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Supervised field trial studies were conducted to understand dissipation kinetics and harvest time residues of a combination product of fluxapyroxad and pyraclostrobin in cumin plant/leaves and seeds at different locations in India. The results showed initial accumulation of fluxapyroxad at the levels of 15.4 and 20.2 mg kg -1 and pyraclostrobin at the level of 21.2 and 33.4 mg kg -1 in cumin leaves/plant in Anand, Gujarat. Fluxapyroxad and pyraclostrobin followed zero-order and first-order dissipation kinetics in cumin plant/leaves samples respectively. The residues translocated to cumin seeds. As the hazard quotient (HQ) was <1 in all cases consumer health risk may be negligible.

Neonicotinoids (NEOs) and fipronil are widely used in pest control, but their spatiotemporal distribution and risk levels in the "river-estuary-bay" system remain unclear. Between 2018 and 2021, 148 water samples from rivers to inshore and offshore seawater in Laizhou Bay, China were collected to investigate the presence of eight NEOs and fipronil and its metabolites (FIPs). Significant seasonal variations in NEOs were observed under the influence of different cultivation practices and climatic conditions, with higher levels in the summer than in the spring. The average concentrations of total neonicotinoids (ΣNEOs) and ∑FIPs decreased from rivers (63.64 ng/L, 2.41 ng/L) to inshore (22.62 ng/L, 0.14 ng/L) and offshore (4.48 ng/L, 0.10 ng/L) seawater of Laizhou Bay. The average concentrations of ΣNEOs decreased by 85.3 % from 2018 to 2021. The predominant insecticides in the study area were acetamiprid, thiamethoxam, imidacloprid, and fipronil sulfone, with a gradual shift toward low-toxicity and environmentally friendly species over time. Influenced by agricultural intensity, ∑NEOs were mostly distributed in the Yellow River, Xiaoqing River, and their estuaries, where they pose chronic ecological risks. However, FIP exhibited high risk in certain rivers and sewage treatment plants owing to the use of animal repellents or landscape gardening insecticides. This study provides evidence of the transfer of NEOs and FIPs from rivers to the ocean and also clarifies their transition dynamics and changes in risk levels from rivers to oceans. Additionally, the study offers data support for identifying critical pesticide control areas., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The first purpose of this study was to determine whether a measurement of the level of direct oral anticoagulants (DOACs) was possible with heparin-calibrated chromogenic anti-factor Xa activity (AXA). The second purpose of this study was to evaluate whether the antidote treatment decision level (30 or 50 ng/mL of DOAC) can be determined by unfractionated heparin (UHF)/low molecular weight heparin (LMWH)-calibrated AXA., Methods: AXA was measured by using two reagents and dedicated analyzers (Sysmex CS-5100 analyzer and STA R Max3). Four types of calibrators were used: 1) Stago DOAC (rivaroxaban, edoxaban, and apixaban)-specific calibrator, 2) Stago LMWH calibrator, 3) Sysmex UHF calibrator, and 4) Sysmex LMWH calibrator. Regression analysis was used between assays. Receiver operating characteristic (ROC) curves were performed, and the concordance rate was calculated., Results: The correlation coefficients were in the range of 0.75 - 0.91 for rivaroxaban and 0.81 - 0.94 for apixaban. The correlation coefficient between edoxaban-calibrated AXA and Sysmex LMWH/Sysmex UHF calibrator-calibrated AXA was low (r = 0.47). Overall correlation between DOAC-calibrated AXA and Stago LMWH-calibrated AXA was linear, at only low concentration in all three DOACs. The concordance rate (89.3 - 100%) is good for de-termining the antidote management level by UFH/LMWH-calibrated AXA, compared with those of DOAC-calibrated AXA in rivaroxaban and apixaban. The concordance rate ranged from 63% to 67% between Sysmex UFH/ LMWH-calibrated AXA and edoxaban-calibrated AXA., Conclusions: The findings of our study suggest limitations in calculating accurate concentrations, when using UFH/LMWH-calibrated AXA to measure DOAC. This study demonstrates that UFH/LMWH-calibrated AXA may be useful in determining the presence of DOACs at the cutoff level for the antidote treatment in rivarovaban and apixaban. However, in edoxaban, UFH/LMWH-calibrated AXA could not accurately measure the presence of DOACs at the cutoff for antidote treatment.

A simple, Accurate, precise method was developed for the estimation of the Lorlatinib in API form and Marketed pharmaceutical dosage form by RP-HPLC. Chromatogram was run through Hypersil C18 (4.6mm×150mm, 5µm) Particle size Column and Mobile phase containing Methanol and Water taken in the ratio of 25: 75% v/v was pumped through column at a flow rate of 1.0 ml/min. Temperature was maintained at 38ºC. Optimized wavelength selected was 310 nm. Retention times of Lorlatinib were found to be 3.513 minutes respectively. The %RSD for the Repeatability and Intermediate Precision of the Lorlatinib were found to be within limits. %Recovery was obtained 98.96% and it was found to be within the limits for Lorlatinib respectively. The LOD, LOQ values obtained from regression equations of Lorlatinib were 0.332µg/ml and 1.0078 µg/ml respectively. Regression equation of Lorlatinib was found to be y = 39948x + 16821 respectively. The Retention times was decreased and run time was decreased, so the method developed was simple and economical that can be adopted in regular Quality control test in Industries.

