Environ Monit Assess (2011) 175:227–238

DOI 10.1007/s10661-010-1508-2

Fluroxypyr biodegradation in soils by multiple factors

Ling Tao · Hong Yang

Received: 2 January 2010 / Accepted: 6 May 2010 / Published online: 28 May 2010
© Springer Science+Business Media B.V. 2010

Abstract Fluroxypyr (4-amino-3,5-dichloro-6- the degradation of fluroxypyr. Analysis by gas

fluoro-2-pyridinyl-1-methylheptyl ester) is a chromatography-mass revealed that the reac-
widely used herbicide for controlling weeds, tion in soils might include the removal of 1-
fungi, and insects. However, extensive use of methylheptyl ester to generate fluroxypyr acid
the herbicide has led to its high accumulation in (4-amino-3,5-dichloro-6-fluoro-2-pyridiny). Our
ecosystems and contamination to soils and crops. results provided initial data that a set of biological
Environmental behaviors and fate of herbicides and physiochemical factors coordinately regulates
are dependent on many physiochemical and the decay of fluroxypyr in soils.
biological factors. Whether fluroxypyr is signifi-
cantly affected and how it is degraded under Keywords Fluroxypyr · Degradation · Soil ·
the environmental conditions is largely unknown. Herbicide
The present study investigated the effects of
soil microbe, soil type, dissolved organic matter
(DOM), temperature, soil moisture, and surfac-
tant on fluroxypyr degradation in soils. Appli- Introduction
cation of DOM derived from sludge and straw
to fluroxypyr-contaminated soils increased deg- Pesticides (or herbicides) are a group of agro-
radation of fluroxypyr. Environmental factors chemicals used extensively in farmland to en-
such as temperature, moisture, soil microbe hance the selective chemical control of wild
and soil type could affect the rate of fluroxypyr weeds, insects, and fungi. Although application
dissipation. Also, the microorganism affected of pesticides to soils has greatly improved food
production, they also bring about substantial con-
tamination to soils and aquatic systems. Unlike
L. Tao · H. Yang (B) other hydrophobic contaminants such as poly-
Department of Applied Chemistry, College of
cyclic aromatic hydrocarbons, most of commer-
Science, Nanjing Agricultural University, Nanjing,
210095, China cial pesticides can easily pass into tissues of
e-mail: [email protected] living organisms and give rise to bioaccumula-
tion. Over the last decade, an increasing amount
L. Tao · H. Yang
of pesticides has been detected in both soils
Jiangsu Key Laboratory of Pesticide Science, College
of Science, Nanjing Agricultural University, Nanjing, and waters (European Environment Agency 1999;
210095, China Barbash et al. 2001; Heath et al. 2010). And
228 Environ Monit Assess (2011) 175:227–238

consequently, there is a need to develop more decay rate of pesticides between different soils are
pesticide-accumulating plants and precise meth- expected due to the differential soil properties.
ods to monitor and remediate contaminated Additionally, several other environmental factors
soils. Hence, a comprehensive understanding of such as temperature, moisture, pH, salinity, and
biodegradation or transformation of pesticides in inorganic nutrients also affect biodegradation of
soils is fundamentally important. anthropogenic compounds in soils (Fonseca et al.
Degradation of pesticides is a complex process 2002; Maheswari and Ramesh 2007).
but is closely related to their presence in the aque- Fluroxypyr (4-amino-3,5-dichloro-6-fluoro-2-
ous phase of soils where the organisms involved in pyridinyl-1-methylheptyl ester) is a herbicide
biodegradation are located. The dominant process widely used in agriculture. It is applied to crop
leading to the dissipation of pesticide residues is fields to control a wide range of broadleaf
associated with soil microorganisms (Walker et al. weeds. Fluroxypyr belongs to Toxicity Category
2001). Some soil microbes may completely de- II (United States Environmental Protection
compose xenobiotic compounds (e.g., pesticides) Agency 1998). Although it is considered as “not
or degrade them to the intermediate state which likely” human carcinogen (with an oral LD50 in
is further utilized by other microbe population rats being over 5,000 mg kg−1 ), subchronic toxicity
(Ohshiro et al. 1996; Shabir et al. 2008). Recently, assays in rats show nephrotoxicity, hispathologi-
import of organic carbon to soils in the form of cal lesions, and decreased renal function (EPA,
compost, sludge, and crop residues has become a Fluroxypyr Fact sheet available at http://www.
very popular practice owning to its low costs and epa.gov/opprd001/factsheets/fluroxypyr.pdf). To
recycling of nutrients (Sigua et al. 2005; Majumdar understand its environmental behavior and fate
and Singh 2007). While the solid organic matter in soils, a comprehensive study was performed to
is applied to soils, a large amount of dissolved investigate the effects of biological and environ-
organic matter (DOM) is introduced. DOM in- mental factors including temperature, moisture,
teracts with organic pollutants and exerts a great soil organic matter (SOM), and surfactant on the
effect on mobilization and other behaviors of pes- degradation of fluroxypyr in soils. The arm of the
ticides in soils (Thom et al. 1997; Esposito et al. study is to assist in understanding of how these
1998; Song et al. 2008). Interaction of dissolved factors modulate bioremediation of fluroxypyr-
organic matter with soil pesticides occurs through contaminated soils.
physicochemical adsorption, solubilization, and
hydrolysis (Dunnivant et al. 1992; Kaiser and
Zech 2000; Huang and Lee 2001; Zsolnay 2003). Materials and methods
DOM may improve pesticide mobility in soils
by increasing its water solubility. Conversely, the Pesticide and soil
hydrophobic fraction of DOM may have a high
affinity to pesticide, thus making it into humified Fluroxypyr (purity of 95%) was obtained from the
surface layers and slowing its mobility within soils. Institute of Pesticide Science, Jiang Su Academy
Low bioavailability of pesticides is a major limit to of Agricultural Sciences, Nanjing, China. Three
bioremediation of contaminated soils. types of fluroxypyr-free soils, Eutric gleysols
Besides, degradation of pesticides also depends (EG), Orthic ferralsols (OF), and Eutric his-
on soil properties (e.g., texture and soil com- tosols (EH) were collected from the surface
position) and several other physical or chemi- layer (0–20 cm) at the Experimental Station of
cal factors (Alexander 1999; Kim et al. 2000; Nanjing Agricultural University (Nanjing), Nan-
Huang and Lee 2001; Bhalerao and Puranik 2007; chang (Jiangxi), and Fushun (Liaoning), respec-
Shukla et al. 2009). It is well known that soil is tively. The soils were air-dried and ground and
a dynamic ecosystem composed of heterogeneous passed through a 2-mm mesh sieve for analysis
materials, including various minerals (quartz, sili- of soil properties and degradation experiments.
cates, carbonates, oxides, and clays) and organic The physicochemical natures of the soils were
compounds (e.g., humic acid). Variations in the summarized in Table 1.
Environ Monit Assess (2011) 175:227–238 229

