Extraction and analysis of moxidectin in wombat plasma and faeces.

Sarcoptic mange in wombats results from a skin infestation by Sarcoptes mites and if untreated, results in a slow and painful death. Moxidectin is a pesticide used to treat internal and external parasites in cattle, but has shown to effectively treat other animals, including wombats. Two methods were developed to analyse wombat plasma, and methods were also developed to analyse faeces and fur. Moxidectin-D3 was used as an internal standard and behaved almost identically to moxidectin, resulting in recoveries of 95-105 % across the three matrices, even when matrix interferences caused signal suppression as high 20 %, or when moxidectin loss was high. This was presumably due to the high binding efficiency of plasma for MOX and MOX-D3. Moxidectin limits of detection were 0.01 ng/mL in plasma, 0.3 ng/g dry weight equivalent for faeces and 0.5 ng/g for fur. This study also developed a method to isolate plasma macromolecules, allowing the extraction of bound moxidectin for quantitative purposes, with an LoQ of 0.05 ng/mL. This method was subsequently used to determine that moxidectin was 97-99.4 % bound to lipoproteins in wombat plasma and 98-99 % bound in sheep, cow and horse plasma. The method reported for plasma was quick, cheap, and conducive to large sample batches, while providing high sensitivity. While faecal samples required additional cleanup steps to reduce the matrix effect, co-extracted matrix components such as undigested chlorophyll continued to result in ionisation suppression in the MS/MS. The methods reported here were used to monitor moxidectin in wombats treated with a single pour-on treatment, and confirmed that the moxidectin concentration in wombat plasma had decreased by more than 90 % by 28 days after application, while providing protection against sarcoptic mites over the majority of their life cycle. Clearance of moxidectin occurred via faecal elimination over the four week period and while moxidectin accumulated on fur due to application as a pour-on, concentrations declined rapidly by the four week period as fur fell out and was replaced by fresh fur that did not contain moxidectin.

Optimization and Comparison of Microwave-Assisted Extraction, Supercritical Fluid Extraction, and Eucalyptus Oil-Assisted Extraction of Polycyclic Aromatic Hydrocarbons from Soil and Sediment.

Polycyclic aromatic hydrocarbons (PAHs) are persistent organic compounds of major concern that mainly accumulate in soils and sediments, and their extraction from environmental matrices remains a crucial step when determining the extent of contamination in soils and sediments. The objective of the present study was to compare the extraction of PAHs (phenanthrene, pyrene, chrysene, and benzo[a]pyrene) from spiked soil and sediment using supercritical fluid extraction (SFE) with ethanol as the modifier, microwave-assisted extraction (MAE), and eucalyptus oil-assisted extraction (EuAE). Recoveries of PAHs were comparable between the three methods, and >80% of applied pyrene, chrysene and benzo[a]pyrene were recovered. The most efficient method of extracting PAHs from naturally incurred soils with different levels of contamination was SFE. A longer extraction time was required for the EuAE method compared with SFE and MAE under optimized conditions. However, EuAE required lower extraction temperatures (15-20 °C) compared with SFE (80 °C) and MAE (110-120 °C), and consumed less solvent than SFE and MAE. Compared with hexane/acetone used in MAE, the use of ethanol in SFE and eucalyptus oil in EuAE can be considered as more sustainable approaches to efficiently extract PAHs from spiked/naturally contaminated soils and sediments. And, although less efficient for matrices containing higher carbon content, EuAE offered a cheap, low-tech approach to extracting PAHs. Environ Toxicol Chem 2023;42:982-994. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Eucalyptus saponin- and sophorolipid-mediated desorption of polycyclic aromatic hydrocarbons from contaminated soil and sediment.