Apixaban (APX) is a novel anti-coagulant drug approved by USFDA. According to referred literature, numerous chromatographic methods such as RP-HPLC and high-performance thin-layer chromatography have been published for the stability study of APX. But these chromatographic methods have been developed using toxic organic solvents that are hazardous to the environment and unsafe for analysts. Hence, green and sustainable analytical chemistry-driven chromatographic method has been developed for the stability study of APX using safe organic solvents for the safety of analysts and the protection of the environment. APX was subjected to forced degradation for the development of a stability-indicating assay method. The method development was carried out by the implementation of chemometric and DoE approaches for minimizing solvent wastage. Principal component analysis was applied for the identification of critical method risk variables (MRVs) and method performance attributes. DoE-based response surface modelling was applied for the optimisation of critical MRVs. The greenness profile scales of published and developed chromatographic methods have been assessed by NEMI and AGREE methods for the estimation of APX. The developed method was found to be more eco-friendly and robust than the published chromatographic methods for the estimation of APX., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: Fipronil is a commonly used pesticide in the agricultural and animal health industries for the protection of crops and control of fleas, ticks, and chewing lice. It is difficult to obtain reproducible retention time and relative retention time (RRT) for a common hydrolytic degradation product of fipronil with the current European Pharmacopeia (EP) monograph for assay and estimation of related substances of fipronil. This situation causes misidentification, mislabeling, and/or false out-of-specification results for this hydrolytic degradation product of fipronil in bulk commercial batches during batch release and/or in the stability samples during the shelf life of a released batch., Objective: This study aimed to develop a reversed-phase ultra performance liquid chromatography (UPLC) method for assay and identification of fipronil including identification and estimation of its related substances in bulk drug substance batches of fipronil and provide consistent retention time of the hydrolytic degradation product., Methods: Fipronil and its related substances were separated by gradient elution on a Halo C18 column (50 mm × 2.1 mm id, 2.0 µm particle size) maintained at 40°C with 0.1% H3PO4 in H2O as mobile phase-A and acetonitrile-methanol (50 + 50, v/v) as mobile phase-B. Fipronil and its related substances were detected and quantified at 280 nm with a quantitation limit of 0.05% of the target (analytical) concentration., Results: The UPLC method was able to separate all analytes of interest by gradient elution with a total run time of 7 min (approximately 40% faster than EP)., Conclusion: In this paper, we report the development and validation of a fast, stability-indicating reversed-phase UPLC method for assay and estimation of related substances of fipronil in stability samples and bulk batches of fipronil., Highlights: The new UPLC method is approximately 40% faster than the current Ph. Eur. monograph for fipronil assay and the new method provides reproducible retention of a common hydrolytic degradation product of fipronil., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: The topical veterinary drug product containing fipronil and permethrin provides an effective repellent protection and high insecticidal efficacy for dogs., Objective: The objective of this study was to develop a stability-indicating high-performance liquid chromatography (HPLC) method for simultaneous detection and quantification of fipronil, permethrin, their key degradation products, and butylated hydroxytoluene (BHT) in a topical drug product., Method: The two active ingredients, their degradation products, and the antioxidant (BHT) were separated by a gradient elution on a Phenomenex Kinetex C18 column (150 × 3 mm, 2.6 µm particle size) maintained at 37°C with H2O acetonitrile isopropyl alcohol 85% H3PO4 (65.5 + 32.5 + 4/0.0053, v/v/v/v) as mobile phase A and acetonitrile (100%) as mobile phase B. The flow rate was 0.9 mL/min, and analytes were detected and quantified at 235 nm., Results: The specificity of the method was demonstrated by adequate separation of fipronil, permethrin, their degradation products, and BHT in the forced degraded finished product. The linearity of the method was demonstrated in the range of 0.2% to 150% of target analytical concentration of both active ingredients and 50% to 150% for BHT. Excellent recoveries of fipronil, permethrin, and BHT in placebo spiked active ingredient solutions in the linearity range showed sufficient accuracy of the method. The LOQ and LOD of the method were determined to be 0.2% and 0.07% of the analytical concentration. A robustness study did not identify any critical parameter that adversely affected the separation and quantification., Conclusions: Here, we report the development and validation of a robust, stability-indicating HPLC method for identification and assay of fipronil, permethrin, and BHT, including estimation of fipronil's and permethrin's degradation products in a topical drug product for dogs., Highlights: The new HPLC method permits the acquisition of data for all analytes of interest for a topical finished drug product containing fipronil, permethrin, and BHT., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Fipronil is a persistent insecticide known to transfer into hen eggs from exposure from animal drinking water and feed, but some questions remain regarding its transfer behavior and distribution characteristics. Therefore, the dynamic metabolism, residue distribution and transfer factor (TF) of fipronil were investigated in 11 edible tissues of laying hens and eggs over 21 days. After a continuous low-dose drinking water exposure scenario, the sum of fipronil and all its metabolites (defined as fipronil T ) quickly transferred to each edible tissue and gradually increased with exposure time. Fipronil T residue in eggs first appeared at 3 days and then gradually increased. After a single high-dose feed exposure scenario, fipronil T residue in edible tissues first appeared after 2 h, quickly peaked at 1 day, and then gradually decreased. In eggs, fipronil T residue first appeared at 2 days, peaked 6-7 days and then gradually decreased. The TF values followed the order of the skin (0.30-0.73) > egg yolk (0.30-0.71) > bottom (0.21-0.59) after drinking water exposure, and the order of the skin (1.01-1.59) > bottom (0.75-1.1) > egg yolk (0.58-1.10) for feed exposure. Fipronil sulfone, a more toxic compound, was the predominant metabolite with higher levels distributed in the skin and bottom for both exposure pathways. Fipronil T was distributed in egg yolks rather than in albumen owing to its lipophilicity, and the ratio of egg yolk to albumen may potentially reflect the time of exposure. The distinction is that the residues after feed exposure were much higher than that after drinking water exposure in edible tissues and eggs. The study highlights the residual characteristics of two exposure pathways, which would contribute to the tracing of contamination sources and risk assessment., Competing Interests: Declaration of competing interest The authors declare no actual or potential conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Fipronil and imidacloprid have been widely detected in UK surface waters in recent years, often at concentrations that ecotoxicological studies have shown can harm aquatic life. Down-the-drain (DTD) passage of pet flea and tick treatments are being implicated as an important source, with many of the UK's 22 million cats and dogs receiving routine, year-round preventative doses containing these parasiticides. The UK Water Industry's 3rd Chemical Investigation Programme (UKWIR CIP3) has confirmed wastewater as a major entry pathway for these chemicals into surface waters, but the routes by which they enter the wastewater system remain unclear. We addressed this knowledge gap by conducting the first quantification of DTD emissions from 98 dogs treated with spot-on ectoparasiticides containing fipronil or imidacloprid, through bathing, bed washing and washing of owners' hands. Both chemicals were detected in 100 % of washoff samples, with bathing accounting for the largest emissions per event (up to 16.8 % of applied imidacloprid and 24.5 % of applied fipronil). Modelled to account for the frequency of emitting activities, owner handwashing was identified as the largest source of DTD emissions from the population overall, with handwash emissions occurring for at least 28 days following product application and an estimated 4.9 % of imidacloprid and 3.1 % of fipronil applied in dog spot-ons passing down-the-drain via this route. The normalised daily per capita emissions for all routes combined were 8.7 μg/person/day for imidacloprid and 2.1 μg/person/day for fipronil, equivalent to 20-40 % of the daily per capita load in wastewater, as estimated from UKWIR CIP3 data. Within the current international regulatory framework adhered to by the UK, the environmental exposure of veterinary medicines intended for use in small companion animals is assumed to be low, and DTD pathways are not considered. We recommend a systematic review of regulations and practices to address this overlooked pollution pathway., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Eltrombopag Olamine is a drug used to treat thrombocytopenia, a disorder where blood platelet counts get lower and severe aplastic anemia. It serves as a thrombopoietin receptor agonist, which give rise to platelet production in the bone marrow., Objectives: The objective of this study is to develop a simple, specific, accurate, precise and economical Ultraviolet spectroscopy method to estimate the amount of Eltrombopag Olamine in bulk and tablet dosage form., Methods: The developed method was performed using methanol for identification and physicochemical characterization of the drug. The validation parameters like linearity, precision, accuracy, robustness limits of detection and quantitation, and specificity were assessed as per ICH Q2 (R2)., Results: The maximum absorbance wavelength (λmax) of the drug was found at 247 nm in methanol. The linearity was found in the concentration range of 2-14 μg/ml with regression equation y = 0.0619x - 0.0123 and r² = 0.999. The standard addition method was used to determine the accuracy of the developed method. The result was found in the % recovery range of 98-99%. The precision was done on λmax with respect to the parameters such as repeatability, intraday, and interday. The method was found to be precise as the % RSD value was found to be <2%. The detection limit value (LOD) and quantitation limit value (LOQ) were 0.0524 μg/ml and 0.1588 μg/ml, respectively., Conclusion: The developed method is simple, economical, accurate and selective. The developed method was adaptable for the estimation of Eltrombopag Olamine analysis in pharmaceutical dosage form and routine quality control laboratory., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The dead body of a 54-year-old man was found at home by his partner. He was off work due to depression. A letter with suicidal intention was present on the scene. He was known to be a heavy drinker, and near the body, an empty bottle of whisky was found. In addition, 2 empty blisters of Eliquis (apixaban) 5 mg, corresponding to 40 tablets, were identified. Apixaban is an oral anticoagulant, acting as a factor Xa inhibitor. Autopsy findings were mostly unremarkable, except numerous bruises and some superficial self-inflected wounds. Histology showed hematomas of calyces and renal pelvis and in the liver, several areas of perivenular haemorrhagic necrosis. Others organs were congestive. Femoral venous blood alcohol was 0.11 g/L. In femoral venous blood, a toxic concentration of apixaban was measured at 1184 ng/mL using LC-MS/MS. Other drugs found at therapeutic concentrations included diazepam (99 ng/mL), nordiazepam (171 ng/mL), flecainide (447 ng/mL), and mianserine (65 ng/mL). Using liquid chromatography coupled to high-resolution mass spectrometry, 2 metabolites were identified, O-desmethyl-apixaban (61.8% of the apixaban response) and hydroxyl-apixaban (4.5% of the apixaban response). Long-term therapy was confirmed by a concentration of 10390 pg/mg in pubic hair., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Fipronil (FIP) is a phenylpyrazole insecticide that, along with neonicotinoid insecticides, is regularly used worldwide. Photodegradation of FIP in aqueous systems is thought mainly to involve the reaction of desulfinylation to give fipronil desulfinyl (FIP-desulfinyl); however, little is known about further degradation reactions. We investigated FIP photodegradation by analyzing photodegradation products by liquid chromatography and liquid chromatography high-resolution tandem mass spectrometry using an Orbitrap instrument. A wide range of products, including dechlorinated compounds, was detected, and the structures were identified. FIP-desulfinyl has previously been found to be an important and persistent FIP photodegradation product; however, we also found that FIP-desulfinyl was photochemically decomposed to a didechlorinated product via a monodechlorinated product. The main photodegradation pathway was probably similar to that of ethiprole, which has a similar skeleton. The photodegradation rate constant was 22.6 times lower for FIP-desulfinyl (0.00372 min -1 ) than FIP (0.0839 min -1 ). The photodegradation rate constant was lower for the newly found didechlorinated product (0.001 min -1 or below) than FIP-desulfinyl, suggesting that the product is persistent in aquatic environments and could be an important indicator of long-term FIP contamination., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A novel approach for trace detection of fipronil with a molecularly imprinted electrochemiluminescence sensor (MIECLS) is proposed. The sensitivity is significantly improved via signal amplification of the enzymatic reaction of horseradish peroxidase (HRP) released from encapsulated liposomes which linked onto the template molecules after rebinding. The molecularly imprinted polymer membrane was prepared through the electropolymerization of monomers with fipronil as a template. After the elution of the template molecules, the analyte fipronil was reabsorbed into the cavities. HRP-encapsulated liposomes were linked to the target molecules by light-triggered click reaction. The higher the concentration of the target was, the more HRP-encapsulated liposomes were present on the molecularly imprinted polymer (MIP) sensor. Then, HRP was liberated from liposomes, and the catalytic degradation of hydrogen peroxide (H 2 O 2 ) by HRP occurs, which changed the electrochemiluminescence intensity of luminol significantly. The change of the ∆ECL intensity was linearly proportional to the logarithm of the fipronil concentration ranging from 1.00 × 10 -14 to 1.00 × 10 -9  mol/L, and the detection limit was 7.77 × 10 -16  mol/L. The recoveries obtained ranged from 95.7 to 105.8% with RSD < 5%. The sensitivity of the detection was significantly improved, and the analysis process was simplified in that the incubation step required in the conventional method was avoided. The sensor proposed provides a feasible platform for ultra-trace amount determination., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Cyantraniliprole is an anthranilic diamide insecticide used for the effective management of diamondback moth in cabbage. Dietary risk assessment of pesticides in food is a major concern now. This study developed a QuEChERS/HPLC-PDA-based highly efficient and reliable method, registering 89.80-100.11% recoveries of cyantraniliprole and its metabolite IN-J9Z38 from cabbage and soil with a relative standard deviation of 0.43-5.77%. Field experiment was conducted to study the residue dissipation of cyantraniliprole in cabbage and soil. Two foliar treatments of 10.26% formulation (Benevia) at 60 (T 1 ) and 120 (T 2 ) gram active ingredient/hectare were applied. The dissipation half-lives of cyantraniliprole in cabbage and soil were determined to be 3.5-4.2 and 3.8-5.3 days at T 1 and 3.9-4.8 and 4.1-4.7 days at T 2 , respectively. The maximum concentrations of IN-J9Z38 at T 1 and T 2 were 0.819 and 1.061 mg/kg, respectively, on the fifth day. A risk quotient value of <1 indicates no dietary risk to the consumers. The residues in the harvested cabbage were below the tolerance level of 2.0 mg/kg established by the regulatory body in India., (© 2021 John Wiley & Sons, Ltd.)