Table 1 Major property of the tested soils used in this study

Soil Organic Texture pH NH4 + –N P K CEC
carbon (%) Sand (%) Silt (%) Clay (%) (mg kg−1 ) (mg kg−1 ) (mg kg−1 ) (cmol kg−1 )
Eutric histosols 16.87 33.74 39.63 26.63 6.49 21.52 37.01 209.78 20.45
Eutric gleysols 2.13 32.12 38.36 29.52 7.58 14.28 34.32 91.45 17.10
Orthic ferralsols 6.42 16.76 27.23 56.01 5.38 7.84 14.81 189.13 6.62

Dissolved organic matter the condition, the retention time for fluroxypyr
was 9.3 min. The recoveries of fluroxypyr spiked
Two types of DOM extracted from sludge (SL) in soils with 0.5–2 mg kg−1 were 91.0–94.1%, with
and straw (ST) were prepared according to the RSD ranging from 1.0% to 1.9%.
previous method (Zhou and Wong 2000). DOM-
ST and DOM-SL were extracted with Milli Q Soil treated with H2 O2 and sterilized
water using a solid: water ratio of 1:10 (w/v) and
shaken at 200 rpm−1 and 4◦ C for 16 h. The sus- To remove SOM, fresh soils (Eutric gleysols,
pension was centrifuged at 6,000×g and 4◦ C for 200 g) were treated with 50 mL 30% H2 O2 based
15 min and filtered through a 0.45-μm sterilized on the method described previously (Khale et al.
membrane (GN-6 Mctrice, Gelman Sciences Ann 2004; Cao et al. 2008). The treated soils were
Arbor, MI). The filtrate was used to analyze total washed with deionized H2 O until the solution
organic carbon (TOC-5000A, Shimadzu, Japan). electric conductivity was below 50 μS (Pu and
DOM-ST and DOM-SL each contain total organic Cutright 2006). For soil sterilization, the same
carbon of 2,103.5 mg DOC L−1 (pH 7.14) and amount (200 g) of fresh Eutric gleysols was sub-
509.8 mg DOC L−1 (pH 6.81), respectively. jected to autoclave treatment. Then, the soils were
pre-incubated at 25 ± 1◦ C for 7 days and thor-
Determination of fluroxypyr in soils oughly mixed with fluroxypyr. Concentration of
fluroxypyr was set at 8 mg kg−1 . The soil moisture
Soils (10 g) in an Erlenmeyer flask were ultra-
sonically extracted with 40 mL acetone–water
(3:1, v/v) solution for 30 min. The extracts were 9 Control
filtrated into a funnel, to which 2 g NaCl was 8 Sterilized
Residual fluroxypyr (mg kg-1 )

SOM-free
added. The mixture was extracted with 40 mL 7
petroleum ether. The water phase was removed, 6
and the organic phase was concentrated to 3 mL
5
on a rotary vacuum evaporator at 40◦ C. The
4
3-mL residue was transferred to a funnel, to
which 2 mL ethanol and 1 mL sulfuric acid 3

