Determination of cyflufenamid residues in 12 foodstuffs by QuEChERS-HPLC-MS/MS.

Agricultural products are often contaminated with trace amounts of pesticide residues. To ensure food safety, a reliable, sensitive, and efficient method that accurately identifies pesticide residues in a wide variety of foodstuffs is needed. This study applied a modified QuEChERS extraction technique, coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to optimize the identification of the cyflufenamid, a new broad-spectrum fungicide with unclear mode of action in toxicity, in 12 foodstuffs. The method has been validated according to the European Union SANTE/12682/2017 guidelines for its linearity, limit of detection (LOD), and limit of quantification (LOQ), indicating an excellent performance. The recovery of cyfluenamid in all matrix were ranged between 70.8% and 117.8%, with relative standard deviation (RSD) <15% which is much lower compared to other methods. The method has been used for determine cyfluenamid in foodstuffs followed the ISO17025:2017 measurement technical note, which demonstrated it is suitable for routine quantitative analysis of cyflufenamid in most of the popular commercial agricultural products.

Detection and quantification of trace airborne transfluthrin concentrations via air sampling and thermal desorption gas chromatography-mass spectrometry.

A rapid thermal desorption-gas chromatography-electron ionization-mass spectrometry (TD-GC-EI-MS) method for airborne transfluthrin detection is studied. Active air sampling of 9 L over 1 h at 23 °C through a Tenax®-loaded tube resulted in efficient capture of airborne transfluthrin. Subsequent thermal desorption was employed to achieve an LOD of 2.6 ppqv (parts per quadrillion by volume). A minimum primary desorption temperature of 300 °C is necessary for optimal recovery of sample from the Tenax® adsorbent. The matrix effects of indoor air lead to an error of 10.9% and 10.5% recovery of sample (10 pg and 100 pg loaded tubes, respectively). The linear range was 74-74,000 ppqv with a correlation coefficient of 0.9981. Active air sampling of a novel passive release device revealed a ∼150 pg/L airborne concentration gradient over 1 m, providing spatial characterization of the device's performance. This efficient method allows for the remote collection of samples and rapid analysis of airborne transfluthrin from industrial applications, optimization studies of commercial products as well as domestic/household monitoring.

Photolysis of cyflufenamid in liquid media.

The photolysis of cyflufenamid (CFA) in different organic solvents and water under ultraviolet irradiation was investigated. The photolytic rate constant and photolytic half-life were measured for the different solutions. Factors influencing the photolysis of CFA were investigated, including initial concentration, types of solvent, pH, occurrence of catalyst (TiO2), and environmental substances (Fe3+, Fe2+, NO3 -, NO2 -). Photolysis of CFA followed first-order kinetics in various systems, and the photolytic rate of CFA decreased with increased initial concentration. Photolytic rates of CFA in different solvents were as follows: n-hexane > methanol > acetonitrile > ultrapure water > ethyl acetate. The pH had a significant effect on the photolysis of CFA, and the photolysis rate reached its peak at pH 9.0. NO2 - and TiO2 had positive effects on the photolysis of CFA, while Fe2+ had an adverse effect. NO3 - in aqueous solution had no effect on the photolysis of CFA. In addition, the rates of photolysis were accelerated at lower concentrations of Fe3+ (0.5-5 mmol L-1) and decreased at higher concentrations (10 mmol L-1). Moreover, a main photolytic product of CFA was confirmed to be N-cyclopropoxy-2,3-difluoro-6-(trifluoromethyl)benzamide, and cleavage of the amido bond was proposed to be the predicted photolysis pathway in n-hexane.

Gas chromatography/negative chemical ionization mass spectrometry of transfluthrin in rat plasma and brain.