Plant uptake and metabolism of pesticides are complex and dynamic processes, which contribute to the overall toxicity of the pesticides. We investigated the metabolic fate of cyantraniliprole, a new diamide class of insecticide, during various growth stages of tomato. Cyantraniliprole was the major residue in leaves, flowers, and fruits, with the relative metabolite-to-parent ratios maintained at < 10% up to 28 days after treatment (DAT). Mature leaves contained consistently higher residues of cyantraniliprole than young leaves throughout the study. Flowers contained the highest cyantraniliprole residues up to 21 DAT, then gradually decreased. Immature green fruits had the highest cyantraniliprole residues (5.3 ± 0.7 ng/g; 42 DAT), and decreased toward red ripening stages (1.4 ± 0.2 ng/g; 84 DAT). Metabolism of cyantraniliprole primarily occurred in the foliage, where 21 metabolites were tentatively identified. Flowers and fruits contained 14 and four of these metabolites, respectively. Major transformation pathways were characterized by ring closure, followed by N-demethylation, and glycosylation. Additionally, plant metabolism of cyantraniliprole was also associated with several minor phase-I, phase-II, and breakdown metabolites. The occurrence of these metabolites in plants varied as a function of tissue types and their developmental stages. Our study highlights a tissue-specific biotransformation and accumulation of metabolites of cyantraniliprole in tomato., (© 2021. The Author(s).)

A modified QuEChERS method for determining cyenopyrafen in strawberries, mandarins and their processed products was established with a good linearity (R 2  > 0.9981), accuracy (recoveries of 83% to 111%) and precision (relative standard deviations of 0.9% to 14%). The limit of quantification (LOQ) was 0.01 mg/kg. Field results showed that the half-lives of cyenopyrafen were 6.8 and 11.8 d in strawberry and mandarin respectively, and that the final residues were within established maximum residue limits (MRLs). The household processing factors (PFs) for cyenopyrafen residues in strawberry and mandarin fruits were also studied: residues increased in strawberry jam (PF 1.51) and mandarin juice (1.31) but decreased in strawberries (0.58) and mandarin pulp (<0.17) after washing and peeling, respectively. A risk assessment showed that the risk from long-term dietary exposures to cyenopyrafen was 73.73%, indicating that consuming these products was unlikely to present a public health concern., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Three rapid, accurate, and ecofriendly processed spectrophotometric methods were validated for the concurrent quantification of remogliflozin (RGE) and vildagliptin (VGN) from formulations using water as dilution solvent. The three methods developed were based on the calculation of the peak height of the first derivative absorption spectra at zero-crossing points, the peak amplitude difference at selected wavelengths of the peak and valley of the ratio spectra, and the peak height of the ratio first derivative spectra. All three methods were validated adapting the ICH regulations. Both the analytes showed a worthy linearity in the concentration of 1 to 60 µg/mL and 2 to 90 µg/mL for VGN and RGE, respectively, with an exceptional regression coefficient (r2 ≥ 0.999). The developed methods demonstrated an excellent recovery (98.00% to 102%), a lower percent relative standard deviation, and a relative error (less than ±2%), confirming the specificity, precision, and accuracy of the proposed methods. In addition, validated spectrophotometric methods were commendably employed for the simultaneous determination of VGN and RGE from solutions prepared in the laboratory and the formulation. Hence, these methods can be utilized for the routine quality control study of the pharmaceutical preparations of VGN and RGE in pharmaceutical industries and laboratories. The ecofriendly nature of the anticipated spectrophotometric procedures was confirmed by the evaluation of the greenness profile by a semi-quantitative method and the quantitative and qualitative green analytical procedure index (GAPI) method.

