82 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 ISSN 1203-8407 2014 Science & Technology Network, Inc.

Abamectin Degradation by Advanced Oxidation Processes: Evaluation

of Toxicity Reduction Using Daphnia similis
Jos Roberto Guimares*
, 1
, Izabela Major Barbosa
1
, Milena Guedes Maniero
1
, Susanne Rath
2

1
School of Civil Engineering, Architecture and Urbanism, University of Campinas - UNICAMP, P.O. Box 6021,
CEP 13083-852, Campinas, SP, Brazil
2
Chemistry Institute, University of Campinas UNICAMP, P.O. Box 6154, CEP 13084-971, Campinas, SP,
Brazil


Abstract:
This work evaluated the toxicity reduction of abamectin in an aqueous solution (500 g L
-1
) undergoing to
photolysis (UV), peroxidation (H
2
O
2
), peroxidation with ultraviolet radiation (UV/H
2
O
2
), Fenton (Fe(II)/H
2
O
2
),
and photo-Fenton (Fe(II)/UV/H
2
O
2
) processes. Toxicity trials were carried out using the microcrustacean Daphnia
similis. Results were based on effective concentration, EC
50
48 h. Using 1.0 mmol L
-1
Fe (II) and 5.0 mmol L
-1
H
2
O
2
and a reaction time of 60 s, the ranking of efficiency for the degradation of abamectin was as follows: photo-
Fenton > Fenton > UV = UV/ H
2
O
2
> H
2
O
2
. However, after a reaction time of 300 s, similar degradation was
obtained for photo-Fenton, UV, and UV/H
2
O
2
(more than 97%), while the Fenton process degraded 80% of the
drug. With the exception of peroxidation, all of the processes evaluated were able to eliminate toxicity in 600 s
of reaction (1.0 mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
).

Keywords: Daphnia similis, Fenton, photo-Fenton, UV/H
2
O
2
, veterinary drug.


Introduction
Abamectin is a macrocyclic lactone in the
avermectin group. It is active antihelmintics, inseticides
and acaricides. Avermectins has been widely employed
in veterinary medicine to treat ecto- and endoparasites
and in agriculture because of these properties. They
are produced by fermentation of an actomycete,
Streptomyces avermitilis, which is a naturally-occurring
soil bacterium (1). Abamectin is composed of
avermectin B
1a
(at least 80%) (C
48
H
72
O
14
)

and
avermectin B
1b
(no more than 20%) (C
47
H
70
O
14
). The
two homologues, B
1a
and B
1b
, form a compound with a
high molar mass (873.08 g mol
-1
). It has low solubility
in water (< 10 g L
-1
) (2) and is soluble in organic
solvents (3).
After abamectin is administered to animals, a
significant amount of the non-metabolized drug is
excreted directly into the environment. Much of the
substance ends up in the soil and water. According to
Tisler and Erzen (4), in the majority of cases, up to
98% of avermectins applied to cattle are excreted in
feces, unaltered or as an active metabolite. Hernando
et al. (5) and Tisler and Erzen (4) reported that
avermectins interfere in the reproduction and survival
of aquatic and terrestrial organisms that have important
roles in the food chain.

*Corresponding author; E-mail address: jorober@fec.unicamp.br
An alternative to degrading organic compounds in
water is the use of Advanced Oxidation Processes
(AOPs). These technologies are efficient at degrading
and removal of recalcitrant organic compounds, even at
low concentrations. AOPs are based on the generation
of hydroxyl radicals (HO

), which are non-selective

and highly reductive species.
The byproducts formed during degradation
processes can be as toxic as or even more toxic than the
original non-degraded substance (6). Even though AOPs
are very efficient at degrading organic compounds, the
toxicity of the treated solutions must be evaluated.
Species of genus Daphnia, phylum Crustacea, order
Cladocera, play a large role in the zooplankton com-
munity and have been widely studied and characterized
in order to evaluate the toxicity of many substances
released into the environment. Many authors have
reported toxic effects of avermectins on some micro-
crustaceans of Genus Daphnia (4, 5, 7-17). According
to Novelli et al. (17), Daphnia similis is sensitive to
avermectins. Thus, Daphnia similis is an appropriate
microcrustacean for monitoring the toxicity of aqueous
abamectin solution submitted to the degradation
processes.
The objective of this study was to evaluate reduction
of toxicity of aqueous abamectin solutions that had
undergone photolysis, peroxidation, UV/H
2
O
2
, Fenton,
and photo-Fenton processes using the Daphnia similis
microcrustacean as a test organism.
J. R. Guimares et al.
J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 83
Material and Methods
Chemical Reagents
Abamectin (>97%) was obtained from Fluka
(Buchs, Switzerland) and methanol (HPLC grade) from
J. T. Baker (Xalostoc, Mexico), while ferrous sulfate
(FeSO
4
.7H
2
O), sulfuric acid, and hydrogen peroxide
(30% m/m) were obtained from Synth (Diadema,
Brazil). Ultra-pure water used in the tests was produced
by a Milli-Q purification system from Millipore
(Billerica, USA).

Stock Solution
The abamectin stock solution (500 mg L
-1
) was
prepared in methanol. The solution was stored in an
amber flask which was protected from light and kept
at -10 C. The working solutions (500 g L
-1
) were
prepared by diluting the stock solution with ultrapure
water.

