Optimization and validation of high-performance liquid chromatography using modified QuEChERS to determine anthelmintic drugs in mutton.

The aim of this study was to rapidly determine the presence of anthelmintic drugs in sheep meat using the optimized high-performance liquid chromatography-ultraviolet (HPLC-UV) method with modified QuEChERS (quick, easy, cheap, effective, rugged, safe) technology. Fifty fresh sheep meat samples from different slaughterhouses were collected. A double extraction procedure (QuEChERS/HPLC-UV technology) was used to extract the target analytes. A multilevel calibration curve from 1 to 1000 g/kg was used to establish instrument linearity for rafoxanide, albendazole, and closantel, whereas 0.1-100 µg/kg was used for ivermectin, levamisole, and oxyclozanide to find the lowest concentration, maximum residue limit (MRL), and occupied range for targeted analytes. The concentration levels were used to investigate the linearity, whereas several certified reference materials were applied to determine accuracy. The process was linear for all combinations, from the limit of quantification (LOQ) to the maximum concentration. The LOQ was established at 0.5 µg/kg for ivermectin, levamisole, and oxyclozanide and 10 µg/kg for rafoxanide, albendazole, and closantel. Recovery values were 70%-120%, and repeatability/reproducibility stated in relative standard deviation was obtained at less than 20%. QuEChERS method revealed that most meat samples contained anthelmintic drug residues, of which the majority exceeded the MRLs. Thus, the drugs should be used correctly in animals to avoid residues in food for human consumption.

Quantitative Analysis of Abamectin, Albendazole, Levamisole HCl and Closantel in Q-DRENCH Oral Suspension Using a Stability-Indicating HPLC-DAD Method.

Combination therapy of many anthelmintic drugs has been used to achieve fast animal curing. Q-DRENCH is an oral suspension, containing four different active drugs against GIT worms in sheep, commonly used in Australia and New Zeeland. The anti-parasitic drugs are Albendazole (ALB), Levamisole HCl (LEV), Abamectin (ABA), and Closantel (CLO). The main purpose of this study is to present a new simultaneous stability-indicting HPLC-DAD method for the analysis of the four drugs. The recommended liquid system was 1 mL of Triethylamine/L water, adjusting the pH to 3.5 by glacial acetic acid: acetonitrile solvent (20:80, v/v). Isocratic elusion achieved the desired results of separation at a 2 mL/min flow rate using Zorbax C-18 as a stationary phase. Detection was performed at 210 nm. The linearity ranges were 15.15 to 93.75 µg/mL for ALB, 25 to 150 µg/mL for LEV, 30 to 150 µg/mL for ABA, and 11.7 to 140.63 µg/mL for CLO. Moreover, the final greenness score was 0.62 using the AGREE tool, which reflects the eco-friendly nature. Moreover, the four drugs were determined successfully in the presence of their stressful degradation products. This work presents the first chromatographic method for simultaneous analysis for Q-DRENCH oral suspension drugs in the presence of their stressful degradation products.

Simple method for the determination of anthelmintic drugs in milk intended for human consumption using liquid chromatography-tandem mass spectrometry.

BACKGROUND: Helminth infections in animals to be consumed by humans are an important medical and public health problem. Pharmaceutical research has focused on developing new anthelmintic drugs for parasite control in these animals. However, the incorrect use of anthelmintics can leave residues in animal products intended for human consumption. Their determination is therefore crucial in terms of food safety. RESULTS: In this work, a simple and sensitive method has been developed for the analysis of anthelmintic drugs in milk. The method involves extraction of the analytes using a QuEChERS (quick, easy, cheap, effective, rugged, and safe) method, and separation and determination by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The use of a core-shell column significantly reduced the analysis time compared with conventional columns. The method was validated and applied to the analysis of different commercial milk samples: whole, semi-skimmed and skimmed cows' milk, and goats' milk. None of the benzimidazoles studied was found in the samples analyzed, so these were spiked with the analytes at three concentration levels (10, 50, and 100 µg kg-1 ). CONCLUSIONS: The proposed method provided high sensitivity compared with other methods for the determination of anthelmintics in milk samples, at concentration levels well below the established maximum residue limit (MRLs) values. The proposed method is simple, easy, precise, accurate, and leads to good recovery levels. It can be used successfully for the routine analysis. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Essential oil of Mentha pulegium induces anthelmintic effects and reduces parasite-associated oxidative stress in rodent model.