The potential for biosurfactant-mediated desorption of polyaromatic hydrocarbons (PAHs) was evaluated using PAH-spiked soil and sediment. PAH desorption behaviors and toxicity of novel saponin biosurfactant extracted from Eucalyptus camaldulensis leaves and sophoro-lipid biosurfactant were investigated. Their PAH desorption efficiencies were compared with rhamnolipid biosurfactant and the industrial-chemical surfactant, Tween 20. Based on the emulsification indices, the salt tolerance of surfactants up to 30 g/L NaCl followed the order of saponin > Tween 20 > sophorolipid > rhamnolipid, while the thermal stability over the range of 15 to 50 °C was in the order of sophorolipid > rhamnolipid > saponin > Tween 20. The saponin biosurfactant emulsion demonstrated the highest stability under a wide range of acidic to basic pHs. PAH extraction percentages of saponin and sophorolipid under the optimized surfactant concentration, volume, and incubation time were 30-50% and 30-70%, respectively. PAH desorption capacities of saponin and sophorolipid were comparable to that of rhamnolipid and Tween 20 for all matrices. Sophorolipid more efficiently desorbed low molecular weight PAHs in soil and sediment compared to the other three surfactants. Microbial respiration was used to determine biosurfactant toxicity to the soil/sediment microbiome and indicated no inhibition of respiration during 60 days of incubation, suggesting that sophorolipid- and saponin-mediated remediation may be sustainable approaches to remove PAHs from contaminated soils and sediments.

Quantification of 3,4-Dimethyl-1H-Pyrazole Using Ion-Pair LC-MS/MS on a Reversed-Phase Column.

BACKGROUND: Few methods exist for the analysis of the soil nitrification inhibitor 3,4-dimethyl-1H-pyrazole (3,4-DMP), which is a pesticide with the ability to reduce the production of nitrogenous greenhouse gases in soils as a result of fertilizer application. Due to its small size and polar nature, 3,4-DMP can be difficult to retain on an LC column, which makes diversion of a co-extracted soil matrix away from the MS/MS impossible. OBJECTIVE: The current study aims to better control the retention time (RT) of 3,4-DMP. Additionally, 3,4-DMP-15N2 was synthesized and used as an internal standard for the soil extraction of 3,4-DMP. METHODS: Perfluoroalkanoic acids were used as ion-pair reagents and were compared for their abilities to improve peak shape and RT, to better separate 3,4-DMP from the soil matrix without the need for cleanup during soil extraction. RESULTS: RTs increased with both the carbon number and the concentration of the perfluoroalkanoic acid, and this improved peak shape and height. Perfluorooctanoic acid performed best, and improved peak height (PH) and shape were obtained by increasing the flow rate, resulting in a better S/N than from formic acid. The method provided a 10-fold improvement limit of quantitation on the most sensitive existing method and the use of 3,4-DMP-15N2 as an internal standard resulted in recoveries of 101-107%. CONCLUSION: Ion-pair reagents drastically increased the retention of 3,4-DMP and allowed the re-use of old LC columns that may otherwise be discarded. Improved separation of 3,4-DMP from the soil matrix allowed much of the matrix to be diverted from the MS/MS spray chamber. HIGHLIGHTS: Greater control of 3,4-DMP retention by the LC column resulting in the ability to separate 3,4-DMP from the soil matrix. The inclusion of ion-pair reagents only in the aqueous phase reduced ionization suppression of the analytes in the MS source.

Whey Protein Peptides Have Dual Functions: Bioactivity and Emulsifiers in Oil-In-Water Nanoemulsion.

Whey protein isolate (WPI)-derived bioactive peptide fractions (1−3, 3−5, 5−10, 1−10, and >10 kDa) were for the first time used as emulsifiers in nanoemulsions. The formation and storage stability of WPI bioactive peptide-stabilized nanoemulsions depended on the peptide size, enzyme type, peptide concentration, and storage temperature. The highly bioactive <10 kDa fractions were either poorly surface-active or weak stabilizers in nanoemulsions. The moderately bioactive >10 kDa fractions formed stable nanoemulsions (diameter = 174−196 nm); however, their performance was dependent on the peptide concentration (1−4%) and enzyme type. Overall, nanoemulsions exhibited better storage stability (less droplet growth and creaming) when stored at lower (4 °C) than at higher (25 °C) temperatures. This study has shown that by optimizing peptide size using ultrafiltration, enzyme type and emulsification conditions (emulsifier concentration and storage conditions), stable nanoemulsions can be produced using WPI-derived bioactive peptides, demonstrating the dual-functionality of WPI peptides.

