Long-term profound investigation of Fenoxaprop-p-ethyl and indirect pesticide pollution in jute-cropped environmental ecosystem for dietary and ecological risk assessment.

Jute is a healthy vegetable due to its high content of carotenoids, vitamins, and minerals, in addition to its industrial utility. The purpose of this study was to look into the dissipation of fenoxaprop-p-ethyl and to track indirect sources of pesticide accumulation in jute ecosystems for three years in a row during tropical monsoon seasons. To avoid the intake of residues over approved limits, a rapid extraction technique was developed to evaluate the residues of this herbicide (Whip-Super 9% EC) in jute leaves, fiber, cropped soil, and water. A modified QuEChERS approach based on liquid chromatography and ultraviolet-visible (UV-VIS) detection was developed and successfully validated in accordance with SANCO requirements. Gas chromatography mass spectrometer (GC-MS) was used to examine fish and water samples from jute fields and retting pond for twenty widely-used multi-class pesticide contaminations from indirect sources. Relative standard deviations (RSD) (≤ 20) and recoveries (100-115%) are observed to be within acceptable ranges. In edible jute leaves, independent of season, first-order kinetics of dissipation was recorded, with half-lives ranging from 0.61 to 0.68 days. Based on the findings of the risk assessment, it is feasible to conclude that the consumer health risks of ingesting jute leaves are insignificant, even on day zero, when used at the recommended amount, and that using jute bags for food packaging and storage is safe. But jute field water was found to be less polluted with indirect pesticides than water from a nearby jute retted pond. Fish samples from this jute retted pond were also analyzed and found to be 26% contaminated. The presence of quinalphos and chlorpyriphos in jute retting water and fish might pose a concern to consumer health and environmental ecosystems.

Determination of quizalofop-p-ethyl in onion: residual dissipation pattern, weed control efficiency, and food safety assessment under field conditions.

Monitoring pesticide residue levels becomes crucial to maintain quality and guarantee food safety as the consumption of onion green leaves and immature and mature bulbs (either raw or processed) rises. A field experiment was conducted for two consecutive seasons with quizalofop-p-ethyl (5% EC) at 50 and 100 g a.i. ha-1 to evaluate weed control efficiency and to determine terminal residues. Post-emergence application of fop herbicide at 100 g a.i. ha-1 kept the weed density and dry weight reasonably at a lower level and enhanced the productivity of onion with higher economic returns. A rapid, sensitive, and analytical method was developed using high-performance liquid chromatography (HPLC) with excellent linearity (r2 > 0.99). The limit of quantification for quizalofop-p-ethyl was established at 0.04 mg kg-1 with signal to noise (S/N) ratio ≥ 10. The method was successfully applied and initial quantified residues were in the range of 2.5-4.4 mg kg-1 irrespective of seasons and doses. Finally, the presence of targeted herbicide residues in harvested samples was confirmed by liquid chromatography tandem mass spectrometry (LC-MS/MS) under optimized operating conditions. Dietary risk assessment assured harvested onions were safe for consumption at the recommended dose. It also can be concluded that quizalofop ethyl did not adversely influence soil micro-organisms at standard rates of application.

Method validation, persistence, and safety evaluation of 2,4-D in tea (Camellia sinensis) by LC-MS/MS.

An analytical method was developed by using LC-ESI(-ve)-MS/MS to investigate the residue dynamics of 2,4-D (2,4-dichlorophenoxyacetic acid) in green tea leaves, processed tea, tea liquor, and tea-cropped soil at Singhiajhora Tea Estate and Putinbari Tea Estate at Terai Region, Darjeeling District, West Bengal, India. In this method, an acidified methanol was used for extraction and subsequent clean-up was done by HLB (hydrophilic lipophilic balanced) cartridges. The method was validated as per SANTE guideline (SANTE/11813/2017). The limit of quantification (LOQ) of 2,4-D was 0.05 mgkg-1 and average % recoveries were in the range from 88.05 to 113.28 with relative standard deviation (RSD) 3.46 to 6.43. The dissipation of 2,4-D followed the 1st-order reaction kinetics with a half-life (T1/2) of 1.51-1.61 day at the recommended dose and 2.50-2.72 day for doubled recommended dose in tea for both locations. This method can be applied successfully for the determination of 2,4 D residues in/on tea matrix and subsequent studies on safety evaluation showed that the use of 2,4-D in tea is safe.