Following the previous findings reported by the present authors on the anthelmintic effect of hydro-ethanolic extract of Mentha pulegium, the volatile constituents of M. pulegium are now assessed in the present study by exploring its anthelmintic and its antioxidant proprieties using in vitro and in vivo assays. Egg hatch assay (EHA) and adult worm's motility assays (AWMA) were used to assess the in vitro activity against Haemonchus. contortus. The in vivo anthelmintic potential was evaluated in mice infected with Heligmosomoides polygyrus using faecal egg count reduction (FECR) and total worm count reduction (TWCR). M. pulegium EO demonstrated 100% inhibition in the EHA at 200 µg/mL (IC50 = 56.36 µg/mL). In the AWM assay, EO achieved total worms paralysis 6 h after treatment exposure. This nematicidal effect was associated to morphological damages observed in the cuticular's worm using environmental scanning electron microscopy (ESEM). At 400 mg/kg, M. pulegium oil showed 75.66% of FECR and 80.23% of TWCR. The antioxidant potential of this plant was also monitored by several in vitro assays: total antioxidant capacity was 205.22 mg GAE/g DW, DPPH quenching effect was IC50 = 140 µg/mL, ABTS activity IC50 = 155 µg/mL and FRAP effect of 660 µg/mL. Regarding the in vivo assay, M. pulegium EO demonstrated a protective effect against oxidative stress by increasing the activity of the endogenous antioxidants (SOD, CAT and GPx) during H. polygyrus infection.

Occurrence and risk assessment of anthelmintics in Tuojiang River in Sichuan, China.

Recently, emerging pollutants, such as anthelmintics have attracted an increasing attention worldwide due to their extensive use and notable stability. However, the information on anthelmintics in the environment of southwest China is scarce. Thus, the occurrence, ecological risk and exposure evaluation of nineteen anthelmintics in Tuojiang River, which is one of the largest tributaries of Yangtze River, and drinking water source of Sichuan, southwest China, were investigated. The result showed that the detection frequency of anthelmintics was relatively high in Tuojiang River, ranging from 65% to 100% in river water. Among the seven kinds of anthelmintics, benzimidazoles are the primary anthelmintics, with concentrations up to 61.12 ng/L and 596.06 ng/g in water and sediment of the Tuojiang river, respectively. The total concentration of 19 anthelmintics in sediment samples from non-agricultural area was higher than that in agricultural area(p = 0.000 < 0.05). This could be attributed to anthropogenic activities, which lead to greater discharge and accumulation of anthelmintics in residential area along the river. It's worth to mention that the highest total concentrations of anthelmintics (109.28 ng/L) was found at the junction of rivers in R31 site. The results could be ascribed to the complexity of junction of Tuojiang River and Yangtze River, which could influence the distribution of pollutant. Besides, the ecological risk assessment showed that the macrocyclic lactones rather than benzimidazoles had relatively high toxicity to non-target organisms in aquatic environment (p = 0.000 < 0.05), with the highest RQEcotox value of 101 for Daphnia magna, while benzimidazoles had relatively high concentrations. The exposure risk could be ignored for both children and adults because the daily intake of anthelmintics via water ingestion were below 10 ng/kg/d. In addition, strong correlations were found between sucralose and most of the selected anthelmintics in Tuojiang River, indicating that sucralose might be a good tracer to evaluated the source of anthelmintics in surface water. This study provides the levels, risks and even some tracer information of pollutants for better understanding of anthelmintics in southwest China.

Determination of Anthelmintic and Antiprotozoal Drug Residues in Fish Using Liquid Chromatography-Tandem Mass Spectrometry.

