This document discusses the analysis of pesticide degradation pathways. It provides background on commonly used pesticides like diuron, carbendazim, mancozeb, 2,4-D, and their toxicity and environmental impacts. It then describes the microbial and abiotic degradation of these pesticides, including common metabolic pathways and products. The document concludes by describing the mass spectrometry equipment available at CEFSER laboratory that can be used to identify unknown degradation products and quantify targeted compounds to study pesticide degradation kinetics by microorganisms.
This document discusses the analysis of pesticide degradation pathways. It provides background on commonly used pesticides like diuron, carbendazim, mancozeb, 2,4-D, and their toxicity and environmental impacts. It then describes the microbial and abiotic degradation of these pesticides, including common metabolic pathways and products. The document concludes by describing the mass spectrometry equipment available at CEFSER laboratory that can be used to identify unknown degradation products and quantify targeted compounds to study pesticide degradation kinetics by microorganisms.
This document discusses the analysis of pesticide degradation pathways. It provides background on commonly used pesticides like diuron, carbendazim, mancozeb, 2,4-D, and their toxicity and environmental impacts. It then describes the microbial and abiotic degradation of these pesticides, including common metabolic pathways and products. The document concludes by describing the mass spectrometry equipment available at CEFSER laboratory that can be used to identify unknown degradation products and quantify targeted compounds to study pesticide degradation kinetics by microorganisms.
This document discusses the analysis of pesticide degradation pathways. It provides background on commonly used pesticides like diuron, carbendazim, mancozeb, 2,4-D, and their toxicity and environmental impacts. It then describes the microbial and abiotic degradation of these pesticides, including common metabolic pathways and products. The document concludes by describing the mass spectrometry equipment available at CEFSER laboratory that can be used to identify unknown degradation products and quantify targeted compounds to study pesticide degradation kinetics by microorganisms.
Analysis of Pesticide Degradation Pathways Prof. dr. Biljana krbi Faculty of Technology, University of Novi Sad, Novi Sad
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Pesticides are widely used for the control of
weeds, diseases, and pests of cultivated plants all over the world, mainly since after the Second World War.
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At present, around 2.5 million tons of
pesticides are used annually and the number of registered active substances is higher than 500.
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However, as pesticides are toxic
substances that may have undesirable effects, their use has to be regulated.
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Risk assessment requires information
of
pesticides
on the toxicological and
ecotoxicological properties of these compounds as well as on their levels in food and environmental compartments
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Therefore, reliable analytical methods are
needed to carry out the monitoring of pesticide residues in those matrices.
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Consumer concerns on food
safety and society awareness of chemical contaminants in the environment have increased in the past few years. As a consequence, more restrictions in the use of chemical products have been imposed at national and international levels.
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DIURON is a systemic herbicide derived
from urea, relatively persistent in soil, with half-lives from 90 to 180 days. It shows slight acute toxicity is a likely carcinogen is a suspected endocrine disruptor
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Diuron has a very slow rate of natural
hydrolysis in a neutral solution at 25 C. However, when hydrolysis occurs the abiotic degradation in water solution is an irreversible reaction giving 3,4-DCA as the only product.
3,4-DCA
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3,4-DCA (3,4-dichloroaniline) is considerably more toxic than diuron itself and has higher water solubility so it can leach out from treated agricultural land. It could condensate in chloroazobenzene which is highly toxic.
3,4-DCA azobenzene
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Biodegradation is a major way of diuron
degradation in the environment. Many studies were conducted and showed that degradation happens under both aerobic and anaerobic conditions, under the influence of Gram positive and negative bacteria as well as some fungi.
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In a major part of the published studies the
accumulation of 3,4-DCA has been reported. There are some individual microbes capable of mineralizing diuron completely without accumulation of aromatic compounds.
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Bacterial degradation pathways of diuron and 3,4dichloroaniline
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Fungal degradation pathways of
diuron and 3,4-dichloroaniline
CO2
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It must be taken into consideration that
NOT all the mechanisms involved are well known.
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Carbendazim is a systemic fungicide widely
used to control a broad range of fungal diseases in agriculture and forestry.
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Carbendazim has slight acute toxicity is a possible carcinogen is a suspected endocrine disruptor
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It is chemically stable and relatively
persistent in the environment It can harm the liver and endocrine system and has mutagenic and teratogenic effects on animals, even at low concentrations. Carbendazim has a half-life (time taken for half the sample to decay) of 3-12 months.
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Microbial metabolism is the main mechanism
responsible for degradation of carbendazim. It is usually hydrolyzed to 2-AB (2-aminobenzimidazole), BZ (benzimidazole) and 2-HB (2-hydroxybenzimidazole).