Experimental System
The photo-reactor was made up of a germicide
lamp (15 W,
max
= 254 nm, 2.0 cm diameter) covered
by a quartz cylinder (5.0 cm diameter, 52.0 cm long)
inserted in the middle of a borosilicate tube (8.0 cm
diameter and 55.0 cm long). The sample volume used
in the reactor was 1.0 L. The solution was homo-
genized by a magnetic stirrer. The tests were run in
batches.
Average irradiance was 3.83 mW cm
-2
. It was
measured by a model VLX 3 W Cole Parmer Series
9811 radiometer calibrated to a 254 nm wavelength.
The radiation dose D (mJ cm
-2
) was calculated using
Equation (1).
(1)

in which D is the UV radiation dose (mJ cm
-2
), E is the
irradiance (mW cm
-2
), and t is the exposure time (s).
For all of the processes evaluated, reaction time
varied from 0 to 600 s. Photolysis, peroxidation, and
peroxidation with UV radiation trials were carried out
at the natural pH of the sample (around 7), while
Fenton and photo-Fenton processes were carried out
after pH was adjusted to 2.85. Experiments carried out
using the Fenton reagent employed Fe(II) at concen-
trations between 0.25 and 1.0 mmol L
-1
, and H
2
O
2
between 1.0 and 10.0 mmol L
-1
. Fe(II) concentrations
greater than H
2
O
2
concentrations were not evaluated

because according to Neyens and Baeyens (18), the
molar ratio of Fe(II):H
2
O
2
must always be less than 1.
In all of the tests carried out with hydrogen peroxide
(peroxidation, peroxidation assisted by ultraviolet
radiation, Fenton, and photo-Fenton), reactions were
stopped and the residual H
2
O
2
concentration was
destroyed by adding sodium bisulfite at a 1:1 molar
ratio with hydrogen peroxide (19).

Analytical Method
The concentration of abamectin in the aqueous
solutions was determined by high performance liquid
chromatography with UV detection. Before quantitation,
the samples were concentrated by solid phase extraction
(SPE) using C
18
cartridges (500 mg/6 mL, Varian) that
had been previously conditioned with methanol (6 mL)
and water (6 mL). A liter of the solution percolated
through the cartridges at a flow rate of 10 mL min
-1
.
The analytes were eluted with 4 mL of methanol. Before
analyses, the solutions were filtered through 0.22 m
Millipore membranes. The extraction efficiency,
evaluated through recovery tests, ranged from 64 to
99% for aqueous solutions containing abamectin with
concentrations ranging from 25 to 500 g L
-1
.
Chromatographic analyses were carried out with a
Waters solvent delivery system (Waters 510), a tunable
absorbance detector (Waters 486), a Rheodyne 7725
injector with a sample loop of 20 L, and a Waters
746 integrator. Abamectin quantitation was performed
at 245 nm. The analytical column was a Waters
XBridge
TM
RP18 (250 mm x 4.6 mm, 5 m) and, as
mobile phase, methanol and water (85:15 v/v) with a
flow rate of 1 mL min
-1
was used. The limit of
detection (LOD) and limit of quantitation (LOQ) were
1.0 g L
-1
and 3.0 g L
-1
, respectively, considering a
concentration factor of 250.
UV-Vis spectra were obtained using a UV-1601PC
spectrophotometer (Shimadzu, Japan). The initial
solution (500 g L
-1
abamectin) and the samples
submitted to the degradation processes were concen-
trated using SPE, as described previously. The analyte
was eluted from the cartridge with methanol (4 mL),
filtered, and diluted 4 times before analysis. Spectra
were registered from 200 to 350 nm, since there was
no observed absorbance in the visible region of the
eletromagnetic spectrum.
Consumption of hydrogen peroxide was evaluated
based on the oxidation-reduction reaction between
residual H
2
O
2
and metavanadate ions. The vanadate
solution (which is yellow) turns red after the reaction
with H
2
O
2
due to the formation of peroxovanadium
cations (Equation 2), which absorb at 450 nm (20).
The limit of detection of the method (LOD) was 0.02
mmol L
-1
.

(2)

Daphnia Similis Toxicity Trials
Acute toxicity trials and cultivation of the organisms
were carried out as specified by the Brazilian National
J. R. Guimares et al.
84 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014
Standards Organization (NBR 12.713) (21). To
evaluate the sensitivity of the organisms, EC
50
48 h
was determined for sodium chloride as described in
NBR 12.713 (21). All toxicity trials were followed
with blank samples, which contained cultivation water
and neonate organisms, and control samples, which
contained all of the reagents present in the sample to
be analyzed, but not abamectin.
The biological trials were carried out for all
processes evaluated. The samples were taken at the
beginning of the test and at 10, 60, and 600 s. They
were diluted between 0.01 and 0.5% (v/v) using the
cultivating water as diluent. Each dilution was evaluated
in four replicates, each sample containing five neonate
organisms. The organisms were incubated at 20 C in
the dark. After 48 h, dead and/or immobile Daphnia
organisms were counted. The dilution corresponding
to the effective median concentration (EC
50
48 h) was
estimated by the Trimmed Spearman-Karber statistical
method (22). All of the trials were carried out in
triplicate and the average values and standard deviations
were calculated. Deviations of up to 20% were
considered acceptable (23).