The objective of this study is to develop a comprehensive and simple method for the simultaneous determination of anthelmintic and antiprotozoal drug residues in fish. For sample preparation, we used the "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) method with a simple modification. The sample was extracted with water and 1% formic acid in acetonitrile/methanol (MeCN/MeOH) (95:5, v/v), followed by phase separation (salting out) with MgSO4 and NaCl (4:1, w/w). After centrifugation, an aliquot of the extract was purified by dispersive solid-phase extraction (d-SPE) prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The method was validated at three concentration levels for all matrices, in accordance with the Codex guidelines (CAC/GL-71). Quantitative analysis was performed using the method of matrix-matched calibration. The recoveries were between 60.6% and 119.9%, with coefficients of variation (CV) <30% for all matrices. The limit of quantitation (LOQ) of the method ranged from 0.02 µg kg-1 to 4.8 µg kg-1 for all matrices. This comprehensive method can be used for the investigation of both anthelmintic and antiprotozoal drugs belonging to different chemical families in fishery products.

Practical High-Throughput Method to Screen Compounds for Anthelmintic Activity against Caenorhabditis elegans.

In the present study, we established a practical and cost-effective high throughput screening assay, which relies on the measurement of the motility of Caenorhabditis elegans by infrared light-interference. Using this assay, we screened 14,400 small molecules from the "HitFinder" library (Maybridge), achieving a hit rate of 0.3%. We identified small molecules that reproducibly inhibited the motility of C. elegans (young adults) and assessed dose relationships for a subset of compounds. Future work will critically evaluate the potential of some of these hits as candidates for subsequent optimisation or repurposing as nematocides or nematostats. This high throughput screening assay has the advantage over many previous assays in that it is cost- and time-effective to carry out and achieves a markedly higher throughput (~10,000 compounds per week); therefore, it is suited to the screening of libraries of tens to hundreds of thousands of compounds for subsequent evaluation and development. The present phenotypic whole-worm assay should be readily adaptable to a range of socioeconomically important parasitic nematodes of humans and animals, depending on their dimensions and motility characteristics in vitro, for the discovery of new anthelmintic candidates. This focus is particularly important, given the widespread problems associated with drug resistance in many parasitic worms of livestock animals globally.

Use of eosin for green spectroscopic determination of mebendazole.

New, sensitive, and reliable spectroscopic methods were constructed for the fast determination of the anthelmintic drug mebendazole. The methods depended on the reaction of the amino group in mebendazole with eosin in acidic medium forming an ion pair complex. The first method, Method I, relied on quenching of the native fluorescence of eosin after reaction with mebendazole at pH 3.7 using acetate buffer. Fluorescence quenching was measured at 538 nm after excitation at 518 nm. This method showed a linear response over the concentration range 5.0-20.0 µg/ml. The second method, Method II, was based on measuring the absorbance of the formed complex at 554 nm; the method showed good linearity from 7.0 to 22.0 µg/ml. Different parameters that influenced the formation of the reaction product were carefully investigated to reach the optimized conditions. A comparison between the proposed methods and a previous spectrophotometric method was carried out and there was no significant difference between them. The methods could be applied successfully to determine mebendazole in its tablet form. Moreover, the methods used water as diluting solvent, which made them compatible with the 'green' analytical chemistry principles. No organic solvents were used throughout the study.

Assessment of the long-acting ivermectin formulation in sheep: Further insight into potential pharmacokinetic interactions.