Fipronil is a broad-spectrum phenyl-pyrazole insecticide that is widely used in agriculture. However, in the environment, its residues are toxic to aquatic animals, crustaceans, bees, termites, rabbits, lizards, and humans, and it has been classified as a C carcinogen. Due to its residual environmental hazards, various effective approaches, such as adsorption, ozone oxidation, catalyst coupling, inorganic plasma degradation, and microbial degradation, have been developed. Biodegradation is deemed to be the most effective and environmentally friendly method, and several pure cultures of bacteria and fungi capable of degrading fipronil have been isolated and identified, including Streptomyces rochei, Paracoccus sp., Bacillus firmus, Bacillus thuringiensis, Bacillus spp., Stenotrophomonas acidaminiphila, and Aspergillus glaucus. The metabolic reactions of fipronil degradation appear to be the same in different bacteria and are mainly oxidation, reduction, photolysis, and hydrolysis. However, the enzymes and genes responsible for the degradation are somewhat different. The ligninolytic enzyme MnP, the cytochrome P450 enzyme, and esterase play key roles in different strains of bacteria and fungal. Many unanswered questions exist regarding the environmental fate and degradation mechanisms of this pesticide. The genes and enzymes responsible for biodegradation remain largely unexplained, and biomolecular techniques need to be applied in order to gain a comprehensive understanding of these issues. In this review, we summarize the literature on the degradation of fipronil, focusing on biodegradation pathways and identifying the main knowledge gaps that currently exist in order to inform future research. KEY POINTS: • Biodegradation is a powerful tool for the removal of fipronil. • Oxidation, reduction, photolysis, and hydrolysis play key roles in the degradation of fipronil. • Possible biochemical pathways of fipronil in the environment are described., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

We developed a sensitive and rapid lateral flow immunochromatographic (LFI) assay for the simultaneous detection of fipronil and its metabolites in eggs and cucumbers using gold nanoparticle (GNP)-labeled monoclonal antibodies (mAbs). Anti-fipronil mAbs (1B6) were produced using two haptens and identified by heterologous indirect competitive enzyme-linked immunosorbent assay (icELISA) with half maximal inhibitory concentration (IC 50 ) and limit of detection (LOD) values of 0.46 ± 0.07 and 0.05 ± 0.01 ng mL -1 , respectively. The developed LFI strip showed high sensitivity and specificity in the detection of fipronil with cut-off and visual limit of detection (vLOD) values of 10 and 0.25 ng mL -1 , respectively. Furthermore, the application of LFI in the detection of fipronil-spiked egg and cucumber samples was validated by liquid chromatography tandem mass spectrometry (LC-MS/MS). Our developed LFI assay is suitable for detection of fipronil and its metabolites in real samples., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

A new member of the pyrazoline family, 3-naphthyl-1-(4-trifluoromethyl)-5-(4-carboxy phenyl)-2-pyrazoline has been evaluated as a precolumn derivatization reagent for the analysis of primary alcohols using HPLC. The simultaneous separation of eight alcohol derivatives (C 1 -C 8 ) within 15 min was achieved on a reverse-phase C 8 column with an isocratic elution mode. The derivatives were detected with fluorescence at an emission wavelength of 470 nm when excited at 360 nm. The identification of the corresponding derivatives was carried out by LC-MS/MS and all showed their characteristic parent peak in negative ion mode. The proposed method was validated using normal analytical tools and was found to be excellent. As a preliminary application, our method was used to determine ethanol concentration in alcohol-containing chocolates and cough syrup., (© 2021 John Wiley & Sons, Ltd.)

Background: During food or feed contamination events, it is of utmost importance to ensure their rapid resolution to minimize impact on human health, animal health and finances. The existing Rapid Alert System for Food and Feed (RASFF) is used by the European Commission, national competent authorities of member countries and the European Food Safety Authority to report information on any direct or indirect human health risk arising from food or feed, or serious risks to animal health or the environment in relation to feed. Nevertheless, no methods exist to to collectively evaluate this vast source of supply chain information., Methods: To aid in the extraction, evaluation and visualization of the data in RASFF notifications, we present the Rapid Alert Supply Network Extractor (RASNEX) open-source tool available from https://doi.org/10.5281/zenodo.4322555 freely. Among RASNEX's functions is the graphical mapping of food and feed supply chain operators implicated in contamination events. RASNEX can be used during ongoing events as a support tool for risk analysis using RASFF notifications as input., Results: In a first use case, we showcase the functionality of RASNEX with the RASFF notification on a 2017/2018 contamination event in eggs caused by the illegal use of fipronil. The information in this RASFF notification is used to visualize nine different flows of main and related food products. In a second use case, we combine RASFF notifications from different types of food safety hazards (Salmonella spp., fipronil and others) to obtain wider coverage of the visualized egg supply network compared to the first use case. Actors in the egg supply chain were identified mainly for Italy, Poland and Benelux. Other countries (although involved in the egg supply chain) were underrepresented., Conclusions: We hypothesize that biases may be caused by inconsistent RASFF reporting behaviors by its members. These inconsistencies may be counteracted by implementing standardized decision-making tools to harmonize decisions whether to launch a RASFF notification, in turn resulting in a more uniform future coverage across European food and feed supply chains with RASNEX., Competing Interests: The authors have declared that no competing interests exist.

Maize seed treatment with chemicals to control underground pests is a common agricultural practice, but inappropriate use of insecticides poses a considerable threat to plant development and soil nontarget organisms. In this study, the availability of tetraniliprole seed dressing to control the black cutworm Agrotis ipsilon (Lepidoptera: Noctuidae) in the maize seeding stage and its safety to earthworms (Eisenia fetida) were investigated. The selective toxicity (ST) of tetraniliprole between E. fetida and A. ipsilon was greater than 4000. No significant adverse effect of tetraniliprole seed treatment on the germination of maize seeds was observed at concentrations of 2.4-9.6 g a.i. /kg seed. Compared with the untreated control, seed treatment with tetraniliprole at 9.6 g a.i. /kg seed greatly reduced the percentage of damaged plants from 88.73% to 26.67%, and achieved the highest control effect of 69.91%. Tetraniliprole of 2.4 g a.i. /kg seed can effectively inhibit A. ipsilon until 14 days after seed germination, with the lowest mortality rate of 44.44%. During the entire exposure period, the maximum residual concentration of tetraniliprole detected in the soil (5.86 mg/kg) was considerably lower than the LC 50 value of tetraniliprole to E. fetida (>4000 mg/kg). According to the low-tier risk assessment, the highest risk quotient (RQ) of tetraniliprole seed treatment to earthworms at test concentrations was 2.8 × 10 -3 , which was evaluated as acceptable. This study provided data support for tetraniliprole seed treatment to control underground pests in maize fields., (Copyright © 2021. Published by Elsevier Inc.)

Cyantraniliprole and chlorantraniliprole are anthranilic diamide insecticides acting on ryanodine receptors. In this study, two camel-derived nanobodies (Nbs, named C1 and C2) recognizing cyantraniliprole as well as chlorantraniliprole were generated. C1-based enzyme-linked immunosorbent assays (ELISAs) for the detection of the two insecticides were developed. The half-maximum signal inhibition concentrations (IC 50 ) of cyantraniliprole and chlorantraniliprole by ELISA were 1.2 and 1.5 ng mL -1 , respectively. This assay was employed to detect these two insecticides in soil and vegetables. The average recoveries of cyantraniliprole from both bok choy (Brassica chinensis L.) and soil samples were 90-129%, while those of chlorantraniliprole were in a range of 89-120%. The insecticide residues in soil and bok choy, which were collected from plots sprayed with cyantraniliprole and chlorantraniliprole, were simultaneously detected by the resulting ELISA and a high-performance liquid chromatography (HPLC) method, showing a satisfactory correlation. Higher concentrations of chlorantraniliprole than cyantraniliprole were detected in soil and vegetables, which indicates the longer persistence of chlorantraniliprole in the environment.

