Metabolism Of Imidacloprid Research Articles

Biodegradation of imidacloprid and diuron by Simplicillium sp. QHSH-33

Imidacloprid (IMI) and diuron (DIU) are widely used pesticides in agricultural production. However, their excessive use and high residues have caused harm to the ecological environment and human health. Microbial remediation as an efficient and low-toxic method has become a research hotspot for controlling environmental pollutants. A fungus QHSH-33, identified as Simplicillium sp., has the ability to degrade neonicotinoids IMI and phenylurea DIU. When QHSH-33 and pesticide were co-cultured in liquid medium for 7 days, the degradation rates of IMI and DIU by QHSH-33 in simulated field soil microenvironment were 50.19 % and 70.57 %, respectively. Through HPLC-MS analysis, it was found that the degradation of IMI mainly involved nitro reduction, hydroxylation and other reactions. Three degradation pathways and eight degradation products were identified, among which two metabolites were obtained by microbial transformation of IMI for the first time. The degradation of DIU mainly involved demethylation and dehalogenation reactions, and two degradation pathways and four degradation products were identified, one of which was a new degradation product of DIU. Toxicity assessment demonstrated that most of the degradation products might be considerably less harmful than IMI and DIU. Whole genome sequencing of QHSH-33 revealed a genome size of 33.2 Mbp with 11,707 genes. The genome of QHSH-33 was annotated by KEGG to reveal 128 genes related to exogenous degradation and metabolism. After local blast with reported IMI and DIU degrading enzymes, seven IMI-degrading related genes and seven DIU-degrading related genes were identified in the QHSH-33 genome. The results of this study will help to expand our knowledge on the microbial decomposition metabolism of IMI and DIU, and provide new insights into the degradation mechanism of IMI and DIU in soil and pure culture system, laying a foundation for QHSH-33 strain applied to the removal, biotransformation or detoxification of IMI and DIU.

Insights into the occurrence, distribution and dissipation of widespread agrochemicals in celery agrosystems for joint risk assessment

Elucidating exposure risks associated with the most widely used agrochemicals and their metabolites in celery agrosystems are vital for food safety and human health. The occurrence, distribution, dissipation and metabolism of imidacloprid (IMI), acetamiprid (ACE), thiamethoxam (THM) and difenoconazole (DIF) in celery tissues reflected by initial depositions, uptake characteristics, half-lives, concentration variations. DIF exhibited unacceptable ecological risk to soil organisms under multi-risk evaluation models, including toxicity exposure ratio, risk quotient, and BITSSD model. The joint dietary risks of target pesticides were 37.273–647.454% and 0.400–2522.016% based on deterministic and probabilistic models, with non-carcinogenic risks of 30.207–85.522% and 1.229–2524.662%, respectively. Children aged 1–6 years suffered the highest exposure, with the leaves posing higher risk than other tissues. Long-term exposure risks should be continuously assessed for ecological sustainability and human health, given the widespread usage and cumulative effects of target pesticides, especially for rural children.

CYP4CE1 Metabolized Nitenpyram through Two Types of Oxidation Reaction, Hydroxylation, and N-Demethylation.

Nitenpyram, taking the place of imidacloprid, is a widely used neonicotinoid insecticide to control Nilaparvata lugens in Asia. Two P450s, CYP4CE1 and CYP6ER1, are key factors in the metabolic resistance against nitenpyram and imidacloprid. In this study, we found that CYP4CE1 expression was strongly associated with nitenpyram resistance in 8 field-collected populations, whereas CYP6ER1 expression correlated with imidacloprid resistance. Hence, we focused on nitenpyram metabolism by CYP4CE1, due to that imidacloprid metabolism by CYP6ER1 has intensively investigated. Mass spectrometry analysis revealed that recombinant CYP4CE1 metabolized nitenpyram into three products, N-desmethyl nitenpyram, hydroxy-nitenpyram, and N-desmethyl hydroxy-nitenpyram, with a preference for hydroxylation. In contrast, CYP6ER1 metabolized nitenpyram into a single product, N-desmethyl nitenpyram. These results provide new insights into the specific catalytic mechanisms of P450 enzymes in neonicotinoid metabolism and underscore the importance of different catalytic reactions in neonicotinoid insecticide resistance.

