Journal of Food Engineering 114 (2013) 404411

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Journal of Food Engineering

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Pesticide residue removal from vegetables by ozonation

J.Y. Chen a, Y.J. Lin b, W.C. Kuo b,
a
Great Eastern Resin Industrial Co., Ltd., Kaohsiung, Taiwan
b
Department of Environmental Science and Engineering, National Pingtung University of Science and Technology, Pingtung, Taiwan

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

Article history: A novel machine was developed to remove pesticide residues from vegetables using ozone. This domestic-
Received 12 March 2012 scale vegetable cleaner consists of a closed cleaning chamber, an ozone generator, a water recirculation pump,
Received in revised form 25 August 2012 and an oxidationreduction potential (ORP) electrode. Two vegetables, Chinese white cabbage and green-
Accepted 29 August 2012
stem bok choy, and three pesticides, permethrin (trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-
Available online 6 September 2012
carboxylate), chloruazuron (1-[3,5-dichloro-4-(3-chloro-5-triuoromethyl-2-pyridyloxy)phenyl]-3-(2,
6-diuorobenzoyl) urea), and chlorothalonil (tetrachloroisophthalonitrile) were used in tests. Cleaning for
Keywords:
15 min with pump recirculation removed 51% of chloruazuron and 53% of chlorothalonil. When the ozone
Pesticide residue
Ozone treatment
production rate was 250 mg/h, removal efciencies were 60% for chloruazuron and 55% for chlorothalonil,
Permethrin increases of 29% over pump recirculation only. When the ozone production rate was 500 mg/h, removal ef-
Chloruazuron ciencies were 75% for chloruazuron and 77% for chlorothalonil; increases of 24% over pump recirculation
Chlorothalonil only. After the ozone treatment, all the pesticide residuals met the Standards for Pesticide Residue Limits
in Foods.
2012 Elsevier Ltd. All rights reserved.

1. Introduction and UV photolysis (Wu et al., 2009). However, these techniques

The worlds population is estimated to be 9.22 billion in 2075
(UN, 2004). Along with this rapid population increase, worldwide
demand for food will increase, as will pesticide use. China applies
an estimated 1.3 Mt of pesticides to agricultural lands annually
(Wu et al., 2007). Taiwans subtropical climate renders many crops
susceptible to damage by diseases and insects, resulting in the use
of large amounts of pesticides to prevent smiting. According to sta-
tistics from Taiwans Bureau of Animal and Plant Health Inspection
and Quarantine, Council of Agriculture, 34,709 tonnes of formu-
lated agro-pesticide were sold in 2010. Notably, Taiwanese farmers
are not the only farmers who spray extremely high pesticide con-
centrations (Zhou and Jin, 2009). Additionally, some harvests are
rushed to market, leaving large amounts of pesticide residues on
plants, vegetables and fruit. Although humans are exposed to small
amounts of pesticide residues after pesticides are metabolized by
plants or decomposed by environmental agents, trace amounts of
pesticide residing in the human body for long periods can cause
chronic diseases and can lead to cancer (Carrozza et al., 2009).
Residual pesticides in water can be destroyed by, say, Fenton
oxidation, electrochemical oxidation, TiO2 catalytic treatment,
Corresponding author. Address: Department of Environmental Science and
Engineering, National Pingtung University of Science and Technology, Pingtung
91207, Taiwan. Tel./fax: +886 8 774 0261.
E-mail address: xwck@mail.npust.edu.tw (W.C. Kuo).
have some disadvantages; for example, catalytic treatments
produce secondary pollutants. Ozone was rst used to disinfect
water supplies in France in the early 1900s. Well over 1000 ozone
disinfection installations now exist worldwide. Ozone, a powerful
oxidant (EH > 2.0 volts), has an oxidation capability exceeded by
uorine only (EH = 3.06 V). Ozones solubility in water at 20 C is
12.07 mg/L. The stability of ozone in air exceeds that in water; how-
ever, both stabilities are within the order of minutes (Metcalf and
Eddy, 2003; Eckenfelder, 2000). Ozone is also used to remove pes-
ticide residues from vegetables or aqueous solutions (Benitez et
al., 2002; Wu et al., 2007; Chelme-Ayala et al., 2009; Ikeura et al.,
2011). Ozone decomposes in water to produce free radicals, and
high pH values are favorable for pesticide degradation (Ong et al.,
1996; Eckenfelder, 2000; Xiong et al., 2011). Based on ozones
strong oxidation capability and its reaction with other inducer
(e.g. OH and hydroxyl radicals (OH)), with high oxidation power
will be produced; these inducers can decompose most organic com-
pounds. As ozone decomposes into oxygen, it does not produce sec-
ondary pollutants. The prevailing ozone treatment method for
removing pesticide residues on vegetables is to pump ozone into
water in a vessel via an ozone generator. Ozone then oxidizes con-
taminants via ozone molecules or its derivatives and removes resi-
dues. The effectiveness of ozone treatment is limited by ozones
poor solubility and the high cost of ozone generators. Increasing
ozone solubility and reducing the cost of ozone production are
key problems associated with ozone treatment. The goals of this
study are as follows.