Results and Discussion
Abamectin Degradation
In all cases in which abamectin was exposed to
UV radiation, that is, for the photolysis, peroxidation
with UV radiation, and photo-Fenton processes (Figure
1), the degradation efficiencies were always higher
than 90% in the first 300 s of the reaction. These
results agree with those reported by Escalada et al.
(24), who verified a high level of degradation of
abamectin by photolysis.
At the very beginning of UV radiation exposure
(60 s) (Figure 1a), at a dose of 229.2 mJ cm
-2

(Equation 1), approximately 70% of the substance was
degraded. After 600 s of UV radiation exposure, at a
dose of 2,292 mJ cm
-2
(Equation 1), more than 95%
was degraded. The photolytic process has already been
shown to be efficient at degrading various organic
compounds (25, 26).
Degradation of abamectin by peroxidation using
5.0 mmol L
-1
H
2
O
2
was less than 10% (Figure 1b).
Increasing the H
2
O
2
concentration from 0.25 to 10.0
mmol L
-1
did not result in an increase in the drug
degradation, remaining below 10%; this shows that
the oxidant has little potential to oxidize abamectin.
Similar results were obtained by Ay and Kargi (27),
Catalkaya and Kargi (28), and Da Silva et al. (25),
who reported that peroxidation was ineffective at
degrading several drugs.














Figure 1. Abamectin degradation by (a) UV; (b) H
2
O
2
(5.0 mmol
L
-1
H
2
O
2
); (c) UV/H
2
O
2
(5.0 mmol L
-1
H
2
O
2
); (d) Fenton (1.0 mmol
L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
); and (e) photo-Fenton (1.0
mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
). C
o,abamectin
= 500 g L
-1
.

For the UV/H
2
O
2
process using 5.0 mmol L
-1

H
2
O
2
,

abamectin was reduced by 34% in 10 s of
reaction, and in 300 s, degradation was greater than
90% (Figure 1c); after this period, there was no increase
in degradation over time. Comparing the UV and
UV/H
2
O
2
processes, it is possible to verify that there
was no increase in the degradation rate of the drug
when H
2
O
2
was added to the photolysis process
(Figures 1a and 1c). In the UV/H
2
O
2
process (5.0
mmol L
-1
), H
2
O
2
consumption was only 1.40 mmol L
-1
;
for degradation by peroxidation

(5.0 mmol L
-1
), the
oxidant was not consumed during the 600 s of reaction
time.
Fenton reagent (1.0 mmol L
-1
Fe(II) and 5.0 mmol
L
-1
H
2
O
2
) (Figure 1d) was efficient at degrading
abamectin in the first 60 s of the experiment, with
levels higher than 70%. Similar results were obtained
by Dal Bosco et al. (29), which reported around 80%
degradation of ivermectin (another avermectin) in
aqueous solution by Fenton reagent using the same
conditions applied in this study.
The fastest degradation rate was verified for photo-
Fenton, attaining degradation efficiencies greater than
90% in the first 60 s of reaction time (Figure 1e).
Similar results were obtained for ivermectin by Dal
Bosco et al. (29) using the photo-Fenton process and
the same initial concentrations of Fe(II) and H
2
O
2,
that
is, 1.0 mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
. It has
been verified that the degradation rate of abamectin
was increased by adding UV radiation to the Fenton
reagent (Figures 1d and 1e). The Fenton reaction
results in Fe(III) (Equation 3) and the increase in the
degradation rate by photo-Fenton is mainly due to
photo-reduction of Fe(III) formed in the Fenton process
(Equations 4 and 5).
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
(a) , (b) , (c) ,
(d) , (e)
J. R. Guimares et al.
J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 85
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.25 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
A
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.25 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
B
H
2
O
2
+ Fe(II) Fe(III) + HO

+
-
OH (3)
[Fe
(III)
(OH)]
2+
Fe(II) + HO

(4)
[Fe
(III)
(RCO
2
)]
2+
Fe(II) + CO
2
+

R (5)

For the photo-Fenton process, H
2
O
2
was quickly
consumed at all oxidant and catalyst concentrations
evaluated. After 600 s of reaction, molar concentrations
of H
2
O
2
were near the methods detection limit (0.02
mmol L
-1
). This was also reported by Dal Bosco et al.
(29) and Elmolla and Chaudhuri (30), who attribute this
to the fact that applied radiation quickly regenerates
Fe(III) to Fe(II), offering greater production of free
hydroxyl radicals (Equations 3 to 5), consuming the
oxidant (Equation 3).

Influence of Fe(II) and H
2
O
2
on the Fenton
Process
When Fe(II) concentration was changed from 0.25
to 1.0 mmol L
-1
, with the H
2
O
2
concentration kept at
5.0 mmol L
-1
(Figure 2A), degradation increased from
40% to around 80%, while for an H
2
O
2
concentration
set at 10.0 mmol L
-1
(Figure 2B), degradation rose
from 20 to 90% in 300 s of reaction.
In the study carried out by Falmann et al. (31) of
the degradation of various pesticides and abamectin
by Fenton reagent, the reaction rate increased with an
increase in the catalyst concentration in the first
minutes, reaching a plateau after 50 min.
Molar ratios of ferrous ions to hydrogen peroxide
(Fe(II):H
2
O
2
) of 0.25:1.0; 0.25:5.0; and 0.25:10 were
also evaluated. It was verified that an increase in
oxidant concentration results in a lower efficiency of
the overall degradation process. In 300 s, a reduction
from 80% to 20% using Fe(II):H
2
O
2
from 0.25:1.0 to
0.25:10 were achieved (Figure 3A). Lower abamectin
degradation when greater concentrations of hydrogen
peroxide were employed could be because the excess
of oxidant in the medium acts as a hydroxyl radical
scavenger (32-33). These results agree with those
published previously in our research group by Dal
Dosco et al. (29), who verified a decrease in degradation
of ivermectin as the oxidant concentration was increased
from 1.0 to 5.0 mmol L
-1
, maintaining the initial con-
centration of Fe(II) (0.25 mmol L
-1
).
When the Fe(II) concentration was maintained at
0.5 mmol L
-1
(Figure 3B), an increase in abamectin
degradation from 40% to 70% was verified when
oxidant concentration was increased from 1.0 to 10.0
mmol L
-1
. For 10 mmol L
-1
Fe(II) (Figure 3C), the
degradation rate was greater in the first 10 s of the
experiment when the concentration of H
2
O
2
was
increased from 5.0 to 10.0 mmol L
-1
.




























