right from its cultivation to its marketing. Its con-
frontations with the challenges begin right from the prime
stage of its cultivation. Several pests such as Fiorinia theae,
Helopeltis sp., Poecilocoris latus, Tropicomyia theae,
Eriophyes theae, and Gracillaria theivora infest the tea crops.
Further, several species of weeds and fungal pathogens also
compete with tea plants for moisture and nutrients
(Seenivasan and Muraleedharan 2011). In order to circumvent
this problem of biotic stress, use of broad-spectrum synthetic
pesticides, such as OCs1
, OPs2
, SPs3
, carbamates, herbicides
and neonicotinoids, are recommended during cultivation as
well as postharvest and storage stage (Gupta et al. 2008;
Cajka et al. 2012). Indiscriminate and imprudent use and
avoidance of waiting period during application of these
chemicals causes the threat of the presence of their residues,
which may result in serious health problems during consump-
tion. Exposure of pesticide residues through food commodi-
ties and environmental component such as water and air lead
to a variety of adverse health effects, ranging from simple
irritation, burning sensation, and itchiness of the skin and eyes
to more severe effects such as affecting the nervous system,
mimicking hormones, causing reproductive problems, gastro-
intestinal problem, and also causing cancer (Kushik and
Chandrabhan, 2006; Chinnachamy and Nair 2009). Tea is
the most consumed drink (6 g/day per individual) in India.
Thus, the presence of pesticide residues in tea may result in
significant potential source of human exposure to pesticide
residues (Jaggi et al. 2001).
Tea is subjected to infusion prior to consumption, which
may result in translocation and loss of pesticides. The subject
of great concern is the injudicious usage of plastics for pack-
aging during production, processing, storage, and transport of
tea; moreover, tea vendors provide hot tea infusion in plastic
bags/pouches and disposable plastic cups to consumers (Wu
et al. 2012; Fierens et al. 2012). This supply of tea infusion in
hot condition may result in leaching of phthalates and phthalic
acid esters into the infusion (Guo and Kannan 2012).
Phthalates are diesters of orthophthalic acid and are used as
plasticizers, so as to increase the flexibility of plastics.
Phthalates are potential endocrine disruptors, which cause se-
rious, acute health hazards either by mimicking or blocking
hormones, thereby disrupting the body’s normal functions
(Roldan et al. 2004). Thus, realizing the adverse implication
associated with pesticides and phthalates has emerged as a
significant and inevitable task for regulatory agencies and
users.
Assessing the nature, behavior, and interaction of chemical
residues with such a complex matrix is the foremost challenge.
There is also a paucity of data on the codetermination of pes-
ticide and phthalate residues in tea samples. Hence, the prime
objective of the present study is the codetermination of organ-
ochlorines (OCs), organophosphates (OPs), synthetic pyre-
throids (SPs), and herbicide and phthalate residues in raw
tea and tea infusion by use of the QuEChERS4
method.
Moreover, translocation of these classes of multi-pesticide
residues from raw tea to tea infusion and their loss during
processing and leaching of phthalate from plastic bags and
disposable cups into tea infusion have also been studied to
assess possible health impacts.
Materials and methods
Chemicals and reagents
All solvents like n-hexane, acetone, ethyl acetate, and aceto-
nitrile of HPLC5
grade and sodium chloride (NaCl) were
Environ Sci Pollut Res

3. procured from Sisco Research Laboratory Pvt. Ltd., India.
Anhydrous magnesium sulfate (MgSO4) was procured from
Sigma Aldrich Chemicals Pvt. Ltd., India. Before use, MgSO4
was baked for 4 h at 600 °C in a muffle furnace to remove
possible phthalate impurities. Primary secondary amine
(PSA)6
bondasil 40 μm part 12213024 from Agilent
Technologies India Pvt. Ltd., India was used for sample
cleanup.