Heterologous expression and characterization of two delta glutathione S-transferases genes involved in imidacloprid metabolism in Grapholita molesta

Glutathione S-transferases (GSTs) are multifunctional enzymes, and insect GSTs play a pivotal role in the metabolism of insecticides. Grapholita molesta is a worldwide pest that causes substantial economic losses to the fruit industry. However, it remains unclear how imidacloprid, a commonly used insecticide in orchards, is metabolized by G. molesta. In the present study, the synergist diethyl maleate (DEM), which inhibits the GST activity, exhibited a 22-fold synergistic ratio against imidacloprid. Two new GST genes, GmGSTD2 (OR096251) and GmGSTD3 (OR096252), were identified and successfully cloned, showing the highest expression in the Malpighian tubes. Knockdown of GmGSTD2 and GmGSTD3 by RNA interference, increased the mortality of G. molesta from 28% to 47% following imidacloprid treatment. Both recombinant GmGSTD2 and GmGSTD3 proteins exhibited 1-chloro-2,4-dinitrobenzene (CDNB) activity and could be inhibited by imidacloprid in vitro, with maximum inhibition was 60% for GmGSTD2 and 80% for GmGSTD3. These results suggested that GSTs participate in the metabolism of imidacloprid with GmGSTD2 and GmGSTD3 playing key roles in this process.

Exogenous tannic acid relieves imidacloprid-induced oxidative stress in tea tree by activating antioxidant responses and the flavonoid biosynthetic pathway

Pesticide stress on plants is receiving increased scrutiny due to its effect on plant secondary metabolism and nutritional quality. Tannic acid (TA) is a natural polyphenolic compound showing excellent antioxidant properties and is involved in alleviating stress. The present study thoroughly investigated the effects and mechanism of exogenous TA on relieving imidacloprid (IMI) stress in tea plants. Our research found that TA(10 mg/L) activated the antioxidant defense system, enhanced the antioxidant ability, reduced the accumulation of ROS and membrane peroxidation, and notably promoted tea plant tolerance to imidacloprid stress. Additionally, TA boosted photosynthetic capacity, strengthened the accumulation of nutrients. regulated detoxification metabolism, and accelerated the digestion and metabolism of imidacloprid in tea plants. Furthermore, TA induced significant changes in 90 important metabolites in tea, targeting 17 metabolic pathways through extensively targeted metabolomics. Specifically, TA activated the flavonoid biosynthetic pathway, resulting in a 1.3- to 3.1-fold increase in the levels of 17 compounds and a 1.5- to 63.8-fold increase in the transcript level of related genes, such as ANR, LAR and CHS in this pathway. As a potential tea health activator, TA alleviates the oxidative damage caused by imidacloprid and improves the yield and quality of tea under pesticide stress.

Ovarian antral follicles metabolize imidacloprid in vitro.

Neonicotinoid insecticides are synthetic nicotine derivatives that have high affinity for invertebrate nicotine receptors and low affinity for mammalian nicotine receptors. However, imidacloprid (IMI), the most commonly used neonicotinoid, can be bioactivated by the liver in mammals to desnitro-imidacloprid, an intermediate metabolite that effectively binds and activates mammalian receptors. However, it is not known if other tissues such as the ovaries can metabolize IMI. Thus, the present study tested the hypothesis that ovarian antral follicles metabolize and bioactivate IMI. Antral follicles were dissected from the ovaries of CD-1 mice and cultured in media containing dimethyl sulfoxide or IMI (0.2-200 µg/ml) for 48 and 96 h. Media were subjected to liquid chromatography-mass spectrometry for detection of phase I IMI metabolites. Follicles from the cultures were used for gene expression analysis of metabolic enzymes associated with IMI metabolism. All IMI metabolites were detected at 48 and 96 h. Oxidized IMI intermediates were detected in media from cultured follicles, but not environmental controls. Reduced IMI intermediates were detected in media from cultured follicles and the environmental controls. At 48 h, IMI did not affect expression of any metabolic enzymes compared with control. At 96 h, IMI induced Cyp2e1 and Cyp4f18 compared with control. These data indicate that mouse ovarian follicles metabolize IMI and that IMI induces ovarian Cyp expression over time.