Polycyclic aromatic hydrocarbon contamination in soils and sediments: Sustainable approaches for extraction and remediation.

Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic environmental pollutants that are extremely hydrophobic in nature and resistant to biological degradation. Extraction of PAHs from environmental matrices is the first and most crucial step in PAH quantification. Extraction followed by quantification is essential to understand the extent of contamination prior to the application of remediation approaches. Due to their non-polar structures, PAHs can be adsorbed tightly to the organic matter in soils and sediments, making them more difficult to be extracted. Extraction of PAHs can be achieved by a variety of methods. Techniques such as supercritical and subcritical fluid extraction, microwave-assisted solvent extraction, plant oil-assisted extraction and some microextraction techniques provide faster PAH extraction using less organic solvents, while providing a more environmentally friendly and safer process with minimum matrix interferences. More recently, more environmentally friendly methods for soil and sediment remediation have been explored. This often involves using natural chemicals, such as biosurfactants, to solubilize PAHs in contaminated soils and sediments to allow subsequent microbial degradation. Vermiremediation and microbial enzyme-mediated remediation are emerging approaches, which require further development. The following summarises the existing literature on traditional PAH extraction and bioremediation methods and contrasts them to newer, more environmentally friendly ways.

Persistence of atrazine and trifluralin in a clay loam soil undergoing different temperature and moisture conditions.

Dissipation kinetics of atrazine and trifluralin in a clay loam soil was investigated in a laboratory incubation experiment under different temperature and moisture conditions. The soil was spiked with diluted atrazine and trifluralin concentrations at 4.50 and 4.25 mg/kg soil, respectively, the moisture content adjusted to 40, 70, and 100% of field capacity (FC) and then incubated in three climatic chambers at 10, 20, and 30 °C. For each of the herbicides, soil samples were collected at 0, 7, 21, 42, 70, and 105 days and analysed by Gas Chromatography-Electron Capture Detector (GC-ECD). A stochastic gamma model was used to model the dissipation of herbicides from the clay loam soil by incorporating environmental factors as covariates to determine half-life and days to complete dissipation. Results showed that temperature played a greater role on atrazine persistence than soil moisture; while the interaction effect of temperature and moisture was significant on the persistence of trifluralin over time. Atrazine dissipated more rapidly at 30 °C compared to 10 and 20 °C, with a half-life of 7.50 days and 326.23 days to reach complete dissipation. Rapid loss of trifluralin was observed at 70% moisture content when incubated at 30 °C, with a half-life of 5.80 days and 182.01 days to complete dissipation. It was observed that the half-life of both herbicides tended to double with every 10 °C decreases of temperature over the range tested. The model indicated that both atrazine and trifluralin have the potential to persist in clay loam soil for several years at temperature ≤20 °C; which could potentially affect following crops in rotation.

Bioaccessibility of Drug Residues on Common Police Station Work Surfaces.

The fraction of any surface-adsorbed contaminant available for absorption is considered the bioaccessible fraction. Applied previously to contaminants such as pesticides and heavy metals on surfaces such as soil, food and cosmetics, the term may also be used to describe the fraction of drug residue bound to work surfaces which may be mobilized via contact transfer with human skin. Police station work surfaces have been shown to commonly contain low levels of drug residues as thin films; however, no information is available on how readily these residues may be transferred to human skin during direct or glancing contact. A bioaccessibility study was undertaken in which jojoba oil and artificial sebum were used to mimic human sebum to identify how readily a mix of six licit and illicit drugs were transferred from three commonly used police station work surfaces. Transfer from surfaces was slightly greater for jojoba oil than sebum when using a direct pressure contact or a wiping motion. Generally, less than 5% of applied residues were recovered via direct contact, and up to 10% when a wiping motion was used to simulate a glancing contact. While swabbing of work surfaces with methanol provides a suitable environmental audit of drug residues present, it does not represent the bioaccessible fraction of residues available for contact transfer, and hence, absorption via skin or unintentional ingestion. The current study indicates that the ability of sebum to mobilize drug residues from thin films on work surfaces via casual contact is limited, and sebum may potentially assist in the preservation of residues on pitted work surfaces and on skin.

