Rapid Analysis of Fentanyl and Fentanyl Analogues from Whole Blood Using SPME Coupled to the Microfluidic Open Interface.

Fentanyl and its analogues are potent opioids that pose a significant threat to society. Over the last several years, considerable focus has been on the concerning trend of increasing fentanyl usage among drug users. Fentanyl analogues are mainly synthesized to evade analytical detection or increase their potency; thus, very low concentrations are sufficient to achieve a therapeutic effect. In an effort to help combat the synthetic opioid epidemic, developing targeted mass spectrometric methods for quantifying fentanyl and its analogues at ultralow concentrations is incredibly important. Most methods used to analyze fentanyl and its analogues from whole blood require manual sample preparation protocols (solid-phase extraction or liquid-liquid extraction), followed by chromatographic separation and mass spectrometric detection. The main disadvantages of these methods are the tedious sample preparation workflows, resulting in lengthy analysis times. To mitigate these issues, we present a targeted method capable of analyzing 96 samples containing fentanyl, several fentanyl analogues, and a common fentanyl (analogue) precursor simultaneously in 2.4 min per sample. This is possible by using a high-throughput solid phase microextraction workflow on the Concept96 autosampler followed by manual coupling of solid-phase microextraction fibers to the microfluidic open interface for tandem mass spectrometry analysis. Our quantitative method is capable of extremely sensitive analysis, with limits of quantification ranging from 0.002 to 0.031 ng mL-1 and linearity ranging from 0.010 to 25.0 ng mL-1. The method shows very good reproducibility (1-18%), accuracy (81-100%) of calibration and validation points, and good interday reproducibility (6-15%).

Monitoring Poly(methyl methacrylate) and Polyvinyl Dichloride Micro/Nanoplastics in Water by Direct Solid-Phase Microextraction Coupled to Gas Chromatography-Mass Spectrometry.

A simple, sustainable, and sensitive monitoring approach of micro/nanoplastics (MNPs) in aqueous samples is crucial since it helps in assessing the extent of contamination and understanding the potential risks associated with their presence without causing additional stress to the environment. In this study, a novel strategy for qualitative and quantitative determination of MNPs in water by direct solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was proposed for the first time. Spherical poly(methyl methacrylate) (PMMA) and irregularly shaped polyvinyl dichloride (PVDC) were used to evaluate the feasibility and performance of the proposed method. The results demonstrated that both PMMA and PVDC MNPs were efficiently extracted by the homemade SPME coating of nitrogen-doped porous carbons (N-SPCs) and exhibited sufficient thermal decomposition in the GC-MS injection port. Excellent extraction performances of N-SPCs coating for MNPs are attributed to hydrophobic cross-linking, electrostatic forcing, hydrogen bonding, and pore trapping. Methyl methacrylate was identified as the marker for PMMA, while 1,3-dichlorobenzene and 1,3,5-trichlorobenzene were the indicators for PVDC. Under the optimal extraction and decomposition conditions, the proposed method exhibited ultrahigh sensitivity, with a limit of detection of 0.0041 µg/L for PMMA and 0.0054 µg/L for PVDC. Notably, a programmed temperature strategy for the GC-MS injector was developed to discriminate and eliminate the potential interferences of intrinsic indicator compounds. Owing to the integration of sampling, extraction, injection, and decomposition into one step by SPME, the proposed method demonstrates exceptional sensitivity, eliminating the necessity for complex sample pretreatment procedures and the use of organic solvents. Finally, the proposed method was successfully applied in the determination of PMMA and PVDC MNPs in real aqueous samples.

Quantitative Determination of Poly(methyl Methacrylate) Micro/Nanoplastics by Cooling-Assisted Solid-Phase Microextraction Coupled to Gas Chromatography-Mass Spectrometry: Theoretical and Experimental Insights.

