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%).

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.

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.

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.

Matrix-Compatible Solid-Phase Microextraction Pin Coupled Directly to Mass Spectrometry using Probe Electrospray Ionization.

A solid-phase microextraction (SPME) pin device with a biocompatible coating on the tip was developed for direct coupling to mass spectrometry (MS) via a vertical dipping-and-spray strategy using an automated probe electrospray ionization (PESI) interface. The developed method provides superior sensitivity compared to standard PESI-MS due to the enrichment effects of SPME and the significant increase in the volume of sample and/or solvent collected during dipping due to the SPME pin's notably larger size. The tips of the SPME pins were coated with a biocompatible coating consisting of small sorbent particles embedded into a polyacrylonitrile (PAN) binder. This coating enables the extraction of small molecules, while preventing larger molecules such as tissue fragments, proteins, and cell matter from coming into the sorbent. The developed SPME pin-PESI-MS method also features much lower matrix effects compared to PESI-MS for the analysis of complex biology samples. When applied for the analysis of 8 drugs of abuse in urine samples, the SPME pin-PESI-MS method provided good linearity (R2 ≥ 0.9997), high sensitivity with limits of detection between 0.003 to 0.03 ng/mL, and good reproducibility with RSD% ≤ 6%. The vertical design of the SPME-PESI-MS direct-coupling interface allows the potential fully automation of the system using a conventional autosampler.

Performance Evaluation of Extraction Coatings with Different Sorbent Particles and Binder Composition.

Binders are critical components used in the preparation of a range of extraction devices, including solid-phase microextraction (SPME) devices. While the main role of a binder is to affix the sorbent particles to the selected support, it is critical to select the optimal binder to ensure that it does not negatively impact the coating's particle sorption capability. This work presents the first comprehensive investigation of the interactions between binders and solid sorbent particles as these interactions can significantly impact the performance of the coating. Specifically, the findings presented herein provide a better understanding of the extraction mechanisms of composite coatings and new rules for predicting the particle adhesion forces and binder distribution in the coating. The influence of binder chemistry on coating performance is investigated by examining a selection of the most used binders, namely, polydimethylsiloxane (PDMS), polyacrylonitrile (PAN), poly(vinylidene difluoride) (PVDF), polytetrafluoroethylene amorphous fluoroplastics (PTFE AF 2400), and polybenzimidazole (PBI). The solid particles (e.g., hydrophilic-lipophilic balanced (HLB) and C18) used in this work were selected for their ability to provide optimal extraction coverage for a broad range of analytes. The results show that PDMS does not change the properties of the solid particles and that the binder occupies a negligible volume due to shrinking after polymerization, resulting in the solid particles making up most of the coating volume. Hence, the coating sorption characteristics correspond closely to the properties of the selected solid particles. On the other hand, the results also showed that PTFE AF 2400 can interact with the active surface of the sorbent, leading to the deactivation of the sorbent particles. Therefore, the extraction performance and permeability coefficients decrease as the size of the penetrant increases, indicating a rigid porous structure. The results of this study can aid in the optimization of SPME devices as they provide reference values that can be used to determine the optimal binder and the sorbent affinity for the targeted compounds. Finally, the present work also provides the broader scientific community with a strategy for investigating the properties of sorbent particle/binder structures and defines the characteristics of a good coating/membrane by analyzing all parameters such as kinetics, thermodynamic equilibria, and morphology.

Sequential Solid-Phase Microextraction with Biocompatible Coating Materials to Address Displacement Effects in the Quantitative Analysis.