Pesticide and phthalate standards
The standard of pesticides α-hexachlorocyclohexane (α-HCH)7
,
β-hexachlorocyclohexane (β-HCH), γ-hexachlorocyclohexane
(γ-HCH), δ-hexachlorocyclohexane (δ-HCH), α-endosulfan,
β-endosulfan, p,p’-dichlorodiphenyldichloroethylene (p,p’-
DDE)8
, p,p’-dichlorodiphenyldichloroethane (p,p’-DDD)9
,
p,p’-dichlorodiphenyltrichloroethane (p,p’-DDT)10
, o,p’-
dichlorodiphenyldichloroethylene(o,p’-DDE)11
, o,p’-dichlorodi-
phenyltrichloroethane (o,p’-DDT)12
, heptachlor, malathion,
chlorpyrifos, 4-bromo-2-chlorophenol, dichlorvos, triazophos,
profenofos, ethion, dimethoate, phosphamidon, fenvalerate-I,
fenvalerate-II, cypermethrin-I, cypermethrin-II, τ-fluvalinate,
fenpropathrin, δ-methrine, β-cyfluthrin-I, β-cyfluthrin-II,
bifenthrin, λ-cyhalothrin, pendimethylene, butachlor, alachlor,
atrazine, and phthalates: dimethyl phthalate (DMP)13
, diethyl
phthalate (DEP)14
, dibutyl phthalate (DBP)15
, di-2(ethylhexyl)
phthalate (DEHP)16
, benzyl butyl phthalate (BzBP)17
, and
dioctyl phthalate (DOP)18
of 97–99 % purity were procured from
Supelco Sigma-Aldrich, USA, Fluka Sigma-Aldrich,
Switzerland, and Rankem Pvt. Ltd., New Delhi, India.
Stock solution and intermediate
Individual stock solutions of pesticide and phthalate stan-
dards were prepared at an approximate concentration of
1000 mg L−1
in n-hexane and acetonitrile, respectively.
Intermediate standards of 10 mg L−1
were prepared by
further dilution of the stock solution. Mixed working stan-
dards were prepared from the individual intermediate so-
lution of pesticides by serial dilution. All the solutions
were stored in refrigerator when not in use.
Sampling
A total of 100 samples of 3 different varieties of raw tea
(20 samples each of loose tea, packed/branded tea and
herbal tea) and tea infusion in 4 different containers (10
samples each of glass, earthen cup, plastic bag, and dis-
posal cup) were collected from the local market of
Lucknow, Uttar Pradesh, India.
QuEChERS sample preparation
Raw tea
20 Twenty grams of raw tea samples was taken for analysis.
After grinding with mortar and pestle, 5 g of the powdered tea
samples was added to 5 ml of distilled water and allowed to
soak for 5 min in a 50-ml polypropylene centrifuge tube. The
samples were mixed with 10 ml ethyl acetate, 4 g of anhy-
drous MgSO4, and 1 g activated NaCl, shaken for 10 min, at
50 rpm, on rotospin and was further centrifuged for 10 min at
8000 rpm. An aliquot of 1-ml tea extracts was cleaned with the
mixture of 50 mg PSA, 150 mg anhydrous MgSO4, and 5 mg
of activated charcoal. The extracts were again shaken for
10 min at 50 rpm on rotospin and centrifuged for 10 min at
8000 rpm. The clean extracts were collected in 2-mlgas chro-
matography (GC) vials for analysis.
Tea infusion
Ten milliliters of tea infusion samples was taken in a 50-ml
polypropylene centrifuge tube and mixed with 10 ml ethyl
acetate, 7 g of anhydrous MgSO4, and 1 g of activated NaCl
for the extraction of pesticides and phthalate residues. After
mixing, the extraction solvent and salt tubes were shaken for
10 min at 50 rpm on rotospin and centrifuged at 8000 rpm for
10 min. One milliliter of extracts was mixed with 50 mg PSA,
150 mg anhydrous MgSO4, and 5 mg of activated charcoal
and then shaken again for 10 min at 50 rpm on a rotospin and
centrifuged for 10 min at 8000 rpm for cleanup. The clean
extracts were collected in 2 ml GC19
vials for analysis.
Phthalate leaching
Phthalate leaching was determined by taking 100 ml of tea
infusion sample (temp. 98 ± 2 °C) in earthen cup, plastic bag,
and disposable cup in three replicates, and a glass cup was
taken as control. The temperature was maintained at 98 ± 2 °C
by using water bath. Ten milliliters of the sample was taken
from all the three containers at time intervals of 10, 30, and
60 min, followed by extraction and analysis.