0260-8774/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jfoodeng.2012.08.033
 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411
(1) Develop a novel domestic-scale ozone cleaner for
vegetables.
(2) Establish the relative concentration curves for liquid ozone
concentrations and ORP.
(3) Conduct experiments to determine the effectiveness of the
proposed ozone vegetable cleaner in removing pesticide res-
idues from vegetables.
(4) Evaluate the toxicity of aqueous pesticide solution before
and after ozone treatment using the Tox Tracer biological
toxicity test.
2. Materials and methods
2.1. Field tests for pesticide residues on vegetables
This experiment had three stages. The rst eld experiment was
conducted during JuneAugust 2004. The planted vegetables were
Chinese white cabbage and green-stem bok choy (foliole vegetables
with short growing periods). Pesticides applied were 10% permeth-
rin emulsion and 5% chloruazuron emulsion. Pesticide doses were
classied into the following three groups: none (no spraying);
standard doses (spraying following plant protection criteria of
50 ppm permethrin and 25 ppm chloruazuron); and, twice the
standard dose (over-spraying). The second eld experiment was
conducted during February-April 2005. The planted vegetable was
Chinese white cabbage, and pesticides applied were 10% permethrin
emulsion and 5% chloruazuron emulsion. Pesticide doses were
classied into the following two groups: standard doses of 50 ppm
permethrin and 25 ppm chloruazuron; and 1.5 times the standard
doses (over-spraying). The third eld experiment was conducted in
November and December 2005. The planted vegetable was Chinese
white cabbage, and pesticides were permethrin, chloruazuron, and
75% chlorothalonil powder (i.e., 50 ppm permethrin, 25 ppm chlor-
uazuron and 100 ppm chlorothalonil). Pesticide doses were 1.5
times the standard dose, simulating general farmer practices.
2.2. Experimental setup
The proposed domestic-scale ozone vegetable cleaner consists of
an airtight cleaning tank, a vegetable basket, an ozone generator, an
ozone concentration controller, a recirculation pump, and a timer.
The cleaner generates and controls the liquid ozone concentration
during the cleaning process for removing pesticide residues. Fig. 1
shows a schematic diagram of the proposed pesticide cleaner.
2.3. Analytical instruments
Analytical instruments used in this study were as follows.
(1) The ozone generator, a Fischer ozone generator (Model 500),
was used in liquid ozone determinations.
(2) The ozone gas analyzer was a Seki electronic ozone UV pho-
tometric analyzer (SOZ-6000, Japan).
(3) The ORP electrode was a pH/ORP controller (Model PC-310,
Suntex, Taiwan).
(4) A UVvisible spectrophotometer (Model UV-160A, Shimadzu,
Japan) was used to determine liquid ozone concentrations.
(5) The following devices were used in pesticide residue
analyses.
(i) Blender: Osterizer (10 speeds).
(ii) Rotary evaporator: Eyela Model N-1000 (Eyela, Japan).
(6) The GC Electron Capture Detector (ECD) used for sample
analysis of pesticide residues and aqueous pesticide solu-
tions was an HP-6890 equipped with an ECD. The carrier
gas was N2 and the analytical column was an HP-5 packed
with Crosslinked 5% PH ME Siloxane.
405
(7) The Tox Tracer Toxicity test used the following devices.
(i) Tox Tracer Luminometer photodetectors (Skalar, Nether-
lands) used a photomultiplier tube, isothermal reaction tank
(15 0.1 C), and a digital screen that displayed% inhibition.
(ii) The Tox Tracer cooling block constant temperature incu-
bator held 30 tubes and 2 microbe index tubes for
microbes cultivation, with automatic temperature con-
trol (15 0.1 C).
2.4. Analytical reagents
(1) Indigo reagent
The Indigo Colorimetric Method was used to determine
liquid ozone concentrations (Hoigne and Bader, 1982).
(2) Pesticides
Three pesticides, permethrin, chloruazuron, and chlorot-
halonil, were chosen based on farmer use and seasonal
variations.
(i) Standard pesticide solutions, 1000 ppm permethrin,
1000 ppm chloruazuron, and 867 ppm chlorothalonil,
were prepared for calibration curves. These standards
were obtained from the Agricultural Chemicals and Toxic
Substances Research Institute, Council of Agriculture,
Taiwan. Suppliers and purities of permethrin, chloru-
azuron, and chlorothalonil were Dr. Ehrenstorfer GmbH,
94%; R.D.H., 99.4%; and R.D.H., 98.5%, respectively.
(ii) Commercial pesticide products: 10% permethrin emul-
sion, 5% chloruazuron emulsion, and 75% chlorothalonil
powder were used as aqueous pesticide solutions for
eld tests and pesticide removal experiments.
(3) Reagents for pesticide residue determination: acetone, NaCl
(30%), dichloromethane, petroleum ether, n-hexane, car-
bonic acid sodium hydride (12%), and anhydrous sodium
sulfate (all were analytical grade).
(4) Reagent for the Tox Tracer Toxicity Test.
(i) Fluorescence biological reagent: a patented Vibrio sheri
was adopted (luminous bacteria were used in this study).
These bacteria were made into biological agent after being
dried and frozen. Each bottle contained about 1 billion
bacteria stored at 20 C. Notably, analysis was completed
within 4 h after a bottle was opened.
(ii) Bacteria activator: a kind of distilled water which can be
added to the biological agent and activate the bacteria.
(iii) Diluent: 2% NaCl solution.
(iv) Osmotic pressure modulation liquid: 22% NaCl solution for
adjusting the NaCl concentration of sample to 2% to balance
the osmotic pressure among biological agents and samples.
2.5. Analytical methods
(1) Preparation of aqueous pesticide solutions.
Human tolerances of pesticide residues are 2 mg/L for per-
methrin, 2 mg/L for chloruazuron, and 1 mg/L for chlorot-
halonil. Initial concentrations were twice the strength of
pesticide residue concentrations, i.e., 4 ppm permethrin,
4 ppm chloruazuron, and 2 ppm chlorothalonil.
(2) Analysis of aqueous pesticide solutions.
The initial pesticide concentration was based on Taiwans
Safety Tolerance Criteria of Pesticide Residues on Farm Prod-
ucts, Department of Health, Republic of China (2012). To
simulate farmer practices in controlling pests within short
periods, doses 1.52.0 times the standard dose were
adopted. Therefore, twice the safety tolerance was used as
the initial pesticide concentration. The safe tolerance for
permethrin, chloruazuron, and chlorothalonil is 2.0 mg/L,
2.0 mg/L, and 1.0 mg/L, respectively. Analytical methods
406 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411