Solid-phase microextraction (SPME) is a simple and effective sample-preparation technique for the analysis of complex samples. However, sample matrices containing high concentrations of nonpolar substances or spiked analytes in free form can cause swelling, saturation, and/or competition phenomena in the coating material. This results in a displacement effect wherein polar analytes with low affinities for the solid coating material are displaced by nonpolar substances in the matrix or spiked analytes with a high affinity. Therefore, the quantitative analysis of polar analytes can be challenging, as the displacement effect causes non-linearity in the calibration curves. This paper presents a comprehensive investigation of the conditions under which the displacement effect occurs and how it influences the quantitative analysis of polar analytes. To remedy this issue, a sequential SPME strategy using two SPME blades with different selectivities is applied. SPME blades offer a large surface area and coating volume─and thus, greater extraction capacity─which may mitigate the displacement effect. In addition, the biocompatible coatings on the SPME blades are comprised of small amounts of sorbent particles embedded by a polyacrylonitrile (PAN) binder, which allows them to be directly immersed into complex matrixes such as biological and food samples, as the PAN acts as a barrier that prevents the adsorption of large macromolecules (e.g., cells and proteins). As such, a C18/PAN-coated blade was applied for the first extraction step, which significantly decreased the concentrations of nonpolar compounds in the sample. In the second step, a hydrophilic-lipophilic balanced (HLB)/PAN-coated blade was employed to extract the polar analytes and any remaining nonpolar analytes. The proposed sequential SPME strategy successfully enabled the quantitative determination of polar and nonpolar drugs of abuse with log P values ranging from 0.16 to 4.98 in biological matrices while also providing good linearities.

High-Throughput and Rapid Screening of Drugs of Abuse in Saliva by Multi-Segment Injection Using Solid-Phase Microextraction-Automated Microfluidic Open Interface-Mass Spectrometry.

There is great demand for analytical methods capable of providing high-throughput and rapid screening, especially for anti-doping and clinical point-of-care applications. In this work, automated microfluidic open interface-mass spectrometry (MOI-MS) was used for coupling with high-throughput, automated solid-phase microextraction (SPME) to achieve this objective. The design of the MOI-MS interface provides a continuous and stable electrospray fluid flow to the MS without introducing any bubble, a feature that we exploit to introduce the concept of multi-segment injection for the determination of multiple samples in a single MS run. By eliminating the need to start a new MS run between sample assays, the developed approach provides significantly simplified protocols controlled by programmed software and increased reproducibility. Furthermore, the biocompatible SPME device, which utilizes coating consisting of hydrophilic-lipophilic balanced particles embedded in a polyacrylonitrile (PAN) binder, can be directly used for biological sample analysis, as the PAN acts as both a binder and a matrix-compatible barrier, thus enabling the enrichment of small molecules while eliminating interferences associated with the presence of interfering macromolecules. The above design was employed to develop a fast, quantitative method capable of analyzing drugs of abuse in saliva samples in as little as 75 s per sample. The findings indicate that the developed method provides good analytical performance, with limits of detection ranging between 0.05 and 5 ng/mL for analysis of 16 drugs of abuse, good calibration linear correlation coefficients (R2 ≥ 0.9957), accuracy between 81 and 120%, and excellent precision (RSD% < 13%). Finally, a proof-of-concept experiment was performed to demonstrate the method's suitability for real-time analysis in anti-doping applications.

Standard Water Generating Vials for Lipophilic Compounds.

The study of non-polar compounds in aqueous environments has always been challenging due to their poor solubility in aqueous media. The low affinity of non-polar compounds toward polar solutions facilitates their attachment to glassware, which results in unstable sample concentrations. To address this challenge, and to enable the preparation of a stable mixture of hydrophobic compounds in an aquatic environment, we introduce an in-vial standard water generating system consisting of a vial containing appropriate aqueous solution and a polydimethylsiloxane thin film spiked with target compounds. In this system, a solution with a stable analyte concentration is attained once equilibrium between the thin-film and aqueous solution has been achieved. The developed standard water system was studied using endocannabinoids and phospholipids as model hydrophobic compounds of biological importance, with results indicating that the concentration of hydrophobic compounds in water can remain stable over multiple days. The results also showed that analytes released from the thin film can compensate for analyte loss due to extractions with solid-phase microextraction fibers, thereby re-establishing equilibrium. Thus, the vial is suitable for the repeatable generation of non-polar standards for routine analysis and quality control. The results of this work show that the developed system is stable and reproducible and therefore appropriate for studies requiring the measurement of free concentrations and accurate quantification.