Translocation of pesticides from raw tea to tea infusion
The translocation of pesticides from raw tea to tea infusion
was determined by spiking the raw tea at a concentration of
20 mg kg−1
. A 2.5-g spiked tea was brewed (temp. 98 ± 2 °C)
in 50 ml of water at different time durations of 2, 5, and
10 min. The tea was then filtered in a sieve and then with
Whatman filter paper no. 1 for complete removal of raw tea
from the filtrate. Ten milliliters of filtrate was subjected for
extraction and analysis. The true concentration was estimated
by applying recovery factor. Concentrations obtained in tea
Environ Sci Pollut Res

4. infusion after analysis was considered as translocated amount
and from this, the percent translocation was calculated.
Analysis
One microliter of clean extract was injected in a gas chroma-
tography (GC) vial for analysis and the detected pesticides
were further confirmed by gas chromatography tandem mass
spectrometry (GC-MS/MS)20
. Clean extracts of the samples
were evaporated in a nitrogen flux evaporator and again made
up with acetonitrile for the phthalate residue analysis using
high-performance liquid chromatography (HPLC) and then
confirmed by liquid chromatography tandem mass spectrom-
etry (LC-MS/MS)21
.
GC-ECD22
The final extracts were analyzed by GC (Shimadzu GC-2010,
Japan) equipped with DB-1 capillary column
(30 m × 0.25 mm × 0.25 μm) and electron-capture detector
(ECD). General operating conditions were as follows: injector
port temperature 280 °C; column temperature program: ini-
tially 165 °C for 1.5 min, increase at 2.70 °C/min to 210 °C
and hold for 2.7 min, increase at 2.70 °C/min to 265 °C and
hold for 2.2 min, and then increase at 8.00 °C/min to 280 °C
and hold for 1.5 min; detector temperature 300 °C; injection
volume 1 μl; split ratio 1:5; column flow rate 0.79 ml/min; and
makeup flow 30.00 ml/min; N2 (99.5 %) was used as carrier
gas.
HPLC-PDA23
An HPLC (Thermo Scientific Dionex Ultimate 3000, USA)
system was equipped with a photo diode array (PDA) detec-
tor, a temperature control module, and an online degasser
system. Analysis were performed on thermo Scientific,
Acclaim-C18 column (5 μm size: 250 mm × 4.5 mm) at
25 °C, using mobile phase of acetonitrile and water. A gradi-
ent program of the mobile phase was as follows: 80:20 v/v
(acetonitrile and water) with flow rate of 1 ml/min, hold for
1.0 min, 90:10 v/v (acetonitrile and water) with flow rate of
1 ml/min, hold for 3.0 min. Only acetonitrile with flow rate of
2 ml/min hold for 9 min, 80:20 v/v (acetonitrile and water)
with the flow rate 1 ml/min hold for 2.1 min. Phthalates were
detected at 230 nm, with injection volume 20 μl.
Gas chromatography tandem mass spectrometry
A Thermo Scientific GC-MS/MS consisting of TSQ Quantum
XLS Mass Spectrometer (Thermo Fisher, USA) equipped with
a TG-5MS capillary column (30 m × 0.25 mm × 0.25 μm) was
used for analysis. Helium was used as carrier gas (purity
99.99 %) with a flow rate of 1.1 ml/min. One-microliter aliquot
of the extract was injected using the splitless mode. The oven
temperature program was initially 70 °C held for 1 min, at the
rate of 15 °C/min, and then increased to 210 °C, then to 230 °C
at the rate of 2 °C/min, and finally maintained to 280 °C at the
rate of 15 °C/min and held for 5.0 min. The selected reaction
monitoring (SRM)26
transitions for malathion were 127/99 and
173/99, for cypermethrin 163/127 and 181/152, and for
fenvalerate 167/125 and 419/225. The injector temperature
was set at 300 °C. The transfer line and source temperature
was set at 280 and 230 °C, respectively. Solvent delay for mass
spectrometry was 5.0 min.
Liquid chromatography tandem mass spectrometry
The LC-MS/MS system was an ultra performance liquid chro-
matography (UPLC)25
(Waters Acquity, USA) equipped with
a triple–quadrupole mass spectrometer (AB SCIEX/API
4000, USA) with electrospray ionization (ESI)26
ion source.