RATIONALE: Transfluthrin is a relatively non-toxic rapid-acting synthetic pyrethroid insecticide. It is widely used in household and hygiene products. A sensitive and accurate bioanalytical method is required for quantification of its concentration in plasma and its potential target organ, the brain for studies to assess its health effects and toxicokinetics in mammals. METHODS: The samples were prepared by liquid-liquid extraction. Gas chromatography mass spectrometry (GC/MS) analysis was performed for the determination of transfluthrin in biological samples with an overall method run time of 15 min. Transfluthrin was quantified using selected-ion monitoring (SIM) in the negative chemical ionization (NCI) mode. Chromatographic separation was achieved using a Zebron® ZB5-MS GC column operating with 1 mL/min constant flow helium. Cis-Permethrin was used as the internal standard. RESULTS: The method was validated to be precise and accurate within the linear range of 1.0-400.0 ng/mL in plasma and 4.0-400.0 ng/mL in brain homogenate, based on a 100 µL sample volume for both matrices. This method was applied to samples following administration of a 10 mg/kg oral dose to male adult rats. The plasma concentrations were observed to be 11.70 ± 5.69 ng/mL and brain concentrations 12.09 ± 3.15 ng/g when measured 2 h post-dose. CONCLUSIONS: A rapid GC/NCI-MS method was demonstrated to be sensitive, specific, precise and accurate for the quantification of transfluthrin in rat plasma and brain. The optimized method was successfully used to quantify the rat plasma and brain concentrations of transfluthrin 2 h after the oral dosing of Sprague-Dawley rats.

Fabrication and evaluation of a fluorophilic adsorbent for multiple monolithic fiber solid-phase microextraction of fluorobenzenes.

A new type of highly fluorinated monolith (HFM) was fabricated and used as adsorbent of multiple monolithic fiber solid-phase microextraction (MMF-SPME). To prepare the HFM, a fluorinated monomer, 2,2,3,3,4,4,5,5,6,6,7,7-dodecafluoroheptyl acrylate was in situ copolymerized with dual cross-linkers (divinylbenzene and ethylenedimethacrylate). The fabrication parameters including the content of monomer and porogenic solvent in the polymerization mixture were optimized to obtain expected extraction performance and life span. The physicochemical properties of the HFM were systematically investigated with elemental analysis, infrared spectroscopy, scanning electron microscopy and mercury intrusion porosimetry. The effective extraction of six fluorobenzenes was selected as a paradigm to demonstrate the fluorophilic characteristic of HFM/MMF-SPME. At the same time, a convenient and effective method for the determination of trace fluorobenzenes in environmental water samples was developed by coupling HFM/MMF-SPME with high performance liquid chromatography/diode array detection (HFM/MMF-SPME-HPLC/DAD). Results indicated that the limits of detection (S/N=3) for targeted compounds were in the range of 1.09-5.88µg/L. The intra-day and inter-day precision (relative standard deviations, n=4, %) at two spiked concentrations were 4.2-10.6% and 6.1-10.8%, respectively. Finally, the developed method was successfully applied to the analysis of fluorobenzenes in spiked real water samples with satisfactory recoveries and repeatability.

Intelligent peak deconvolution through in-depth study of the data matrix from liquid chromatography coupled with a photo-diode array detector applied to pharmaceutical analysis.

Multivariate curve resolution-alternating least squares (MCR-ALS) method was investigated for its potential to accelerate pharmaceutical research and development. The fast and efficient separation of complex mixtures consisting of multiple components, including impurities as well as major drug substances, remains a challenging application for liquid chromatography in the field of pharmaceutical analysis. In this paper we suggest an integrated analysis algorithm functioning on a matrix of data generated from HPLC coupled with photo-diode array detector (HPLC-PDA) and consisting of the mathematical program for the developed multivariate curve resolution method using an expectation maximization (EM) algorithm with a bidirectional exponentially modified Gaussian (BEMG) model function as a constraint for chromatograms and numerous PDA spectra aligned with time axis. The algorithm provided less than ±1.0% error between true and separated peak area values at resolution (Rs) of 0.6 using simulation data for a three-component mixture with an elution order of a/b/c with similarity (a/b)=0.8410, (b/c)=0.9123 and (a/c)=0.9809 of spectra at peak apex. This software concept provides fast and robust separation analysis even when method development efforts fail to achieve complete separation of the target peaks. Additionally, this approach is potentially applicable to peak deconvolution, allowing quantitative analysis of co-eluted compounds having exactly the same molecular weight. This is complementary to the use of LC-MS to perform quantitative analysis on co-eluted compounds using selected ions to differentiate the proportion of response attributable to each compound.