(98%) were added. The sulfuric acid phase was 2

discarded, and 2% sodium sulfate was mixed 1
with the organic phase. The mixture was evapo- 0
rated in vacuum to dryness at 40◦ C. Fluroxypyr 0 3 6 9 12 15 18 21 24
in extracts was analyzed using external standard Incubation time (d)
calibration by HPLC (Waters 515, Waters Tech-
Fig. 1 Effect of microorganism and SOM on the degra-
nologies Co. Ltd., USA) with ultraviolet detec- dation of fluroxypyr in Eutric gleysols. Control: fluroxypyr
tor under the operating conditions: wavelength, at 8 mg kg−1 in soil; Sterilized: fluroxypyr at 8 mg kg−1
210 nm; Hypersil reversed phase C18 column (μ in sterilized soil; SOM-free: fluroxypyr at 8 mg kg−1 in
soil treated with H2 O2 . Soils were incubated at 25 ± 1◦ C
Bondpark, 250 mm × 4.6 mm i.d.); mobile phase,
and 60% moisture. The fluroxypyr residues in soils were
methanol/water (85:15; v/v), 0.8 mL min−1 ; injec- quantified at the time intervals of 0, 1, 2, 4, 6, 8, 11, 15, and
tion volume, 20 μL; the temperature, 25◦ C. Under 21 days. Values are the means ± SD (n = 3)
230 Environ Monit Assess (2011) 175:227–238

Table 2 Degradation kinetic parameters of fluroxypyr at 8 mg kg−1 in Eutric gleysols subjected to treatments
Soil treatment Ct = C0 e−kt R2 k DT50 (day)
Control (unsterilized) Ct = 7.7865e−0.1073 t 0.9919 0.1073 6.46
Sterilized Ct = 7.5078e−0.0313 t 0.9606 0.0313 22.14
Microbe Ct = 7.3086e −0.0243 t 0.8139 0.0243 28.52
H2 O2 Ct = 7.7317e−0.0184 t 0.9717 0.0184 37.68
Organic matter Ct = 8.0828e −0.0078 t 0.8439 0.0078 88.85
DOM-ST Ct = 7.5387e−0.1243 t 0.9929 0.1243 5.57
DOM-SL Ct = 7.4742e−0.1318 t 0.9838 0.1318 5.25
1% TX-100 Ct = 7.8366e−0.1057 t 0.9892 0.1057 6.56
2% TX-100 Ct = 7.7095e−0.1051 t 0.9904 0.1051 6.59
Soils were incubated at 25 ± 1◦ C and 60% moisture
DT50 50% dissipation time; R2 correlation coefficient; Ct concentration at a certain moment; C0 initial concentration;
k degradation rate constant

content was maintained at 60% water holding ypyr concentration in soils was set at 8 mg kg−1 ,
capacity during the experiment. Concentrations of and DOM concentrations were set at 20 mg (20,
fluroxypyr in soils were determined at the time 40, 80, and 160 for dose-dependent experiment)
points of 0, 1, 2, 4, 6, 8, 11, 15, and 21 days. DOC kg−1 . The treated soils were incubated in
Each treatment was repeated in triplicate for all darkness at 25 ± 1◦ C for 0, 1, 2, 4, 6, 8, 11, 15, and
experiments. 21 days and fluroxypyr in soils was quantified.

Soil treated with DOM Soil treated with surfactant TX-100

Soils (Eutric gleysols) were pre-incubated at Fresh soils (Eutric gleysols) were pre-incubated at
25 ± 1◦ C for 7 days and mixed with fluroxypyr 25 ± 1◦ C for 7 days. Then, the soils were exposed
and/or DOM (DOM-ST and DOM-SL). Flurox- to fluroxypyr at 8 mg kg−1 and simultaneously

a 8
b 8 0
Control
20
Residual fluroxypyr (mg kg -1 )

7 DOM-ST 7
Residual fluroxypyr (mg kg -1 )

DOM-SL 40
6 6 80
5 5 160

4 4

3 3
2 2
1 1
0 0
0 3 6 9 12 15 18 21 24 0 3 6 9 12 15 18 21 24
Incubation time (d) Incubation time (d)

Fig. 2 Effect of DOM on the degradation of fluroxypyr in soil containing DOM-SL at 0, 20, 40, 60, 80, and 160 mg
in Eutric gleysols. a control: fluroxypyr at 8 mg kg−1 in DOC kg−1 . The soils were incubated at 25 ± 1◦ C and
soil; ST fluroxypyr at 8 mg kg−1 in soil with DOM-ST at 60% moisture. The fluroxypyr residues in the soils were
20 mg DOC kg−1 ; SL fluroxypyr at 8 mg kg−1 in soil with quantified at the time intervals of 0, 1, 2, 4, 6, 8, 11, 15,
DOM-SL at 20 mg DOC kg−1 ; b fluroxypyr at 8 mg kg−1 and 21 days. Values are the means ± SD (n = 3)
Environ Monit Assess (2011) 175:227–238 231