Acetonitrile was chosen as the mobile phase, and the total
flow rate was 0.3 ml/min. The ESI interface in the positive
multiple reaction monitoring (MRM)27
mode was chosen for
the identification and quantification of the compounds. The
set of parameters used is shown as follows: ion source (IS)28
spray voltage 5500 eV, source temperature (TEM) 0.0 °C;
atomization gas pressure (GS1)-10 Psi (nitrogen); heated aux-
iliary gas (GS2)-0 Psi (nitrogen); curtain gas pressure (CUR)-
10 Psi (nitrogen); collision flow (CAD)-4 Psi. MRM transi-
tions of DEP 223/177, DBP 279.2/205.2, DEHP 391/149, and
DOP 391.1/261.2 were noticed for analysis.
Results and discussion
Quality control
Separation profiles of OCs, OPs, SPs, and herbicides on GC
and phthalates on HPLC are given in Figs. 1 and 2, respec-
tively. Quality control of the analysis was done by recovery,
precision, limit of detection (LOD)2
, and limit of quantifica-
tion (LOQ)30
. Calculation of pesticide and phthalate was done
by comparing with the matrix match standard in raw tea and
tea infusion. In each batch solvent, blank and matrix match
standard were run. Table 1 shows the LOD, LOQ, and per-
centage of relative standard deviation (%RSD)31
of data asso-
ciated with recovery estimation. The recovery was determined
at spiking level of 0.1 mg kg−1
(OCs, SPs, and herbicides) and
0.5 mg kg−1
(OPs and phthalates). Recovery of pesticides was
ranged for OCs (72.4–99.1), OPs (66.1–98.1), SPs (73.3–
96.9), herbicides, (79.0–94.7), and phthalates (70.1–101.3).
The LOD was in the range of 0.004–0.018 mg kg−1
and
LOQ in the range 0.010–058 mg kg−1
. The precision of the
method was determined by RSD <5 % during recovery esti-
mation (European Commission 2007; SANCO 2013).
Environ Sci Pollut Res

5. QuEChERS sample preparation for the extraction of pesticide
and phthalate residues showed good recovery and precision
and lower LOD and LOQ and better compatibility with the
gas and liquid chromatography systems (EURACHEM,
Guide 1998; Xu et al. 2014; Srivastava et al. 2014; Yina et al.
2014; Cho et. al. 2014). Tea is a complex matrix having large
contents of catechins, caffeine, flavonoids, fluoride, xanthine,
theaflavin, theophylline, and essential amino acids, etc., which
Fig. 1 The chromatogram of 36 pesticides by GC: a matrix match standard; b detected sample
Fig. 2 The chromatogram of six phthalates by HPLC: a matrix match standard; b deteceted sample
Environ Sci Pollut Res

6. Table 1 Recovery (%), %RSD, LOD, and LOQ of pesticides and phthalates in tea infusion and raw tea
Pesticides Spiked conc.
(mg kg−1
)
Tea infusion Raw tea
Recovery
(%)
RSD(%) LOD
(mg kg−1
)
LOQ
(mg kg−1
)
Recovery
(%)
RSD
(%)
LOD
(mg kg−1
)
LOQ
(mg kg−1
)
α-HCH 0.1 90.93 2.327 0.007 0.021 82.10 3.150 0.008 0.026
β-HCH 0.1 95.23 1.683 0.005 0.016 87.43 2.348 0.006 0.021
γ-HCH 0.1 92.94 3.073 0.010 0.028 83.13 3.197 0.008 0.027
δ-HCH 0.1 91.70 3.221 0.010 0.029 85.43 1.757 0.005 0.015
α-Endosulfan 0.1 97.89 1.473 0.005 0.014 88.20 3.356 0.009 0.030