Quantitation of sulfobutyl ether-ß-cyclodextrin (Captisol™) in Vestipitant IV solution by liquid chromatography with ultraviolet (UV) detection.

This work describes a simple, sensitive and fast liquid chromatographic method using ultraviolet (UV) detection for the quantitation of Captisol™ (sulfobutyl ether-ß-cyclodextrin, SBE-ß-CD) in Vestipitant (GW597599) IV formulation. The chromatographic system consists of a cyano-modified silica stationary phase column with 0.5mM copper(II) acetate in 50/50 (v/v) water/acetonitrile and 0.05% (v/v) of trifluoroacetic acid as the mobile phase. Due to the fact that SBE-ß-CD does not possess a chromophore suitable for UV detection, copper(II) acetate is used as a detection reagent. At low pH copper(II) acetate interacts with SBE-ß-CD and produces mixed copper(II) [Cu(2+)] chelate and copper(II) mono acetate [CuOAc(+)] complexes, while displacing sodium ions [Na(+)] from the sulfobutyl ether (SBE) group. The copper(II)-SBE-ß-CD interaction has optical properties that allow its detection by UV. This novel method is highly reproducible and reliable for accurate quantitation of SBE-ß-CD content in the Vestipitant IV solution, and in the solution without the Vestipitant matrix.

From Spill to Sequestration: The Molecular Journey of Contamination via Comprehensive Multiphase NMR.

Comprehensive multiphase NMR is a novel NMR technique that permits all components (solutions, gels, and solids) to be studied in unaltered natural samples. In this study a wide range of CMP-NMR interaction and editing-based experiments are combined to follow contaminants (pentafluorophenol (PFP) and perfluorooctanoic acid (PFOA)) from the solution state (after a spill) through the gel-state and finally into the true solid-state (sequestered) in an intact water-swollen soil. Kinetics experiments monitoring each phase illustrate PFOA rapidly transfers from solution to the solid phase while for PFP the process is slower with longer residence times in the solution and gel phase. Interaction-based experiments reveal that PFOA enters the soil via its hydrophobic tails and selectively binds to soil microbial protein. PFP sorption shows less specificity exhibiting interactions with a range of gel and solid soil components with a preference toward aromatics (mainly lignin). The results indicate that in addition to more traditional measurements such as Koc, other factors including the influence of the contaminant on the soil-water interface, specific biological interactions, soil composition (content of lignin, protein, etc.) and physical accessibility/swellability of soil organic components will likely be central to better explaining and predicting the true behavior of contaminants in soil.

Sensitive simultaneous determination of 19 fluorobenzoic acids in saline waters by solid-phase extraction and liquid chromatography-tandem mass spectrometry.

A solid-phase extraction (SPE) procedure using C18 stationary phase was optimized for the preconcentration of 19 fluorinated derivatives of benzoic acid (FBA): mono-, di-, tri-, and tetrafluorosubstituted in the ring, trifluoromethylbenzoic acid and 3,5-bistrifluoromethyl benzoic acid from undiluted salt-rich (>20%) reservoir waters. Quantitative (>90%) retention/elution of 16 out of 19 analyte compounds was achieved allowing a fourfold preconcentration factor accompanied by the elimination of >99% of salt. For the three most polar compounds (2,6-dFBA, 2,3,6-tFBA, and 2,4,6-tFBA) the non-quantitative recoveries (>70%) were corrected by dedicated custom-synthesized deuterated internal standards. The FBAs were determined by HPLC - MS/MS revisited in terms of a choice of column, elution conditions and MS/MS signal acquisition parameters allowing the baseline separation and a gain in sensitivity. For a sample intake of 4 mL, detection limits for all the compounds in a reservoir water sample containing more than 20% salt were between 0.01 and 0.05 ng/mL which represents a gain of a factor of 10-20 in comparison with the state-of the art LC-MS/MS procedures for samples of similar complexity.

Bacteria detection based on the evolution of enzyme-generated volatile organic compounds: determination of Listeria monocytogenes in milk samples.