Bioefficacy, residue dynamics and dietary risk assessment of gibberellic acid in improving the potential yield of tomato (Solanum lycopersicum L.).

The increasing use of gibberellic acid (GA3) to promote fruit growth and yield has necessitated research into its trace level determination and estimation in harvested product. The phytohormone has increased the tomato yield (tonne ha-1) up to 24.7% with uniform fruit shape, size colour and lustre. A fast, simple, high-throughput analytical method was standardised based on electrospray ionisation - liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted using acidified (1% formic acid) methanol. The method was validated as per the SANTE/12682/2019 guidelines. The limits of detection (LOD) and quantification (LOQ) were 0.01 and 0.05 mg kg-1. The average recoveries at LOQ and higher levels were in the range of 86-108% with relative standard deviation (RSD) < 20%. The validated method was successfully applied under field condition by following first-order kinetics with half-lives (T1/2) 1.76 days (recommended dose) and 1.99 days (double dose). The estimated pre-harvest intervals (PHIs) were 6 days (recommended dose) and 8 days (double dose). Studies on dietary risk assessment concluded that even after spray of GA3 at recommended dose, the harvested produce (tomato) could be consumed safely.

Corrigendum to "Curcumin protects rat liver from streptozotocin-induced diabetic pathophysiology by counteracting reactive oxygen species and inhibiting the activation of p53 and MAPKs mediated stress response pathways" [Toxicol. Rep. 2 (2015) 365-376].

[This corrects the article DOI: 10.1016/j.toxrep.2014.12.017.].

Review of extraction and detection techniques for the analysis of pesticide residues in fruits to evaluate food safety and make legislative decisions: Challenges and anticipations.

Fruits are vital parts of the human diet because they include necessary nutrients that the body needs. Pesticide use has increased dramatically in recent years to combat fruit pests across the world. Pesticide usage during production, on the other hand, frequently results in undesirable residues in fruits after harvest. Consumers are concerned about pesticide residues since most of the fruits are directly consumed and even recommended for the patients as dietary supplements. As a result of this worry, pesticide residues in fruits are being randomly monitored to re-assess the food safety situation and make informed legislative decisions. To assess the degree of pesticide residues in fruits, a simple and quick analytical procedure is usually required. As a result, pesticide residue detection (using various analytical techniques: GC, LC and Biosensors) becomes critical, and regulatory directives are formed to regulate their amounts via the Maximum Residue Limit (MRL). Over the previous two decades, a variety of extraction techniques and analytical methodologies for xenobiotic's efficient extraction, identification, confirmation and quantification have been developed, ranging from traditional to advanced. The goal of this review is to give readers an overview of the evolution of numerous extraction and detection methods for pesticide residue analysis in fruits. The objective is to assist analysts in better understanding how the ever-changing regulatory landscape might drive the need for new analytical methodologies to be developed in order to comply with current standards and safeguard consumers.

Simultaneous Monitoring and Decontamination of Pesticide Residues in Phytomedicine-Enriched Betel Leaf Utilizing QuEChERS-GC-MS/MS Technology to Safeguard Public Health.

BACKGROUND: The presence of undesirable substances, including pesticides (xenobiotics) in betel leaf (Piper betel), is a great concern for consumers because it is chewed and consumed directly. To protect the consumer's health, a modified QuEChERS method for monitoring purposes and subsequent decontamination process has been developed. OBJECTIVE: The goal of this work was to establish a multi-residue analytical method for monitoring nonpermitted organophosphorus pesticide residues in betel leaf, as well as cost-effective cleaning strategies. METHOD: The homogenized 15 g samples (20 betel leaf samples collected in West Bengal, India) were extracted with a modified QuEChERS method using acetonitrile, reconstituted to acetone, and finally analyzed by GC-MS/MS. Possible decontamination techniques (such as tap water washing, 2% saltwater washing, and lukewarm water washing) were evaluated. RESULTS: The limit of detection ranged from 0.003 to 0.005 mg/kg, and limit of quantification was 0.01 mg/kg. Recoveries ranged from 80 to 120% with RSDr 9%. One sample was found to contain three pesticides 4 to 7 times higher than MRLs. Suggested decontamination methods allowed reducing toxic traces below European limits. CONCLUSIONS: The suggested approach is useful for determining pesticide residues in betel leaves quickly. Traditional techniques of processing betel leaves may reduce pesticide residues below regulatory limits. HIGHLIGHTS: A multi-residue method and decontamination of pesticides in betel leaf using QuEChERS-GC-MS/MS technology with satisfactory method performance was achieved. Domestic decontamination techniques have a high efficacy in reducing pesticide residues from betel leaves, making them safe for human consumption.