Figure 2. Influence of Fe(II) concentration on abamectin degradation
by Fenton reagent. C
o,abamectin
= 500 g L
-1
. (A) C
H2O2
= 5.0 mmol
L
-1
, (B) C
H2O2
= 10.0 mmol L
-1
.

Abamectin degradation efficiency was around
90% in 300 s of reaction using molar ratios of 0.25:1.0
and 1.0:10 Fe(II):H
2
O
2
, and 65% for 0.5:10.0 mmol L
-1

Fe(II):H
2
O
2
. However, in 10 s of reaction, degradation
rate was ranked in the following order: 1.0:10 > 0.5:10
> 0.25:1.0 Fe(II) to H
2
O
2
. The efficiency at a ratio of
1.0:10 was 77%; for 0.5:10 it was 53%, and for 0.25:10
it was 29%. Therefore, in order to achieve a high level
of efficiency of drug removal, lower reagent concen-
trations can be used with a longer reaction time, or
larger concentrations of oxidant can be used with
shorter test times.
The concentration of abamectin reached a plateau
in the Fenton process after a certain period of time,
which has also been observed by other authors (29,
35). This can take place due to inherent oxidant and
Fe(II) consumption in the process or due to the
reaction between hydroxyl radicals and excess ferrous
ions resulting in ferric ions, as shown in Equation 6.
Formation of organic compounds with smaller chains
reduces the activity of Fenton reagent, and complexation
of intermediaries formed with ferric cations resulting
h
h
J. R. Guimares et al.
86 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.25 mmol L
-1
Fe(II)/ 1.0 mmol L
-1
H
2
O
2
0.25 mmol L
-1
Fe(II)/ 5.0 mmol L
-1
H
2
O
2
0.25 mmol L
-1
Fe(II)/ 10.0 mmol L
-1
H
2
O
2
A
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.5 mmol L
-1
Fe(II)/1.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
B
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
A
0.25 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.25 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
B











































Figure 3. Influence of H
2
O
2
concentration on abamectin degradation
by Fenton reagent. C
o,abamectin
= 500 g L
-1
. (A) C
Fe(II)
= 0.25 mmol
L
-1
, (B) C
Fe(II)
= 0.5 mmol L
-1
, (C) C
Fe(II)
= 1.0 mmol L
-1
.

in recalcitrant compounds could be the cause of the
stagnation of abamectin concentration. It is also worth
emphasizing that the Fenton reagent is not an advanced
oxidation process that yields a large amount of hydroxyl
radicals.

HO

+ Fe(II) Fe(III) +
-
OH (6)





























Figure 4. Influence of Fe(II) concentration on the efficiency of the
photo-Fenton process. C
o,abamectin
= 500 g L
-1
. (A) C
H2O2
= 5.0
mmol L
-1
, (B) C
H2O2
= 10.0 mmol L
-1
.

Influence of Fe(II) Concentration on the Photo-
Fenton Process
For photo-Fenton, an increase in Fe(II) concen-
tration from 0.25 to 1.0 mmol L
-1
(Figure 4A and 4B)
resulted in a higher abamectin degradation rate in the
first 10 s of exposure. Degradation efficiency increased
from 50% to about 80% when 5.0 mmol L
-1
was used
(Figure 4A), and from 30% to around 90% when 10.0
mol L
-1
was used (Figure 4B). However, after 300 s of
reaction, in all of the conditions evaluated, the same
degradation efficiency was reached, around 98%. A
greater degradation efficiency of the target compound
with the increased concentration of catalyst is the result
of a greater number of hydroxyl radicals in the medium
formed by the Fenton reaction (Equation 3) (36).

Influence of H
2
O
2
Concentration on the photo-
Fenton Process
When the catalyst concentration was kept at 0.25
mmol L
-1
, an increase in oxidant concentration from 1.0
mmol L
-1
to 10.0 mmol L
-1
reduced the degradation of
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
1.0 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
C
J. R. Guimares et al.
J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 87
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.25 mmol L
-1
Fe(II)/1.0 mmol L
-1
H
2
O
2
0.25 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0.25 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
A
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
C
/
C
o
Time (s)
0.5 mmol L
-1
Fe(II)/1.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
0.5 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
B
0 100 200 300 400 500 600
0.0
0.2
0.4
0.6
0.8
1.0
1.0 mmol L
-1
Fe(II)/5.0 mmol L
-1
H
2
O
2
1.0 mmol L
-1
Fe(II)/10.0 mmol L
-1
H
2
O
2
C
C
/
C
o
Time (s)
abamectin from 50% to 34% in 10 s, and from 90% to
77% in 60 s (Figure 5A). It was due to the scavenger
effect of hydroxyl radicals by hydrogen peroxide when
low Fe(II) concentrations are used.
When the initial concentration of Fe(II) was
doubled to 0.5 mmol L
-1
, increasing the H
2
O
2
level to
10.0 mmol L
-1
resulted in greater degradation efficiency,
from 49% to 67% in 10 s of reaction (Figure 5B). For
Fe(II) concentration of 1.0 mmol L
-1
, after 10 s of
reaction, degradation increased from 77% to 88% with
an increase in H
2
O
2
concentration from 5.0 to 10.0
mmol L
-1
; after this time, no difference was verified
between degradation for the two concentrations of
H
2
O
2
(Figure 5C).
For tests carried out with Fenton reagent or with
the photo-Fenton process, note that when catalyst and
oxidant are used at concentration ratios of 0.25:10.0,
there was a reduction in degradation rate. This may be
because of the low concentration of the catalyst
compared to the oxidant, or to the formation of
hydroperoxyl radicals (Equation 7), which are produced
in reactions with excess oxidant and are less reactive
and have less reduction potential (1.70 V) than
hydroxyl radicals (2.8 V) (37).