Insights into Free and Conjugated Forms of Neonicotinoid Insecticides in Human Serum and Their Association with Oxidative Stress.

Following exposure, neonicotinoid insecticides (NEOs) can be metabolized by both Phase I and Phase II reactions catalyzed by human cytochrome P450 enzymes. However, toxicities of parent NEOs and their metabolites are still unclear, and little is known about biotransformation rates and pathways of NEOs in humans. In this study, 98 serum samples collected in China were analyzed for free, conjugated and total forms of six parent NEOs (i.e., acetamiprid (ACE), imidacloprid (IMI), clothianidin (CLO), thiacloprid (THD), thiamethoxam (THM), and dinotefuran (DIN)) and four metabolites (i.e., N-desmethyl-acetamiprid (N-dm-ACE), 1-methyl-3-(tetrahydro-3-furylmethyl) (DIN-U), 5-hydroxy-imidacloprid (5-OH-IMI), olefin-imidacloprid (Of-IMI)). NEOs and their metabolites were detected in all serum samples, and the total median concentrations of free, conjugated, and total forms of 10 NEOs were 2.04, 2.01, and 5.12 ng/mL, respectively. Conjugated forms of NEOs accounted for only half (53%) of the total forms of NEOs. Based on the profiles of Phase I and Phase II metabolites of NEOs in serum, it was found that age is a determinant in Phase I metabolism of DIN and Phase II metabolism of IMI. The Phase II metabolites of NEOs are associated with oxidative DNA damage, and the conjugated forms of IMI, DIN, and 5-OH-IMI in serum were significantly positively correlated with oxidative stress. Overall, the amount of NEOs present in conjugated forms in human serum was determined to document the existence of a considerable proportion of free forms of these insecticides.

Connecting the Pipes: Agricultural Tile Drains and Elevated Imidacloprid Brain Concentrations in Juvenile Northern Leopard Frogs (Rana pipiens).

Neonicotinoids are neurotoxic insecticides and are often released into nearby wetlands via subsurface tile drains and can negatively impact nontarget organisms, such as amphibians. Previous studies have indicated that imidacloprid, a commonly used neonicotinoid, can cross the amphibian blood-brain barrier under laboratory conditions; however, little is known about the impact of low concentrations in a field-based setting. Here, we report aqueous pesticide concentrations at wetland production areas that were either connected or not connected to agricultural tile drains, quantified imidacloprid and its break down products in juvenile amphibian brains and livers, and investigated the relationship between imidacloprid brain concentration and brain size. Imidacloprid concentrations in brain and water samples were nearly 2.5 and 5 times higher at tile wetlands (brain = 4.12 ± 1.92 pg/mg protein; water = 0.032 ± 0.045 μg/L) compared to reference wetlands, respectively. Tile wetland amphibians also had shorter cerebellums (0.013 ± 0.001 mm), depicting a negative relationship between imidacloprid brain concentration and cerebellum length. The metabolite, desnitro-imidacloprid, had liver concentrations that were 2 times higher at tile wetlands (2 ± 0.3 μg/g). Our results demonstrate that imidacloprid can cross the amphibian blood-brain barrier under ecological conditions and may alter brain dimensions and provide insight into the metabolism of imidacloprid in amphibians.

Do differentially charged nanoplastics affect imidacloprid uptake, translocation, and metabolism in Chinese flowering cabbage?

Micro(nano)plastics are ubiquitous in the environment. Among the microplastics, imidacloprid (IMI) concentration has been increasing in some intensive agricultural regions, thus receiving increased attention. However, only a few studies have investigated the interaction of nanoplastics (polystyrene (PS)) and IMI in vegetable crops. We studied the effects of positively (PS-NH2) and negatively (PS-COOH) charged nanoplastics on the uptake, translocation, and degradation of IMI in Chinese flowering cabbage grown in Hoagland solution for 28 days. PS-NH2 co-exposure with IMI inhibited plant growth, resulting in decreased plant weight, height, and root length. Translocation of IMI from the roots to the shoots was significantly lower in the presence of PS-NH2, whereas PS-COOH accelerated the accumulation and translocation of IMI in plants, thus potentially affecting IMI metabolism in plants. Notably, IMI-NTG and 5-OH-IMI were the two dominant metabolites. PS-NH2 co-exposure with IMI induced significant oxidation stress and considerably affected the activities of superoxide dismutase (SOD) and peroxidase (POD), indicating that the antioxidant defense system was the main mechanism for reducing oxidative damage. Notably, both positively and negatively charged nanoplastics can accumulate in Chinese flowering cabbage. Plants in the PS-COOH alone treatment group had the highest concentration of nanoplastics in both roots and shoots. The accumulation of nanoplastics, IMI, and its metabolites in plants raises concerns about their combined potential toxicity because it compromises food safety.

