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.

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.

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.