Electrochemical detection of benzodiazepines, following liquid chromatography, for applications in pharmaceutical, biomedical and forensic investigations

Benzodiazepines are an important class of drugs commonly administered with a potential for abuse and environmental pollution. This review focuses on the liquid chromatographic electrochemical detection of the benzodiazepine class of drugs. These are characterised by a readily electrochemically reducible azomethine group, with a number also substituted by other electrochemically active groups. Liquid chromatography employing both single and dual electrode detection has been reported for a variety of benzodiazepines and their metabolites in biological, pharmaceutical, biomedical and forensic investigations. Recently, electrochemistry has been utilised to mimic biological oxidation processes and has been combined with liquid chromatography/mass spectroscopy for their identification and quantification of the products generated. The present review focuses on recent developments in liquid chromatographic- electrochemical determination of benzodiazepines reported since 2006, with earlier reports given in summary

Gray Wolves

Wolf conflicts are primarily related to predation on livestock, pets and other domestic animals, as well as their direct and indirect impacts on native ungulates (i.e., big game). Economic losses vary widely with some livestock producers facing high levels of depredation in some areas. This publication focuses on wolf ecology, damage, and management, particularly as it relates to wolf depredation on livestock and other conflicts with people. Wolves and people share the same environments more than people realize. In the U.S., wolves are not confined to wilderness areas. Though curious, wolves generally fear people and rarely pose a threat to human safety. Wolf attacks on people are, and always have been, very rare compared to other wildlife species. However, there have been several cases of human injuries and a few deaths due to wolves in North America over the past 100 years. The main factors contributing to these incidents were habituation to people, rabies infections, conditioning to human foods, and the presence of domestic dogs. It is unusual for wild wolves to associate or interact with people, linger near buildings, livestock, or domestic dogs, but it does occur especially in areas of high wolf densities in and around rural communities. This type of behavior may be more prevalent in areas where wolves are not legally harvested. This “bold” behavior is more typical of a habituated or food- conditioned animal, a released captive wolf, or a released wolf-dog hybrid. The scale and scope of wolf depredation on livestock depends on local wolf density; numbers and kinds of livestock; livestock husbandry practices; availability and vulnerability of alternative prey; human density; road density; severity of winters; and local hunting pressure. In many instances, wolves live around livestock without causing damage or only occasional damage. Wolf pack size has been shown to increase the likelihood of depredations on domestic animals, with larger packs more likely to cause damage. Most losses occur between April and October when livestock are on summer pastures or grazing allotments. Cattle, especially calves, are the most common livestock killed by wolves. When wolves kill sheep or domestic poultry, often multiple individuals are killed or injured. As of 2019, stable wolf populations exist in many regions in the U.S., including Alaska, Minnesota, Wisconsin, Michigan, Wyoming, Montana, and Idaho, with growing populations in parts of Oregon and Washington (Figures 19, 20). Wolves have recently been documented in northern California and northwestern Colorado. A small population of introduced Mexican wolves exists in Arizona and New Mexico, and a small population of red wolves exists in eastern North Carolina. Both the Mexican and red wolf populations are considered more vulnerable to extinction than other North American wolf populations

A novel electroanalytical approach to the measurement of B vitamins in food supplements based on screen-printed carbon sensors