The aim of the current study was to evaluate the in vivo pharmacokinetic of ivermectin (IVM) after the administration of a long-acting (LA) formulation to sheep and its impact on potential drug-drug interactions. The work included the evaluation of the comparative plasma profiles of IVM administered at a single therapeutic dose (200 µg/kg) and as LA formulation at 630 µg/kg. Additionally, IVM was measured in different gastrointestinal tissues at 15 days posttreatment with both IVM formulations. The impact of the long-lasting and enhanced IVM exposure on the disposition kinetics of abamectin (ABM) was also assessed. Plasma (IVM and ABM) and gastrointestinal (IVM) concentrations were analyzed by HPLC with fluorescent detection. In plasma, the calculated Cmax and AUC0-t values of the IVM-LA formulation were 1.47- and 3.35-fold higher compared with IVM 1% formulation, respectively. The T1/2ab and Tmax collected after administration of the LA formulation were 2- and 3.5-fold longer than those observed after administration of IVM 1% formulation, respectively. Significantly higher IVM concentrations were measured in the intestine mucosal tissues and luminal contents with the LA formulation, and in the liver, the increase was 7-fold higher than conventional formulation. There was no drug interaction between IVM and ABM after the single administration of ABM at 15 days post-administration of the IVM LA formulation. The characterization of the kinetic behavior of the LA formulation to sheep and its potential influence on drug-drug interactions is a further contribution to the field.

Development and Optimisation of a Multiresidue Method for the Determination of 40 Anthelmintic Compounds in Environmental Water Samples by Solid Phase Extraction (SPE) with LC-MS/MS Detection.

A comprehensive multiresidue method was developed and validated for the determination of 40 anthelmintic compounds, including 13 transformation products, in surface and groundwater samples at sub nanogram per litre (ng L-1) levels. Anthelmintic residues were extracted from unfiltered water samples using polymeric divinylbenzene solid phase extraction (SPE) cartridges, and eluted with methanol: acetone (50:50, v/v). Purified extracts were concentrated, filtered and injected for UHPLC-MS/MS determination. The method recovery (at a concentration representative of realistic expected environmental water levels based on literature review) ranged from 83-113%. The method was validated, at three concentration levels, in accordance to Commission Decision 2002/657/EC and SANTE/11813/2017 guidelines. Trueness and precision, under within-laboratory reproducibility conditions, ranged from 88-114% and 1.1-19.4%, respectively. The applicability of the method was assessed in a pilot study whereby 72 different surface and groundwater samples were collected and analysed for the determination of these 40 compounds for the first time in Ireland. This is the most comprehensive method available for the investigation of the occurrence of both anthelmintic parent compounds and their transformation products in raw, unfiltered environmental waters.

Effect of nutraceuticals on acanthocephalan Neoechinorhynchus buttnerae and its toxicity to the host tambaqui Colossoma macropomum.

The production of tambaqui Colossoma macropomum has been undergoing financial losses due to parasitic infection by the acanthocephalan Neoechinorhynchus buttnerae, raising an alert for aquaculture in South America. The lack of adequate treatment and use of unlicensed chemicals encourages research for alternative solutions with minimal side effects. The objectives of this study were to evaluate the in vitro antiparasitic potential of commercial nutraceutical products (Natumix® and BioFish®) against N. buttnerae and to assess the respective in vivo toxic effects on the host tambaqui. For in vitro assays, parasitized fish were necropsied for acanthocephalans sampling. The parasites were exposed to three concentrations (0.078, 0.313 and 1.25 mg/ml) of each product, as well as controls (one without product and another with a solubilizer). For the in vivo acute toxicity test, juvenile fish (<0.1 g) were exposed to five increasing concentrations of each product. Mortality of tambaqui was recorded at 24, 48, 72 and 96 h. The estimated lethal concentration (LC) for 10, 50, 90 and 99% of fish was determined to classify the toxicity of the products on the target species. After in vitro efficacy tests, the highest concentrations (1.25 mg/ml) caused 100% mortality of the parasites in both products, but only Natumix® caused 100% mortality using the intermediate concentration (0.313 mg/ml) after 24 h. According to the acute toxicity result, the LC50 classified the nutraceutical products as slightly toxic for tambaqui. The tested products had a parasiticidal effect on N. buttnerae, and the toxicity test showed that both products have therapeutic potential when added to the diet.

Fluorescence polarization immunoassay for rapid screening of the pesticides thiabendazole and tetraconazole in wheat.