Method development, validation, monitoring, seasonal effect and risk assessment of multiclass multi pesticide residues in surface and ground water of new alluvial zone in eastern India.

A liquid-liquid extraction (LLE) method was validated as per SANTE/12682/2019 guidelines for gas chromatography-mass spectrometric (GC-MS) determination of thirty-six multiclass pesticides in environmental waters. Seasonal (summer, monsoon, and winter) effects on the magnitude of pesticide residues in environmental water (river, pond, and tube well) of six different urban areas of Nadia and North 24 Parganas districts (New alluvial zone, Eastern India) was monitored for subsequent risk assessment. Total 288 water samples (96 each of river, pond, and tube-well) irrespective of locations and seasons were monitored for multiclass multi pesticide residues during the experiment. Each sample (750 mL) was extracted with ethyl acetate/dichloromethane (8:2) liquid-liquid partitioning and filtration (0.22 µm nylon filter paper) and total residue was reconstituted in acetone (1 mL) for GC-MS analysis with developed and validated method resulting satisfactory recovery percentages (77.84-118.15%). The maximum no. of organochlorine (OC) and organophosphorus (OP) pesticide residues were dominated in river and pond water irrespective of seasons and monitoring sites. About 74% of river water samples were found to be contaminated with concerned pesticides in variable magnitudes. Monsoon (July to October) season was highly alarming with the highest presence of total pesticide residues in different types of environmental waters. Risk quotient (RQ) [acute and chronic] was also evaluated in pond and river water as sometimes used for drinking purposes. RQ value (5900) of total endosulfan indicates the highest risk of chronic toxicity to river fishes. Seven water samples from tube-wells were also monitored and found to be contaminated with butachlor and chlorpyriphos in non-significant amounts (< 0.1 ng mL-1), irrespective of seasons and sites, thus safe for consumption.

Phytomedicinal Role of Pithecellobium dulce against CCl(4)-mediated Hepatic Oxidative Impairments and Necrotic Cell Death.

Present study investigates the beneficial role of the aqueous extract of the fruits of Pithecellobium dulce (AEPD) against carbon tetrachloride (CCl(4))-induced hepatic injury using a murine model. AEPD has been found to possess free radical (DPPH, hydroxyl and superoxide) scavenging activity in cell-free system. CCl(4) exposure increased the activities of various serum maker enzymes and intracellular reactive oxygen species (ROS) production. In line with these findings, we also observed that CCl(4) intoxication increased the lipid peroxidation and protein carbonylation accompanied by decreased intracellular antioxidant defense, activity of cytochrome P450 and CYP2E1 expression. DNA fragmentation and flow cytometric analyses revealed that CCl(4) exposure caused hepatic cell death mainly via the necrotic pathway. Treatment with AEPD both pre- and post-toxin exposure protected the organ from CCl(4)-induced hepatic damage. Histological findings also support our results. A well-known antioxidant vitamin C was included in this study to compare the antioxidant potency of AEPD. Combining all, results suggest that AEPD protects murine liver against CCl(4)-induced oxidative impairments probably via its antioxidative property.

Multiclass Multipesticide Residue Analysis in Fish Matrix by a Modified QuEChERS Method Using Gas Chromatography with Mass Spectrometric Determination.