The rapid detection of Listeria monocytogenes contamination in food is essential to prevent food-borne illness in humans. The aim of this study was to differentiate non-contaminated milk from milk contaminated with L. monocytogenes using enzyme substrates coupled with the analysis of volatile organic compounds (VOCs). The method is based on the activity of ß-glucosidase and hippuricase enzymes and the detection of a specific VOC i.e. 2-nitrophenol and 3-fluoroaniline, respectively. VOCs were extracted, separated and detected by headspace-solid phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME GC-MS). This approach required the inclusion of the selective agent's cycloheximide, nalidixic acid and acriflavine HCl in the growth medium to inhibit interfering bacteria. The VOCs were liberated by L. monocytogenes provided that samples contained at least 1-1.5×10(2) CFU ml(-1) of milk prior to overnight incubation. This approach shows potential for future development as a rapid method for the detection of L. monocytogenes contaminated milk.

19F DOSY NMR analysis for spin systems with nJFF couplings.

NMR is a powerful method for identification and quantification of drug components and contaminations. These problems present themselves as mixtures, and here, one of the most powerful tools is DOSY. DOSY works best when there is no spectral overlap between components, so drugs containing fluorine substituents are well-suited for DOSY analysis as (19)F spectra are typically very sparse. Here, we demonstrate the use of a modified (19)F DOSY experiment (on the basis of the Oneshot sequences) for various fluorinated benzenes. For compounds with significant (n) JFF coupling constants, as is common, the undesirable J-modulation can be efficiently suppressed using the Oneshot45 pulse sequence. This investigation highlights (19)F DOSY as a valuable and robust method for analysis of molecular systems containing fluorine atoms even where there are large fluorine-fluorine couplings.

Development of novel chiral capillary electrophoresis methods for the serotonin receptor (5-HT2A) antagonist MDL 100,907 (volinanserin) and for its key intermediate compound.

Enantioselective capillary electrophoretic methods were elaborated for the determination of the enantiomeric purity of (R)-MDL 100,907 and its preparatively resolved key intermediate compound during the synthesis route. The pKa values of the intermediate compound and the end product determined by CE were 10.5±0.1 and 9.0±0.1, respectively. The enantiopurity of the intermediate compound can be monitored in fully protonated state by applying 15mM sulfobutylether-ß-cyclodextrin at pH 5 when the peak belonging to the impurity migrates before the main component. The fact that the consecutive steps of the synthesis do not affect the enantiomeric purity was verified by the other, newly developed CE method. The enantiomers of rac-MDL 100,907 were resolved by 15mM carboxymethyl-γ-cyclodextrin at pH 3. The applicability (selectivity, LOD, LOQ, repeatability, precision and accuracy) of the methods was studied as well.

Transfluthrin indoor air concentration and inhalation exposure during application of electric vaporizers.

Different household insecticide applications via two electric vaporizers emitting transfluthrin were realized in a full-scale experimental room under controlled air exchange rate conditions. On-line high-time resolved measurements of the gas-phase concentrations of the active substance during and immediately after the spreading periods were performed with a High Sensitivity Proton-Transfer-Reaction Mass Spectrometer (HS-PTR-MS). Experimental and modelled data from the ConsExpo 4.0 software were also compared to evaluate the sources of differences. Different application scenarios were also compared. Averaged inhaled concentrations over 1h, 1week, and 5months were estimated to be 8.3, 1.8, and 1.8µg.m(-3), respectively. Corresponding margins of exposures range from 1000 to 10,000, claiming for the absence of effect. Dermal and dust ingestion pathways, although roughly estimated, seems being non-negligible. This claims for a more in-depth integrated risk assessment.

Analysis of 5-HT(2A) receptor binding with [(11)C]MDL 100907 in rats: optimization of kinetic modeling.

PURPOSE: Preclinical positron emission tomography studies are important to follow disease progression and develop new pharmacological agents. We investigated whether kinetic modeling of 5-HT2A tracer [(11)C]MDL 100907 is possible in rats. PROCEDURES AND RESULTS: Kinetic modeling with either metabolite-corrected plasma curve or with the cerebellum as a reference tissue resulted in a good correlation of nondisplaceable binding potential (BPND) calculated from a two-tissue compartment model (2TCM) or different reference tissue models. Injecting the tracer by a slower bolus decreases the variation in 2TCM outcome parameters and results in a good correlation between k3/k4 and the other models. Application of 0.2 mg/kg cold MDL 100907 resulted in almost complete occupancy of 5-HT2A receptors. CONCLUSIONS: A reference tissue model can be used for [(11)C]MDL kinetic modeling in rats, which is preferable in pharmacological or longitudinal studies.