2
3

5 4

7 6

1. Cleaning chamber ; 2.Chamber cover; 3.Vebetable basket; 4.Recirculation pump; 5.

Recirculation pipe; 6.Ozone generator; 7.Ozone discharge tube.; 8.Vent.

Fig. 1. Experimental setup of the the pesticide residue cleaning machine.

followed those recommended by Taiwans Council of Agri-
culture (2011). These tests were applied to investigate the
effects of ozone oxidation and decomposition on a pesticide.
Experimental procedures were as follows. (1) In total, 4 ppm
permethrin, 4 ppm chloruazuron, and 2 ppm chlorothalonil
were combined into an 8L aqueous solution, and poured into
the ozone reaction tank. (2) The initial pesticide solution
concentration was then determined. (3) The ozone generator
output was adjusted to the pre-set value and ozone was then
pumped into the reaction tank. (4) Timer settings were
15 min, 30 min, and 45 min and GC analyses were conducted
at these time intervals.
Determination of pesticide residue from aqueous solutions
followed the method promulgated by Council of Agriculture,
Republic of China (2011).
Preparation of working solutions of permethrin, chloruazu-
ron and chlorothalonil followed procedures described
below. A volume of 10 mL sample was added to acetone
and the nal volume was increased to 100 mL. The sample
was vibrated in an ultrasonic oscillator for 10 min and
10 mL was obtained, mixed with 10 g anhydrous sodium
sulfate, and then ltered through a 0.2 lm membrane lter;
1 mL was used for nal analysis.
(3) Analysis of pesticide residues on vegetables.
Determination of pesticide residue on vegetables followed
CNS Method of Test for Pesticide Residues in Food-Multi-res-
idue Analysis (I), Ministry of Economic Affairs, Republic of
China (2012):
Sample extraction and purication procedures were modi-
ed from those in previous studies and are described
briey. Roughly 50 g of a vegetable sample was mixed with
80 mL acetone for 1 min in a high-speed blender. In total,
80 mL acetone was added and then mixed for 1 min at high
speed. The resulting mixture was then poured into a
Buchner funnel with a lter paper for ltration. The residue
was rinsed with acetone and acetone was added to a nal
volume of 160 mL. Then 40 mL of the sample mixture
was added with 50 mL petroleum ether in a separating
funnel. This study then used 50 mL dichloromethane to
extract the mixture twice; each extraction took 1 min.
The dichloromethane layer was collected and 20 g anhy-
drous sodium sulfate was added to the dehydrate and l-
tered. A rotary evaporator was used to dry the sample at
40 C. The sample was further dissolved in 5 mL acetone.
In total, 1 mL sample from the previous step was obtained
and injected into a Florisil cartridge pre-rinsed with 10 mL
n-hexane followed by rinsing with 5 mL n-hexane/dichloro-
methane = 1:2 (v/v) three times; all ltrates were then
collected. Nitrogen gas was utilized to purge the sample
until almost dry; 1 mL acetone was added for GC-ECD
analysis.
(4) Tox Tracer Toxicity Test (Lin et al., 2006).
The Tox Tracer toxicity test was applied to determine
biological toxicity before and after ozone treatment of
aqueous pesticide solutions. Test procedures were as
follows.
 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411
(i) Sample preparation.
a. In total, 8 L of 4 ppm permethrin and 4 ppm chloruazuron
were prepared. Pesticides solutions were then poured into
the ozone reaction tank.
b. A pre-treatment sample was obtained before the ozone gen-
erator started.
c. The ozone generator and water recirculation pump were
then started and ozone output rate was set at 250 mg/h or
500 mg/h. The ozone generator was preheated for 1 min to
stabilize the output concentration before connecting the
gas and reaction tank.
d. Ozone treatments lasted 15 min, 30 min, and 45 min; a
sample was taken at the end of each duration.
e. The pH of samples was 6.08.0.
f. In total, 100 lL osmotic pressure modulation liquid (22%
NaCl) was added to 1 mL sample and mixed.
g. At minimum 50 mL diluent was used to dilute samples.
(ii) Reagent preparation (bacterial activation).
a. A bottle of reconstituted solution was stored in a refrigera-
tor at 4 C.
b. In total, 12 mL diluent was poured into a special tube, which
was then placed in a constant-temperature incubator
(15 0.1 C) until its temperature stabilized.
c. A bottle of Microtox Reagent was removed from the refrig-
erator and mixed with 1 mL reconstituted solution at 4 C.
d. The mixture was poured into the tube (step (b)) and incu-
bated for 20 min.
(iii) Sample analysis followed the operational step of the Tox
Tracer toxicity analyzer.
3. Results and discussion
3.1. The proposed ozone vegetable cleaner and ozone-orp calibration
curve
As shown in Fig. 1, the ozone concentration controller deter-
mines the ozone concentration in water during the cleaning process,