Table 3 Degradation kinetic parameters of fluroxypyr at 8 mg kg−1 in Eutric gleysols with varied amounts of DOM-SL at
25 ± 1◦ C and 60% soil moisture
Content of DOM (mgDOC kg−1 ) Ct = C0 e−kt R2 k DT50 (day)
0 Ct = 7.7865e−0.1075 t 0.9919 0.1073 6.46
20 Ct = 7.4742e−0.1318 t 0.9838 0.1318 5.25
40 Ct = 7.2393e−0.1395 t 0.9703 0.1395 4.95
80 Ct = 7.0592e−0.1400 t 0.9744 0.1400 4.93
160 Ct = 7.1173e−0.1548 t 0.9936 0.1548 4.46
DT50 50% dissipation time; R2 correlation coefficient; Ct concentration at a certain moment; C0 initial concentration;
k degradation rate constant

treated with surfactant TX-100 at 1% and 2% Soil treated with different moisture
(w/w), respectively. The treated soils were incu-
bated in darkness at 25 ± 1◦ C for 0, 1, 2, 4, 6, 8, 11, Soils (Eutric gleysols) were incubated at 25 ± 1◦ C
15, and 21 days, and the fluroxypyr concentration for a week with moisture contents of 20%, 40%,
in soils was determined. 60%, and 80%, respectively. The soils then were
treated with 8 mg kg−1 fluroxypyr and incubated
under the same conditions (20%, 40%, 60%, and
Soil treated with different temperature 80% soil moisture contents, respectively) in dark-
ness at 25 ± 1◦ C for 0, 1, 2, 4, 6, 8, 11, 15, and
Soils (Eutric gleysols) were pre-incubated at 25 21 days. Finally, the soil samples were used for
± 1◦ C for 7 days. Afterwards, the soils were fluroxypyr determination.
treated with 8 mg kg−1 fluroxypyr and incubated
in darkness at 15 ± 1◦ C, 25 ± 1◦ C, and 35 ±
Degradation of fluroxypyr in different soil types
1◦ C, respectively. The fluroxypyr concentrations
in soils were determined at the time intervals of 0,
The fresh Eutric gleysols, Orthic ferralsols, and
1, 2, 4, 6, 8, 11, 15, and 21 days.
Eutric histosols were pre-incubated at 25 ± 1◦ C

8
15 ˚C
8 0
7 25˚C
1%
Residual fluroxypyr (mg kg -1 )

7
Residual fluroxypyr (mg kg-1 )

35˚C
2% 6
6
5
5
4 4

3 3

2 2

1 1

0 0
0 3 6 9 12 15 18 21 24 0 3 6 9 12 15 18 21 24
Incubation time (d) Incubation time (d)

Fig. 3 Effect of surfactant TX-100 on the degradation of Fig. 4 Effect of temperature on the degradation of
fluroxypyr in Eutric gleysols. Soils contained fluroxypyr fluroxypyr in Eutric gleysols. Soil containing fluroxypyr at
at 8 mg kg−1 and TX-100 at 0%, 1%, and 2% and were 8 mg kg−1 was incubated with 60% moisture and 15 ±
incubated at 25 ± 1◦ C and 60% moisture. The fluroxypyr 1◦ C, 25 ± 1◦ C, and 35 ± 1◦ C, respectively. The fluroxypyr
residues in the treated soils were quantified at the time residues in treated soils were quantified at the time inter-
intervals of 0, 1, 2, 4, 6, 8, 11, 15, and 21 days. Values are vals of 0, 1, 2, 4, 6, 8, 11, 15, and 21 days. Values are the
the means ± SD (n = 3) means ± SD (n = 3)
232 Environ Monit Assess (2011) 175:227–238

Table 4 Degradation kinetic parameters of fluroxypyr at 8 mg kg−1 in Eutric gleysols at different temperature
Temperature (◦ C) Ct = C0 e−kt R2 k DT50 (day)
15 Ct = 7.3686e−0.0860 t 0.9781 0.0860 8.06
25 Ct = 7.7865e−0.1075 t 0.9919 0.1073 6.46
35 Ct = 7.5945e−0.1191 t 0.9569 0.1191 5.81
Soils were constantly incubated with 60% moisture
DT50 50% dissipation time; R2 correlation coefficient; Ct concentration at a certain moment; C0 initial concentration;
k degradation rate constant

for 7 days. After that, the soils were exposed to diluted sample of 1 μL was injected in the splitless
fluroxypyr 8 mg kg−1 and incubated in darkness mode. The parameters of mass detector were set
at 25 ± 1◦ C for 0, 1, 2, 4, 6, 8, 11, 15, and 21 days. at ion mass/charge ratio of 20–550 m/z, with a
Following the treatment, fluroxypyr concentra- scan model. The identity of some selected volatile
tions soils were determined. compounds was verified by comparing their mass
spectra and retention time with those obtained for
FT-IR spectral analysis of soils authentic standard compounds.