© 2017 Elsevier B.V. This paper describes the development of a novel electrochemical assay for the measurement of water-soluble vitamins in food and pharmaceutical products. The optimum conditions for the determination of vitamin B1 (thiamine), B2 (riboflavin) and B6 (pyridoxine) in phosphate buffer were established using cyclic voltammetry in conjunction with screen printed carbon electrodes (SPCEs). The optimum current response for all three vitamins was achieved in 0.1 M phosphate buffer pH 11 using an initial potential of −1.0 V. Using square wave voltammetry, the linear ranges for thiamine, riboflavin, and pyridoxine were found to be: 15–110 µg/ml, 0.1–20 µg/ml, and 2–80 µg/ml respectively. The application of the method to a commercial food product yielded a recovery of 95.78% for riboflavin, with a coefficient of variation (CV) of 3.38% (n = 5). The method was also applied to a multi-vitamin supplement for the simultaneous determination of thiamine, riboflavin and pyridoxine. In both cases only simple dilution with buffer followed by centrifugation was required prior to analysis. The resulting square wave voltammetric signals were completely resolved with Ep values of −0.7 V, +0.2 V, and +0.6 V respectively. The recoveries determined for the vitamin B complex in a commercial supplement product were found to be 110%, 114%, and 112% respectively (CV = 7.14%, 6.28%. 5.66% respectively, n = 5)

Design and development of novel screen-printed microelectrode and microbiosensor arrays fabricated using ultrafast pulsed laser ablation

© 2016 Elsevier B.V. All rights reserved. A new generic platform for the development of microbiosensors combining screen-printing and ultrafast pulsed laser technologies has been developed, characterised and evaluated. This new platform consists of a layer of screen-printed carbon ink containing the enzyme and mediator, covered with an insulating layer formed from a dielectric screen printed ink. Microholes were drilled through the insulated layer by ultrafast pulsed laser ablation to generate the microbiosensor array. The geometry of the microelectrode array was evaluated by optical microscopy, white light surface profiling and scanning electron microscopy. The electrochemical behaviour of the microelectrode array was characterised by cyclic voltammetry and compared with macroelectrodes. The analytical performance of the microbiosensor array was evaluated with external counter and reference electrodes for hydrogen peroxide and glucose determination showing linearity up to 4 mmol L-1 and 20 mmol L-1 (360 mg dL-1) respectively. The full screen printed three-electrode configuration shows linearity for glucose determination up to 20 mmol L-1 (360 mg dL-1). This study provides a new fabrication method for microelectrode and microbiosensor arrays capable for the first time to retain the activity of the enzymatic system after processing by pulse laser ablation

Novel reductive-reductive mode electrochemical detection of Rohypnol following liquid chromatography and its determination in coffee

© 2014 Elsevier B.V. Rohypnol (flunitrazepam) has been successfully determined in coffee by high performance liquid chromatography dual electrode detection (LC-DED) in the dual reductive mode. Initial studies were performed to optimise the chromatographic conditions and these were found to be 50% acetonitrile, 50% 50mM pH 2.0 phosphate buffer at a flow rate of 0.75mLmin-1, employing a Hypersil C18, 5μm, 250mm×4.6mm column. Cyclic voltammetric studies were made to ascertain the redox behaviour of Rohypnol at a glassy carbon electrode over the pH range 2-12. Hydrodynamic voltammetry was used to optimise the applied potential at the generator and detector cells; these were identified to be -2.4V and +0.8V for the redox mode and -2.4V and -0.1V for the dual reductive mode respectively. A linear range of 0.5-100μgmL-1, with a detection limit of 20ngmL-1 was obtained for the dual reductive mode. Further studies were then performed to identify the optimum conditions required for the LC-DED determination of Rohypnol in beverage samples. A convenient and rapid method for the determination of Rohypnol in beverage samples was developed using a simple sample pre-treatment procedure. A recovery of 95.5% was achieved for a sample of white coffee fortified at 9.6μgmL-1 Rohypnol

Sequence-Specific Recognition and Cooperative Dimerization of N-Terminal Aromatic Peptides in Aqueous Solution by a Synthetic Host