and then transmits a signal to the controller, whose output terminal
is connected to the ozone generator that is functioning or stopped
according to concentration variance to maintain ozone solubility
in the optimal state in the water tank, and reduce energy usage.
Ozone in water generally reacts with OH and H2O to generate
OH, which has a high oxidation power. ORP is a common indicator
to show the oxidation reduction status of a solution. Thus, we ar-
gue that ozone decomposition in liquid changes the ORP of water.
Therefore, the ORP is utilized to detect potential change by ozone
decomposition in water. A sample of an aqueous solution is ob-
tained and its ozone concentration is determined using the Indigo
Method. The ORP (mV) is plotted on the x-axis and liquid ozone
concentration (mg/L) is plotted on the y-axis, such that the rela-
tional curve after regression can be obtained as y = ax2 + bx + c,
where a, b, and c are constants. By replacing the liquid ozone con-
centration, ORP can represent variations in ozone concentrations in
water. Measurement results were used when developing the pro-
posed ozone cleaning device. A Fischer Ozone Generator 500 was
used to produce ozone using pure oxygen as the feed gas rate of
850 L/min; the produced ozone concentration was 24.3 g/m3.
Experiment duration was 20 min and the liquid ozone concentra-
tion and gas-phase ozone concentration were determined every
3 min. With measurement time on the x-axis and liquid ozone con-
centration (mg/L) on the y-axis, the calibration curve between time
and liquid ozone concentration was obtained. Again, with mea-
surement time on the x-axis and ORP (mV) on the y-axis, the rela-
tionship between time and ORP was established. Through
regression analysis, a relational curve between liquid ozone
concentration and ORP was calibrated (Fig. 2).
407
y = -74.668x2 + 402.06x + 508.38
1200
1000
800
ORP (mV)
600
400
200
0
0.00 1.00 2.00 3.00 4.00
Liquid ozone concentration (mg/L)
Fig. 2. Relationship between liquid ozone concentration and ORP.
The liquid concentration of ozone dissolved in water is related
to pH, gas ozone concentration (mg/m3), airow, the diffuser, and
aqueous solution volume; the gas ozone concentration is the most
critical factor. Commercially available ozone generators determine
ozone output (mg/h) by multiplying gas ozone concentration (mg/
m3) by gas ow rate (m3/min). Under a constant ozone production
rate within a given period, as gas ozone concentration increases,
airow decreases, and vice versa. Therefore, the key tasks in ozone
applications are to control the gas ozone concentration, ow rate,
and diffuser (as the size of bubbles decreases, the surface area of
ozone bubbles contacting water increases, and ozone solubility in
water increases). When the ozone concentration in unit time can
be increased and distributed evenly in water within a short period,
treatment effectiveness is enhanced.
3.2. Ozone treatment of aqueous pesticide solutions
In pesticide analyses, the R2 value of calibration curves were
higher than 0.99. The standard curves were recalibrated every other
week. The PCB congener 209 (1 mg/L) was spiked as the internal
standard with the detection limit of 0.1 lg/L. The recovery rates
of 100 10% were typically observed, including the errors of extrac-
tion and GC analysis. Table 1 shows experimental results. At an
ozone production rate of 250 mg/h, pesticide removal efciencies
improved over time. Taking 15 min as the time a household would
likely use the proposed ozone vegetable cleaner, average removal
efciencies for the three pesticides were poor at roughly 10%.
Furthermore, average time-weighted removal efciencies for per-
methrin, chloruazuron, and chlorothalonil were 24.5%, 12.7%,
and 14.2%, respectively. When ozone production rate was
500 mg/h (Table 2), removal efciencies by ozone treatment for
the three pesticide solutions increased as treatment time increased.
We conclude that removal efciency for these pesticide solutions
improved as ozone treatment time increased. Further, average
time-weighted removal efciencies for permethrin, chloruazuron,
and chlorothalonil were 33.0%, 15.0%, and 24.2%, respectively.
3.3. Effects of ozone treatment on pesticide residues on vegetables
Chinese white cabbages sprayed with excessive amounts of per-
methrin were harvested in the rst stage of the eld experiment.
Samples were picked on day 5 (safe harvest day is day 4) after
spraying for a preliminary two-group comparison. Samples were
soaked for 15 min, followed by operation of the ozone generator
and water pump recirculation for 15 min under an ozone output
rate of 160 mg/h. This test compares the removal efciency of
conventional water cleaning and the proposed ozone vegetable
cleaner. Vegetables in the soaking group were cut into pieces and
408 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411