Trace determination of imidacloprid and its major metabolites in wheat-soil system.

Trace analysis method is a reliable basis for studying the translocation and metabolism of imidacloprid used as an insecticide in wheat, and it clarifies whether biologically active metabolites including residual imidacloprid, have long-lasting insecticidal potency against wheat aphids under seed treatment during the entire growth period. In this study, a highly sensitive analytical method was established to determine the residues of imidacloprid and its six metabolites (5-hydroxy imidacloprid, imidacloprid olefin, imidacloprid guanidine, imidacloprid urea, 6-chloronicotinic acid, and imidacloprid nitrosimine) in wheat-soil systems, such as in wheat leaves, wheat ears, wheat grains, roots, and soil. All the compounds were extracted using an ACN:water (8:2, v/v) mixture and purified by dispersive solid-phase extraction. The average recoveries ranged from 74.4% to 109.5% for all matrices, with intra- and inter-day variations of less than 14.9%. The limit of quantitation was in the range of 0.001-0.005 mg/kg. The method is demonstrated to be sensitive and accurate for monitoring imidacloprid and its metabolites at trace levels during the entire growth period under field conditions.

Mechanism of the distinct toxicity level of imidacloprid and thiacloprid against honey bees: An in silico study based on cytochrome P450 9Q3

The honey bee, Apis mellifera, shows variation in sensitivity to imidacloprid and thiacloprid, which does not reside at the target site but rather in the rapidly oxidative metabolism mediated by P450s (such as a single P450, CYP9Q3). An in silico study was conducted to investigate the various metabolism of imidacloprid and thiacloprid. The binding potency of thiacloprid was stronger and a stable π-π interaction with Phe121 and the N–H⋯N hydrogen bond with Asn214 are found in the CYP9Q3-thiacloprid system but absent in imidacloprid, which might affect the potential metabolic activity. Moreover, the values of highest occupied molecular orbit (HOMO) energy and the vertical ionization potential (IP) of two compounds demonstrated that thiacloprid is more likely to oxidation. The findings revealed the probable binding modes of imidacloprid and thiacloprid with CYP9Q3 and might facilitate future design of the low bee toxicity neonicotinoid insecticides.

Uptake, translocation and metabolism of imidacloprid loaded within fluorescent mesoporous silica nanoparticles in tomato (Solanum lycopersicum)

Fluorescence-labeling technology has been widely used for rapid detection of pesticides in agricultural production. However, there are few studies on the use of this technology to investigate pesticide uptake and transport in plants with fluorescent nanoparticle formulations. Here, we investigated uptake, transport, accumulation and metabolism of imidacloprid loaded in fluorescent mesoporous SiO2 nanoparticles (Im@FL-MSNs) in tomato plants, and compared the results with the pesticide application in granular formulation. The results revealed that Im@FL-MSNs applied via root uptake and foliar spray both could effectively transport in tomato plants and carry the imidacloprid to plant tissues. Neither Im@FL-MSNs nor imidacloprid was detected inside of tomato fruits from root uptake or foliar spray applications. Compared with the foliar application of granular formulation, imidacloprid in Im@FL-MSNs demonstrated a higher concentration in leaves (1.14 ± 0.07 mg/kg > 1.08 ± 0.04 mg/kg, 1.13 ± 0.09 mg/kg > 1.11 ± 0.02 mg/kg), longer half-life (0.84 d < 1.31 d, 0.90 d < 1.36 d) and small numbers of metabolites formed. These results suggest that mesoporous silica nanoparticles could serve as an effective and efficient pesticide carrier for achieving the high use efficiency in plant protection. The information is also helpful to guide the pesticide applications and assess the risks associated with environmental quality and dietary consumption of vegetables.