Dried Eutric gleysols, Orthic ferralsols, and Eu-

tric histosols were analyzed by a Bruker Tensor- Results and discussion
27 infrared spectrophotometer (Bruker Optics
Inc., Germany). FT-IR spectra of three soils were Effects of microbe and organic matter
recorded ranging from 4,000 to 400 cm−1 , and data on fluroxypyr degradation in soils
were obtained from a 200-mg KBr disk containing
1 mg of the sample. Eutric gleysols were treated with 30% H2 O2 to
remove SOM and subjected to sterilization by
autoclave, respectively. The soil microbe and or-
Analysis of fluroxypyr degradation products
ganic matter affected significantly the degradation
of fluroxypyr (Fig. 1). Compared to the treat-
Soils (10 g) were exposed to fluroxypyr
(8 mg kg−1 ) for 6 days and transferred to an
Erlenmeyer flask. The sample was ultrasonically
extracted with 40 mL acetone–water (3:1, v/v) 8 20%
40%
solution for 30 min. The extract was filtrated 7
Residual fluroxypyr (mg kg -1 )

60%
into a funnel, to which 2 g NaCl was added. The 6
80%
mixture was extracted with 40 mL petroleum 5
ether. The organic phase was concentrated to
4
dryness on a rotary vacuum evaporator at 40◦ C.
3
The degradation products were analyzed by
Shimadzu 2010 gas chromatography-mass detec- 2
tor system (Shimadzu, Japan). An RTX-5MS cap- 1
illary column (30 m × 0.125 mm i.d., 0.25 μm 0
film thickness) was used under the following in- 0 3 6 9 12 15 18 21 24
strumental conditions: helium flow, 50 mL min−1 ; Incubation time (d)

injector temperature, 250◦ C; transfer line tem- Fig. 5 Effect of moisture on the degradation of fluroxypyr
perature, 280◦ C; energy of electron, 70 eV. The in Eutric gleysols. Soil containing fluroxypyr at 8 mg kg−1
temperature program for SPME procedure was was incubated at 25 ± 1◦ C, each with 20%, 40%, 60%,
and 80% moisture, respectively. The fluroxypyr residues
set as follow: initial 80 (1 min) to 140◦ C at ramp
in treated soils were quantified at the time intervals of 0, 1,
rate of 20◦ C min−1 ; from 130◦ C to 280◦ C at ramp 2, 4, 6, 8, 11, 15, and 21 days. Values are the means ± SD
rate of 10◦ C min−1 and 280◦ C for 3 min. The (n = 3)
Environ Monit Assess (2011) 175:227–238 233

Table 5 Degradation kinetic parameters of fluroxypyr at 8 mg kg−1 in Eutric gleysols at 25 ± 1◦ C with different moisture
Moisture (%) Ct = C0 e−kt R2 k DT50 (day)
20 Ct = 7.2127e−0.0657 t 0.9934 0.0657 10.50
40 Ct = 7.3686e−0.0860 t 0.9981 0.0860 8.02
60 Ct = 7.7865e−0.1075 t 0.9919 0.1073 6.46
80 Ct = 7.4606e−0.1181 t 0.9961 0.1181 5.84
DT50 50% dissipation time; R2 correlation coefficient; Ct concentration at a certain moment; C0 initial concentration;
k degradation rate constant

ment with H2 O2 or sterilization, the untreated DOM-SL (Table 3). There was a tendency that
fluroxypyr (control) decay in soils was more pro- treatment with high amounts of DOM resulted
nounced. By the end of the incubation (21 days), in more fluroxypyr degradation (Fig. 2b), sug-
the concentration of fluroxypyr residues in control gesting that DOM was able to affect fluroxypyr
soils was 0.77 mg kg−1 , whereas the contents of dissipation. It was possible that DOM modulated
fluroxypyr residues in H2 O2 -treated and sterilized microbial activities through their active organic
soils were 5.12 and 3.78 mg kg−1 , respectively. fractions (Said-Pullicino and Gigliotti 2007; Jiang
Also, the 50% dissipation time (DT50 ) for the et al. 2008; Song et al. 2008). Therefore, addition
control soils was 6.46 days, whereas DT50 values of DOM to soils could facilitate the decay of
for H2 O2 - and autoclave-treated soils were 37.68 fluroxypyr.
and 22.14 days, respectively, showing the more
rapid decay rate of fluroxypyr in control soils Effect of surfactant on fluroxypyr
(0.1073; Table 2). These results indicated that soil degradation in soils
fluroxypyr degradation largely depended on the
availability of soil bacteria. Microorganisms such Recent studies have demonstrated that surfac-
as Pseudomonas and Actinomycetes species are tants enhance the solubilization of organic conta-
the predominant microbe for degradation of toxic minants in polluted soils and thus improve their
organic substances because these bacteria use the bioavailability (Mulligan et al. 2001; Yu et al.
compounds as a source of nutrients (Gupta and 2007; Cao et al. 2008). In this study, however,
Baummer 1996; Jung et al. 2005). In this study,
fluroxypyr dissipated very slowly when soils were
treated with H2 O2 , suggesting that removal of soil 8 EG
microbe and organic matter considerably affected 7 OF
Residual fluroxypyr (mg kg-1 )