Polybenzimidazole: a novel, fluorocarbon-free, SPME sorbent binder with good thermal and solvent resistance properties for GC and LC analysis.

A novel solid-phase microextraction (SPME) coating is presented that uses polybenzimidazole (PBI) as a binder to immobilize micro-size sorbent particles onto a support. An evaluation of the developed binder's thermal and solvent desorption capabilities demonstrated its compatibility with both gas and liquid chromatography (GC and LC). The incorporation of hydrophilic-lipophilic balanced (HLB) particles provided optimal extraction coverage for an array of chemically diverse analytes possessing a range of hydrophobicities and molecular weights. The developed binder's performance was assessed by comparing it to a selection of binders commonly used in the literature, including polydimethylsiloxane (PDMS) and polyacrylonitrile (PAN), as well as the more recently developed polyvinylidene fluoride (PVDF) and polytetrafluoroethylene amorphous fluoroplastic (PTFE AF 2400). The results revealed that PBI provides better performance compared to PVDF and PTFE AF 2400 in terms of its environmental impact, while also being convenient for use in coating preparation and offering good matrix compatibility. The thermal analysis revealed that PBI exhibited more than 93% weight retention at 550 °C, which is superior to PVDF's 80.07% weight retention at 393.78 °C. To the best of our knowledge, this work is the first to use PBI as a particle binder in SPME coatings. The PBI coating maintained high extraction efficiencies under extreme conditions with pH values of 3 and 12. The performance of PBI in combination with HLB was assessed by employing it to extract several drugs of abuse and McReynolds compounds for LC and GC analysis, respectively. The results indicated that PBI performs similarly to PAN for LC but is outperformed by PDMS in GC applications with respect to extraction and desorption kinetics. Nonetheless, the thermal and solvent desorption results indicated that PBI can be used for both applications, as it remains stable at temperatures over 350 °C and is stable when solvent desorption is applied.

Coated Blade Spray with a Barrier: Improving Negative Electrospray Ionization and Sample Preparation.

Substrate-based electrospray ionization (ESI) techniques like paper, wooden tip, plastic tip, and metal-needle-based spray suffer from corona discharge, high background noise, and unstable spray in negative ionization mode, especially for the analysis of complex biological matrices, such as blood and urine. Coated blade spray coupled with mass spectrometry (CBS-MS) combines solid-phase microextraction's (SPME) efficient sample clean-up and enrichment and ambient MS's fast analysis and has proven to be an appealing alternative tool for the fast screening of target analytes in complex matrices. This paper documents the development of a new CBS blade design that features a barrier at the far end of the ESI tip. The findings of this work show that the addition of this simple barrier enabled the total RSD% to be reduced to less than 10% for sample preparation, ionization, and the MS detection of several drugs of abuse in negative mode, without compensation using internal standards. The improved stability of ESI in negative mode was investigated by observing the ESI process with a microscope camera and testing via CBS-MS. The new design was applied for the analysis of three drugs of abuse in urine, with the calibration curve correlation coefficient (R2 ≥ 0.9997) being calculated without the use of internal standards. The overall RSD% of the peak area for one compound in 42 samples was 6.9%, which highlights the method's incredible reproducibility compared to other ambient MS techniques for analyzing real samples. The CBS device with a barrier was also applied for the on-blade sampling of 14 drugs of abuse in 20 µL of plasma spot in positive ionization mode. The results of these tests yielded a calibration curve correlation coefficient of R2 ≥ 0.9883 and limits of quantification (LOQs) between 0.25 and 25 ng/mL. The obtained results provide guidance on CBS device design optimization and the effective automation of the protocol.

Profiling of Unsaturated Lipids by Raman Spectroscopy Directly on Solid-Phase Microextraction Probes.