BACKGROUND: Pesticide residue in fish is a global food safety concern. However, very few validated methods are available targeting simultaneous analysis of multiple classes of pesticides. OBJECTIVE: The aim of this study was to validate a quick, easy, cheap, effective, rugged, and safe workflow-based method for the quantitative determination of multiclass pesticides in fish matrix using GC-MS determination. METHOD: The sample was extracted with acetonitrile, and the cleanup method involved dispersive solid-phase extraction with C-18 sorbent, which effectively scavenged the coextracted matrix components and removed those from the extract. The data on recovery and precision of the method satisfied the criteria of SANTE/11813/2017 guidelines. Average recoveries of pesticides were in the range of 80-120% with precision RSDs ≤20%. The LOD and LOQ were in the ranges of 0.001-0.029 and 0.005-0.125 µg/mL, respectively, for all pesticides. The expanded uncertainty was in the range of 14-20%, based on the single-laboratory validation data (coverage factor, k = 2, confidence level, 95%). CONCLUSIONS: The validation data prove that the method is convenient and acceptable for the routine analysis of multiclass pesticide residues in fish matrices for regulatory compliance. HIGHLIGHTS: The study achieves multiresidue analysis of pesticides in fish matrix with MS-based confirmation. The method combines the advantages of nontarget analysis based on National Institute of Standards and Technology library matching in full scan mode with selected-ion monitoring-based sensitivity.

A 35 kDa Phyllanthus niruri protein suppresses indomethacin mediated hepatic impairments: Its role in Hsp70, HO-1, JNKs and Ca2+ dependent inflammatory pathways.

The present study has been conducted to explore a novel strategy to modulate the unfavourable effects of indomethacin by Phyllanthus niruri protein (PNP) and the underlying mechanism PNP exploits for the amelioration of that pathophysiology. In hepatocytes, indomethacin enhanced reactive oxygen species (ROS), reduced intracellular antioxidant capacity, up regulated mitogen activated protein kinase (MAPKs), disrupted mitochondrial membrane potential, activated apoptotic pathways and there by reduced the viability of the hepatocytes. Simultaneous treatment with PNP modulated these detrimental actions of the drug and retained cell viability. Similarly, in mice, indomethacin elevated serum marker enzymes (e.g. Alanine Transaminase), decreased antioxidant enzyme activities, elevated oxidations of lipids and proteins, increased intracellular calcium overload mediated endoplasmic reticular stress (ER stress) pathways, up regulated the pro-inflammatory cytokines and there by leading to the mitochondrial dependent caspase-3 activation and poly-ADP ribose polymerase (PARP) cleavage. Moreover investigation of these inherent molecular pathways exhibited that these alterations are associated with up regulation of MAPKs, inducible nitric oxide synthase (iNOS), heme oxygenase-1 and down regulation of survival proteins. However, PNP suppressed those apoptotic indices as evidenced from histopathological studies and DNA fragmentation analysis. Combining, results suggest that PNP could possibly provide a protection against indomethacin-induced hepatic pathophysiology.

Silymarin Protects Mouse Liver and Kidney from Thioacetamide Induced Toxicity by Scavenging Reactive Oxygen Species and Activating PI3K-Akt Pathway.

Silymarin (SMN) has been shown to possess a wide range of biological and pharmacological effects. Besides, SMN has antioxidant and free radical scavenging activities. Thioacetamide (TAA) is a well-documented liver toxin that requires oxidative bioactivation to elicit its hepatotoxic effect which ultimately modifies amine-lipids and proteins. Our study has been designed in a TAA exposed mouse model to investigate whether SMN could protect TAA-induced oxidative stress mediated hepatic and renal damage. Results suggest that TAA generated reactive oxygen species (ROS), caused oxidative stress and induced apoptosis in the liver and kidney cells via JNK as well as PKC and MAPKs signaling. All these detrimental effects of TAA could, however, be suppressed by SMN which not only scavenged ROS but also induced PI3K-Akt cell survival pathway in the liver and prevented apoptotic pathways in both the organs. Histological studies, collagen staining and DNA fragmentation analysis also supported our results. Combining, we say that SMN possess beneficial role against TAA mediated hepatic and renal pathophysiology.