Table 1
Ozone treatment of aqueous pesticide solution (250 mg/h).

Test Permethrin Chloruazuron Chlorothalonil

Conc. (ppm) Removal eff. (%) Conc. (ppm) Removal (%) Conc. (ppm) Removal (%)
Before treatment 3.76 4.59 2.05
15 min treatment 3.15 16 4.23 8 1.92 7
30 min treatment 2.93 22 4.15 10 1.81 12
45 min treatment 2.66 29 3.84 16 1.69 18

Table 2
Ozone treatment of aqueous pesticide solution (500 mg/h).

Test Permethrin Chloruazuron Chlorothalonil

Conc. (ppm) Removal eff. (%) Conc. (ppm) Removal (%) Conc. (ppm) Removal (%)
Before treatment 3.46 5.29 1.92
15 min treatment 2.76 20 5.03 5 1.61 19
30 min treatment 2.45 29 4.55 14 1.56 21
45 min treatment 2.07 40 4.29 19 1.43 28

soaked in a water tank to investigate dissolution efciency of water
for pesticide residues on vegetables. The proposed ozone vegetable
cleaner was applied for the group with the ozone generator and
pump recirculation. Un-rinsed vegetables in the group were cut
into pieces and placed in a cleaning tank with 8 L water. Ozone out-
put rate was 160 mg/h and pump water ow rate was 12 L/min to
recirculate water in the tank, facilitating removal of impurities on
vegetables and distributing ozone evenly in the water, thereby
improving removal efciency for pesticide residues. Table 3 shows
experimental results. The background concentration (before treat-
ment) of pesticide residue on Chinese white cabbage was
3.98 ppm, and was reduced to 2.65 ppm when the cabbage was
cut into pieces and soaked in water for 15 min, resulting in a per-
methrin removal efciency of 33.5% by water soaking. When vege-
tables were treated with ozone for 15 min, the concentration of
permethrin declined to 1.92 ppm for a removal efciency of
51.8%, below the Standards for Pesticide Residue Limits in Foods
of 2.0 ppm. Thus, the permethrin removal efciency of the proposed
ozone vegetable cleaner was 18.3% higher than that of conventional
water soaking for the same period. Samples of green-stem bok choy,
which were picked after they were sprayed with chloruazuron,
were not analyzed due to an improper analytical process, resulting
in an unsuccessful calculation of ozone treatment efciency. To
compensate, green-stem bok choy was replanted in a pot in Novem-
ber 2004, and permethrin was applied as the pesticide. To simulate
overdosing by farmers, the green-stem bok choy was sprayed with
permethrin at twice the standard dose and harvested 1 day before
the safe harvest day (day 4). Permethrin residue removal efciency
was not calculated in this experiment due to a lack of a background
concentration value for permethrin. However, analytical results
demonstrate that ozone treatment outperformed water soaking
for removal of permethrin on green stem bok choy.
In the second stage, two pesticides, permethrin and chloruazu-
ron, were sprayed on Chinese white cabbage in March 2005 at the
dosage typically used by farmers (1.5 times the standard dose).
Samples were picked on day 5 after spraying (safe harvest day is
day 4). Cabbage sprayed with permethrin underwent ozone treat-
