Developing and Evaluating the Greenness of a Reliable, All-in-One Thin-Film Microextraction Protocol for Determining Fentanyl, Methadone, and Zolpidem in Plasma, Urine, and Oral Fluid.

This paper proposes an all-in-one microextraction-based protocol capable of determining and quantifying fentanyl, methadone, and zolpidem in plasma, urine, and saliva at concentrations below those required by international regulatory organizations. A homemade thin-film microextraction device featuring an octyl-cyanopropyl stationary phase was coupled with LC-MS/MS. The proposed method was developed and validated according to FDA criteria, providing extraction efficiency values ranging from 26.7% to 76.2% with no significant matrix effects (2.6% to 15.5% signal suppression). The developed protocol provided low limits of quantification (mostly equal to 1 ng mL-1) and good reproducibility (intra- and inter-day RSDs of less than 9.6% and 12.0%, respectively) and accuracy (89% to 104% of the test concentration). An assessment of the protocol's environmental impact indicated that attention must be devoted to eliminating the use of toxic reagents and developing its capability for in situ sampling and in-field analysis using portable instruments. The proposed TFME-based protocol provides clinical laboratories with a versatile, one-step tool that enables the simultaneous monitoring of fentanyl, methadone, and zolpidem using the most popular biological matrices.

Quantification of prohibited substances and endogenous corticosteroids in saliva using traditional, alternative microextraction-based, and novel 3D printed sample-preparation methods coupled with LC-MS.

Oral fluid has gained significant interest as an alternative matrix for drug testing due to its easy and non-invasive collection. Despite these advantages, achieving suitably low limits of detection remains a clear challenge in the use of oral fluids for drug screening. In this study, we demonstrate that the application of commercially available SPME fibers followed by liquid chromatography tandem mass spectrometry can enable the comprehensive detection and confirmation of drugs in oral fluid samples. To this end, we develop and test a sample-preparation protocol for a panel of 46 drugs covering the most popular drugs of abuse and doping agents available worldwide. Human saliva samples were collected using a Salivette® device (CE IVD certified) and sampled using SPME devices coated with a C18 extraction phase. The proposed protocol was validated with respect to its lower limits of quantification (LLOQ), linearity, matrix effects, precision, and extraction recovery. Linearity was confirmed for all compounds (R2 > 0.97), except for testosterone (R2 = 0.953) and metandrostenolon (R2 = 0.958). Furthermore, 4 compounds suffered from matrix effects, with less than 10 % deviation from acceptance criteria. After analytical validation, saliva samples from volunteers were analyzed to determine free concentrations of cortisol at different times after awaking. Finally, a 3D-printed prototype device was designed and successfully applied to extract small molecules, thus demonstrating a new modern low-cost approach for bioanalysis.

Innovative, simple, and green: A sample preparation method based on 3D printed polymers.

The present study evaluates the capability of fifteen 3D printed thermoplastic polymers as novel stationary phases for the extraction of forty-three physicochemically diverse analytes from fortified human oral fluid samples. Prototype extraction devices were prepared in 96-well plate-compatible format using fused deposition modeling 3D printer. The sample preparation was performed with 5-step protocol utilizing 96-well plates and semiautomated benchtop shaker. All resulting extracts were analyzed via high-performance liquid chromatography (operated in reversed-phase gradient elution mode) and tandem mass spectrometry (with electrospray ionization and triple quadrupole mass spectrometer). Exceptionally favorable results were observed for three polymer types: polyamide 6 (reinforced with 15% carbon fiber), LAYFOMM-60 (polyurethane with water-soluble polyvinyl alcohol), and S-FLEX 90A (thermoplastic polyurethane). Furthermore, this study also introduces an automated and repeatable 3D printing method for the fast fabrication of high-throughput, and highly selective sample preparation devices, most of which are ready-to-use without any additional processing or chemical functionalization. As such, the proposed printing method represents a significant step towards the introduction of novel polymeric stationary phases for analytical sample preparation, thus providing laboratory personnel with a method that is safer and more convenient, while minimizing negative environmental impacts.

Finding the best hardware configuration for 2D SLAM in indoor environments via simulation based on Google Cartographer.

One of the most challenging topics in robotics is simultaneous localization and mapping (SLAM) in the indoor environments. Due to the fact that Global Navigation Satellite Systems cannot be successfully used in such environments, different data sources are used for this purpose, among others light detection and ranging (LiDARs ), which have advanced from numerous other technologies. Other embedded sensors can be used along with LiDARs to improve SLAM accuracy, e.g. the ones available in the Inertial Measurement Units and wheel odometry sensors. Evaluation of different SLAM algorithms and possible hardware configurations in real environments is time consuming and expensive. In our study, we evaluate the accuracy of mapping and localization (based on Absolute Trajectory Error and Relative Pose Error). Our use case is a robot used for room decontamination. The results for a small room show that for our robot the best hardware configuration consists of three LiDARs 2D, IMU and wheel odometry sensors. On the other hand, for long hallways, a configuration with one LiDAR 3D sensor and IMU works better and more stable. We also described a general approach together with tools and procedures that can be used to find the best sensor setup in simulation.