An integrated approach, based on mass spectrometry, for the assessment of imidacloprid metabolism and penetration into mouse brain and fetus after oral treatment

Imidacloprid is an insecticide belonging to neonicotinoids, a class of agonists of the nicotinic acetylcholine receptors that shows higher affinities in insects compared to mammals. However, recent evidence show that neonicotinoids can bind to the mammalian receptors, leading to detrimental responses in cultured neurons. We developed an analytical strategy which uses mass spectrometry with multiple reaction monitoring (targeted approach) and high-resolution acquisitions (untargeted approach), which were applied to quantify imidacloprid and to identify its metabolites in biological tissues after oral treatments of mice. Mouse dams were treated with doses from 0.118 mg/kg bw day up to 41 mg/kg day between gestational days 6–9. Results showed quantifiable levels of imidacloprid in plasma (from 30.48 to 5705 ng/mL) and brain (from 20.48 to 5852 ng/g) of treated mice, proving the passage through the mammalian blood-brain barrier with a high correspondence between doses and measured concentrations. Untargeted analyses allowed the identification of eight metabolites including imidacloprid-olefin, hydroxy-imidacloprid dihydroxy-imidacloprid, imidacloprid-nitrosimine, desnitro-imidacloprid, 6-chloronicotinic acid, 5-(methylsulfanyl)pyridine-2-carboxylic acid and N-imidazolidin-2-ylidenenitramide in plasma and brain. Moreover, analysis of embryonic tissues after oral treatment of mouse dams showed detectable levels of imidacloprid (816.6 ng/g after a dose of 4.1 mg/Kg bw day and 5646 ng/g after a dose of 41 mg/Kg bw day) and its metabolites, proving the permeability of the placenta barrier.Although many studies have been reported on the neurotoxicity of neonicotinoids, our study paves the way for a risk assessment in neurodevelopmental toxicity, demostrating the capability of imidacloprid and its metabolites to pass the biological barriers.

Uptake, distribution and translocation of imidacloprid-loaded fluorescence double hollow shell mesoporous silica nanoparticles and metabolism of imidacloprid in pakchoi

Fluorescent labeling techniques have been increasingly used to study the transport of pesticides in crops instead of radioisotope techniques due to the high cost, low resolution and handling and disposal of radioisotope materials. Nevertheless, the use of fluorescent labeling techniques to study the uptake and translocation mechanisms of pesticides in crops has rarely been reported. The distribution, translocation and metabolism of imidacloprid in the vegetable pakchoi were studied via a combination of fluorescence microscopy and UPLC-MS/MS. The translocation rate of imidacloprid-loaded fluorescence double hollow shell mesoporous silica nanoparticles (FL-MSNs@Im) under sunlight conditions were observed by fluorescence microscopy to be higher than that under the dark. The FL-MSNs@Im was mainly transported from the ducts to the stem and leaf parts through transpiration under hydroponic conditions. In contrast, FL-MSNs@Im was transported downward mainly through the sieve tubes of the phloem driven by photosynthesis after the foliar spray. Moreover, fewer metabolites of imidacloprid from FL-MSNs@Im were produced than those from 70% Im granules. This work revealed how FL-MSNs@Im was absorbed, distributed and translocated as well as how the released imidacloprid was metabolized in pakchoi plants.

Peroxiredoxin alleviates the fitness costs of imidacloprid resistance in an insect pest of rice

Chemical insecticides have been heavily employed as the most effective measure for control of agricultural and medical pests, but evolution of resistance by pests threatens the sustainability of this approach. Resistance-conferring mutations sometimes impose fitness costs, which may drive subsequent evolution of compensatory modifier mutations alleviating the costs of resistance. However, how modifier mutations evolve and function to overcome the fitness cost of resistance still remains unknown. Here we show that overexpression of P450s not only confers imidacloprid resistance in the brown planthopper, Nilaparvata lugens, the most voracious pest of rice, but also leads to elevated production of reactive oxygen species (ROS) through metabolism of imidacloprid and host plant compounds. The inevitable production of ROS incurs a fitness cost to the pest, which drives the increase or fixation of the compensatory modifier allele T65549 within the promoter region of N. lugens peroxiredoxin (NlPrx) in the pest populations. T65549 allele in turn upregulates the expression of NlPrx and thus increases resistant individuals’ ability to clear the cost-incurring ROS of any source. The frequent involvement of P450s in insecticide resistance and their capacity to produce ROS while metabolizing their substrates suggest that peroxiredoxin or other ROS-scavenging genes may be among the common modifier genes for alleviating the fitness cost of insecticide resistance.