ment. Cabbage sprayed with chloruazuron was picked on day 8
after spraying (safe harvest day is day 9). Samples were classied
into the following three groups: rinsed; treated with ozone for
15 min; and, treated with ozone for 30 min. Cabbage was washed
under owing water uncut; the cabbage was then cut into pieces.
The proposed ozone vegetable cleaner was used for ozone treat-
ment groups. Unwashed vegetables were cut into pieces and placed
in a cleaning tank lled with 8 L water, ozone output rate was
500 mg/h with a recirculation ow rate of 20 L/min for 15 min
and 30 min. Table 4 shows experimental results. The removal ef-
ciencies for permethrin and chloruazuron were 6% and 4%, respec-
tively, by rising under water. Another study obtained very low
removal efciencies for pesticide residues by washing (for
20 min) under tap water 17.6%, 17.1%, 19.1%, and 15.2% for chlor-
pyrifos, p,p-DDT, cypermethrin, and chlorothalonil, respectively
(Zhang et al., 2007). When treated with the proposed ozone vegeta-
ble cleaner for 15 min and 30 min, permethrin removal efciencies
were 51% (residue = 1.51 ppm) and 58% (residue = 1.29 ppm),
respectively, and chloruazuron removal efciencies were as high
as 46% (residue = 0.31 ppm). We conclude that under a 500 mg/h
ozone output rate, permethrin and chloruazuron removal efcien-
cies increased as ozone treatment time increased, and ozonation
with water recirculation improved the pesticide removal efcien-
cies signicantly and all could meet the Standards for Pesticide Res-
idue Limits in Foods.
In the third experimental stage, chloruazuron and chlorothalo-
nil were sprayed on Chinese white cabbage in November 2005 at

Table 3
The rst stage ozone treatment on pesticide residue (Permethrin).

Test 1st time (Aug., 2004) 2nd time (Nov., 2004)

Permethrin-cabbage Permethrin-green stem bok choy
Conc. (ppm) Rem. eff.(%) Conc.(ppm) Rem. eff. (%)
Before treatment 3.98 NA NA
Soaking for 15 min 2.65 33.5 7.18 NA
O3 treatment for15 min 1.92 51.8 4.79 NA
(O3 output @160 mg/h)

NA: due to experimental errors that no results were available.

 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411 409

Table 4
The 2nd stage ozone treatment on pesticide residue (O3 output @500 mg/h).

Test Permethrin Chloruazuron

Conc. (ppm) Rem. eff. (%) Conc. (ppm) Rem. eff. (%)
No rinse (before treatment) 3.10 0.57
Tap water rinse (whole pack) 2.90 6 0.55 4
15 min O3 treatment 1.51 51 0.31 46
30 min O3 treatment 1.29 58 N.D. >91a
a
The chloruazuron residue on the Chinese cabage, after 30 min ozone treatment, was found to below detection limit (0.05 ppm). Based on this, removal efciency was
calculated to be greater than 91%.

Table 5
The 3rd stage ozone treatment on pesticide residue.

Test Chlorothalonil Chloruazuron

Conc. (ppm) Rem. eff. (%) Conc.(ppm) Rem. eff. (%)
No rinse (before treatment) 6.37 0.51
Tap water rinse (whole pack) 5.13 19 0.42 18
Pump recirculation for 15 min 3.00 53 0.25 51
250 mg/h 15 min treatment 2.85 55 0.20 60
30 min treatment 2.32 64 0.17 67
500 mg/h 15 min treatment 1.47 77 0.13 75
30 min treatment 0.84 87 0.09 83