Direct detection of glucuronide metabolites of lidocaine in sheep urine.

The anaesthetic lidocaine is metabolised quickly to produce a series of metabolites, including several hydroxylated metabolites, which are further metabolised by addition of a glucuronic acid moiety. Analysis of these glucuronide metabolites in urine is performed indirectly by cleaving the glucuronic acid group using ß-glucuronidase. However, direct analysis of intact glucuronide conjugates is a more straightforward approach as it negates the need for long hydrolysis incubations, and minimises the oxidation of sensitive hydrolysis products, while also distinguishing between the two forms of hydroxylated metabolites. A method was developed to identify three intact glucuronides of lidocaine in sheep urine using LC-MS/MS, which was further confirmed by the synthesis of glucuronide derivatives of 3OH-MEGX and 4OH-LIDO. Direct analysis of urine allowed the detection of the glucuronide metabolites of hydroxylidocaine (OH-LIDO), hydroxyl-monoethylglycinexylidide (OH-MEGX), and hydroxy-2,6-xylidine (OH-XYL). Analysis of urine before and after ß-glucuronidase digestion showed that the efficiency of hydrolysis of these glucuronide metabolites may be underestimated in some studies. Analysis of urine in the current study from three different sheep with similar glucuronide metabolite concentrations resulted in different hydrolysis efficiencies, which may have been a result of different levels of substrate binding by matrix components, preventing enzyme cleavage. The use of direct analysis of intact glucuronides has the benefit of being less influenced by these matrix effects, while also allowing analysis of unstable metabolites like 4OH-XYL, which rapidly oxidises after hydrolysis. Additionally, direct analysis is less expensive and less time consuming, while providing more information about the status of hydroxylated metabolites in urine.

Air Quality Inside Police Drug Safes and Drug Storage Areas.

Storage of drug-based evidence inside sealed safes may allow chemical vapors to accumulate, creating concerns of drug exposure by inhalation, or the possibility of cross-contamination of drug evidence. Air samples were taken from inside eight drug safes and one small storage room at nine city and country police stations, as well as a large centralized drug evidence storage vault, in New South Wales (NSW), Australia. Sorbent tubes containing charcoal were used to determine whether any drug residues could be detected in the air, and to identify the types of chemicals present. Carbon traps were extracted and analyzed by LC-MS-MS for a suite of 22 licit and illicit drug residues and 2 metabolites. Carbon traps and SPME fibers were also analyzed by GC-MS for general volatile organic compound (VOC) residues. No detectable drug residues, either as airborne dust or vapor, were found in the safes, the storage room or the large central repository vault. No drugs were detected in any of the 34 urine samples collected at 8 of the 10 sampling locations, while only one of the five hair samples was positive for cocaine (9 pg/mg) provided by police exhibit officers at 3 of the 10 sampling locations. VOC analysis identified a variety of solvents associated with drug manufacture, plasticisers, personal care products and volatiles associated with plants such as cannabis. The results indicate that strong chemical odours emanating from drug safes are unlikely to be drug residues due to low volatility of drugs, and are more likely VOCs associated with their manufacture or from plant growing operations. Consideration should be given to the quality of air flow in rooms in which safes are housed and the use of air filtering inside safes to reduce the likelihood of VOC accumulation, and therefore the risk of human exposure.

The presence of licit and illicit drugs in police stations and their implications for workplace drug testing.

The presence of licit and illicit drug residues on surfaces was studied in 10 police stations and a central drug evidence store in New South Wales, Australia, with the results compared to similar surfaces in four public buildings (to establish a community baseline). The results of almost 850 workplace surface swabs were also compared to the outcome of drug analysis in urine and hair samples volunteered by police officers. Surfaces were swabbed with alcohol and the swabs were extracted and analysed by LC-MS/MS. Low level concentrations of the more commonly used drugs were detected at four public sites and one restricted access police office facility. Surface swabs taken in 10 city and country police stations yielded positive results for a broader suite of drugs than at background sites however 75-93% of the positive drug results detected in police stations were below 40ng, which is only slightly greater than the largest background result measured in the current study. This study indicates that contamination issues are more likely to be focussed in higher risk areas in police stations, such as counters and balances in charge areas, and surfaces within drug safes although front reception counters also returned surface contamination. All 64 urine samples collected in this study were negative, while only 2 of the 11 hair samples collected from donors resulted in trace concentrations for cocaine, but not its metabolite benzoylecgonine. Positive hair samples were only obtained from police donors in very high risk jobs, indicating that the exposure risk is low. Minor changes to the materials used as work surfaces, and some procedural changes in police stations and large evidence stores are suggested to decrease the likelihood of drugs contaminating work surfaces, thereby reducing the potential exposure of police officers to drugs in the workplace.