β-Endosulfan 0.1 91.25 2.897 0.009 0.026 84.57 3.548 0.009 0.030
p,p’-DDE 0.1 90.33 3.404 0.010 0.030 79.07 2.841 0.007 0.023
p,p’-DDD 0.1 94.49 3.216 0.010 0.030 83.10 2.324 0.006 0.019
p,p’-DDT 0.1 91.06 1.208 0.004 0.011 86.17 2.853 0.008 0.025
o,p’-DDE 0.1 90.42 3.541 0.009 0.028 79.40 2.847 0.007 0.023
o,p’-DDT 0.1 88.47 1.248 0.010 0.030 78.23 3.654 0.009 0.029
Heptachlor 0.1 99.10 1.203 0.004 0.011 72.40 3.683 0.008 0.027
Malathion 0.5 86.84 2.686 0.009 0.025 79.02 0.758 0.010 0.031
Chlorpyrifos 0.5 95.30 2.970 0.008 0.024 79.54 0.736 0.009 0.029
4-bromo-2-
chlorophenol
0.5 82.89 1.812 0.006 0.017 81.80 0.340 0.004 0.013
Dichlorvos 0.5 98.07 1.307 0.004 0.012 78.73 0.917 0.012 0.038
Triazophos 0.5 67.46 1.102 0.004 0.010 79.45 0.866 0.011 0.034
Profenofos 0.5 96.20 3.356 0.009 0.028 79.45 0.926 0.012 0.037
Ethion 0.5 84.91 1.932 0.006 0.018 75.14 0.558 0.007 0.022
Dimethoate 0.5 85.31 2.123 0.005 0.015 77.46 0.818 0.010 0.032
Phosphamidon 0.5 66.13 3.693 0.004 0.011 76.12 0.865 0.011 0.034
Fenvalerate-I 0.1 94.40 3.137 0.010 0.029 76.10 3.949 0.009 0.030
Fenvalerate-II 0.1 93.83 2.687 0.008 0.023 75.17 3.424 0.008 0.026
Cypermethrin-I 0.1 88.95 3.463 0.010 0.030 73.30 3.739 0.009 0.027
Cypermethrin-II 0.1 87.72 1.813 0.006 0.017 78.67 3.698 0.009 0.029
τ-Fluvalinate 0.1 96.86 3.255 0.010 0.030 76.13 3.794 0.009 0.029
Fenpropathrin 0.1 93.85 2.930 0.009 0.026 80.43 3.200 0.008 0.026
δ-Methrin 0.1 90.97 2.152 0.007 0.020 78.50 2.880 0.007 0.023
β-Cyfluthrin-I 0.1 94.35 3.016 0.010 0.030 76.50 3.736 0.009 0.029
β-Cyfluthrin-II 0.1 91.49 2.457 0.009 0.027 75.00 3.762 0.009 0.028
Bifenthrin 0.1 89.33 3.442 0.010 0.030 74.93 3.873 0.009 0.029
λ-Cyhalothrin 0.1 84.33 2.718 0.008 0.022 79.80 1.807 0.005 0.014
Pendimethylene 0.1 93.31 3.182 0.010 0.029 78.97 3.620 0.009 0.029
Butachlor 0.1 87.50 2.368 0.007 0.020 80.37 3.531 0.009 0.028
Alachlor 0.1 88.57 2.431 0.007 0.021 82.60 4.178 0.011 0.035
Atrazine 0.1 94.72 2.225 0.007 0.021 80.17 3.254 0.008 0.026
Phthalates
DMP 0.5 90.10 0.71 0.010 0.031 70.13 1.106 0.012 0.039
DEP 0.5 84.35 0.65 0.009 0.027 73.10 1.543 0.018 0.056
DBP 0.5 85.61 1.21 0.016 0.051 94.14 0.807 0.012 0.038
DEHP 0.5 86.44 0.64 0.009 0.027 101.34 0.713 0.012 0.040
BzBP 0.5 89.24 0.77 0.011 0.034 75.88 1.312 0.016 0.050
DOP 0.5 93.14 1.25 0.018 0.058 93.04 1.168 0.017 0.054
RSD relative standard deviation, LOD limit of detection, LOQ limit of quantification
Environ Sci Pollut Res

7. cause matrix interferences and affect the recovery of analytes
(Chen et al. 2014; Liu et al. 2014). In this method, an efficient,
clean practice was done by using charcoal, PSA, and magnesium
sulfate (Srivastava et al. 2011; Chen et al. 2011). Charcoal use as
sorbent and may reduce the recovery of pesticides to minimize
this risk; different amounts of charcoal were optimized and it
was found that 5 mg of charcoal results to a minimum effect on
recovery with efficient pigment removal (Sharma et al. 2008).