the dosage typically used by farmers (1.5 times the standard). Sam-
ples were subjected to ozone treatment on day 6, 1 day before the
safe harvest day to simulate a rushed harvest. Cabbage samples
sprayed with chlorothalonil and chloruazuron were picked on
day 6 (safe harvest day is day 7), while cabbage samples sprayed
with chloruazuron were picked on day 8 (safe harvest day is
day 7) after spraying.
In this experiment, recirculation cleaning with a pump for
15 min (without ozone) was utilized to investigate the removal ef-
ciency of the ozone vegetable cleaner, recirculation pump, and
ozone treatment for pesticide residues on vegetables. Moreover,
ozone treatment of aqueous pesticide solutions indicates
that ozone oxidation and decomposition efciency of pesticide
increased as the ozone output rate increased. Therefore, ozone out-
put rates of 250 mg/h and 500 mg/h were applied under the same
condition to determine whether an increased ozone output rate im-
proved removal efciency for pesticide residues on vegetables.
Experimental samples were classied into the following six
groups: rinsed with water; pump recirculation wash for 15 min;
250 mg/h ozone treatment for 15 min; 250 mg/h ozone treatment
for 30 min; 500 mg/h ozone treatment for 15 min; and 500 mg/h
ozone treatment for 30 min. Table 5 lists experimental results.
The removal efciency for pesticide residues on vegetables was
low with whole-pack rinsing at 19% for chlorothalonil and 18%
for chloruazuron. Only pesticide residue on leaves was removed,
while pesticides in leaves were not removed.
When the proposed cleaner was used without ozone injection
for 15 min, pesticide removal from leaf surfaces by water recircu-
lation was effective because the leaves were cut into small pieces
and soaked in water; pesticide residue was removed from leaves
via pump recirculation. Therefore, pesticide residue removal ef-
ciencies of 53% for chlorothalonil and 51% for chloruazuron by
pump recirculation were higher than those by rinsing alone. When
the ozone production rate of 250 mg/h was combined with pump
recirculation and ozone treatment for 15 min, removal efciencies
improved slightly to 55% for chlorothalonil and 60% for chloru-
azuron. When ozone treatment duration was increased to
30 min, removal efciencies increased to 64% for chlorothalonil
and 67% for chloruazuron. When the ozone production rate was
increased to 500 mg/h under the same experimental conditions
and combined with a 15 min ozone treatment, removal efciency
was 77% for chlorothalonil and 75% for chloruazuron. Removal
efciency increased to 87% for chlorothalonil and 83% for chloru-
azuron when ozone treatment duration was increased to 30 min. In
both ozone production rates, the pesticide residues were all below
the Standards for Pesticide Residue Limits in Foods of chlorothalo-
nil (5.0 ppm) and chloruazuron (2.0 ppm).
Overall, removal efciency by pump recirculation washing ex-
ceeded 50%. When ozone treatment was combined with an ozone
production rate of 250 mg/h, removal efciency increased by
roughly 10% (range, 216%). When the ozone production rate
was increased to 500 mg/h, removal efciency increased an addi-
tional 20% (range, 2434%). We conclude that increasing the ozone
production rate increases removal efciency for pesticide residues
on vegetables.
3.4. Toxicity of wastewater from pesticides after ozone treatment
The primary function of the proposed ozone vegetable cleaner is
to remove or oxidize the pesticide residues on vegetables. Pesticide
residue is removed from the leaf surfaces into water and oxidized
or decomposed via pump recirculation and ozone oxidation by the
generator when ozone is injected into the reaction tank. From an
environmental protection perspective, pesticides are toxic com-
pounds that can be decomposed by ozone, and partially trans-
formed into wastewater or dissolved into water (depends on
pesticide solubility), or oxidized by ozone into another derivative.
The Tox Tracer test was selected as the biological toxicity test to
determine whether the toxicity of wastewater increased after
ozonation. Table 6 shows Tox Tracer biological toxicity test results.
The principle of the Tox Tracer biological toxicity test is similar
to that of the Microtox biological toxicity test; that is, both take
advantage of the fact that toxicity makes the indicator organisms
light weaken. A photomultiplier tube was used to detect light var-
iance and calculate the percentage light inhibition and 50% effect
concentration (EC50). When a toxic compound reacts with uores-
cent bacteria, their metabolism is inhibited or terminated, making
uorescence decrease or disappear. Variance in uorescent light
intensity can reect the toxicity of a toxic compound to uorescent
bacteria (Sotelo et al., 1989).
The test index of the Microtox toxicity test is light intensity and
its unit of toxicity is EC50, which is the effective concentration of a
410 J.Y. Chen et al. / Journal of Food Engineering 114 (2013) 404411
Table 6
Toxicity test of wastewater after ozone treatment.
O3 output Test Permethrin (%) Chloruazuron (%)
250 mg/h Before treatment 57.1 49.3
15 min treatment 32.7 12.4
30 min treatment 44.7 56.7
45 min treatment 49.8 57.6
500 mg/h Before treatment 57.1 49.3