EH
the fluroxypyr decay in soils.
6

5
Effect of DOM on fluroxypyr degradation in soils 4

3
Fluroxypyr in soils with 20 mg DOC kg−1 of
2
DOM from straw and sludge was degraded more
quickly than the control (Fig. 2a). Both DOM 1
had a similar effect on the decay of fluroxypyr. 0
The dissipative DT50 for fluroxypyr in the con- 0 3 6 9 12 15 18 21 24
Incubation time (d)
trol soil, and soils with DOM-ST and DOM-
SL were 6.47, 5.57, and 5.25 days, respectively Fig. 6 Effect of soil type on the degradation of fluroxypyr.
(Table 2). To understand further, the effect of EG Eutric gleysols with 8 mg kg−1 fluroxypyr; OF Orthic
DOM on fluroxypyr degradation, a concentration- ferralsols with 8 mg kg−1 fluroxypyr; EH Eutric histosols
with 8 mg kg−1 fluroxypyr. The soils were incubated at
dependent experiment with DOM (0, 20, 40, 80,
25 ± 1◦ C and 60% moisture. The fluroxypyr residues in
and 160 mg DOC kg−1 ) was performed. The soils were quantified at the time intervals of 0, 1, 2, 4, 6,
fluroxypyr DT50 decreased with concentrations of 8, 11, 15, and 21 days. Values are the means ± SD (n = 3)
234 Environ Monit Assess (2011) 175:227–238

Table 6 Degradation kinetic parameters of fluroxypyr at 8 mg kg−1 in the three types of soils at 25 ± 1◦ C and 60% moisture
Soil type Ct =C0 e−kt R2 k DT50 (day)
Eutric histosols Ct = 7.9644e−0.1404 t 0.9869 0.1404 4.49
Eutric gleysols Ct = 7.7865e−0.1075 t 0.9919 0.1073 6.46
Orthic gerralsols Ct = 7.9701e−0.1275 t 0.9874 0.1275 5.44
DT50 50% dissipation time; R2 correlation coefficient; Ct concentration at a certain moment; C0 initial concentration;
k degradation rate constant

application of surfactant TX-100 at 1% and 2% At 15◦ C, fluroxypyr degradation was relatively

did not appear to affect the fluroxypyr dissipation slow, but an increase of temperature up to 25–
in soils (Fig. 3). Also, no significant difference was 35◦ C resulted in enhanced fluroxypyr degrada-
observed in DT50 of fluroxypyr in soils with TX- tion. Under the conditions, soils at 25◦ C or 35◦ C
100 at 0%, 1%, and 2%, respectively (Table 2), had a relatively lower DT50 for fluroxypyr dissipa-
indicating that degradation of fluroxypyr in soils tion (Table 4). Notably, the DT50 for fluroxypyr
was not changed by TX-100. decay showed only a small change (from 6.46 to
5.81 days) when the temperature changed from
Effect of temperature on fluroxypyr 25◦ C to 35◦ C, which were supposed to be the
degradation in soils optimal temperature for soil microbe activity.

In Eutric gleysols incubated at 15◦ C, 25◦ C, and Effect of soil moisture on fluroxypyr degradation
35◦ C, no difference of fluroxypyr degradation in
the treatments was observed during the first 2 days Fluroxypyr degradation in soils was also affected
(Fig. 4). Following that time, however, the soils by the soil water capacity. The dissipation rate
with 35◦ C displayed a rapid decay of fluroxypyr of fluroxypyr in Eutric gleysols with 80% mois-
relative to soils with 15◦ C or 25◦ C. Also, soils ture content was higher than any other moisture
incubated at 25◦ C had greater fluroxypyr dissi- treatments (Fig. 5). By the end of the experiment
pation than soils at 15◦ C after 8 days, when a (21 days), a negative relationship between the
positive correlation between the fluroxypyr degra- residual concentrations of fluroxypyr and mois-
dation and temperature was observed. The ob- ture treatments could be established. The nega-
servation was supported by the data of DT50 , tive correlation was also found between DT50 and
which showed 8.06, 6.46, and 5.81 days for 15◦ C, moisture contents (Table 5), with a correlation
25◦ C, and 35◦ C, respectively (Table 4); also, the coefficient of 0.9332. Apparently, soil with 20%
residual fluroxypyr concentrations in soils with moisture content had the longest half time for
15◦ C, 25◦ C, and 35◦ C for 21 days were 1.21, 0.77, fluroxypyr decay, indicating that a low water hold-
and 0.59 mg kg−1 , respectively. These results in- ing capacity is a limit to fluroxypyr degradation.
dicated temperature is one of the most impor-
tant factors that influence microbial growth and Effect of soil type on fluroxypyr degradation
biological reactions (Zavala et al. 2004; Kurola
and Salkinoja-Salonen 2007; Guo et al. 2008) and Three types of soils, Eutric histosols, Orthic fer-
consequently the fluroxypyr degradation (Fig. 1). ralsols, and Eutric gleysols, were tested for in-