Fluorescence polarization immunoassays (FPIAs) for thiabendazole and tetraconazole were first developed. Tracers for FPIAs of thiabendazole and tetraconazole were synthesized and the tracers' structures were confirmed by HPLC-MS/MS. The 4-aminomethylfluorescein-labeled tracers allowed achieving the best assay sensitivity and minimum reagent consumption in comparison with aminofluorescein-labeled and alkyldiaminefluoresceinthiocarbamyl-labeled tracers. Measurements of fluorescence polarization were performed using a portable device. The developed FPIA methods were applied for the analysis of wheat. Fast and simple sample preparation technique earlier developed by authors for pesticides was adapted for thiabendazole and tetraconazole. The limits of detection of thiabendazole and tetraconazole in wheat were 20 and 200 µg/kg, and the lower limits of quantification were 40 and 600 µg/kg, respectively. The recovery test was performed by two methods-FPIA and HPLC-MS/MS. The results obtained by FPIA correlated well with those obtained by HPLC-MS/MS (r2 = 0.9985 for thiabendazole, r2 = 0.9952 for tetraconazole). Average recoveries of thiabendazole and tetraconazole were 74 ± 4% and 72 ± 3% by FPIA, and average recoveries of thiabendazole and tetraconazole were 86 ± 2% and 74 ± 1% by HPLC-MS/MS (n = 15). Graphical abstract ᅟ.

Virtual screening of natural anti-filarial compounds against glutathione-S-transferase of Brugia malayi and Wuchereria bancrofti.

Glutathione-S-transferase also referred as GST is one of the major detoxification enzymes in parasitic helminths. The crucial role played by GST in various chronic infections has been well reported. The dependence of nematodes on detoxification enzymes to maintain their survival within the host established the crucial role of GST in filariasis and other related diseases. Hence, this well-established role of GST in filariasis along with its greater nonhomology with its human counterpart makes it an important therapeutic drug target. Here in this study, we have tried to explore the inhibitory potential of some of the well-reported natural ant-filarial compounds against the GST from Wuchereria bancrofti (W.bancrofti) and Brugia malayi (B.malayi). In silico virtual screening, approach was used to screen the selected natural compounds against GST from W.bancrofti and B.malayi. On the basis of our results, here we are reporting some of the natural compounds which were found to be very effective against GSTs. Along with we have also revealed the characteristic of the active site of BmGST and WbGST and the role of important active site residues involve in the binding of natural compounds within the active site of GSTs. This information will oped doors for using natural compounds as anti-filarial therapy and will also be helpful for future drug discovery.

Albendazole treatment in laying hens: Egg residues and its effects on fertility and hatchability.

This work characterized the egg residual concentrations of albendazole (ABZ) and its sulphoxide (ABZSO) and sulphone (ABZSO2 ) metabolites and evaluated their effect on egg fertility and hatchability after ABZ treatments to laying hens. Seventy hens were allocated in groups: Group-1 was the control without treatment; Group-2 received a single ABZ oral dose (10 mg/kg); Group-3, -4 and -5 were treated with ABZ in medicated feed over 7 days at 10, 40, or 80 mg kg-1  day-1 , respectively. Eggs were analyzed to determine the ABZ/metabolite level by HPLC or subjected to incubation to evaluate the fertility and hatchability. Only ABZSO and ABZSO2 metabolites were quantified in egg after ABZ single oral administration with maximum concentrations of 0.47 ± 0.08 and 0.30 ± 0.07 µg/ml, respectively. ABZ and its metabolites were found in eggs after 7-day ABZ treatments. The egg residue exposure estimated as AUCs (areas under the concentration vs. time curve) were 100.5 (ABZ), 56.3 (ABZSO) and 141.3 µg hr g-1 (ABZSO2 ). ABZ administration did not affect the egg fertility at any dosages. Egg hatchability was not affected by ABZ treatment at 10 mg/kg in medicated feed, but it decreased when the dose was 4-8 times higher. These results should be considered when ABZ is used for deworming laying hens.

Determination of 4-Hexylresorcinol in Shrimp Samples by Solid Phase Extraction Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry.