Building a Real-Time Testing Platform for Unmanned Ground Vehicles with UDP Bridge.

Perception and vehicle control remain major challenges in the autonomous driving domain. To find a proper system configuration, thorough testing is needed. Recent advances in graphics and physics simulation allow researchers to build highly realistic simulations that can be used for testing in safety-critical domains and inaccessible environments. Despite the high complexity of urban environments, it is the non-urban areas that are more challenging. Nevertheless, the existing simulators focus mainly on urban driving. Therefore, in this work, we describe our approach to building a flexible real-time testing platform for unmanned ground vehicles for indoor and off-road environments. Our platform consists of our original simulator, robotic operating system (ROS), and a bridge between them. To enable compatibility and real-time communication with ROS, we generate data interchangeable with real-life readings and propose our original communication solution, UDP Bridge, that enables up to 9.5 times faster communication than the existing solution, ROS#. As a result, all of the autonomy algorithms can be run in real-time directly in ROS, which is how we obtained our experimental results. We provide detailed descriptions of the components used to build our integrated platform.

Modifying current thin-film microextraction (TFME) solutions for analyzing prohibited substances: Evaluating new coatings using liquid chromatography.

For identifying and quantifying prohibited substances, solid-phase microextraction (SPME) continues to arouse interest as a sample preparation method. However, the practical implementation of this method in routine laboratory testing is currently hindered by the limited number of coatings compatible with the ubiquitous high-performance liquid chromatography (HPLC) systems. Only octadecyl (C18) and polydimethylsiloxane/divinylbenzene ligands are currently marketed for this purpose. To address this situation, the present study evaluated 12 HPLC-compatible coatings, including several chemistries not currently used in this application. The stationary phases of SPME devices in the geometry of thin film-coated blades were prepared by applying silica particles bonded with various functional ligands (C18, octyl, phenyl-hexyl, 3-cyanopropyl, benzenesulfonic acid, and selected combinations of these), as well as unbonded silica, to a metal support. Most of these chemistries have not been previously used as microextraction coatings. The 48 most commonly misused substances were selected to assess the extraction efficacy of each coating, and eight desorption solvent compositions were used to optimize the desorption conditions. All samples were analyzed using an HPLC system coupled with triple quadrupole tandem mass spectrometry. This evaluation enables selection of the best-performing coatings for quantifying prohibited substances and investigates the relationship between extraction efficacy and the physicochemical characteristics of the analytes. Ultimately, using the most suitable coatings is essential for trace-level analysis of chemically diverse prohibited substances.

Polyamide Noncoated Device for Adsorption-Based Microextraction and Novel 3D Printed Thin-Film Microextraction Supports.

Polyamide noncoated device for adsorption-based microextraction (PANDA microextraction) is a brand new, easy to prepare, environmentally friendly, inexpensive, and efficient sample preparation method created entirely with the use of 3D printing. The proposed method is based on the extractive proprieties of the unmodified polyamide and carbon fiber blends and is compared with the highly selective thin-film microextraction (TFME). In addition, 3D printing was used to simplify the process of TFME. Prototype sample preparation devices were evaluated by the extraction of oral fluid spiked with 38 small molecules with diverse chemical natures, such as lipophilicity in the log P range of 0.2-7.2. The samples were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry. The results indicate that chemically and thermally resistant 3D printed supports can be successfully used as a cost-saving, environmentally friendly solution for the preparation of TFME devices, alternative to the conventional metal supports, with only marginal differences in the extraction yield (mean = 4.0%, median = 1.8%, range = 0.0-22.3%, n = 38). Even more remarkably, in some cases, the newly proposed PANDA microextraction method exceeded the reference TFME in terms of the extraction efficacy and offered excellent sample cleanup as favorable matrix effects were observed (mean = -8.5%, median = 7.5%, range = -34.7-20.0%, n = 20). This innovative approach paves the road to the simplified sample preparation with the use of emerging extractive 3D printing polymers.

Benefits of Innovative and Fully Water-Compatible Stationary Phases of Thin-Film Microextraction (TFME) Blades.