Vibrational spectra and trace determination of p-difluorobenzene and ethylbenzene.

In this work, the structures and normal vibrations of p-difluorobenzene and ethylbenzene in the first excited state have been studied using resonant two-photon ionization spectrum and ab initio quantum chemical calculations. A vibronic spectrum of p-difluorobenzene has been reinvestigated in some detail and three new vibrational modes, 260(1), 130(1) and 30(1), are assigned respectively. The band origin of ethylbenzene of the S1←S0 transition appeared at 37,586 cm(-1), and more vibrational modes are observed. These appropriate structures and vibrational frequencies of the S0 and S1 states of both molecules are calculated using Hartree-Fock and configuration interaction singles methods with 6-311++G(2d, 2p) basis set. All spectral bands of both molecules have been successfully assigned with the help of our computed results and analogy with the reported spectra for similar molecules. The strongest transition 50(1) of p-difluorobenzene is selected to measure a concentration detective limit of the setup system and the achieved concentration detective limit is 3.125 ppb.

The pH-dependence of organofluorine binding domain preference in dissolved humic acid.

In this study we explore the relationship between solution pH and the distribution of the binding interactions at different domains of a dissolved humic acid (HA) for three xenobiotics: pentafluoroaniline (PFA), pentafluorophenol (PFP), and hexafluorobenzene (HFB). The components of HA where xenobiotic interactions occur are identified using the (1)H{(19)F} Reverse Heteronuclear Saturation Transfer Difference (RHSTD) Nuclear Magnetic Resonance (NMR) spectroscopy experiment. At low pH, PFA and PFP interact preferentially with aromatic components of HA. Increasing pH reduces this preference. Conversely, HFB interacts with all components of HA equally, across the entire pH range. The possible roles of both aromatic-specific interactions and conformational changes of HA behind these observations are explored. It is shown that T-oriented π-π interactions at π-electron accepting HA structures are slightly stronger for PFA and PFP than for HFB. Using DOSY NMR it is shown that the pH-dependence of the interactions is correlated with changes in the conformation of the carbohydrate components of HA rather than with the aromatic components. It is argued that the observed preference for aromatic HA is caused by restricted access to the non-aromatic components of HA at low pH. These HA components form tightly bound hydrophobic domains due to strong inter- and intra-molecular hydrogen bonds. At high pH, these structures open up, making them more available for interactions with polar compounds.

Identifying the effective concentration for spatial repellency of the dengue vector Aedes aegypti.

BACKGROUND: Current efforts are underway to quantify the chemical concentration in a treated air space that elicits a spatial repellent (deterrent) response in a vector population. Such information will facilitate identifying the optimum active ingredient (AI) dosage and intervention coverage important for the development of spatial repellent tools--one of several novel strategies being evaluated for vector-borne disease control. This study reports initial findings from air sampling experiments conducted under field conditions to describe the relationship between air concentrations of repellent AIs and deterrent behavior in the dengue vector, Aedes aegypti. METHODS: Air samples were taken inside and outdoors of experimental huts located in Pu Tuey Village, Kanchanaburi Province, Thailand in conjunction with mosquito behavioral evaluations. A mark-release-recapture study design using interception traps was used to measure deterrency of Ae. aegypti against 0.00625% metofluthrin coils and DDT-treated fabric (2g/m2) within separate experimental trials. Sentinel mosquito cohorts were positioned adjacent to air sampling locations to monitor knock down responses to AI within the treated air space. Air samples were analyzed using two techniques: the U.S. Environmental Protection Agency (USEPA) Compendium Method TO-10A and thermal desorption (TD). RESULTS: Both the USEPA TO-10A and TD air sampling methods were able to detect and quantify volatized AIs under field conditions. Air samples indicated concentrations of both repellent chemicals below thresholds required for toxic responses (mortality) in mosquitoes. These concentrations elicited up to a 58% and 70% reduction in Ae. aegypti entry (i.e., deterrency) into treated experimental huts using metofluthrin coils and DDT-treated fabric, respectively. Minimal knock down was observed in sentinel mosquito cohorts positioned adjacent to air sampling locations during both chemical evaluations. CONCLUSIONS: This study is the first to describe two air sampling methodologies that are appropriate for detecting and quantifying repellent chemicals within a treated air space during mosquito behavior evaluations. Results demonstrate that the quantity of AI detected by the mosquito vector, Ae. aegypti, that elicits repellency is far lower than that needed for toxicity. These findings have important implications for evaluation and optimization of new vector control tools that function through mosquito behavior modification as opposed to mortality.