Pesticide residues in raw tea and tea infusion samples
Among the total analyzed pesticides, only three, malathi-
on, cypermethrin, and fenvalerate, were detected in tea
samples (Table 2). Loose tea and branded tea were detect-
ed to have these pesticides; mean levels of fenvalerate
residues were the same in loose tea (0.032 mg kg−1
(BDL32
−0.484)) and branded tea (0.030 mg kg−1
(BDL
−0.601)). Compared to that in branded tea (0.118 mg kg−1
(BDL −1.236)), the residual level of malathion was higher
in loose tea (0.257 mg kg−1
(BDL −3.450)). On the other
hand, the residual level of cypermethrin (0.065 mg kg−1
(BDL −1.315)) was higher in loose tea compared to that
in branded tea (0.030 mg kg−1
(BDL −0.612)). In herbal
tea, cypermethrin (0.053 mg kg−1
(BDL −1.063)) and
fenvalerate (0.045 mg kg−1
(BDL −0.551)) were detected.
None of the detected pesticide residues were above max-
imum residue limit or MRL33
(PFA 1954/Codex 2005).
However, no pesticide was detected in the tea infusion
samples.
Studies on pesticide residue determination in tea showed
the presence of cypermethrin in different varieties of tea
(green tea, dark tea, black tea, oolong tea, scented tea) with
the concentration ranging from 20.13 to 187.65 μg kg−1
(Hua et al. 2010). In this study, cypermethrin was also de-
tected in varieties of tea (loose, branded, herbal) with resid-
ual level of 0.612–1.315 mg kg−1
. The occurrence of
cypermethrin residues in tea evidenced their indiscriminate
use and the minimum effect of tea manufacturing processes
(leaf harvesting, withering, rolling, fermentation, microwave
heating, and drying) on the cypermethrin sprayed on the tea
bushes (Yang et al. 2009; Sood et al. 2004). Fenvalerate was
also detected in a variety of tea (loose, branded and herbal)
with residual level ranging from 0.030 to 0.045 mg kg−1
as
compared to that detected only in dark tea with a concentra-
tion of 13.18 μg kg−1
(Hua et al. 2010).
Phthalate residues in raw tea and tea infusion samples
Analysis showed that only tea infusion samples contained
in plastic bags and disposable cups were contaminated
with phthalate residues (Table 2). Only DBP with a resi-
due level 0.038 mg kg−1
(BDL −0.183) was reported in
the tea infusion sample in plastic bag. In the tea infusion
sample in disposable cup, phthalate residues of DBP
0.026 mg kg−1
(BDL −0.159) and DEP 0.004 mg kg−1
(BDL −0.045) were detected. Commission directive
2007/19/EC is related to plastic materials and migration
of their constituents set the limit of phthalates on the basis
of toxicological evaluation and exposure. None of the
phthalate residues were above the specific migration limit
(SML)34
set by the European Union.
Table 2 Level of pesticide and phthalate residues in raw tea and tea infusion samples
Sample Detected pesticides/phthalates Number of sample No of samples
> MRLa
(mg kg−1
)
Level of residues (mg kg−1
)
Analyzed Detected
Loose tea Malathion 20 2 – 0.257 (BDL −3.450)
Cypermethrin 20 1 – 0.065 (BDL −1.315)
Fenvalerate 20 2 – 0.032 (BDL −0.484)
Branded tea Malathion 20 2 – 0.118 (BDL −1.236)
Cypermethrin 20 1 – 0.030 (BDL −0.612)
Fenvalerate 20 1 – 0.030 (BDL −0.601)
Herbal tea Cypermethrin 20 1 – 0.053 (BDL −1.063)
Fenvalerate 20 2 – 0.045 (BDL −0.551)
Tea Infusion in glass cup – 10 – – –
Tea Infusion in earthen cup – 10 – – –
Tea Infusion in plastic bag DBP 10 3 – 0.038 (BDL −0.183)
Tea Infusion in disposable cup DBP 10 2 – 0.026 (BDL −0.159)
DEP 10 1 – 0.004 (BDL −0.045)
MRL maximum residue limit
Environ Sci Pollut Res

8. Translocation of pesticides from raw tea to tea infusion
and their loss during processing
All the analyzed pesticides showed less than 5 % of
translocation with all boiling durations (2, 5, and
10 min) except malathion, 4-bromo-2-chlorophenol, di-
chlorvos, and dimethoate (Fig. 3). The percent translo-
cation of malathion was 8.1, 10.9, and 9.0 for boiling
durations of 2, 5, and 10 min, respectively. The percent
translocation of 4-bromo-2-chlorophenol was 7.3, 10.0,
and 8.2 for boiling durations of 2, 5, and 10 min, re-
spectively. The percent translocation of dichlorvos and
dimethoate were 28.8, 38.6, and 36.7 and 37.4, 49.2,
and 34.9, respectively, for boiling durations of 2, 5,
and 10 min. In making of tea infusion, residues of pes-
ticides face boiling temperature which may result in mi-
gration (from raw tea to infusion) or loss due to evapo-
ration, hydrolysis, and thermal breakdown. Evaluation
of behavior of 36 pesticides of different classes showed
that organophosphate pesticides have maximum translo-
cation (Gupta et al. 2008). Higher translocation of di-
methoate and dichlorvos was due to their higher solu-
bility in water (39 g L−1
dimethoate and 10 g L−1
di-
chlorvos by WHO).