The treatment of inflammatory skin conditions relies on a deep understanding of how drugs and tissue behave and interact. Although numerous methods have been developed that aim to follow and quantify topical drug pharmacokinetics, these tools can come with limitations, assumptions, and trade-offs that do not allow for real-time tracking of drug flow and flux on the cellular level in situ. We have developed a quantitative imaging toolkit that makes use of stimulated Raman scattering microscopy and deep learning-based computational image analysis to quantify the uptake of specific drug molecules in skin without the need for labels. Analysis powered by trained convolutional neural networks precisely identified features such as cells, cell junctions, and cell types within skin to enable multifactorial analysis of skin pharmacokinetics. We imaged and quantified the flow and flux of small molecule drugs through the layers and structures of ex vivo nude mouse ear skin and extracted pharmacokinetic parameters through convolutional neural network-based image processing, including relative area under the curve accumulation, time of maximum drug concentration, and in situ partition ratios. This approach, which facilitates the direct observation and quantification of pharmacokinetics, can be used to glean mechanistic insight into underlying phenomena in skin pharmacokinetics., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Environmental risk assessment of complex chemical mixtures has increasingly been prioritized as a management goal, especially in the regulatory sector. Although fipronil and its three degradates (-sulfone, -sulfide and -desulfinyl) have been frequently quantified in waterways, little information is available about the likelihood and magnitude of ecological risk posed by these chemical mixtures - collectively known as fiproles - in surface water. In the present study, a probabilistic risk assessment of mixtures of fipronil and its three degradates was conducted for three effluent-dominated southern California rivers: Los Angeles River (LAR), San Gabriel River (SGR) and Santa Clara River (SCR), California, USA. The assessments, which used fiproles as an integrated proxy, were based on three levels of toxicity endpoints: median lethal concentration (LC50), half-maximal effective concentration (EC50), and lowest observed effect concentration (LOEC), to gain comprehensive assessment information. Probabilistic approaches based on species sensitivity distribution (SSD) and exposure concentration distribution (ECD) were developed with the log-logistic model by pooling the toxicity and occurrence data, respectively. The 5th percentile hazardous concentrations (HC5s) were calculated to be at low parts per billion levels, enabling these values to be used to estimate the chemical-specific benchmarks for components that lack ecotoxicity data. The single substance potentially affected fraction (ssPAF) of fiproles revealed risk levels for the three rivers in descending order: LAR ≥ SGR > SCR. The overall risk probability estimated from the joint probability curve (JPC) by Monte Carlo simulation was 1.13 ± 0.20% (LC50), 9.31 ± 1.46% (EC50), and 6.58 ± 1.43% (LOEC) for the three rivers collectively. These results derived from the fiproles indicates that fipronil and its degradates pose risks to the aquatic organisms in the surface water of the three rivers. The present study provides a methodology for the use of a proxy in the risk assessment of chemical mixtures., Competing Interests: Declaration of Competing Interest The author declares that he has no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

A rapid, simple, precise and accurate high performance liquid chromatographic (HPLC) analytical method was developed and validated for the determination of the active substances (a.s.) azoxystrobin, topramezone, acetamiprid, fluometuron and folpet in their respective commercially available formulations. The method was used for the analysis of samples under the frame of the national quality control program of plant protection products in the Greek market. Chromatographic separation of the active substances from additives and co-formulants is achieved using isocratic elution with acetonitrile and 0.1% phosphoric acid solution (60:40 v/v) at a flow rate of 0.4 mL min -1 on a C 18 monolithic column (Chromolith Performance-RP18e 100 × 4.6 mm) and UV detection at 230 nm. Validation parameters of the method fulfilled acceptability criteria. Recovery of all individual compounds was in the range 97.8-100.9%. Precision expressed as relative standard deviation was lower than the theoretical values of the modified Horwitz equation. Correlation coefficients of linearity of response were better than 0.999. The benefits of the proposed method are significant reduction in analysis time and total cost since all analytes were determined with the same extraction procedures and chromatographic system. Analysis of real samples for all active ingredients confirmed fitness for purpose of the suggested method.

Pydiflumetofen is a novel and efficient broad-spectrum chiral fungicide consisting of a pair of enantiomers. A simple and sensitive chiral analytical method was established to determine the enantiomers of this chiral fungicide in food and environmental samples by ultra-high-performance liquid chromatography tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) using QuEChERS method coupled with octadecylsilane-dispersive solid-phase extraction (C 18 -dSPE) as extraction procedure. The specific optical rotation and the absolute configuration of the enantiomers were identified by polarimetry and electronic circular dichroism (ECD). The elution order of the pydiflumetofen enantiomers on Lux Cellulose-2 was S-(-)-pydiflumetofen and R-(+)-pydiflumetofen. The average recoveries of eleven matrices ranged from 71.3% to 107.4%. The intraday relative standard deviations (RSDs) were less than 11.8%, and the interday RSDs were less than 12.6% for the two enantiomers. Stereoselective dissipation in pakchoi and soil were observed: S-(-)-pydiflumetofen was degraded faster than R-(+)-pydiflumetofen in pakchoi, causing the enantiomer fraction (EF) of the enantiomers to change from 0.50 to 0.42 in 7 days. However, R-(+)-pydiflumetofen was degraded faster than S-(-)-pydiflumetofen in soil, causing the EF of the enantiomers to change from 0.49 to 0.52 in 21 days. This study provides a method for monitoring pydiflumetofen enantiomer residues, which is crucial for improving risk assessments and the development of chiral pesticides., (Copyright © 2020 Elsevier Inc. All rights reserved.)

In this study, a new method for simultaneous determination of cyantraniliprole, chlorantraniliprole, tetrachlorantraniliprole, cyclaniliprole and flubendiamide in edible mushrooms by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) combined with a modified QuEChERS procedure. The samples were extracted using acetonitrile and then cleaned up by primary secondary amine (PSA) and octadecylsilane (C18). The determination of these insecticides was achieved in less than 5 min using an electrospray ionization source in positive mode (ESI+) for cyantraniliprole and chlorantraniliprole, while negative mode (ESI-) for tetrachlorantraniliprole, cyclaniliprole and flubendiamide. The linearities of the calibrations for all target compounds were acceptable (R 2  ≥ 0.9922). The limits of detection and quantification were 0.05-2 μg kg -1 and 5 μg kg -1 , respectively. Acceptable recoveries (73.5-110.2%) were acquired for these insecticides with RSDs less than 12.7%. The results demonstrated that the proposed method was effective and convenient for the determination of these insecticides in edible mushrooms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Advances in genomic analyses enable the identification of new proteins that are associated with disease. To validate these targets, tool molecules are required to demonstrate that a ligand can have a disease-modifying effect. Currently, as tools are reported for only a fraction of the proteome, platforms for ligand discovery are essential to leverage insights from genomic analyses. Fragment screening offers an efficient approach to explore chemical space. Presented here is a fragment-screening platform, termed PhABits (PhotoAffinity Bits), which utilizes a library of photoreactive fragments to covalently capture fragment-protein interactions. Hits can be profiled to determine potency and the site of crosslinking, and subsequently developed as reporters in a competitive displacement assay to identify novel hit matter. The PhABit platform is envisioned to be widely applicable to novel protein targets, identifying starting points in the development of therapeutics., (© 2020 Wiley-VCH GmbH.)

Oxathiapiprolin is a chiral fungicide used in China for the prevention and treatment of grape downy mildew, but its potential risk could be inaccurately assessed without distinguishing its enantiomers. In this study, an effective and sensitive chiral analytical method was first established for quantification of oxathiapiprolin enantiomers using supercritical fluid chromatography tandem mass spectrometry. Baseline separation for oxathiapiprolin enantiomers was achieved for less than 3 min by using a Lux Cellulose-2 chiral column with the resolution of 1.51. The elution order of the eluting enantiomers was identified as (-)-oxathiapiprolin and (+)-oxathiapiprolin by an optical rotation detector. The grape samples were extracted by QuEChERS method, with the average recoveries of each enantiomer in grapes were in the range of 88.1-111.8% and the relative standard deviations were less than 18.9%. The enantioselective analysis of the dissipation of oxathiapiprolin in field grape samples showed that (-)-oxathiapiprolin was dissipated faster than (+)-oxathiapiprolin. The results indicate that this proposed method could provide data support for the risk assessment of oxathiapiprolin in agricultural produces in a more accurate way., (© 2020 Wiley-VCH GmbH.)