Curcumin protects rat liver from streptozotocin-induced diabetic pathophysiology by counteracting reactive oxygen species and inhibiting the activation of p53 and MAPKs mediated stress response pathways.

Curcumin (CUR) is a highly pleiotropic molecule and possesses anti-inflammatory, hypoglycemic, antioxidative, wound-healing and antimicrobial activities. The present study was carried out to investigate whether CUR plays any beneficial role in streptozotocin (STZ) induced hepatic pathophysiology in diabetic rats. STZ exposure increased hepatic damage associated serum markers (ALT, ALP and LDH) as well as NO production in the liver tissue. Moreover, the same exposure enhanced ROS generation and lipid peroxidation; reduced GSH levels and antioxidant enzyme activities. Hyperglycemia induced hepatic pathophysiology also activated stress response pathways (involving phosphorylation of p38, ERK1/2 MAPKs and p53) and reduced mitochondrial membrane potential which in turn led to cellular apoptosis as evidenced from increased hepatic DNA fragmentation as well as FACS analysis. However, treatment with CUR effectively counteracts diabetes-induced, oxidative stress mediated hepatic damage and could act as a therapeutic in lessening liver dysfunction in diabetic subjects.

Atorvastatin induced hepatic oxidative stress and apoptotic damage via MAPKs, mitochondria, calpain and caspase12 dependent pathways.

Atorvastatin (ATO), a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor, is used widely for the treatment of hypercholesterolemia and hypertriglyceridemia. Application of this drug has now been made somehow limited because of ATO associated several acute and chronic side effects. The present study has been carried out to investigate the dose-dependent hepatic tissue toxicity in ATO induced oxidative impairment and cell death in mice. Administration of ATO enhanced ALT, ALP level, increased reactive oxygen species (ROS) production and altered the pro oxidant-antioxidant status of liver by reducing intracellular GSH level, anti-oxidant enzymes activities and increasing intracellular lipid peroxidation. Our experimental evidence suggests that ATO markedly decreased mitochondrial membrane potential, disturbed the Bcl-2 family protein balance, enhanced cytochrome c release in the cytosol, increased the levels of Apaf1, caspase-9, -3, cleaved PARP protein and ultimately led to apoptotic cell death. Besides, ATO distinctly increased the phosphorylation of p38, JNK, and ERK MAPKs, enhanced Caspase12 and calpain level. Histological studies also support the dose-dependent toxic effect of ATO in these organs pathophysiology. These results reveal that ATO induces hepatic tissue toxicity via MAPKs, mitochondria and ER dependent signaling pathway, in which calcium ions and ROS act as the pivotal mediators of the apoptotic signaling.

Amelioration of aspirin induced oxidative impairment and apoptotic cell death by a novel antioxidant protein molecule isolated from the herb Phyllanthus niruri.

Aspirin has been used for a long time as an analgesic and anti-pyretic drug. Limitations of its use, however, remain for the gastro-intestinal side effects and erosions. Although the role of aspirin on gastro-intestinal injury has been extensively studied, the molecular mechanisms underlying aspirin-induced liver and spleen pathophysiology are poorly defined. The present study has been conducted to investigate whether phyllanthus niruri protein (PNP) possesses any protective role against aspirin mediated liver and spleen tissue toxicity, and if so, what signaling pathways it utilizes to convey its protective action. Aspirin administration in mice enhanced serum marker (ALP) levels, reactive oxygen species (ROS) generation, reduced antioxidant power and altered oxidative stress related biochemical parameters in liver and spleen tissues. Moreover, we observed that aspirin intoxication activated both the extrinsic and intrinsic apoptotic pathways, as well as down regulated NF-κB activation and the phosphorylation of p38 and JNK MAPKs. Histological assessments and TUNEL assay also supported that aspirin induced tissue damages are apoptotic in nature. PNP treatment after aspirin exposure effectively neutralizes all these abnormalities via the activation of survival PI3k/Akt pathways. Combining all results suggest that PNP could be a potential protective agent to protect liver and spleen from the detrimental effects of aspirin.

Cytochrome P450s: mechanisms and biological implications in drug metabolism and its interaction with oxidative stress.