Lipids play a critical role in cellular signaling, energy storage, and the construction of cellular membranes. In this paper, we propose a novel on-site approach for detecting and differentiating enriched unsaturated lipids based on the direct coupling of SPME probes with Raman spectroscopy. To this end, different SPME particles, namely, hydrophilic-lipophilic balanced (HLB), mixed-mode (C8-SCX), and C18, were embedded in polyacrylonitrile (PAN) and tested for their efficacy as biocompatible coatings. The C18/PAN coating showed less background interference compared to the other sorbent materials during the analysis of unsaturated lipids. In addition, different SPME parameters that influence extraction efficiency, such as extraction temperature, extraction time, and washing solvent, were also investigated. Our results indicate a clear dependence between the Raman band intensity related to the number of double bonds in fatty acids mixture and the number of double bonds in a fatty acid. Our findings further show that Raman spectroscopy is especially useful for the analysis of lipid unsaturation, which is calculated as the ratio of n(C═C)/n(CH2) using the intensities of the Raman bands at 1655/1445 cm-1. Furthermore, the developed protocol reveals great SPME activity and high detection ability for several unsaturated lipids in different complex matrixes, such as cod liver oil. Finally, the applicability of this technology was demonstrated via the characterization of cod liver oil and other vegetable oils. Thus, the proposed SPME-Raman spectroscopy approach has a great future potential in food, environmental, clinical, and biological applications.

Understanding the effect of spatial positioning of coated blade spray devices relative to the mass spectrometry inlet on different instrument platforms and its application to quantitative analysis of fentanyl and related analogs.

RATIONALE: We evaluated the effect that the spatial positioning of coated-blade spray (CBS) devices with respect to the mass spectrometry (MS) inlet has when coupling to diverse MS platforms (i.e., triple quadrupole, linear ion trap and time of flight). Furthermore, as a proof of concept, we evaluated CBS-MS as a tool for quantitation of fentanyl and four analogues on said instruments. METHODS: Custom-made MS interfaces were made to accurately position the blade in front of the MS inlet. CBS devices, coated with hydrophilic-lipophilic balanced particles, were used for both the optimization of the CBS position and the quantitation of fentanyl and analogues in urine and plasma samples on all instruments. RESULTS: The SCIEX triple quadrupole instrument was the most sensitive to the position of the blade due to the presence of a curtain gas flowing laminarly out of the MS inlet. After optimization, the analytical capabilities of CBS on each instrument were assessed and the results obtained on both SCIEX and Waters platforms matched the performance obtained using a more advanced instrument by ThermoFisher Scientific. Furthermore, excellent figures of merit were attained for the quantitation of fentanyl and analogues on both triple quadrupole and linear ion trap platforms. CONCLUSIONS: We demonstrated that optimization of MS parameters on different instrument vendors and front ends, such as the position of the CBS tip regarding the MS inlet, is vital to exploit the full quantitative potential of this technology. Application of the technology to screen and quantify fentanyl and analogues showed great potential when considering its coupling with portable mass spectrometers for therapeutic drug monitoring and point-of-care applications.

Effect of household air pollutants on the composition of exhaled breath characterized by solid-phase microextraction and needle-trap devices.

Exposure to household air pollutants is becoming a serious environmental health risk. Various methods can be applied to assess humans' exposure status to indoor pollutants, with breath monitoring being among the best options. Breath sampling is fast and non-invasive, and contains compounds that can be used as markers for evaluating exposure length and estimating internal concentrations of pollutants. However, the distribution of compounds between gas and droplets in breath samples represents one of the key challenges associated with this analytical method. In this work, a needle-trap device (NTD) was prepared by packing the needle with a porous filter, divinyl benzene, and Carboxen to enable the exhaustive capture of both droplet-bound and gaseous components. Furthermore, fiber-based solid-phase microextraction (SPME) was also applied to extract compounds from only the gas phase to distinguish this portion of analytes from the total concentration in the sample. Dynamic, real-time breath sampling was enabled via a new sampling tube equipped with 2 one-way valves, which was specially designed for this work. Both methods provided satisfactory reproducibility, repeatability, and sensitivity, with detection limits as low as 0.05 ng mL-1. To investigate the real-world applicability of the proposed devices, breath samples were obtained from volunteers who had been exposed to candle and incense smoke and aerosol sprays, or had smoked cannabis. The results revealed the high concentration of organic air pollutants in inhaled air (maximum of 215 ng mL-1) and exhaled breath (maximum of 14.4 ng mL-1) and a correlation between the components in inhaled air and exhaled breath. Significantly, the findings further revealed that the developed NTD has enhanced breath-sample determinations, especially for polar compounds, which tend to remain trapped in breath droplets.