Determinations of micro/nanoplastics (MNPs) in environmental samples are essential to assess the extent of their presence in the environment and their potential impact on ecosystems and human health. With the aim to provide a sensitive method with simplified pretreatment steps, cooling-assisted solid-phase microextraction (CA-SPME) coupled to gas chromatography-mass spectrometry (GC-MS) is proposed as a new approach to quantify mass concentrations of MNPs in water and soil samples. The herein proposed CA-SPME method offers the unique advantage of integrating the thermal decomposition of MNPs and enrichment of signature compounds into one step. Poly(methyl methacrylate) (PMMA) was used as a model substance to verify the method performance in this work. Theoretical insights demonstrated that pyrolysis is the rate-determining step during the extraction process and that PMMA is effectively decomposed at 350 °C with an estimated incubation time of 13 min. Eight compounds were identified in the pyrolysis products by CA-SPME-GC-MS with the use of a DVB/CAR/PDMS coating, wherein methyl methacrylate was considered as the best indicator and dimethyl 2-methylenesuccinate was selected as the confirmation compound. Under the optimized conditions, the proposed method exhibited wide linearity (0.5-2000 µg for water and 5-1000 µg for soil) and high sensitivity, with limits of detection of 0.014 and 0.28 µg for water and soil, respectively. Finally, the proposed method was successfully applied for determinations of PMMA MNPs in real water and soil samples with satisfactory recoveries attained. The method only required the employment of a filter membrane for water analysis, while soil samples were analyzed directly without any pretreatment. The solvent-free approach, straightforward operation, and high sensitivity of the proposed method show great potential for the analysis of MNPs in different environmental samples.

Proteomic Analysis of Human Saliva via Solid-Phase Microextraction Coupled with Liquid Chromatography-Mass Spectrometry.

Proteomics of human saliva samples was achieved for the first time via biocompatible solid-phase microextraction (bio-SPME) devices. Upon introduction of a porogen to a conventional C18 coating, porous C18/polyacrylonitrile (PAN) SPME blades were able to extract peptides up to 3.0 kDa and more peptides than commercial SPME blades. Following Trypsin digestion, salivary proteomic analysis was achieved via SPME-LC-MS/MS. Seven endogenous proteins were consistently identified in all saliva samples via bio-SPME. Taking advantage of this strategy, untargeted peptidomics was applied for the comparison of saliva samples between healthy and SARS-CoV-2 positive individuals. The results showed clear peptidomic differences between the viral and healthy saliva samples. This proof-of-concept study demonstrates the potential of bio-SPME-LC-MS/MS for peptidomics and proteomics in biomedical applications.

Embedding Mixed Sorbents in Binder: Solid-Phase Microextraction Coating with Wide Extraction Coverage and Its Application in Environmental Water Analysis.

Solid-phase microextraction (SPME) is a simple and highly effective sample-preparation technique for water analysis. However, the extraction coverage of a given SPME device with a specific coating can be an issue when analyzing multiple environmental contaminants. Therefore, instead of synthesizing one sorbent material with dual or multiple functions, we investigated a new strategy of preparing SPME blades using a homogeneous slurry made by mixing three different sorbent particles─namely, hydrophobic/lipophilic balanced (HLB), HLB-weak cationic exchange (HLB-WCX), and HLB-weak anionic exchange (HLB-WAX)─with a polyacrylonitrile (PAN) binder. The developed coating is matrix compatible, as the binder functions not only as a glue for immobilizing the sorbent particles but also as a porous filter, which only allows small molecules to enter the pores and interact with the particles, thus avoiding contamination from large elements. The results confirmed that the proposed mixed-coating SPME device provides good extraction performance for polar and nonpolar as well as positively and negatively charged compounds. Based on this device, three comprehensive analytical methodologies─high-throughput SPME-LC-MS/MS (for the quantitative analysis of targeted drugs of abuse and artificial sweeteners), in-bottle SPME-LC-high resolution MS (HRMS) (for the untargeted screening of organic contaminants), and on-site drone sampling SPME-LC-HRMS (for on-site sampling and untargeted screening)─were developed for use in environmental water analysis. The resultant data confirm that the proposed strategies enable comprehensive water quality assessment by using a single SPME device.

Evaluation of thin film microextraction based on graphene oxide/ polymer composite: Experimental and theoretical insights.