Table 7 Correlation Soil properties Linear equation Correlation coefficient R2

between the degrading
half-life of fluroxypyr and Organic carbon y = −0.1257x + 6.5282 0.9353
soil properties pH y = 0.4596x + 2.4836 0.2633
CEC y = −0.029x + 5.8899 0.0450
NH4 + y = −0.0736x + 6.5343 0.2520
Phosphorus y = −0.0074x + 5.6749 0.0028
R2 correlation coefficient Potassium y = −0.0147x + 7.8661 0.8894
Environ Monit Assess (2011) 175:227–238 235

Fig. 7 Differential FT-IR

spectra of three soils:
Eutric gleysols (EG),
Orthic Ferralsols (OF),
and Eutric Histosols
(EH). FT-IR spectra of
three soils were recorded
at room temperature
within the range
4,000–400 cm−1 . The
spectra of the samples
were obtained from a
200-mg KBr disk
containing 1 mg of the
sample

vestigating the possible effect on the degradation properties and fluroxypyr degradation revealed
of fluroxypyr. Soils were exposed to 8 mg kg−1 that the soil organic carbon was closely related
fluroxypyr for 21 days. The residual concentra- to the fluroxypyr decay (correlation coefficient
tions of fluroxypyr in Eutric histosols, Orthic fer- 0.9353; Table 7). On the other hand, several other
ralsols, and Eutric gleysols were shown to be 0.42, factors such as pH, CEC, NH4 + , and phosphorus
0.55, and 0.77 mg kg−1 , respectively (Fig. 6), and nutrition showed the least relevance to the degra-
the degradation DT50 for the soils were 4.49, 5.44, dation of herbicide.
and 6.46 days, respectively (Table 6), indicating To get insights into the properties of the soils
that Eutric histosols had the highest capability for that affected the fluroxypyr degradation, we ana-
fluroxypyr dissipation. lyzed the FT-IR spectra of the soils, which showed
Because Eutric histosols had a relatively higher an adsorption area within 4,000∼3,000 cm−1 and
level of organic carbons than the other soils a peak at 1,634 cm−1 . Eutric histosols had the
(Table 1), it was possible that the abundance higher adsorption in the areas than Orthic ferral-
of organic carbons would be responsible for sols and Eutric gleysols (Fig. 7), suggesting that
the rapid decay of fluroxypyr (Haberhauer and the EH contained more –OH functional groups
Gerzabek 1999; Jung et al. 2008). In addition, and unsaturants compared to the OF and EG. We
the short dissipation half time of fluroxypyr in also found several other differences of absorption
Eutric histosols was 4.49 days, whereas the DT50 among the soils. For instance, the absorption peak
for Eutric gleysols and Orthic ferralsols were 6.46 at 3,622∼3,621 cm−1 represents the N–H stretch-
and 5.44 days, respectively (Table 6), suggesting ing vibration signal; however, only in EG and
that the soil organic carbon would predominantly OF was the peak detected (Table 8). Also, the
affect the fluroxypyr decay in soils. Taken to- absorption peak at 1,384 cm−1 representing C–
gether, analysis of the correlation between the soil H bending vibration signal was detected only in

Table 8 Major infrared absorption peaks in the FT-IR spectra of soils

Soil type Main infrared absorption frequencies (cm−1 )
Eutric histosols – 3,422 1,634 1,384 1,033 – 779 531 469
Eutric gleysols 3,622 3,442 1,635 1,384 1,033 913 796 534 470
Orthic gerralsols 3,621 3,430 1,632 1,033 – 778 532 470
236 Environ Monit Assess (2011) 175:227–238

EH and EG but not in OF. Additionally, EG was by GC-MS and its total ion chromatogram of
tested to have a weak signal at 913 cm−1 , which GC-MS was shown in Fig. 8. The standard
suggests the C–H bend vibration absorption peak fluroxypyr chromatogram displayed a retention
on aromatic structure (Guo et al. 2008). time at 18.041 min (Fig. 8a), which was confirmed
by the presence of ions with m/z = 366 (molecular
Fluroxypyr degradation pathway in soils ion peak), m/z = 254, m/z = 209, and m/z = 181
(Fig. 8b). Compared to the standard fluroxypyr
Extracts from Eutric gleysols exposed to total ion chromatogram, the chromatographic
8 mg kg−1 fluroxypyr for 6 days were determined peak at 16.402 min could be the signal of