This article describes the selective recognition and noncovalent dimerization of N-terminal aromatic peptides in aqueous solution by the synthetic host compound, cucurbit[8]uril (Q8). Q8 is known to bind two aromatic guests simultaneously and, in the presence of methyl viologen, to recognize N-terminal tryptophan over internal and C-terminal sequence isomers. Here, the binding of Q8 to aromatic peptides in the absence of methyl viologen was studied by isothermal titration calorimetry (ITC), 1H NMR spectroscopy, and X-ray crystallography. The peptides studied were of sequence X-Gly-Gly, Gly-X-Gly, and Gly-Gly-X (X = Trp, Phe, Tyr, and His). Q8 selectively binds and dimerizes Trp-Gly-Gly (1) and Phe-Gly-Gly (4) with high affinity (ternary K = 109−1011 M-2); binding constants for the other 10 peptides were too small to be measured by ITC. Both peptides bound in a stepwise manner, and peptide 4 bound with positive cooperativity. Crystal structures of Q8·1 and Q8·42 reveal the basis for selective recognition as simultaneous inclusion of the hydrophobic aromatic side chain into the cavity of Q8 and chelation of the proximal N-terminal ammonium group by carbonyl groups of Q8. The peptide sequence selectivity and positively cooperative dimerization reported here are, to the best of our knowledge, unprecedented for synthetic hosts in aqueous solution. Specific peptide recognition and dimerization by synthetic hosts such as Q8 should be important in the study of dimer-mediated biochemical processes and for the separation of peptides and proteins

An electrocatalytic screen-printed amperometric sensor for the selective measurement of thiamine (Vitamin B1) in food supplements

An electrocatalytic screen-printed sensor has been investigated for the measurement of the biologically important biomolecule vitamin B1 (thiamine) for the first time in food supplements. Under basic conditions, the vitamin was converted to its electrochemically active thiolate anion species. It was shown that an electrocatalytic oxidation reaction occurred with the screen-printed carbon electrode containing the mediator cobalt phthalocyanine (CoPC-SPCE). This had the advantage of producing an analytical response current at an operating potential of 0 V vs. Ag/AgCl compared to +0.34 V obtained with plain SPCEs. This resulted in improved selectivity and limit of detection. Detailed studies on the underlying mechanism occurring with the sensor are reported in this paper. A linear response was obtained between 0.1 and 20 µg mL −1 , which was suitable for the quantification of the vitamin in two commercial products containing vitamin B1. The mean recovery for a multivitamin tablet with a declared content of 5 mg was 101% (coefficient of variation (CV) of 9.6%). A multivitamin drink, which had a much lower concentration of vitamin B1 (0.22 mg/100 mL), gave a mean recovery of 93.3% (CV 7.2%). These results indicate that our sensor holds promise for quality control of food supplements and other food types

The Wide Field Spectrograph (WiFeS)

This paper describes the Wide Field Spectrograph (WiFeS) under construction at the Research School of Astronomy and Astrophysics (RSAA) of the Australian National University (ANU) for the ANU 2.3m telescope at the Siding Spring Observatory. WiFeS is a powerful integral field, double-beam, concentric, image-slicing spectrograph designed to deliver excellent thoughput, wavelength stability, spectrophotometric performance and superb image quality along with wide spectral coverage throughout the 320-950 nm wavelength region. It provides a 25x38 arcsec. field with 0.5 arcsec. sampling along each of twenty five 38X1 arcsec slitlets. The output format is optimized to match the 4096x4096 pixel CCD detectors in each of two cameras individually optimized for the blue and the red ends of the spectrum, respectively. A process of "interleaved nod-and-shuffle" will be applied to permit quantum noise-limited sky subtraction. Using VPH gratings, spectral resolutions of 3000 and 7000 are provided. The full spectral range is covered in a single exposure at R=3000, and in two exposures in the R=7000 mode. The use of transmissive coated optics, VPH gratings and optimized mirror coatings ensures a throughput (including telescope atmosphere and detector) > 30% over a wide spectral range. The concentric image-slicer design ensures an excellent and uniform image quality across the full field. To maximize scientific return, the whole instrument is configured for remote observing, pipeline data reduction, and the accumulation of calibration image libraries.Comment: Accepted for publication in Astrophysics & Space Science, 16 pages, 14 figure