HO

+ H
2
O
2
HO
2

+ H
2
O (7)

UV-Visible Spectrometry
The maximum absorbance of abamectin is at 245
nm and the low-pressure mercury lamp emits a
maximum dose at 254 nm. This property could have
contributed to the high degradation efficiency of
abamectin by the processes with added UV light.
Figure 6 presents the UV-Vis spectra of abamectin
solutions submitted to peroxidation (Figure 6A),
photolysis (Figure 6B), peroxidation combined with
UV radiation (Figure 6C), Fenton (Figure 6D), and
photo-Fenton (Figure 6E) processes.
As degradation by UV radiation and UV/H
2
O
2

progressed, a reduction at the 245 nm absorbance band
took place (Figure 6B and 6C), showing that abamectin
was degraded over time. In 600 s of reaction, no
absorbance peak was seen, showing that the parent
molecule was no longer in the solution. Similar results
were reported by Escalada et al. (24) for abamectin
using an exposure time of 300 s of UV radiation at
254 nm. The authors attributed this to the opening of
rings by cleaving double bands, mainly in carbons 3,
14, and 22.
The UV-spectra recorded for abamectin solutions
that underwent the photo-Fenton process (Figure 6E)
showed a complete degradation of the molecule after
600 s. However, when Fenton reagent was used, a







































Figure 5. Influence of H
2
O
2
concentration on degradation of
abamectin by the photo-Fenton process. C
o,abamectin
= 500 g L
-1
.
(A) C
Fe(II)
= 0.25 mmol L
-1
, (B) C
Fe(II)
= 0.5 mmol L
-1
, (C) C
Fe(II)
=
1.0 mmol L
-1
.

reduction at the peak of the absorption band was only
seen with an increase in reaction time (Figure 6D).
During the peroxidation trials, there was no
reduction in the characteristic peak of absorption of
abamectin at 245 nm (Figure 6A), proving that this
process is not efficient at degrading this drug.

Monitoring Toxicity Reduction
The Daphnia similis microcrustacean used as a
test organism in the toxicity trials meets the official
J. R. Guimares et al.
88 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014
200 225 250 275 300 325 350
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
t = 600 s
t = 300 s
t = 60 s
t = 30 s
t = 10 s
t = 0 s
A
b
s
o
r
b
a
n
c
e
(nm)
200 225 250 275 300 325 350
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
t = 600 s
t = 300 s
t = 60 s
t = 30 s
t = 10 s
t = 0 s
A
b
s
o
r
b
a
n
c
e
(nm)
200 225 250 275 300 325 350
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
t = 600 s
t = 300 s
t = 60 s
t = 30 s
t = 10 s
t = 0 s
A
b
s
o
r
b
a
n
c
e
(nm)
200 225 250 275 300 325 350
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
t = 600 s
t = 300 s
t = 60 s
t = 30 s
t = 10 s
t = 0 s
A
b
s
o
r
b
a
n
c
e
(nm)
200 225 250 275 300 325 350
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
t = 600 s
t = 300 s
t = 60 s
t = 30 s
t = 10 s
t = 0 s
A
b
s
o
r
b
a
n
c
e
(nm)

A B












C D












E










Figure 6. UV-Vis spectra of abamectin solutions submitted to (A) peroxidation (1.0 mmol L
-1
H
2
O
2
), (B) photolysis, (C) peroxidation
combined with UV radiation (1.0 mmol L
-1
H
2
O
2
), (D) Fenton (0.25 mmol L
-1
Fe(II) and 1.0 mmol L
-1
H
2
O
2
), and (E) photo-Fenton (0.25
mmol L
-1
Fe(II) and 1.0 mmol L
-1
H
2
O
2
).

requirements to be considered acceptable for trials, as
recommended in NBR 12.713/2009 (21). No sign of
immobility of organisms was observed when they
were exposed to the control (which contained all of
the reagents present in the sample to be analyzed, but
not abamectin) and blank solutions (which contained
cultivation water and neonate organisms). In addition,
the average effective concentration for the reference
substance (sodium chloride), EC
50
48 h, was 2.68 g L
-1

(n = 11), with variations of less than 2SD (twice the
standard deviation), as recommended in NBR
12.713/2009 (21).
Average EC
50
48 h for Daphnia similis was 0.16
g L
-1
(n = 4), with a confidence interval (95%)
between 0.15 and 0.20 g L
-1
. Authors such as Tisler
and Erzen (4) found EC
50
48 h for abamectin of 0.25
g L
-1
in trials using Daphnia magna. Ivermectin, a
synthetic derivative of abamectin, was shown to be
more toxic than abamectin for different genera of
Daphnia: EC
50
48 h of 70 ng L
-1
for D. similis (29) and
5.7 ng L
-1
for D. magna (11).
Sensitivities of D. magna and D. similis can be
considered similar from the ecotoxicological point of
view, with acceptable variance (a correlation coefficient
J. R. Guimares et al.
J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 89
Table 1. Toxicity results for UV and UV/H
2
O
2
processes (C
o,abamectin
= 500 g L
-1
).