Experimental and theoretical assessments of a graphene oxide-based polymer as adsorbent for thin film microextraction (TFME) were conducted as part of this research. Graphene oxide (GO) was embedded in the organic polymer poly(styrene-co-divinylbenzene) (PS-DVB) to prepare a sorbent suitable for direct-immersion TFME. A TFME membrane coating prepared with the GO/PS-DVB sorbent and polydimethylsiloxane (PDMS) as binder was then applied for extraction of organic pollutants from aqueous and gaseous samples. The surface morphology of the TFME coating was examined by scanning electron microscopy (SEM). Various TFME parameters influencing extraction efficiency, such as extraction time and temperature, desorption temperature, and ionic strength, were investigated and optimized. In a comparison of TFME membranes, the GO/PS-DVB/PDMS TFME membrane was shown to yield higher extraction efficiencies for the targeted analytes than the pure PDMS and DVB/PDMS TFME membranes. The calibration graphs of the organic pollutants displayed linearity for most of the target analytes within the 10-2000 ng L-1 concentration range. The repeatability (RSD %, n = 5) and reproducibility (RSD %, n = 3) of the method were in the ranges of 2.2-5.9 %, and 3.2-8.5 %, respectively, at a concentration level of 500 ng L-1, whereas accuracy (%) ranged between 79.8 and 119 %. The developed method was successfully applied for determinations of organic pollutants in tap water, lake water, and wastewater samples. Furthermore, the impact of mass transfer kinetics on extractions by the GO/PS-DVB/PDMS TFME membrane from gaseous samples was theoretically discussed and experimentally verified. The results of this work demonstrate that the GO/PS-DVB/PDMS TFME method is a simple, efficient, and environmentally friendly method for pre-treatment of organic pollutants.

Perspective on SPME-MS: Green and high-performance methods for rapid screening.

The direct coupling of solid-phase microextraction (SPME) with mass spectrometry (MS) offers rapid analysis with high sensitivity and low matrix effects by benefiting from the integration of sampling, high enrichment, and clean-up functions of SPME. Eliminating chromatographic separation reduces the amount of gas/solvent needed for analysis, while direct desorption in SPME-MS consumes none or few microliters of organic solvents per sample, further enhancing the greenness of the SPME technology. Over the past two decades, the rapid evolution of SPME-MS has given rise to numerous novel technologies that employ diverse ionization techniques and interfaces, several of which have already been commercialized. Drawing from an extensive review published earlier this year and our research experience, we provide perspectives on three aspects of these technologies: interface design and automation, integration with state-of-art MS instrumentation, and anticipated future developments.

Sequential thin film microextraction and overcoated thin film microextraction devices for characterization of sparkling wine aroma profiles and partitioning equilibria.

Solid Phase Microextraction (SPME) is a commonly used, robust method for characterization of aroma profiles in food matrices. However, challenges such as saturation, swelling, and competition can occur when sampling such complex matrices, resulting in decreased accuracy in the quantitation of polar compounds. In this study, sequential thin film micro-extraction (TFME) was employed to study the aroma profile of sparkling wine, with a focus to evaluate the displacement of polar analytes at extraction times longer than their corresponding equilibrium time. This investigation also describes advancements in the production of TFME devices, specifically the overcoating of hydrophilic-lipophilic balance/polydimethylsiloxane (HLB/PDMS) thin films to increase their matrix compatibility. Sequential thin film micro-extraction and overcoated HLB/PDMS thin films were evaluated for characterization of sparkling wine samples. The results were encouraging, showing that these advancements can decrease competition phenomena and increase the calibration linearity range compared to traditional micro-extraction approaches more commonly used for the characterization of such samples. In addition, multiphase equilibria investigation involving micellar systems enabled by the microextraction technology provides better understanding between wine aroma and its composition.

Perspective on in vivo SPME for human applications: Starting from monitoring doxorubicin during lung chemo-perfusion.

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Mapping the metabolic responses to oxaliplatin-based chemotherapy with in vivo spatiotemporal metabolomics.