Coated Blade Spray-Mass Spectrometry as a New Approach for the Rapid Characterization of Brain Tumors.

Brain tumors are neoplasms with one of the highest mortality rates. Therefore, the availability of methods that allow for the quick and effective diagnosis of brain tumors and selection of appropriate treatments is of critical importance for patient outcomes. In this study, coated blade spray-mass spectrometry (CBS-MS), which combines the features of microextraction and fast ionization methods, was applied for the analysis of brain tumors. In this approach, a sword-shaped probe is coated with a sorptive material to enable the extraction of analytes from biological samples. The analytes are then desorbed using only a few microliters of solvent, followed by the insertion of the CBS device into the interface on the mass spectrometer source. The results of this proof-of-concept experiment confirmed that CBS coupled to high-resolution mass spectrometry (HRMS) enables the rapid differentiation of two histologically different lesions: meningiomas and gliomas. Moreover, quantitative CBS-HRMS/MS analysis of carnitine, the endogenous compound, previously identified as a discriminating metabolite, showed good reproducibility with the variation below 10% when using a standard addition calibration strategy and deuterated internal standards for correction. The resultant data show that the proposed CBS-MS technique can be useful for on-site qualitative and quantitative assessments of brain tumor metabolite profiles.

Needle-Trap Device Containing a Filter: A Novel Device for Aerosol Studies.

Particles in an aerosol sample contain a portion of the total available analytes. Therefore, particle trapping is required to fully characterize a gaseous sample. Needle-trap devices (NTDs) are highly useful to this end, as they allow sampling and preconcentration of free analytes, as well as the trapping of particles. Packing sorbents into the needle creates a filter that traps solid particles or liquid droplets. However, the particle-trapping efficiency of sorbent-packed NTDs is limited, especially for nanoparticles. To address this issue, an aerogel based on electrospun polyacrylonitrile (PAN) was prepared for trapping small particles to analyze particle-bound analytes. The PAN aerogel filter was fabricated by cutting electrospun PAN fibers and removing the remaining solvent via freeze-drying to obtain a light porous fibrous structure. The PAN aerogel was heated (H-PAN) prior to packing to ensure stability during thermal desorption. The trapping efficiency of the H-PAN-packed NTD was measured using a range of conditions, with high filtration efficiencies (>99%) being obtained in all cases. The mechanical stability of the H-PAN aerogel was tested using multiple extraction/desorption cycles with and without solid sorbent particles, with results indicating high repeatability (n = 94, relative standard deviation (RSD) <6%). The developed NTD was compared to thin-film microextraction with respect to their ability to characterize breath samples obtained with or without face masks; the NTD was able to trap both free and droplet-bound analytes, while thin-film microextraction was only able to extract free analytes, which is fully reflected in concentrations obtained with these two methods.

The Effect of Sorbent Particles in a Binder on the Mass Transfer Kinetics in Separation Media: In Silico Study and Experimental Verification.