Process Time (s) Degradation (%)
Toxicity
reduction (%)
Standard
deviation (%)
UV
0 0.0 0 -
10 33.6 38 9
60 68.1 67 19
600 97.5 NT -
UV/H
2
O
2
(0.25 mmol L
-1
)
0 0.0 0 -
10 32.5 59 8
60 66.1 74 5
600 98.6 89 3
UV/H
2
O
2
(5.0 mmol L
-1
)
0 0.0 0 -
10 34.7 44 5
60 66.8 70 5
600 99.1 NT -
NT non-toxic

of R
2
= 0.99). In toxicity studies with D. magna and D.
similis on industrial effluents and different chemical
compounds, such as phenol and potassium dichromate,
it was concluded that the two species have similar
sensitivity (38).
It is worth restating that there are frequently
differences in EC
50
results because many of the
variables are linked to the trials, such as: reagents from
different lots; use of different solvents; and the actual
handling by operators, which can interfere in the
uniformity of the results obtained.
During the degradation of contaminants in water,
it is very important to investigate the toxicity profile,
since the intermediates generated might be more toxic
than the parent molecule. For the degraded samples,
toxicity results were expressed as toxicity reduction.
The relationship between EC
50
(dilution needed for
50% of organisms to become motionless) and toxicity
reduction is presented in Equations 8 and 9. Toxicity
reduction was calculated from relative toxicity,
according to Equation 8. Relative toxicity was calculated
using Equation 9 in which EC
50,0
is the dilution
needed for 50% of organisms to become motionless at
the beginning of the degradation test (t = 0 min) and
EC
50,t
is the dilution needed for 50% of organisms to
become motionless at 10, 60, or 600 s.

Toxicity reduction (%) = 100 Relative toxicity (%)
(8)
(9)

Toxicity Reduction During UV/H
2
O
2
Process
The results presented on Table 1, referring to the
bioassays carried out using UV and UV/H
2
O
2
, show
that abamectin concentration and toxicity decrease
over reaction time for both processes evaluated. There
was a relationship between the toxicity parameter and
the abamectin degradation. It might suggest that the
toxicity of the solution is directly related to the level
of abamectin present and the byproducts of degradation
are not highly toxic. Therefore, for the complete
detoxification of abamectin solutions, almost complete
degradation of the drug is necessary.
For UV and UV/H
2
O
2
(5.0 mmol L
-1
H
2
O
2
)
processes, in 600 s of reaction time, the minimum
number of immobilized organisms in the different
solutions (varied dilutions) was not reached when
toxicity tests were carried out. So, it was impossible to
calculate the decrease in toxicity. In these cases, the
solutions were described as not toxic (NT), at least in
the experimental conditions used in this work.
For photolysis, the toxicity removal was 38% and
67% at 10 s and 60 s, respectively. For the same
reaction times, the peroxidation combined with UV
radiation was able to reduce 44% and 70% of the
toxicity. Therefore, both processes showed similar
efficiency regarding toxicity removal.

Toxicity Reduction During Fenton Process
Fenton reagent led to a reduction in toxicity that
was directly proportional to the removal of abamectin
from the solution (Table 2). For example, when the
best conditions were used (1.0 mmol L
-1
Fe(II) and 5.0
mmol L
-1
H
2
O
2
), the values were statistically equal.
Dal Bosco et al. (29) got similar results in a study of
ivermectin degradation by Fenton reagent. They
reported a toxicity decrease of 78% and the ivermectin
degradation of 81% using the same conditions applied
in the present work.
It was verified that when low concentrations of
Fe(II) and H
2
O
2
were used (Fe(II) 0.25 mmol L
-1
and
H
2
O
2
1.0 mmol L
-1
, a molar ratio of 1:4), the reduction
J. R. Guimares et al.
90 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014
Table 2. Toxicity results for the Fenton process (C
o,abamectin
= 500 g L
-1
).

Process Time (s) Degradation (%)
Toxicity
reduction (%)
Standard
deviation (%)
Fenton
(1.0 mmol L
-1
Fe(II)/
5.0 mmol L
-1
H
2
O
2
)
0 0.0 0 -
10 64.1 65 20
60 76.3 71 9
600 89.0 NT -
Fenton
(0.25 mmol L
-1
Fe(II)/
1.0 mmol L
-1
H
2
O
2
)
0 0.0 0 -
10 29.0 36 12
60 67.2 55 17
600 94.7 75 10
Fenton
(0.25 mmol L
-1
Fe(II)/
5.0 mmol L
-1
H
2
O
2
)
0 0.0 0 -
10 29.6 8 7
60 34.6 27 8
600 46.5 29 10
NT non-toxic

Table 3. Toxicity results for the photo-Fenton process (C
o,abamectin
= 500 g L
-1
).