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The further degradation of 2-AB or BZ
was not elucidated in earlier studies. Some bacterium strains capable of complete degradation were reported in recent studies.
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Proposed degradation pathway of carbendazim by a bacterium strain
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Mancozeb is a fungicide from a subclass of carbamate pesticides called dithiocarbamates.
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In the soil it negatively affects microbial
decomposition activities, hence the nitrogen and carbon mineralization are affected. Mancozeb is of low soil persistence with half lives from 2 to 8 days. It has moderate mobile potential in soils due to its high adsorption coefficient. In water it can be quickly hydrolyzed.
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The main hydrolysis degradates are
ethylenethiourea (ETU), ethyleneurea (EU) and ethylenebisisothiocyanate sulfide (EBIS).
ETU
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Ethylenethiourea has greater tendency to
be mobile due to its high water solubility and weak adsorption in soil. While mancozeb has a relatively low toxicity, ETU is a suspected human carcinogen is on EU Priority list of endocrine disruptors
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shows toxicity to aquatic organisms and
when administered under various toxicological experimental conditions has been shown or suspected to have caused tumorogenic and teratological effects
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ETU is only weakly adsorbed to soil particles
and therefore its high soil mobility makes it a potential contaminant for groundwater.
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ETU can be degraded by microorganisms
to ethyleneurea (EU), which is further degraded to CO2
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2,4-Dichlorophenoxyacetic acid, more
commonly referred to as 2,4-D, is one of the most widely used herbicides throughout the world.
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It effectively controls unwanted and invasive
weeds across agricultural fields, lawns, public parks, lakes and more. Introduced in 1946, 2,4-D is among the most rigorously researched and regulated molecules of all time.
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It is classified as a moderately hazardous
pesticide when its acute toxicity is concerned. Direct exposure can cause serious eye and skin irritation weakness, diarrhea, respiratory tract irritation... It is considered possibly carcinogenic to humans and an endocrine disruptor.
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2,4-D has been included on the EPA list of
compounds that are likely to leach from soil. Its half life is 1.5 to 16 days. Soil microbes are primarily responsible for its disappearance in soil.
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A number of 2,4-D-degrading bacteria
have been isolated and characterized from a variety of environmental habitats. Incomplete degradation can result in creation of dichlorphenol which is more toxic than 2,4-D.
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In order to be able to elucidate the degradation
pathways and kinetics of pesticides in the soil by selected microorganisms, it is necessary to identify all the products of degradation present in the soil sample.
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Since not all the products can be predicted,
this analytical task presents a screening of unknown contaminants. Highly accurate mass and high resolution data acquisition are necessary requirements for this type of analysis
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CEFSER laboratory for Mass Spectrometry is
equipped with The Exactive-Thermo Fisher Scientific new OrbitrapTM mass spectrometer coupled with Accela U-HPLC chromatograph.
The pre-requisite for the analysis of low concentration compounds in the presence of higher abundant ones is resolution higher than 40 000.
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Pesticide Mixture in Horse Feed Matrix
239.15181 C 11 H 19 O 2 N 4 6.50 ppm
Relative Abundance
100
R = 15,000 Error = 6.50 ppm
80 60 40 20 0 239.00
239.05
239.10
239.20
239.25
239.15033 C 11 H 19 O 2 N 4 0.32 ppm
100 Relative Abundance
239.15 m/z
80
R = 80,000 Error = 0.32 ppm
60 40 20 0 239.00
239.05
239.10
239.15 m/z
239.20
239.25
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Thus, the new benchtop Orbitrap mass
spectrometer we posses in the CEFSER Lab demonstrates high resolving power (up to 100,000), providing precise mass accuracy for complex sample analysis, and
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excellent sensitivity, linearity and selectivity in multi-residue screening of complex matrices
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while a triple quadrupole mass spectrometer TSQ
VantageTM (Thermo Scientific) is capable of highly sensitive full-scan as well as Selected Reaction Monitoring (SRM) scan functions.
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The MS/MS fragmentation pattern is a
powerful tool for obtaining confidence in the identification of the studied compounds.
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With these two instruments, CEFSER lab is
equipped to identify possible unknown metabolites of biodegradation of pesticides by employing the Exactive high resolution MS quantify targeted compounds by TSQ Vantage
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Quantification of the original pesticide
before and after bioaugmentation will reveal the degree of its degradation, and
give
information
degradation kinetics.
about
the
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By untargeted high resolution screening
of the samples, degradation products present in the samples will be identified and the degradation pathways of studied pesticides will be elucidated.