The widespread use of neonicotinoids (NEOs) and fipronils (FIPs) as insecticides in modern agriculture has been widely investigated because of their adverse effects on human health. This study aimed to develop an accurate quantitative approach to analyze NEOs and FIPs in tea by using a modified QuEChERS method based on dispersive solid-phase extraction procedure coupled with liquid chromatography-high resolution mass spectrometry. To minimize matrix interferences largely, we selected polyvinylpolypyrrolidone (PVPP) and strong cationic exchange adsorbent (PCX) as dispersive adsorbents to eliminate polyphenols and caffeine from tea extract, respectively. Under optimal conditions, a slight matrix effect was observed for NEOs and FIPs due to the highest reductions of polyphenols and caffeine (94% and 88%, respectively). The method was validated by the European Medicines Agency and Eurachem guidelines and was successfully applied to tea samples for NEOs and FIPs analysis. NEOs and FIPs were commonly detected in tea., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Fipronil, a widely used insecticide and pesticide, with its toxic metabolite fipronil sulfone was detected in fipronil-contaminated eggs as a result of inappropriate use. However, little is known about whether fipronil and fipronil sulfone transfer into fetus through the blood-placenta barrier. Our objectives were to investigate the transplacental transfer and the pharmacokinetics of fipronil and fipronil sulfone in rats. Male and female (with 13 days of gestation) Sprague-Dawley rats were used in pharmacokinetics and transplacental transfer experiments, respectively. Biologic samples were collected at each time point after fipronil intravenous or oral administration. To monitor fipronil and fipronil sulfone in the plasma, placenta, amniotic fluid, and fetus, a validated liquid chromatography tandem mass spectrometry method was developed. After fipronil administration in male rats, the oral bioavailability decreased, whereas the biotransformation increased as the dose increased, revealing an enhancement of first-pass effect and a fast metabolism in vivo. The results of fipronil transplacental transfer in pregnant rats demonstrated that the concentration of fipronil and fipronil sulfone varied in the following order, respectively: placenta > plasma > fetus > amniotic fluid and plasma > placenta > fetus > amniotic fluid. This is the first direct evidence that fipronil and fipronil sulfone cross the blood placental barriers and enter the fetus. The amount of fipronil distributed to the fetus was greater than that of fipronil sulfone in the short term, but by contrast, pharmacokinetic data showed that the latter stayed longer in the body. These findings provide constructive information for public health alarm. SIGNIFICANCE STATEMENT: Fipronil and fipronil sulfone interfere with the GABAergic system. Fipronil can cause thyroid dysfunction, which may affect brain growth and nerve development. Although we knew that fipronil and fipronil sulfone could enter eggs, there was no direct evidence that they would enter fetuses. This research provided evidence on the pharmacokinetics and transplacental transfer of fipronil and fipronil sulfone, confirming our hypothesis., (Copyright © 2020 by The American Society for Pharmacology and Experimental Therapeutics.)

In the summer of 2017, several European Union Member States were involved in a food alert caused by the presence of fipronil pesticide residues in chicken eggs. The food alert became a major news and received wide coverage both in the mass media and on the Internet. This article describes a study that analyzed how the Italian online information sources represented the fipronil alert, using web monitoring techniques and both manual and automatic content analysis methods. The results indicate that the alert was amplified because general news media could represent the alert within the frame of a political scandal, and because different social actors exploited the case. However, online information sources correctly communicated that the risks for consumers were low, reporting mainly what was officially communicated by the Italian health authorities. The study provides empirical evidence on how the online information sources represent food risks and food alerts and offers useful indications for health authorities in charge of the public communication of food risks., (© 2020 Society for Risk Analysis.)

Oxathiapiprolin is the first chiral piperidinyl thiazole isoxazoline fungicide developed to control downy mildew and other diseases, and there were no prior reports on its enantiomeric residue. In this study, a modified quick, easy, cheap, effective, rugged, and safe extraction and purification method followed by ultra-high performance liquid chromatography-tandem mass spectrometry determination was first developed and validated for the residue analysis of oxathiapiprolin enantiomers and its metabolite IN-E8S72 in green tea and other crops. Oxathiapiprolin enantiomers and IN-E8S72 were separated on a chiral Lux Cellulose-3 column with the use of 0.1% formic acid in acetonitrile and 5 mmol/L ammonium acetate in water as mobile phases. IN-E8S72 was eluted first, followed by (-)-oxathiapiprolin, and then (+)-oxathiapiprolin. The recoveries ranged from 53.3 to 125.3% with relative standard deviations ranging from 1.4 to 16.0%. The limits of quantification for (-)-oxathiapiprolin and (+)-oxathiapiprolin were 0.005 mg/kg in romaine lettuce, head cabbage, potato, grape, and garlic, 0.01 mg/kg in soybean and pea, and 0.025 mg/kg in green tea and dry pepper. The limits of quantification of IN-E8S72 were twice those of (-)-oxathiapiprolin. Screening results with real market samples indicated that there was no enantiomeric excess in the oxathiapiprolin residue in romaine lettuce., (© 2020 Wiley-VCH GmbH.)

New degradation impurities at m/z 327.15 and m/z 311.16 using gradient UHPLC method with UV detection and highly selective QDa mass detection were observed during the ruxolitinib hydrobromide (RUX.HBr) : excipient binary mixture degradation study. High mass resolution LC-MS and nuclear magnetic resonance (NMR) techniques were employed to identify and fully characterize the degradation compounds. The degradation impurities were unambiguously identified as (R)-4-amino-6-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)pyrimidine-5-carboxylic acid and (R)-3-(4-(6-amino-5-formylpyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile and mechanism of their formation was proposed. It has been confirmed that the degradation products are formed in mixtures of RUX.HBr with some excipients in the presence of oxygen. Based on the forced degradation study, the chemically stable of pharmaceutical formulations were prepared to eliminate the formation of these impurities., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Fipronil (FP) is an emerging insecticide, however, its occurrence in drinking water and source water is limited. In this work, a total of 789 tap water and 95 ground water samples were collected from China in June 2019 in order to assess occurrence of FP and its derivatives (FPs). FPs were also analyzed in source, treated (n = 10, July), and tap water samples (n = 81, July and October 2019) originating from the central Yangtze River and its tributary, the Hanshui River in Wuhan. The sum concentrations of FPs (ΣFPs) in the tap water in China ranged from not detected (ND) to 5.07 (median: 0.03 ng/L), with FP found in 55.3% of the samples, and other targets ≤ 50.0%. Significant regional variations in the ΣFPs values were found between East China (75th percentile: 0.31 ng/L) and Northwest China (0.04), as well as between East China and North China (0.04). Similar ΣFPs values were found for ground water and tap water. The estimated daily intake of ΣFPs via water ingestion was below 200 pg/kg-bw/day for all age groups and was lower than the reference dose for FP (0.2 μg/kg-bw/day). Additionally, FPs were found in all of the source water samples collected in Wuhan with concentrations in the range of 0.84-2.72 ng/L for ΣFPs (median: 2.39). Most of these FPs were removed during water treatment. Higher concentration of ΣFPs in tap water was observed in July (median: 0.04 ng/L) compared to that in October (ND). This is the first study on the occurrence of FPs in the Yangtze River, the fate of FPs during the tap water treatment, and the regional distribution of FPs in tap water from China., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Selpercatinib and pralsetinib are potent and selective tyrosine kinase inhibitors targeting the rearranged during transfection (RET) receptor in various types of cancer. In this study, a bioanalytical assay was developed and fully validated for selpercatinib and pralsetinib in mouse plasma and partially in eight mouse tissue homogenates using liquid chromatograph-tandem mass spectrometry. Samples were pre-treated by protein precipitation with acetonitrile using erlotinib as internal standard. Separation of the analytes was performed on an ethylene bridged octadecyl silica C18 column by gradient elution using ammonium hydroxide (in water) and methanol. Analytes were detected by positive electrospray ionization in selected reaction monitoring mode. A linear concentration range of 2-2000 ng/ml was used for the validation of the assay for both inhibitors. The precision values (within-day and between-day) ranged between 3.4 and 10.2% for selpercatinib and 3.1-14.6% for pralsetinib in all matrices. Furthermore, data obtained for accuracy were between 91.7 and 109.3% and 85.1-114.1% for selpercatinib and pralsetinib, respectively. No significant matrix effects or extraction losses were observed and both analytes were stable under all investigated conditions. Finally, a pilot study for selpercatinib in mice was conducted employing this method, followed by a successful incurred sample reanalysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