Fig. 8 Analysis of
fluroxypyr degradation
products by total ion
chromatogram of
GC-MS. The materials
extracted from Eutric
gleysols were exposed to
fluroxypyr at 8 mg kg−1
and incubated at 25 ±
1◦ C and 60% moisture
content for 6 days (a).
The MS spectra of
fluroxypyr (b) and
fluroxypyr degradation
product (c), and the
degradation pathway (d)
were presented. The
retention times of
fluroxypyr and
degradation product of
were 18.041 and
16.402 min, respectively.
The molecular ion peak
of fluroxypyr was m/z =
366. Degradation product
was fluroxypyr acid
(m/z = 254, molecular ion
peak)
Environ Monit Assess (2011) 175:227–238 237

fluroxypyr degradation product (A), fluroxypyr isolate, Aspergillus niger. International Biodeteriora-
acid. Its structure was confirmed by the presence tion and Biodegradation, 59, 315–321.
Cao, J., Guo, H., Zhu, H. M., Jiang, L., & Yang, H.
of molecular ion peak m/z = 254 (molecular (2008). Effects of SOM, surfactant and pH on the
ion peak) and some fragment ions: m/z = 209 sorption-desorption and mobility of prometryne in
and m/z = 181 (Fig. 8c). The fluroxypyr in soil soils. Chemosphere, 70, 2127–2134.
might be degraded to fluroxypyr acid by losing Dunnivant, F. M., Jardine, P. M., Taylor, D. L., &
Mccarthy, J. F. (1992). The cotransport of cadmium,
1-methylheptyl ester (Fig. 8d).
2,2 ,4,4 ,5,5 -hexachlorobiphenyl, and 2,2 ,4,4 ,6,6 -
hexachlorobiphenyl by dissolved organic carbon
in laboratory columns containing aquifer material.
Conclusions Environmental Science and Technology, 26, 360–368.
Esposito, E., Paulillo, S. M., & Manfio, G. P. (1998).
Biodegradation of the herbicide diuron in soil by in-
Effects of soil microbe, SOM, temperature, digenous actinomycetes. Chemosphere, 37, 541–548.
moisture, and surfactant on the degradation of European Environment Agency (1999). Environmental as-
fluroxypyr in soils were investigated over the sessment report no.3, Groundwater quality and quan-
tity in Europe. Copenhagen: European Environment
times of 0, 1, 2, 4, 6, 8, 11, 15, and 21 days, respec- Agency.
tively. Application of DOM of sludge and straw to Fonseca, J. C., Marques, J. C., & Madeira, V. M. C. (2002).
fluroxypyr-contaminated soils induced the decay Degradability and sediment sorption of an alcohol
of fluroxypyr. The dissipation rate was increased polyglycol ether surfactant putatively useful for the
control of red swamp crayfish in rice fields. Environ-
with the amount of DOM added. Environmen- mental Monitoring and Assessment, 75, 1–10.
tal factors such as temperature, moisture, and Guo, H., Zhu, H. M., & Yang, H. (2008). Degradation and
soil microbe and soil type also affected the rate adsorption behavior of napropamide in soils. Environ-
of fluroxypyr decomposition. The microorganism mental Science, 29, 1729–1736, (Chinese).
Gupta, G., & Baummer, J. (1996). Biodegradation of
was effective on the degradation of fluroxypyr. atrazine in soil using poultry litter. Journal of Haz-
SOM played an important role in affecting degra- ardous Materials, 45, 185–192.
dation of fluroxypyr. Finally, the degradation re- Haberhauer, G., & Gerzabek, M. H. (1999). Drift
action for fluroxypyr showed that 1-methylheptyl and transmission FT-IR spectroscopy of forest
soils: An approach to determine decomposition
ester was removed from fluroxypyr to generate processes of forest litter. Vibrational Spectroscopy, 19,
fluroxypyr acid (4-amino-3,5-dichloro-6-fluoro-2- 413–417.
pyridiny). Our research provided the evidence Heath, E., Ščančar, J., Zuliani, T., & Milačič, R. (2010).
that a set of multiple biological and physio- A complex investigation of the extent of pollution in
sediments of the Sava River: Part 2: Persistent or-
chemical determinants can affect the degrada- ganic pollutants. Environmental Monitoring and As-
tion of fluroxypyr, which is necessary to develop sessment, 163, 277–293.
cost-effective techniques for bioremediation of Huang, X., & Lee, L. S. (2001). Effects of dissolved
fluroxypyr-contaminated soils. organic matter from animal waste effluent on chlor-
pyrifos sorption by soils. Journal of Environmental
Quality, 30, 1258–1265.
Acknowledgement This study was financially supported Jiang, L., Huang, J., Liang, L., Zheng, P. Y., & Yang, H.
by the National Natural Science Foundation of China (No. (2008). Mobility of prometryne in soil as affected by
20777037) and the Fundamental Research Funds for the dissolved organic metter. Journal of Agricultural and
Central Universities (KYZ200918). Food Chemistry, 56, 11933–11940.
Jung, H., Ahn, Y., Choi, H., & Kim, I. S. (2005). Effects
of in-situ ozonation on indigenous microorganisms
in diesel contaminated soil: Survival and regrowth.
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