A method for the rapid determination of 4-hexylresorcinol (4-HR) residue in shrimp by solid phase extraction (SPE) ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. 4-HR was extracted twice with methanol, and the extract was formulated into methanol-water solution (1:1). After being cleaned up and concentrated by a PRIME HLB solid phase extraction column, the sample was analyzed by UPLC-MS/MS and quantitatively determined by an external standard method. The separation was performed with a gradient system consisting of water and acetonitrile as the mobile phase. Monitoring was performed by electrospray ionization (ESI) in negative ion mode using multiple ion reaction monitoring (MRM). Good linearity was obtained in the concentration range of 1.0⁻100.0 µg/L, with correlation coefficients larger than 0.999. The limit of detection (LOD) was 0.25 µg/kg and the limit of quantification (LOQ) was 0.80 µg/kg. The average recoveries of 4-HR at spiked concentrations of 2.40, 6.40 and 16 µg/kg ranged from 81.35% to 94.68% with the relative standard deviations (n = 6) from 3.57% to 6.86%. The results showed that the method is simple, fast, sensitive, reliable, and reproducible; thus, it could be used as a rapid confirmation and quantitative analysis method of 4-HR residue in aquatic products.

Anthelmintic efficacy of hydro-methanolic extracts of Larrea tridentata against larvae of Haemonchus contortus.

An in vitro study was conducted to determine the anthelminthic activity of hydro-methanolic extracts of Larrea tridentata on sheathed and exsheathed larvae of Haemonchus contortus. Larvae of the parasite were incubated at 20-25 °C in hydro-methanolic extracts at concentrations of 12.5, 25, 50, 100, and 200 mg/mL for 24, 48, or 72 h. Ivermectin and water were the positive and negative controls, respectively. Total phenolic compounds of leaves of L. tridentata were 97.88 ± 10.45 mg/g of dry matter. Other compounds detected in this shrub by HPLC-mass spectrometry were sesamin, galocatechin, peonidin 3-O rutinoside, methyl galangin, epigallocatechin 7-O-glucuronide, and epigalocatechin. Mortality rate of sheathed and exsheathed H. contortus was low (16-34%) with doses ≤ 100 mg/mL of the extracts. At 200 mg/ml, the hydro-methanolic extracts of L. tridentata killed 32.1 and 68.4% of sheathed and exsheathed larvae, respectively, regardless of incubation time. The effective concentration of the L. tridentata extract for 50% larvae mortality (EC50) after 24 h of incubation was 36 mg/mL (CI = 6-94). Microscopic observations revealed damage to the cuticle of this parasite exposed to extracts of L. tridentata. These in vitro results provided evidence that L. tridentata extracts possess anti-Haemonchus contortus properties, particularly during the exsheathed stage of this nematode. It would be necessary to assess the safety of this shrub in vivo and also to carry out in vivo efficacy studies.

Anthelmintic effects of indigenous multipurpose fodder tree extracts against Haemonchus contortus.

Condensed tannins (CT) extracted from Balanites aegyptiaca, Tamarindus indica, and Celtis toka browses were used to evaluate their anthelmintic effect on different developmental stages of Haemonchus contortus. To achieve this objective, various serial concentrations of each CT extract of the foliages were used to test adult motility, inhibition of egg hatchability, and larval development. The fodders were selected based on their multipurpose advantage and accessibility to use as fodder for livestock in the low lands of the Gambella region. The fastest and slowest adult motility rate was observed in 2-ml (4 min) and 0.125-ml dose of C. toka, respectively, which is better than that in ivermectin. Egg hatchability inhibition was observed with dose difference within species, but there is no difference between B. aegyptiaca and T. indica. The foliage extracts of the studied browses were observed to inhibit the larvae by 100% at 2 ml, which is similar to ivermectin. There is no significant difference observed in larvae development inhibition between the species and ivermectin (p > 0.05). CT extracts of studied plants have found to own significant anthelmintic activity in a dose-dependent manner. They could serve as anthelmintic economically and eco-friendly after further and series of in vivo experiments.