As part of a new generation of diamide insecticides, cyantraniliprole has broad application prospects. In the present study, a QuEChERS-UPLC-MS/MS method was established to determine the residues of cyantraniliprole and its main metabolite J9Z38 in soil and earthworms. Moreover, the accumulation and toxicity of cyantraniliprole and J9Z38 in earthworms were evaluated. The present results show that the detection method of cyantraniliprole and J9Z38 has high sensitivity and accuracy, which could be used for the accurate quantification of cyantraniliprole and J9Z38 residues in soil and earthworms. Additionally, cyantraniliprole degraded faster than its main metabolite J9Z38 in the artificial soil. Moreover, the bioenrichment efficiency of cyantraniliprole was higher than J9Z38. The toxicity test result showed that cyantraniliprole and J9Z38 could induce oxidative stress effect in earthworms from 5.0 mg/kg, finally resulting in cellular damage. Moreover, the oxidative damage degree induced by cyantraniliprole was higher than J9Z38. Combining the results of residue test and toxicity test, although cyantraniliprole degraded faster than its main metabolite J9Z38 in the artificial soil, its risk to earthworms was higher than J9Z38., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The 18 kDa translocator protein (TSPO) is the main molecular target to image neuroinflammation by positron emission tomography (PET). However, TSPO-PET quantification is complex and none of the kinetic modelling approaches has been validated using a voxel-by-voxel comparison of TSPO-PET data with the actual TSPO levels of expression. Here, we present a single case study of binary classification of in vivo PET data to evaluate the statistical performance of different TSPO-PET quantification methods. To that end, we induced a localized and adjustable increase of TSPO levels in a non-human primate brain through a viral-vector strategy. We then performed a voxel-wise comparison of the different TSPO-PET quantification approaches providing parametric [ 18 F]-DPA-714 PET images, with co-registered in vitro three-dimensional TSPO immunohistochemistry (3D-IHC) data. A data matrix was extracted from each brain hemisphere, containing the TSPO-IHC and TSPO-PET data for each voxel position. Each voxel was then classified as false or true, positive or negative after comparison of the TSPO-PET measure to the reference 3D-IHC method. Finally, receiver operating characteristic curves (ROC) were calculated for each TSPO-PET quantification method. Our results show that standard uptake value ratios using cerebellum as a reference region (SUV CBL ) has the most optimal ROC score amongst all non-invasive approaches.

Herein, a cold-induced aqueous two-phase system (CI-ATPS)-based concurrent sample enrichment and cleanup strategy was developed for the analysis of fipronil and its metabolites in dietary samples by liquid chromatography-high resolution mass spectrometry. By optimizing the conditions in CI-ATPS, the analytes were extracted to a small volume acetonitrile-rich phase with the enrichment factor of three-folds. Additionally, the lipids could be efficiently precipitated between the lower and upper layer to complete the removal of lipids. In detection analysis, a target single ion monitoring mode was employed to enhance the detection capability of fipronil and its metabolites. Generally, this established method could detect the target analytes in dietary samples at ng/kg level. Finally, this method was validated and certificated by a proficiency test, then was also successfully applied to dietary samples in the Total Diet Study, and the results showed that 56.3% of samples were detected with fipronil or its metabolites., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Direct oral anticoagulants (DOACs) have been commonly used for the treatment of venous thromboembolism and for the prevention of stroke in patients with atrial fibrillation. Despite not being initially recommended, monitoring DOACs plasma concentrations is now recognized as essential in emergency situations and in special populations. Moreover, the inter-individual variability found in real studies as well as the high reported non-adherence are corroborating the importance of determining the individual relationship between administered doses, plasma concentrations and pharmacological effects. Therefore, accurate but user-friendly bioanalytical techniques are required to monitor DOACs plasma concentrations in routine clinical practice and phase IV clinical trials. Herein, a fast and simple high performance liquid chromatography (HPLC) method coupled to diode array detection (DAD) was developed, validated and applied to quantify the four currently marketed DOACs (apixaban, edoxaban, dabigatran and rivaroxaban). Sample preparation was performed by solid phase extraction followed by evaporation and concentration of the analytes. Chromatographic separation was accomplished within 6 min on a reversed-phase column (octadecyl-silica packing material; 55 mm × 4 mm, 3 μm particle size), applying a mobile phase composed of an aqueous solution of formic acid (0.1 %, v/v) and acetonitrile, pumped with a gradient elution at 30 °C. The proposed method was linear (r 2 ≥ 0.993) within the concentration ranges of 0.017-5.28 μg mL -1 , 0.066-5.28 μg mL -1 , 0.033-5.28 μg mL -1 and 0.017-5.28 μg mL -1 for apixaban, dabigatran, edoxaban and rivaroxaban, respectively, all of them including the expected range of therapeutic concentrations. Overall, intra- and inter-day trueness of quality control samples, including at the lower limit of quantification (LLOQ), varied between -12.98 to 5.79 %, while imprecision was lower than 16.43 %, supporting that the method is accurate and precise in accordance to international guidelines. Recovery and stability were also assessed and allowed the method to be applied in clinical practice, during therapeutic drug monitoring., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: The aim of the study was to perform analytical verification and comparison of chromogenic assays for determination of dabigatran, rivaroxaban and apixaban concentration on BCSXP and STA Compact Max analysers., Materials and Methods: Precision, linearity, measurement uncertainty estimation and determination of limit of blank, limit of determination and limit of quantification were calculated. Analytical performance specifications were set according to manufacturer specifications and literature data on between laboratory variability. Comparison of the methods was done using Bland-Altman and Passing-Bablok regression analysis., Results: Obtained results have shown acceptable precision on STA Compact Max only for dabigatran (CV = 3.5%) at lower concentration level comparing to manufacturer declaration (CV = 3.6%). On BCSXP, the highest coefficient of variation has been shown for apixaban (6.1%) at lower concentration level. Within laboratory precision was not met on STA Compact Max for all assays. Bland-Altman analysis has shown statistically significant bias for dabigatran (23.2%, 95%CI 11.2 - 35.3; P < 0.001) and apixaban (8.4%, 95%CI 1.2 - 15.6; P = 0.023). Passing-Bablok regression analysis has shown systematic and proportional deviation between methods for rivaroxaban (y = 6.52 (2.94 to 11.83) + 0.84 (0.80 to 0.89) x., Conclusion: Chromogenic assays for dabigatran, rivaroxaban and apixaban on BCSXP and STA Compact Max analysers are shown as methods with satisfactory long-term analytical performance specifications for determination of direct oral anticoagulants in clinical laboratories. However, we cannot recommend interchangeable use because of the significant bias between assays., Competing Interests: Potential conflict of interest: None declared., (Croatian Society of Medical Biochemistry and Laboratory Medicine.)