Process
Time
(s)
Degradation (%)
Toxicity
reduction (%)
Standard
deviation (%)
photo-Fenton
(1.0 mmol L
-1
Fe(II)/
5.0 mmol L
-1
H
2
O
2
)
0 0.0 0 0
10 77.3 NT -
60 91.5 NT -
600 96.9 NT -
photo-Fenton
(0.25 mmol L
-1
Fe(II)/
1.0 mmol L
-1
H
2
O
2
)
0 0.0 0 0
10 49.6 41 13
60 90.7 71 14
600 98.6 NT -
photo-Fenton
(0.25 mmol L
-1
Fe(II)/
5.0 mmol L
-1
H
2
O
2
)
0 0.0 0 0
10 52.1 47 15
60 78.9 NT -
600 99.4 NT -
NT non-toxic

in toxicity was different from the degradation efficiency.
In 600 s of reaction, toxicity was reduced by 75%,
while degradation of abamectin was 94%. This may
occurred due to the formation of toxic byproducts.
In the assay using 0.25 mmol L
-1
Fe(II) and 5.0
mmol L
-1
H
2
O
2
, a molar ratio of 1:20, 46% of the drug
was degraded in 600 s and toxicity was reduced by
around 29%. The least efficient removal of the target
compound, as well as reduction of toxicity, can be
attributed to the use of greater H
2
O
2
concentrations,
which led to an excess of hydrogen peroxide in the
medium. In excess, H
2
O
2
acts as an OH radical
scavenger (Equation 8).

Toxicity Reduction During Photo-Fenton Process
The data presented in Figure 6 and on Table 3
shows that the photo-Fenton process was highly
efficient and abamectin was quickly degraded. Clearly,
this is due to the greater concentration of hydroxyl
radicals in the medium, mainly resulting from the
photo regeneration of Fe(III) to Fe(II). The toxicity of
the solution reduced at the beginning of the trials
(Table 3).
The greatest initial concentrations of catalyst and
oxidant (1.0 mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
)
offered the greatest reduction in toxicity: after 10 s of
exposure, the solution was non-toxic to Daphnia
similis. When lower concentration of catalyst was used
(0.25 mmol L
-1
Fe(II)), the toxicity reduction was 41%
and 47% in 10 s, using 1.0 mmol L
-1
and 5.0 mmol L
-1

H
2
O
2
, respectively.
In general, from the results obtained in practically
all of the toxicity trials, it is possible to suggest that all
of the solutions formed by the target compound
(abamectin) and by the intermediates formed during the
degradation processes are less toxic to the Daphnia
similis microcrustacean than the original abamectin
solution.
J. R. Guimares et al.
J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014 91
Conclusions
For the UV and UV/H
2
O
2
processes, there was no
difference in degradation rate and toxicity reduction
for Daphnia similis in 600 s of radiation exposure.
When 1.0 mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
were used for 600 s, Fenton reagent proved to be
efficient at reducing toxicity, resulting in non-toxic
aqueous solution for Daphnia similis. Excess H
2
O
2
in
relation to the amount of catalyst (0.25 mmol L
-1

Fe(II) and 5.0 mmol L
-1
H
2
O
2
) interfered in toxicity
reduction and slowed degradation rates. This happened
because hydroperoxyl radicals were produced, which
compete with the hydroxyl radicals in the reaction.
The photo-Fenton process was the fastest at
reducing toxicity and degrading abamectin. Excessive
concentrations of oxidant and catalyst reduced the
degradation rate. After 600 s of reaction, at all of the
concentrations evaluated, approximately 99% of
abamectin was degraded and the remaining solution
was not toxic to Daphnia similis. Increasing catalyst
and oxidant concentrations in the photo-Fenton
reactions increased reduction of toxicity. Among the
conditions evaluated, the best results were obtained
using 1.0 mmol L
-1
Fe(II) and 5.0 mmol L
-1
H
2
O
2
.
Finally, AOPs and photolysis are efficient at
degrading abamectin in aqueous matrices, reducing
the toxicity of the solutions.

Acknowledgements
The authors would like to thank FAPESP for
financial support (2008/06470-0), and FAPESP (2009/
12641-5) and CAPES (PNPD0233080) for providing
scholarships to I.M. Barbosa and M.G. Maniero.