Evaluation of co-nanoencapsulation process on the toxicity and biochemical metabolism of imidacloprid and lambda-cyhalothrin in Myzus persicae (Sulzer)

The aphid Myzus persicae (Sulzer) is one of the most economically important plant pests worldwide. Excessive use of chemical pesticides to control this pest affects its successful management. In this study, we used a novel technology to improve insecticide efficacy while reducing doses through the combination of insecticides. Therefore, the joint action of pyrethroid lambda-cyhalothrin (LAM) and neonicotinoid imidacloprid (IMI) was evaluated on the aphid M. persicae using leaf-dipping method. Co-nanoencapsulated formulation of these insecticides was prepared using phosphatidylcholine and chitosan as carrier and surface-coating material, respectively. The results of toxicological studies revealed a considerable synergistic effect at 2:1 ratio of technical grade IMI plus LAM and their co-encapsulated formulation. Nanoparticles produced were spherical with size of 67.56 nm. The LC50 values for the technical grade of IMI, LAM and their mixture were 12.65, 89.82 and 4.07 mg ai l−1; for co-encapsulated formulation, the value was 7.06 mg ai l−1. To evaluate the effect of co-nanoencapsulation process on detoxification enzymes, the activities of general esterases and glutathione S-transferases and the amount of cytochrome P450s were measured in M. persicae. The technical grade mixture of IMI and LAM caused pronounced changes in the levels of all tested enzymes compared with the control. In contrast, co-nanoencapsulation protected IMI and LAM from cytochrome P450s- and glutathione S-transferase-induced enzymatic degradation. Considering the toxicological and biochemical findings, co-nanoencapsulation can be valuable as an eco-friendly method for improving the efficacy of the insecticides while preserving the synergistic effects and protecting the active ingredients against detoxification enzymes.

Antibiotics increased host insecticide susceptibility via collapsed bacterial symbionts reducing detoxification metabolism in the brown planthopper, Nilaparvata lugens

Symbionts participate in various physiological activities of their insect hosts, including detoxification metabolism. Emerging evidence has revealed that the bacterial symbiont Arsenophonus is involved in insecticide detoxification metabolism of Nilaparvata lugens, which harbors diverse symbionts. However, it is still unknown whether other bacterial symbionts have a functional role in this process. This study showed that pretreatment with antibiotics significantly increased N. lugens susceptibility to imidacloprid, chlorpyrifos, and clothianidin, and the detoxifying enzyme activities of the cytochrome P450 enzyme (P450) and glutathione S-transferase (GST) were significantly inhibited. Notably, the P450 genes NlCYP6ER1 and NlCYP4CE1, which are related to imidacloprid metabolism, were dramatically downregulated in ciprofloxacin- and tetracycline-pretreated N. lugens, respectively. Furthermore, the expression levels of various detoxifying genes (GSTs and P450s) were significantly positively correlated with Wolbachia, Arsenophonus, Acinetobacter, and Staphylococcus. These results indicated that bacterial symbionts may affect insecticide metabolism by regulating the expression of the insect host’s GST and P450 genes, and provide a foundation for further study on the mechanism of symbiont-mediated host detoxification metabolism in insect pests.