Simultaneous determination of timolol maleate, rosuvastatin calcium and diclofenac sodium in pharmaceuticals and physiological fluids using HPLC-UV.

A novel HPLC-UV method was developed for the simultaneous determination of timolol (TM), rosuvastatin (RST), and diclofenac sodium (DS) in pharmaceuticals, human plasma and aqueous humor using naproxen sodium as internal standard (IS). The target compounds were analyzed on Hypersil BDS C(18) column (250 mm × 4.6 mm, 5 µm), applying 0.2% triethylamine (TEA) and acetonitrile (ACN) (40:60, v/v), in isocratic mode as mobile phase, pH 2.75 adjusted with 85% phosphoric acid at a flow rate of 1 ml/min. The column oven temperature was kept at 45°C and the peak response was monitored at 284 nm after injecting a 50 µl sample into HPLC system. The direct liquid-liquid extraction procedure was applied to human plasma and bovine aqueous humor samples using mobile phase as an extraction solvent after deproteination with methanol. The different HPLC experimental parameters were optimized and the method was validated according to standard guidelines. The recoveries of the suggested method in human plasma were 98.72, 96.04, and 95.14%, for TM, RST, and DS, while in aqueous humor were 94.99, and 98.23%, for TM, and DS, respectively. The LOD values were found to be 0.800, 0.500, and 0.250 ng/ml, for TM, RST, and DS, respectively, while their respective LOQ values were 2.00, 1.50, and 1.00 ng/ml. The co-efficient of variation (CV) were in the range of 0.1492-1.1729% and 1.0516-4.0104%, for intra-day and inter-day studies, respectively. The method was found accurate in human plasma and bovine aqueous humor and will be applied for the quantification of these compounds in plasma, and aqueous humor samples using animal models and in pharmaceuticals.

Direct, metal-free amination of heterocyclic amides/ureas with NH-heterocycles and N-substituted anilines in POCl3.

A POCl(3)-mediated, direct amination reaction of heterocyclic amides/ureas with NH-heterocycles or N-substituted anilines is described. Compared to the existing methods, this operationally simple protocol provides unique reactivity and functional group compatibility because of the metal-free, acidic reaction conditions. The yields are generally excellent.

Determination of rosuvastatin and ezetimibe in a combined tablet dosage form using high-performance column liquid chromatography and high-performance thin-layer chromatography.

This paper describes validated HPLC and HPTLC methods for the simultaneous determination of rosuvastatin (ROS) and ezetimibe (EZE) in a combined tablet dosage form. The isocratic RP-HPLC analysis was performed on a Chromolith C18 column (100 x 6 mm id) using 0.1% (v/v) orthophosphoric acid solution (pH 3.5)-acetonitrile (63 + 37, v/v) mobile phase at a flow rate of 1 mL/min at ambient temperature. Quantification was carried out using a photodiode array UV detector at 245 nm over the concentration range of 0.5-10 microg/mL for ROS and EZE. The HPTLC separation was carried out on an aluminum-backed sheet of silica gel 60F(254) layers using n-butyl acetate-chloroform-glacial acetic acid (1 + 8 + 1, v/v/v) mobile phase. Quantification was achieved with UV densitometry at 245 nm over a concentration range of 0.1-0.9 micro/spot for ROS and EZE. The analytical methods were validated according to International Conference on Harmonization guidelines. Low RSD values indicated good precision. Both methods were successfully applied for the analysis of the drugs in laboratory-prepared mixtures and commercial tablets. No chromatographic interference from the tablet excipients was found. These methods are simple, precise, and sensitive, and are applicable for simultaneous determination of ROS and EZE in pure powder and tablets.