Work place drug testing of police officers after THC exposure during large volume cannabis seizures.

Police officers responsible for the seizure and removal of illegally grown cannabis plants from indoor and outdoor growing operations face the prospect of THC exposure while performing their work duties. As a result, a study investigating the amount of THC on hands and uniforms of officers during raids on cannabis growing houses (CGHs) and forest cannabis plantations (FCPs) and in the air at these sites was conducted. Swabs of gloves/hands, chests, and heads/necks were collected and analysed for THC. Results of hand swabs indicated that officers removing plants from FCPs were exposed to THC concentrations up to 20 times those involved in raids at CGHs, which was mainly associated with the number and size of plants seized. Air samples collected inside cannabis houses showed no detectable THC. Air samples collected inside the cargo area of the storage trucks used during FCP raids indicated that THC can be volatilised when lush plants are compressed by other seized plants loaded on top of them in the truck over a period of several days, allowing composting of plants at the bottom of the load to commence. The elevated temperature and humidity inside the truck may assist the decarboxylation of THCA to THC, as well as increasing the rate of volatilisation of THC. More than 100 urine samples were collected from officers in raids on both CGHs and FCPs and all tested negative for THC. Removal of cannabis plants by officers often resulted in cuts, abrasions and ruptured blisters on exposed skin surfaces, particularly at FCPs. The results in this study suggest that even when small areas of damaged skin are directly exposed to THC by contact transfer, the likelihood of showing a positive THC urine test is low.

Quantitation of the anaesthetic xylazine in ovine plasma by LC-MS/MS.

Removal of the wool-bearing skin around a young lamb's rump (mulesing) provides long term health benefits for the animal, and the use of a sedative and analgesic agent such as xylazine may assist with pain relief to reduce discomfort and stress. Sensitive analytical methods are essential for monitoring pharmaceuticals and their metabolites in animals destined for human consumption. The following work reports a method that is 200 times more sensitive for xylazine detection than previously published methods, with lower limits of quantitation for xylazine and its primary metabolite in animals of 0.5pg and 2pg on-column, respectively. The use of a square wave solvent gradient immediately prior to analyte elution resulted in larger MS/MS peaks and a reduction in baseline noise, allowing reliable detection of lower analyte concentrations. The method uses as little as 1mL of plasma which allows replication within a sample if required, and requires simple sample preparation, minimising the introduction of matrix components into the MS/MS.

Pharmacokinetics of pergolide after intravenous administration to horses.

OBJECTIVE: To determine the pharmacokinetics of pergolide after IV administration to horses. ANIMALS: 8 healthy adult horses. PROCEDURES: Pergolide mesylate was administered IV at a dose of 20 µg/kg (equivalent to 15.2 µg of pergolide/kg) to each horse, and blood samples were collected over 48 hours. Pergolide concentrations in plasma were determined by means of high-performance liquid chromatography-tandem mass spectrometry, and pharmacokinetic parameters were determined on the basis of noncompartmental methods. RESULTS: After IV administration of pergolide, mean ± SD clearance, elimination half-life, and initial volume of distribution were 959 ± 492 mL/h/kg, 5.64 ± 2.36 hours, and 0.79 ± 0.32 L/kg, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: With an elimination half-life of approximately 6 hours, twice-daily dosing may be more appropriate than once-daily dosing to reduce peak-trough fluctuation in pergolide concentrations. Further pharmacodynamic and pharmacokinetic studies of pergolide and its metabolites will be necessary to determine plasma concentrations that correlate with clinical effectiveness to determine the therapeutic range for the treatment of pituitary pars intermedia dysfunction.