Leaching of phthalates in tea infusion from plastic bag
and disposable cup
Residues of phthalate were detected only in tea infusion sam-
ple in the plastic bag and disposal cup. None of phthalate
residues were detected in raw tea and tea infusion taken in
the glass and earthen cups. In this experiment, leaching of
phthalate residues was determined with different retention
times in disposal cups and plastic bags. Phthalate concentra-
tions observed in tea infusions in plastic bag and disposable
cup at different time intervals are summarized in Table 3. As
shown, the leaching of phthalate residues in tea infusions in
plastic bags and disposal cups increased as retention time
prolonged. BzBP and DEHP showed minor loss as retention
time was extended. Leaching of DMP and DEHP was started
after 10 min of retention time. Leaching of DBP has maxi-
mum compares to other types of phthalate in each type of
containers (plastic bag and disposal cup).
Various challenges were associated with the analysis of
phthalates as there is a possibility of contamination with the
reagent and solvent used in the extraction and cleanup,
which may contain phthalates. To minimize this risk, pro-
cedural blanks were used with every batch of samples in
exactly the same way that the samples were prepared,
0
1
2
3
4
5
2 min 5min 10 min
OC Pesticides
noitacolsnarT%
0
1
2
3
4
5
2 min 5min 10 min
SP Pesticides
noitacolsnarT%
0
1
2
3
4
5
2 min 5min 10 min
Herbicides
%Translocation
0
10
20
30
40
50
2 min 5min 10 min
OP Pesticides
%Translocation
Fig. 3 Percent translocation of OCs, OPs, SPs, and herbicides from raw tea to tea infusion at different time intervals
Environ Sci Pollut Res

9. except that the blanks do not contain the sample matrix but
do use equal amounts/volumes of all solvents, reagents,
chemicals, and glassware that come into contact with the
samples (Guo and Kannan. 2012). Various studies discuss
that food material packed in plastic bags and environmental
water samples show detection of DMP, DEP, and DEHP
(Fierens et al. 2012; Roldan et al. 2004). In this study, tea
infusions kept in plastic bags and disposable cups also
show the detection of DMP, DEP, and DEHP.
Conclusion
An analytical result of the QuEChERS sample preparation
showed that the method was suitable for the extraction of
pesticide and phthalate residues. The evaluation of 36 pesti-
cides, covering the 4 major groups (OCs, OPs, SPs and her-
bicides), and 6 phthalate residues in raw tea (loose pack and
branded and herbal tea) and tea infusion samples (glass,
kulhad, plastic bag, and disposable cup) showed that only
15 % of the raw tea samples were found with residues of
malathion, cypermethrin, and fenvalerate, which were below
MRL values (PFA 1954/Codex 2005). Only 30 % of the tea
infusion samples contained in plastic bag and 20 % of the
samples contained in disposable cups were contaminated with
DBP and DEP. During processing, residues of pesticide face
the boiling temperature which may result in migration (from
raw tea to brew) or loss. Carrying and drinking tea infusion in
plastic bags and disposal cups may increase health risk due to
phthalate leaching.
Acknowledgments The authors are grateful to Director, CSIR-IITR,
Lucknow, for his keen interest and providing the research facilities. I
am also thankful to Dr. M.K.R. Mudiam for his support. The financial
assistance of all India network project on monitoring of pesticides resi-
dues at national level (GAP-168) is also acknowledged. Manuscript based
CSIR-IITR communication no.3183.
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