BACKGROUND: Cytochrome monooxygenases P450 enzymes (CYPs) are terminal oxidases, belonging to the multi-gene family of heme-thiolate enzymes and located in multiple sites of ER, cytosol and mitochondria. CYPs act as catalysts in drugs metabolism. AREAS COVERED: This review highlights the mitochondrial and microsomal CYPs metabolic functions, CYPs mediated ROS generation and its feedback, bioactivation of drugs and related hypersensitivity, metabolic disposition as well as the therapeutic approaches. WHAT THE READERS WILL GAIN: CYPs mediated drugs bioactivation may trigger oxidative stress and cause pathophysiology. Almost all drugs show some adverse reactions at high doses or accidental overdoses. Drugs lead to hypersensitivity reactions while metabolic predisposition to drug hypersensitivity exaggerates it. Mostly different intermediate bioactive products of CYPs mediated drug metabolism is the principal issue in this respect. On the other hand, CYPs are the main source of ROS. Their generation and feedback are of major concern of this review. Besides drug metabolism, CYPs also contribute significantly to carcinogen metabolism. Ultimately other enzymes in drug metabolism and antioxidant therapy are indispensible. Importance of this field: In a global sense, understanding of exact mechanism can facilitate pharmaceutical industries' challenge of developing drugs without toxicity. Ultimate message: This review would accentuate the recent advances in molecular mechanism of CYPs mediated drug metabolism and complex cross-talks between various restorative novel strategies evolved by CYPs to sustain the redox balance and limit the source of oxidative stress.

A 35 kD Phyllanthus niruri protein modulates iron mediated oxidative impairment to hepatocytes via the inhibition of ERKs, p38 MAPKs and activation of PI3k/Akt pathway.

It has been reported that the herb, Phyllanthus niruri, possess antioxidant, anti-infection, anti-asthmatic, anti-diuretic, anti-soresis and many more beneficial activities. The goal of our present study was to evaluate the protective role of a 35 kD protein (PNP) isolated from this herb against iron-induced cytotoxicity in murine hepatocytes. Exposure of hepatocytes to iron (FeSO4) caused elevation of reactive oxygen species (ROS) production, enhanced lipid peroxidation and protein carbonylation, depleted glutathione levels, decreased the antioxidant power (FRAP) of the cells and reduced cell viability. Iron mediated cytotoxicity disrupted mitochondrial membrane potential (Δψm) and thereby caused apoptosis mainly by the intrinsic pathway via the down-regulation of IκBα with a concomitant up-regulation of NF-kB as well as the phosphorylation of ERKs and p38 MAP kinases. In addition, iron-induced cytotoxicity disrupted the normal balance of Bcl-2 family proteins in hepatocytes. Incubation of hepatocytes with PNP, however, protected the cells from apoptosis by stabilizing the mitochondria and arresting the release of cytochrome c. It also suppressed caspase activation and cleavage of PARP. Moreover, this protein has strong free radical scavenging activity and thereby scavenged ROS extensively. Combining all, results suggest that simultaneous treatment with PNP might suppress the iron-induced cytotoxicity in hepatocytes.

Iron induces hepatocytes death via MAPK activation and mitochondria-dependent apoptotic pathway: beneficial role of glycine.

In the present study we investigated the beneficial role of glycine in iron (FeSO4) induced oxidative damage in murine hepatocytes. Exposure of hepatocytes to 20 µM FeSO4 for 3 hours enhanced reactive oxygen species (ROS) generation and induced alteration in biochemical parameters related to hepatic oxidative stress. Investigating cell signalling pathway, we observed that iron (FeSO4) intoxication caused NF-κB activation as well as the phosphorylation of p38 and ERK MAPKs. Iron (FeSO4) administration also disrupted Bcl-2/Bad protein balance, reduced mitochondrial membrane potential, released cytochrome c and induced the activation of caspases and cleavage of PARP protein. Flow cytometric analysis also confirmed that iron (FeSO4) induced hepatocytes death is apoptotic in nature. Glycine (10 mM) supplementation, on the other hand, reduced all the iron (FeSO4) induced apoptotic indices. Combining, results suggest that glycine could be a beneficial agent against iron mediated toxicity in hepatocytes.