References
(1) Seelanan, P.; Srisa-Art, M.; Petsom, A.; Nhujak, T.
Anal. Chim. Acta 2006, 570, 8-14.
(2) Disenrens, H.; Henzelin, M. J. Chromatogr. A 1999,
833, 13-18.
(3) Borges, F.A.; Silva, H.C.; Buzzulini, C.; Soares, V.E.;
Santos, E.; Oliveira, G.P.; Costa, A.J. Vet. Parasitol.
2008, 155, 299-307.
(4) Tisler, T.; Erzen, N. K. Ecotoxicology 2006, 15, 495-
502.
(5) Hernando, M.D.; Mezcua, M.; Fernndez-Alba,
A.R.; Barcel, D. Talanta 2006, 69, 334-342.
(6) Vogna, D.; Marotta, R.; Andreozzi, R.; Napolitano,
A.; Ischia, M. Chemosphere 2004, 54, 497-505.
(7) Boonstra, H.; Reichman, E.P.; Brink, P.J. Arch.
Environ. Contam. Toxicol. 2011, 60, 77-89.
(8) Holmstrup, M.; Bindesbol, A.M.; Oosting, G.J.;
Duschl, A.; Scheil, V.; Kohler, H.R.; Loureiro, S.;
Soares, A.M.V.M.; Ferreira, A.L.G.; Kienle, C.;
Gerhardt, A.; Laskowski, R.; Kramarz, P.E.; Bayley,
M.; Svendsen, C.; Spurgeon, D.J. Sci. Total Environ.
2010, 408, 3746-3762.
(9) Schweitzer, N.; Fink, G.; Ternes, T.A.; Duis, K.
Aquat. Toxicol. 2010, 97, 304-313.
(10) Pitterna, T.; Cassayre, J.; Huter, O.F.; Jung, P.M.J.;
Maienfish, P.; Kessabi, F.M.; Quaranta, L.; Tobler,
H. Bioorg. Med. Chem. 2009, 17, 4085-4095.
(11) Garric, J.; Vollat, B.; Duis, K.; Pry, A.; Junker, T.,;
Ramil, M.; Fink, G.; Ternes, T.A. Chemosphere
2007, 69, 903-910.
(12) Sanderson, H.; Laird, B.; Pope, L.; Brain, R.;
Wilson, C.; Johnson, D.; Bryning, G.; Peregrine,
A.S.; Boxall, A.; Solomon, K. Aquat. Toxicol. 2007,
58, 229-240.
(13) Jencic, V.; Cerne, M.; Erzen, N.K.; Kobal, S.; Flajs,
V.C. Ecotoxicology 2006, 15, 249-257.
(14) Erzen, N.K; Kolar, L.; Flajs, V.C.; Kuzner, J.; Marc,
I.; Pogacnik, M. Ecotoxicity 2005, 14, 627-635.
(15) Brewer, B.N.; Armbrust, K.L.; Mead, K.T.; Holmes,
W.E. Rapid Commun. Mass Spectrom. 2004, 18,
1693-1696.
(16) Novelli, A.; Vieira, B.H.; Vasconcelos, A.M.; Peret,
A.C.; Espndola, E.L.G. Ecotoxicol. Environ. Safety
2012, 75, 87-93.
(17) Novelli, A.; Vieira, B.H.; Cordeiro, D.; Cappelini,
L.T.D.; Vieira, E.M.; Espndola, E.L.G. Chemosphere
2012, 86, 36-40.
(18) Neyens, E.; Baeyens, J. J. Hazard. Mater. 2003, 98,
33-50.
(19) Xu, X.R.; Li, H.B.; Wang, W.H.; Gu, J.D. Chemo-
sphere 2004, 57, 595-600.
(20) Nogueira, R.F.P.; Oliveira, M.C.; Paterlini, W.C.
Talanta 2005, 66, 86-91.
(21) ABNT (Associao Brasileira de Normas Tcnicas)
NBR 12713 - Aquatic ecotoxicology - Acute Toxicity
- Test with Daphnia spp (Cladocera, Crustacea) 2009.
(22) Hamilton, M.A.; Russo R.C.; Thurston, R.V.
Environ. Sci. Technol. 1977, 11, 714-719.
(23) Ben, W.; Qiang, Z; Pan, X.; Chen, M. Water Res.
2009, 43, 4392-4402.
(24) Escalada, J.P.; Gianotti, J.; Pajares, A.; Massad,
W.A; Amat-Guerri, F.; Garcia, N.A. J. Agric. Food
Chem. 2008, 56, 7355-7359.
(25) Da Silva, C.R., Maniero, M.G., Rath, S. Guimares,
J.R. J. Adv. Oxid. Technol. 2011, 14, 106-114.
(26) Yuang, F.; Hu, C.; Hu, X.; Qu, J.; Yang, M. Water
Res. 2009, 43, 1766-1774.
(27) Ay, F.; Kargi, F. J. Hazard. Mater. 2010, 179, 622-627.
(28) Catalkaya, E.C.; Kargi, F. J. Hazard. Mater. 2009,
168, 688-694.
(29) Dal Bosco, S.M.; Barbosa, I.M.; Candello, F.P.;
Maniero, M.G.; Rath, S.; Guimares, J.R. J. Adv.
Oxid. Technol. 2011, 14, 292-301.
J. R. Guimares et al.
92 J. Adv. Oxid. Technol. Vol. 17, No. 1, 2014
(30) Elmolla, E.; Chaudhuri, M. J. Hazard. Mater. 2009,
172, 1476-1481.
(31) Fallmann, H.; Krutzler, T.; Bauer, R.; Malato, S.;
Blanco, J. Catal. Today 1999, 54, 309-319.
(32) Fan, C.; Tsui, L.; Liao, M.C. Chemosphere 2011, 82,
229-236.
(33) Nogueira, R.F.P.; Trov, A.G.; Silva M.R.A.; Villa
R.D. Quim. Nova 2007, 30, 400-408.
(34) Klamerth, N.; Rizzo, L.; Malato, R.S.; Maldonado,
M.I.; Aguera, A.; Frnandez-Alba, A.R. Water Res.
2010, 44, 545-554.
(35) Guimares, J.R.; Gasparini, M.C.; Maniero, M.G.;
Mendes, C.G.N. J. Braz. Chem. Soc. 2012, 23, 1680-
1687.
(36) Lu, L.A.; Ma, Y.S.; Kumar, M.; Lin, J.G. Chem.
Eng. J. 2011, 166, 150-156.
(37) Legrini, O.; Oliveros, E.; Braun, A.M. Chemosphere
1993, 93, 698-671.
(38) Buratini, S.V.; Bertoletti, E.; Zagatto, P.A. Bull.
Environ. Contam. Toxicol. 2004, 73, 878-882.
Received for review May 20, 2013. Revised manuscript
received November 6, 2013. Accepted November 10, 2013.