15 min treatment 48.8 31.8
30 min treatment 47.0 49.3a
45 min treatment 52.1 45.6
a A proportional relationship existed between liquid ozone
Due to a broken vial in the analytical process, with a possibility of contami-
nation the result was for reference only.
toxic compound when light intensity is reduced by 50%. The EC50 of
uorescent bacteria is represented by a Gamma (c) function, which
is the ratio of lost uorescence to residual uorescence (Eq. (1)).
c Loss of fluorescence=Residual of fluorescence
Initial fluorescence
 Residual fluorescence=Residual fluorescence
Io  Ir=Ir Io=Ir  1
where, Io is the Initial uorescence, Ir is the Residual uorescence
after testing time t
When c is 1, i.e., uorescence loss equals residual uorescence
(or Ir = Io/2), uorescence is reduced by 50%, and the correspond-
ing toxic compound concentration is EC50.
Toxicity variance of a pesticide solution with illumination be-
fore and after ozone treatment is expressed as% inhibition, indicat-
ing that as the value increases, toxicity increases.
The toxicity inhibition percentage of the permethrin solution be-
fore ozonation was 57.14% under an ozone production rate of
250 mg/h (Table 6). However, after ozonation for 15 min, 30 min,
and 45 min, the toxicity inhibition percentage of the permethrin
solution declined to 32.7%, 44.7%, and 49.8%, respectively. Toxicity
inhibition percentages for the permethrin wastewater after ozona-
tion were all lower than those before ozonation. However, this per-
centage increased as ozonation duration increased, likely resulting
from derivatives in pesticide oxidation or decomposition processes.
As ozonation duration increased, the concentration of derivatives
also increased, causing the inhibition percentage to rst decrease
and then increase. Overall, the toxicity inhibition percentage of per-
methrin wastewater rst declined and then increased as ozone
treatment duration increased, and were lower than those of the
permethrin solution. We conclude that permethrin biological toxic-
ity after ozonation was lower than before ozonation.
When the ozone production rate was 250 mg/h for ozonation of
chloruazuron, the toxicity inhibition percentage was 49.3%. When
treated with ozone for 15 min, 30 min, and 45 min, inhibition per-
centages reduced to 12.4%, 56.7%, and 57.6%, respectively.
According to the denition of the gamma function, a negative inhi-
bition percentage means that residual uorescence exceeds initial
uorescence. Thus, chloruazuron wastewater after ozone treat-
ment did not cause inhibition, but rather increased the bacterial
uorescence. We conclude that biological toxicity of chloruazu-
ron wastewater after ozonation was lower than that of the chlor-
uazuron solution before ozonation.
4. Conclusions
The proposed domestic-scale ozone vegetable cleaner over-
comes the disadvantage of low ozone solubility. The prototypes
efciency for ozone treatment of pesticide solutions and pesticide
residues on vegetables was assessed. The following conclusions are
based on experimental results.
(1) Gas-phase production capacity is the critical factor when
dissolving ozone in water, and is the most important factor
during ozone treatment of pesticides. When ozone produc-
tion rate increased, ozone injected into water dissolved
instantly and evenly, and the optimal treatment efciency
was achieved.
(2) The liquid ozone concentration increased rapidly within
1 min after ozone was injected into water, and the ORP lev-
els increased rapidly. When ORP peaked, it then decreased
slightly and stabilized at the ozone saturation concentration.
concentration and ORP in this study.
(3) Ozone treatments of liquid permethrin, chloruazuron, and
chlorothalonil solutions indicate that removal efciency
improved as ozone production rate increased. Under the
same ozone production rate, removal efciency increased
as treatment duration increased.
(4) Soaking for 15 min removed 33.5% of permethrin residue on
vegetables; whole-pack rinsing removed roughly 6% of per-
methrin, 18% of chloruazuron and 19% of chlorothalonil.
The proposed ozone vegetable cleaner removed over 50%
1 of these pesticides with water recirculation for 15 min. After
adding ozone treatment with an ozone production rate of
250 mg/h for 15 min, removal efciency for pesticide residue
increased by about 10%. When ozone production rate
increased to 500 mg/h for a 15 min ozone treatment,
removal efciency improved by an additional 20%. Removal
efciency of the proposed cleaner for pesticide residue was
18.3% better than that of soaking alone, and 4252% better
than that of whole-package rinsing.
(5) Under the ozone production rate of 250 or 500 mg/h, all pes-
ticide residues of the Chinese white cabbage after ozone
treatment could meet the Standards for Pesticide Residue
Limits in Foods.
Acknowledgement
The authors would like to thank the National Science Council of
the Republic of China, Taiwan, for nancially supporting this re-
search under Contract No. NSC 93-2622-E-020-006-CC3. The
authors are also grateful for the technical assistance in the gas
ozone analyses provided by Professor Young Ku, Department of
Chemical Engineering, National Taiwan University of Science and
Technology, Taipei, Taiwan.
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