In this paper a dispersive magnetic-solid phase extraction (MSPE) using a graphene nanocomposite (rG/Fe 3 O 4 ) followed by ultra high performance liquid chromatography with photodiode array detection has been developed for the simultaneous analysis of new class of oral anticoagulants (NOAs) in human plasma. The performance of the nanocomposite graphene@Fe 3 O 4 on the magnetic solid phase extraction of apixaban, rivaroxaban and dabigatran has been optimized using a Box-Behnken design of experiment. The amount of graphene nanocomposite, the sample pH and the adsorption time were the investigated parameters as a function of the extraction recovery. The analytical method was fully validated based on linearity, limit of detection (LOD), limit of detection (LOQ), inter- and intra-day precision and trueness, and extraction yield. Under optimal condition, excellent linearity (R 2 > 0.9987) over the range (0.001-5.0 μg/mL), limit of detection (0.003 μg/mL), precision (0.81-8.97% RSD) and trueness (-5 to 9 % BIAS%) were observed for the target drugs. The average extraction recovery under optimal from plasma samples ranged between 96.6-98.6% for apixaban, rivaroxaban and dabigatran and the internal standard. The proposed method was developed, validated and successfully applied to the measurement of these NOAs in patients. The new approach offers an attractive alternative for the simultaneous analysis of the selected NOAs from plasma samples, providing several advantages including fewer sample preparation steps, ease of performance, and higher recoveries compared to traditional methodologies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Nonvitamin K antagonist oral anticoagulants (NOACs) have emerged as the preferred choice for the treatment of atrial fibrillation (AF). The establishment of a therapeutic range to minimize bleeding and thrombosis is important for personalized treatment of NOACs. The importance of dried blood spots (DBSs) has increased in medical care. An efficient and effective DBS analytical method could facilitate the concentration management of NOACs. The postcolumn infused internal standard (PCI-IS) method was applied to estimate spot volume and quantify dabigatran, rivaroxaban, and apixaban concentrations on DBS cards. The extraction solvent contented 0.1% formic acid and 70% ACN with a successive extraction procedure. Paired DBS and plasma samples from patients undergoing NOAC therapy ( n = 269) were used to calculate conversion factors. [ 13 C 6 ]-Rivaroxaban was selected as the PCI-IS. The quantification accuracy for the three NOACs was within 88.9-104.3%. The RSDs of the repeatability and intermediate precision were below 10%. The obtained conversion factors of DBS to plasma concentrations of dabigatran, apixaban, and rivaroxaban were 1.81, 1.59, and 1.31, respectively. Bland-Altman analysis showed that the % differences between predicted and measured plasma concentrations were within a bias of ±20%. The result showed that PCI-IS was an accurate and efficient LC-MS/MS method to simultaneously estimate blood volume and NOAC concentrations on DBS cards. The stability results revealed that the DBS sampling strategy could improve compound stability. The developed method offers a new strategy for the therapeutic drug monitoring of NOACs and may improve the safe use of these drugs.

Fipronil is an insecticide that is not approved in the European Union in food. In 2017, fipronil was involved in a European health alert due to its presence in fresh hen eggs because of an illicit use in poultry farms, so reliable methods are needed to determine fipronil and its main metabolites in these matrixes. In this work, we report the first approach to the study of fipronil and two metabolites, fipronil-sulfone and fipronil-sulfide by CE. MEKC mode was employed using a solution of 50 mM ammonium perfluorooctanoate pH 9.0 with 10% (v/v) methanol as background electrolyte. The proposed method was combined with a simple sample treatment based on salting-out assisted LLE (SALLE) using acetonitrile as extraction solvent and ammonium sulfate as salt. The SALLE-MEKC-UV method allowed the simultaneous quantification of fipronil and fipronil-sulfone. Validation parameters yielded satisfactory results, with precision, expressed as relative SD, below 14% and recoveries higher than 83%. Limits of detection were 90 µg/kg for fipronil and 150 µg/kg for fipronil-sulfone, so in terms of sensitivity further studies of sample treatments allowing extra preconcentration or the use of more sensitive detection, such as MS, would be needed., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Larotrectinib, is an orally active novel small molecule approved for the treatment of solid tumors in pediatrics and adult patients. It acts by inhibiting tropomyosin receptor kinase. In this paper, we report the development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of larotrectinib in mice plasma as per the FDA regulatory guideline. Plasma samples processing was accomplished through simple protein precipitation using acetonitrile enriched with internal standard (IS, enasidenib). The chromatographic analysis was performed using a gradient mobile phase comprising 10 mM ammonium acetate and acetonitrile at a flow-rate of 0.8 mL/min on an X-Terra Phenyl column. The UV detection wave length was set at λ max 262 nm. Larotrectinib and the IS eluted at 3.85 and 6.60 min, respectively with a total run time of 8.0 min. The calibration curve was linear over a concentration range of 0.20-5.00 μg/mL (r 2 =≥0.992). The intra- and inter-day precision and accuracy results were within the acceptable limits. Results of stability studies indicated that larotrectinib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycles and long-term storage at -80°C. The validated HPLC method was successfully applied to a pharmacokinetic study in mice., Competing Interests: There are no conflict of interests to declare in connection with the contents of this manuscript., (© Georg Thieme Verlag KG Stuttgart · New York.)

Neonicotinoids (NEOs) and fipronil (FIP) are insecticides that are widely used in modern agriculture and have received considerable attention in recent years due to their adverse effects on non-target organisms in the environment. In the present study, a new method to simultaneously detect eight common NEO insecticides and FIP and its three transformation products (FIPs) in sediments was developed using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) based on a combined pretreatment of continuous solvent extraction (CSE) and solid phase extraction (SPE). Under optimized conditions, 5.0 g of freeze-dried sediment samples were initially extracted with methanol (20 mL)-methanol (15 mL)-water (20 mL) in sequence, and then the extract was cleaned with hydrophilic-lypophilic balance SPE cartridges, and HPLC-MS/MS analysis was conducted. The established method was validated to be sensitive, linear, accurate, and precise. The limits of detection (LOD) and limits of quantification (LOQ) of target compounds were 0.012-0.055 μg/kg d.w and 0.031-0.091 μg/kg d.w, respectively. Good linearity (R 2  > 0.990) was observed between 4.0 × 10 -2 and 20.0 μg/kg d.w. The recovery rates of all target insecticides were between 75.5% and 98.5%, and the relative standard deviations (RSD) were all less than 15.0% at the low, medium, and high spiked levels. Finally, the optimized method was applied to analyze 12 target insecticides in the sediments obtained from Jiaozhou Bay of China and its main inflow rivers. Acetamiprid, thiamethoxam, fipronil sulfide, and fipronil sulfone were detected in the river sediment samples at the concentration from

Introduction: Larotrectinib (VITRAKVI) is an orally potent tropomyosin receptor kinase (Trk) inhibitor that acts by competitive inhibition of all corresponding receptor kinases. It demonstrated a marked response rate (75%) and robust anticancer activity in Trk fusion-positive patients. This response is independent of cancer type, age and gender., Methods: In this study, an efficient and accurate LC-MS/MS analytical method was developed for Larotrectinib (LRB) quantification in addition to evaluation of its metabolic stability. LRB and lapatinib (LTP) (which is chosen as an internal standard; IS) were eluted utilizing an isocratic mobile phase with a reversed phase elution system (C 18 column)., Results and Discussion: The linearity range of the established method was 5-500 ng/mL ( r 2 ≥ 0.9999) in the human liver microsomes (HLMs) matrix. Various parameters were calculated to validate the method sensitivity (limit of quantification was 5 ng/mL) and reproducibility (inter and intra-day accuracy and precision were below 3% in all samples) of our methodology. For evaluation of LRB metabolic stability in HLMs matrix, in vitro half-life (48.8 min) and intrinsic clearance (14.19 µL/min/mg) were computed., Conclusion: Accordingly, we can conclude that LRB is a moderate extraction ratio drug when compared with other tyrosine kinase inhibitors (TKIs). According to our knowledge, the discussed procedure in this study is the first LC-MS/MS analytical method for evaluating LRB metabolic stability., Competing Interests: The authors declare no conflicts of interest in this work., (© 2020 Attwa et al.)