Octadecyl (C18) groups are arguably the most popular ligands used for preparation of solid phase microextraction (SPME) devices. However, conventional C18-bonded silica particles are not fully compatible with the nearly 100% aqueous composition of typical biological samples (e.g., plasma, saliva, or urine). This study presents the first evaluation of thin-film SPME devices coated with special water-compatible C18-bonded particles. Device performance was assessed by extracting a mixture of 30 model compounds that exhibited various chemical structures and properties, such as hydrophobicity. Additionally, nine unique compositions of desorption solvents were tested. Thin-film SPME devices coated with C18-bonded silica particles with polar end-capping groups (10 µm) were compared with conventional trimethylsilane end-capped C18-bonded silica particles of various sizes (5, 10, and 45 µm) and characteristics. Polar end-capped particles provided the best extraction efficacy and were characterized by the strongest correlations between the efficacy of the extraction process and the hydrophobicity of the analytes. The results suggest that the original features of octadecyl ligands are best preserved in aqueous conditions by polar end-capped particles, unlike with conventional trimethylsilane end-capped particles that are currently used to prepare SPME devices. The benefits associated with this improved type of coating encourage further implementation of microextractraction as greener alternative to the traditional sample preparation methods.

LiDAR Point Cloud Generation for SLAM Algorithm Evaluation.

With the emerging interest in the autonomous driving level at 4 and 5 comes a necessity to provide accurate and versatile frameworks to evaluate the algorithms used in autonomous vehicles. There is a clear gap in the field of autonomous driving simulators. It covers testing and parameter tuning of a key component of autonomous driving systems, SLAM, frameworks targeting off-road and safety-critical environments. It also includes taking into consideration the non-idealistic nature of the real-life sensors, associated phenomena and measurement errors. We created a LiDAR simulator that delivers accurate 3D point clouds in real time. The point clouds are generated based on the sensor placement and the LiDAR type that can be set using configurable parameters. We evaluate our solution based on comparison of the results using an actual device, Velodyne VLP-16, on real-life tracks and the corresponding simulations. We measure the error values obtained using Google Cartographer SLAM algorithm and the distance between the simulated and real point clouds to verify their accuracy. The results show that our simulation (which incorporates measurement errors and the rolling shutter effect) produces data that can successfully imitate the real-life point clouds. Due to dedicated mechanisms, it is compatible with the Robotic Operating System (ROS) and can be used interchangeably with data from actual sensors, which enables easy testing, SLAM algorithm parameter tuning and deployment.

Evaluation of swabs from 15 commercially available oral fluid sample collection devices for the analysis of commonly abused substances: doping agents and drugs of abuse.

Oral fluid testing is steadily building its position as a valuable complement or alternative to plasma and urine analyses in everyday laboratory practice. However, the great significance of the sample collection process in the attainment of representative results is not always paralleled by the attention given to its informed selection. Few evaluations of commercially available sample collection devices have been published until now, and the current work intends to fill this gap by presenting an evaluation of swabs from 15 different devices for the analysis of 49 popular drugs. Swabs, derived from sample collection devices, were used to collect a drug-fortified mixture. Then, swab-retrieved samples were subjected to instrumental analysis with the high-performance liquid chromatography coupled with tandem mass spectrometry method. Results within the 80-120% range were considered to have no significant impact on analyte concentration (thus satisfactory) and were observed in 44.1% of all results. Out of the 15 evaluated swabs, 7 provided results in the aforementioned range for more than half of the substances under study. The possibility of matrix effects originating from swab materials was also investigated. The selection of an appropriate oral fluid sample collection method plays a critical role in the success of the analytical procedure, a fact that is well-illustrated by the tremendous differences between analyte concentrations observed in this research. Perhaps, the tedious labour of improving sample preparation and analysis methods already in-use could be spared if only greater emphasis were to be put on the improvement and better selection of suitable solutions for oral fluid collection.

Pharmacological Aspects of Over-the-Counter Opioid Drugs Misuse.

Several over-the-counter (OTC) drugs are known to be misused. Among them are opioids such as codeine, dihydrocodeine, and loperamide. This work elucidates their pharmacology, interactions, safety profiles, and how pharmacology is being manipulated to misuse these common medications, with the aim to expand on the subject outlined by the authors focusing on abuse prevention and prevalence rates. The reviewed literature was identified in several online databases through searches conducted with phrases created by combining the international non-proprietary names of the drugs with terms related to drug misuse. The results show that OTC opioids are misused as an alternative for illicit narcotics, or prescription-only opioids. The potency of codeine and loperamide is strongly dependent on the individual enzymatic activity of CYP2D6 and CYP3A4, as well as P-glycoprotein function. Codeine can also be utilized as a substrate for clandestine syntheses of more potent drugs of abuse, namely desomorphine ("Krokodil"), and morphine. The dangerous methods used to prepare these substances can result in poisoning from toxic chemicals and impurities originating from the synthesis procedure. OTC opioids are generally safe when consumed in accordance with medical guidelines. However, the intake of supratherapeutic amounts of these substances may reveal surprising traits of common medications.