Interspecies differences in cytochrome P450-mediated metabolism of neonicotinoids among cats, dogs, rats, and humans

Neonicotinoid insecticides are used for agricultural and non-agricultural purposes worldwide. Pets are directly exposed to neonicotinoids in veterinary products and through environmental contamination. Cytochrome P450 (CYP) is among the most significant xenobiotic metabolizing enzymes that oxidizes several chemicals, including neonicotinoids. However, CYP activities and metabolite compositions of neonicotinoid metabolites are unknown in most domesticated pet species. Our objectives were to reveal the differences in metabolites of neonicotinoids (imidacloprid, clothianidin, and acetamiprid) and CYP activities among common pet species (cats and dogs), humans, and rats. The results indicated that the CYP-mediated neonicotinoid metabolism was different depending on species and each neonicotinoid. Among these four species, the kinetics of imidacloprid metabolism indicated that rats have the highest rate of oxidation of imidacloprid to 4OH-imidacloprid, while the greatest enzyme kinetics of imidacloprid metabolism to 5OH-imidacloprid were found in rats and humans. Clothianidin was rapidly metabolized to 1-methyl-3-nitroguanidine and dm-clothianidin in rats, but cats and humans showed the lowest formation of dm-clothianidin. CYP activities in metabolism of acetamiprid to dm-acetamiprid and N-acetyl-acetamiprid were determined to be significantly higher in humans compared to other species. However, further studies should be targeted at identifying the differences in hepatic metabolism of neonicotinoids in these species using recombinant CYP enzymes.

Translocation and metabolism of imidacloprid in cabbage: Application of 14C-labelling and LC-QTOF-MS

Imidacloprid (IMI) is a widely used neonicotinoid insecticide effective against sucking and some chewing insects. Translocation and metabolism of IMI in plants are related to food safety. In this study, 14C-labeled IMI was used to investigate its translocation, transformation, radioactive IMI metabolites and possible metabolic pathways in cabbage. The amount of IMI accumulated in the edible part of cabbage accounted for 80.3–95.4% of the applied amounts by foliar application. There was a tendency to transport from edible parts to inedible parts. The proportions of extractable IMI decreased gradually from 92.4% to 83.0% in edible parts, greater than that in inedible parts over the experiment (0–19 days), while the bound residues showed an opposite trend. The half-life of IMI was determined as 33.0 and 63.0 days in the edible parts and whole plant, respectively. Five radioactive components including the parent IMI were detected by HPLC-LSC. The relative content of M1 was less than 0.01 mg kg−1, which was not required to identify according to the metabolic scheme proposed by the US Environmental Protection Agency. The metabolites N-nitro(1-6-chloro-3-pyridylmethyl)-4,5-dihydroxyimidazol-2-imine (M2), N-nitro(1-6-chloro-3-pyridylmethyl)-4/5-hydroxyimidazole-2-imine (M3) and 1/3-(1-6-chloro-3-pyridylmethyl)-2,4-imidazodione (M4) were identified by LC-QTOF-MS. The primary metabolism of IMI in cabbage included hydrolysis and oxidation. The residue level and daily intake values of IMI in cabbage were estimated to be 0.033–0.078 mg kg−1 and 9.56–20.01 ng d−1 kg−1, respectively, which were far below the maximum residue level and allowable daily intake values.

In vitro metabolism of imidacloprid and acetamiprid in rainbow trout and rat

Providing an alternative to pyrethroids, organophosphates, and carbamates, the neonicotinoids are now the most widely used insecticides in the world. They are water soluble and relatively stable in soil and water which allows for run-off through surface waters and thus potentially impacting aquatic species and environments.While the mammalian metabolism of neonicotinoids has been studied extensively, there is a lack of understanding of their metabolism in fish species. The current study constitutes the first report of the metabolism of imidacloprid (IMI) and acetamiprid (AC) in rainbow trout.Formation of respective metabolites 5-hydroxy-imidacloprid and N-desmethyl-acetamiprid was conserved across orders of biological organization in both microsomal and liver slice assays.Michaelis–Menten kinetics were determined for the microsomal conversion of IMI to 5-hydroxy-imidacloprid in rainbow trout (Km = 79.2 µM; Vmax = 0.75 pmole/min/mg) and rat (Km = 158.7 µM; Vmax = 38.4 pmole/min/mg). Kinetics for the microsomal demethylation of AC to N-desmethyl-acetamiprid were determined in the rat (Km = 70.9 µM; Vmax = 10 pmoles/min/mg). N-desmethyl-acetamiprid was found in detectable but below quantifiable levels across the range of test concentrations which precluded a calculation of kinetic rate constants in rainbow trout (RBT).Ultimately, the formation of the metabolites 5-hydroxy-imidacloprid and N-desmethyl-acetamiprid was conserved across RBT and rat species.