Adjuvant chemotherapy improves the survival outlook for patients undergoing operations for lung metastases caused by colorectal cancer (CRC). However, a multidisciplinary approach that evaluates several factors related to patient and tumor characteristics is necessary for managing chemotherapy treatment in metastatic CRC patients with lung disease, as such factors dictate the timing and drug regimen, which may affect treatment response and prognosis. In this study, we explore the potential of spatial metabolomics for evaluating metabolic phenotypes and therapy outcomes during the local delivery of the anticancer drug, oxaliplatin, to the lung. 12 male Yorkshire pigs underwent a 3 h left lung in vivo lung perfusion (IVLP) with various doses of oxaliplatin (7.5, 10, 20, 40, and 80 mg/L), which were administered to the perfusion circuit reservoir as a bolus. Biocompatible solid-phase microextraction (SPME) microprobes were combined with global metabolite profiling to obtain spatiotemporal information about the activity of the drug, determine toxic doses that exceed therapeutic efficacy, and conduct a mechanistic exploration of associated lung injury. Mild and subclinical lung injury was observed at 40 mg/L of oxaliplatin, and significant compromise of the hemodynamic lung function was found at 80 mg/L. This result was associated with massive alterations in metabolic patterns of lung tissue and perfusate, resulting in a total of 139 discriminant compounds. Uncontrolled inflammatory response, abnormalities in energy metabolism, and mitochondrial dysfunction next to accelerated kynurenine and aldosterone production were recognized as distinct features of dysregulated metabolipidome. Spatial pharmacometabolomics may be a promising tool for identifying pathological responses to chemotherapy.

The impact of normothermic and hypothermic preservation methods on kidney lipidome-comparative study using chemical biopsy with microextraction probes.

Introduction: Normothermic ex vivo kidney perfusion (NEVKP) is designed to replicate physiological conditions to improve graft outcomes. A comparison of the impact of hypothermic and normothermic preservation techniques on graft quality was performed by lipidomic profiling using solid-phase microextraction (SPME) chemical biopsy as a minimally invasive sampling approach. Methods: Direct kidney sampling was conducted using SPME probes coated with a mixed-mode extraction phase in a porcine autotransplantation model of the renal donor after cardiac death, comparing three preservation methods: static cold storage (SCS), NEVKP, and hypothermic machine perfusion (HMP). The lipidomic analysis was done using ultra-high-performance liquid chromatography coupled with a Q-Exactive Focus Orbitrap mass spectrometer. Results: Chemometric analysis showed that the NEVLP group was separated from SCS and HMP groups. Further in-depth analyses indicated significantly (p < 0.05, VIP > 1) higher levels of acylcarnitines, phosphocholines, ether-linked and longer-chain phosphoethanolamines, triacylglycerols and most lysophosphocholines and lysophosphoethanolamines in the hypothermic preservation group. The results showed that the preservation temperature has a more significant impact on the lipidomic profile of the kidney than the preservation method's mechanical characteristics. Conclusion: Higher levels of lipids detected in the hypothermic preservation group may be related to ischemia-reperfusion injury, mitochondrial dysfunction, pro-inflammatory effect, and oxidative stress. Obtained results suggest the NEVKP method's beneficial effect on graft function and confirm that SPME chemical biopsy enables low-invasive and repeated sampling of the same tissue, allowing tracking alterations in the graft throughout the entire transplantation procedure.

Rapid screening of drugs in environmental water using metal organic framework/Ti3C2Tx composite coated blade spray-mass spectrometry.

Simultaneous rapid screening of multiple drugs of abuse in environmental water facilitates effective monitoring and trend assessments. Herein, a novel porphyrin-based metal organic frameworks modified Ti3C2Tx nanosheets (Cu-TCPP/Ti3C2Tx) composite was prepared and utilized as solid-phase microextraction (SPME) coating for the simultaneous analysis of 21 drugs from water samples. The composite was embedded with matrix-compatible polyacrylonitrile binder to prepare a coated blade with thin and uniform coating layer. Ambient mass spectrometry (MS) technique was used to create a coated blade spray-MS (CBS-MS) method for the quantitative determination of drugs in water samples. High throughput and automated sample preparation were achieved with the use of a Concept 96-well plate system, enabling analysis of 21 drugs of abuse within 1 min per sample, while using only 8 µL of organic solvent for desorption and CBS-MS detection. The developed method showed favorable linearity (R2 ≥ 0.9983) in the range of 0.05 to 10 ng mL-1, low limits of detection (1.5-9.0 ng L-1), sufficient recovery (67.6-133.2%), as well as satisfactory precision (RSDs≤13.5%). This study not only delivers a novel and efficient SPME coating composite, but also demonstrates the excellent performance of a high-throughput, efficient, and green analytical method for determination of drugs in environmental water.