Selecting the optimal binder and the sorbent affinity for selected compounds can cause the composite to behave either as an efficient extraction coating, as a permeable membrane, or as an impermeable barrier. If the compound partitions onto the sorbent with high preference, it becomes stationary and the composite behaves as an impermeable barrier, while appropriately optimized affinity will result in effective permeation. To understand this phenomenon, we utilize solid-phase microextraction to characterize the mass transfer attributes of different separation composites. Our results indicate that for strong sorbents, the extraction rate is primarily controlled by the diffusion in the extraction phase rather than the sample matrix, even if it is relatively thin. Low analyte diffusion is caused by the retarding force generated by the partitioning of analytes into the sorbent, as migration through the composite is driven by the unbound form of the compound in the binder. One of the main contributions of this work is that an understanding of the extraction composite parameters that control mass transfer during extraction enables better optimization of binder/sorbent extraction phase composition for a given application. Another contribution of this work shows how a heterogeneous coating model can be simplified into a homogeneous coating model. The developed models enable an enhanced understanding of mass transfer kinetics, and they provide insight into how to optimize the extraction phase parameters for a given method involving sorbent particles in polymeric media, including membranes and paints, in addition to extraction coatings.

Optimizing a High-Throughput Solid-Phase Microextraction System to Determine the Plasma Protein Binding of Drugs in Human Plasma.

Plasma protein binding refers to the binding of a drug to plasma proteins after entering the body. The measurement of plasma protein binding is essential during drug development and in clinical practice, as it provides a more detailed understanding of the available free concentration of a drug in the blood, which is in turn critical for pharmacokinetics and pharmacodynamics studies. In addition, the accurate determination of the free concentration of a drug in the blood is also highly important for therapeutic drug monitoring and in personalized medicine. The present study uses C18-coated solid-phase microextraction 96-pin devices to determine the free concentrations of a set of drugs in plasma, as well as the plasma protein binding of drugs with a wide range of physicochemical properties. It should be noted that the extracted amounts used to calculate the binding constants and plasma protein bindings should be measured at respective equilibrium for plasma and phosphate buffer. Therefore, special attention is placed on properly determining the equilibration times required to correctly estimate the free concentrations of drugs in the investigated systems. The plasma protein binding values obtained with the 96-pin devices are consistent with those reported in the literature. The 96-pin device used in this research can be easily coupled with a Concept96 or other automated robotic systems to create an automated plasma protein binding determination protocol that is both more time and labor efficient compared to conventional equilibrium dialysis and ultrafiltration methods.

Immuno-Enriched Microspheres - Magnetic Blade Spray-Tandem Mass Spectrometry for Domoic Acid in Mussels.

Paramagnetic microspheres can be used in planar array fluorescence immunoassays for single or multiplex screening of food contaminants. However, no confirmation of the molecular identity is obtained. Coated blade spray (CBS) is a direct ionization mass spectrometry (MS) technique, and when combined with triple quadrupole MS/MS, it allows for rapid confirmation of food contaminants. The lack of chromatography in CBS, though, compromises the specificity of the measurement for unequivocal identification of contaminants, based on the European Union (EU) regulation. Therefore, a rapid and easy-to-use immuno-magnetic blade spray (iMBS) method was developed in which immuno-enriched paramagnetic microspheres replace the coating of CBS. The iMBS-MS/MS method was fully optimized, validated in-house following the EU 2021/808 regulation, and benchmarked against a commercial lateral flow immunoassay (LFIA) for on-site screening of DA. The applicability of iMBS-MS/MS was further demonstrated by analyzing incurred mussel samples. The combination of immunorecognition and MS/MS detection in iMBS-MS/MS enhances the measurement's selectivity, which is demonstrated by the rapid differentiation between the marine toxin domoic acid (DA) and its structural analog kainic acid (KA), which cannot be achieved with the LFIA alone. Interestingly, this first-ever reported iMBS-MS/MS method is generic and can be adapted to include any other immuno-captured food contaminant, provided that monoclonal antibodies are available, thus offering a complementary confirmatory analysis approach to multiplex immunoassay screening methods. Moreover, thanks to its speed of analysis, iMBS-MS/MS can bridge the logistics gap between future large-scale on-site testings using LFIAs and classical time-consuming confirmatory MS analysis performed in official control laboratories.