Evaluation of drug uptake and deactivation in plant: Fate of albendazole in ribwort plantain (Plantago laceolata) cells and regenerants.

Albendazole (ABZ) is a benzimidazole anthelmintic widely used especially in veterinary medicine. Along with other drugs, anthelmintics have become one of a new class of micro-pollutants that disturb the environment but the information about their fate in plants remains limited. The present study was designed to test the uptake and biotransformation of ABZ in the ribwort plantain (Plantago lancelota), a common meadow plant, which can come into contact with this anthelmintic through the excrements of treated animals in pastures. Two model systems were used and compared: cell suspensions and whole plant regenerants. In addition, time-dependent changes in occurrence of ABZ and its metabolites in roots, basal parts of the leaves and tops of the leaves were followed up. Ultrahigh-performance liquid chromatography coupled with high mass accuracy tandem mass spectrometry (UHPLC-MS/MS) led to the identification of 18 metabolites of ABZ formed in the ribwort. In both model systems, the same types of ABZ biotransformation reactions were found, but the spectrum and abundance of the ABZ metabolites detected in cell suspensions and regenerants differed significantly. Cell suspensions seem to be suitable only for qualitative estimations of drug biotransformation reactions while regenerants were shown to represent an adequate model for the qualitative as well as quantitative evaluation of drug uptake and metabolism in plants.

Structural characterization of product ions of regulated veterinary drugs by electrospray ionization and quadrupole time-of-flight mass spectrometry. Part 3: Anthelmintics and thyreostats.

RATIONALE: Previously, we have reported a liquid chromatography/tandem mass spectrometry method for the identification and quantification of regulated veterinary drugs in food animals. The method uses three selected transition ions per analyte but structural characterization is also needed. This work is a continuation of two previous publications in which we propose structures of the selected transition ions of 130 veterinary drugs altogether. METHODS: In this work, 24 additional veterinary drugs were analyzed by infusion into a high-resolution quadrupole time-of-flight (QTOF) mass spectrometer using electrospray ionization (ESI) in positive or negative mode. The TOF analyzer was calibrated to achieve low error mass accuracy in the MS and MS/MS modes. Also, the MS(2) and MS(3) spectra were obtained by using a Q-Trap mass spectrometer to further determine the possible pathways of ion formation. RESULTS: The low error mass spectrometry analysis allowed the elucidation of the ion formulae of selected transition ions for qualitative identification. The rational interpretation of data including a review of the published literature led to the proposed structures of the MS/MS product ions of 24 compounds covering two classes of regulated veterinary drugs (anthelmintics and thyreostats). In addition, the use of MS(2) and MS(3) experiments led to the establishment of fragmentation patterns. CONCLUSIONS: The identification and quantification of veterinary drug residues is helpful information for regulatory monitoring programs in defense of regulatory enforcement actions. Published in 2016. This article is a U.S. Government work and is in the public domain in the USA.

Study on the interaction between three benzimidazole anthelmintics and eosin Y by high performance liquid chromatography associating with resonance light scattering and its application.

A novel, highly selective, and sensitive resonance light scattering (RLS) detection approach coupled with high performance liquid chromatography (HPLC) was researched and developed for the synchronous analysis of three kinds of benzimidazole anthelmintics, including mebendazole (MBZ), albendazole (ABZ), and fenbendazole (FBZ) for the first time. In the pH range of 3.5-3.7 Britton-Robinson buffer medium, three kinds of anthelmintics, which were separated by HPLC, reacted with eosin Y (EY) to form 1:1 ion-association complexes, resulting in significantly enhanced RLS signals and the maximum peak located at 335 nm. The enhanced RLS intensity was in proportion to the MBZ, ABZ, and FBZ concentration in the range 0.2-25, 0.2-23, and 0.15-20 µg/mL, respectively. The limit of detection was in the range of 0.064-0.16 µg/mL. In addition, human urine was determined to validate the proposed method by spiked samples and real urine samples. Satisfactory results were obtained by HPLC-RLS method. Graphical Abstract The diagram mechanism of generating resonance between emitted light and scattered light.