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.

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.

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.

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.

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.

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.

Automated sequential SPME addressing the displacement effect in food samples.

The displacement effect can be an issue for the quantitation of analytes with low affinity towards the extraction phase in solid-phase microextraction (SPME) for food samples that have low level of binding matrix or high level of hydrophobic compounds. In this communication, automated sequential SPME-GC-MS strategy was developed for addressing the displacement issue. The SPME thin film with PDMS coating was firstly used for the extraction of hydrophobic components in the sample which cause displacement and then SPME fiber with DVB/CAR/PDMS coating was applied in the second step for the extraction of the remain compounds. This new strategy was investigated by using 10 key food odorants as target analytes and tested in commercial beer samples. The results suggested that sequential SPME can decrease the displacement effect and improve the extraction efficiency for polar analytes.

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.

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%).

In vivo solid phase microextraction for therapeutic monitoring and pharmacometabolomic fingerprinting of lung during in vivo lung perfusion of FOLFOX.

In vivo lung perfusion (IVLP) is a novel isolated lung technique developed to enable the local, in situ administration of high-dose chemotherapy to treat metastatic lung cancer. Combination therapy using folinic acid (FOL), 5-fluorouracil (F), and oxaliplatin (OX) (FOLFOX) is routinely employed to treat several types of solid tumours in various tissues. However, F is characterized by large interpatient variability with respect to plasma concentration, which necessitates close monitoring during treatments using of this compound. Since plasma drug concentrations often do not reflect tissue drug concentrations, it is essential to utilize sample-preparation methods specifically suited to monitoring drug levels in target organs. In this work, in vivo solid-phase microextraction (in vivo SPME) is proposed as an effective tool for quantitative therapeutic drug monitoring of FOLFOX in porcine lungs during pre-clinical IVLP and intravenous (IV) trials. The concomitant extraction of other endogenous and exogenous small molecules from the lung and their detection via liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) enabled an assessment of FOLFOX's impact on the metabolomic profile of the lung and revealed the metabolic pathways associated with the route of administration (IVLP vs. IV) and the therapy itself. This study also shows that the immediate instrumental analysis of metabolomic samples is ideal, as long-term storage at -80 °C results in changes in the metabolite content in the sample extracts.

Determination of free concentration of endocannabinoids in brain tissue.

The release of metabolites from their bound to free forms is the main regulatory path in living species. Therefore, the ability to determine the free concentrations of small molecules is highly critical in many biological samples. The main challenges in achieving this task are the interferences inherent to complex matrices and the ability to distinguish between the free and total concentrations. This paper presents a non-invasive microextraction method that enables the determination of endocannabinoids in brain tissue. The proposed method is based on two key principles: the availability of the free concentration of endocannabinoids for partitioning to the solid-phase microextraction (SPME) fiber; and negligible depletion enabled by the small volume of extraction phase on the fiber. These features allow the presented SPME method to provide information about the free concentration of analytes without disturbing the binding equilibrium between the analytes and the matrix. The determination of spiked samples with known concentrations enables the percentage of analyte bound to the tissue to be calculated, which can then be applied to calculate the total concentration from the determined free concentration. This manuscript focuses on the determination of the free concentration and tissue binding percentages of endocannabinoids in brain tissue. Significantly, SPME's small size and potential for non-invasive sampling enable its application in live animal subjects with minimal tissue damage.

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.

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.

High throughput and automated solid-phase microextraction and determination by liquid chromatography-mass spectrometry for the analysis of mycotoxins in beer.

There is high demand for rapid screening of toxics in food analysis. In this study, a new high-throughput and automated solid-phase microextraction (SPME) system was employed for the sample preparation of mycotoxins in beers. Matrix compatible SPME blades with thin coating layer were used, which significantly decreased the matrix effects in beer samples (≤ 12%). This SPME system allows 96 samples to be processed automatically and simultaneously with average preparation time of 57 s per sample. After sample preparation, the 96-well plate with desorption solution was sealed with a thin film and put into the LC-MS sampler for analysis via positive/negative ESI switching mode. The results also showed good sensitivity (limits of detection between 0.02 and 3 ng/mL) with R2≥ 0.9971, reproducibility (intra- and inter-day ≤ 8% and ≤ 13%, respectively), and accuracy (recoveries between 79% and 121%).

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.

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.

Coupling of Solid-Phase Microextraction Directly to Mass Spectrometry via an Improved Microfluidic Open Interface to Facilitate High-Throughput Determinations.

Mass spectrometry analysis can be performed by introducing samples directly to mass spectrometry, allowing the increase of the analysis throughput; however, some disadvantages of direct-to-mass spectrometry analysis include susceptibility to matrix effects and risk of instrument contamination from inadequate sample preparation. Solid-phase microextraction is one of the most suitable sample preparation methods for direct-to-mass spectrometry analysis, as it offers matrix-compatible coatings which ensure analyte enrichment with minimal or no interference from matrix. One of the ways solid-phase microextraction can be coupled directly to mass spectrometry is via a microfluidic open interface. This manuscript reports improvements made to the initial microfluidic open interface design, where the system components have been simplified to mostly commercially available materials. In addition, the analysis of samples has been automated by implementing software that fully controls the analysis workflow, where the washing procedure is optimized to completely reduce the carryover. Herein, the extraction and desorption time profiles from thin and thick SPME devices was studied where the overall workflow consisted of high-throughput sample preparation of 1.3 min per 96 samples and <1 min per sample instrumental analysis.

Comparison of different approaches for direct coupling of solid-phase microextraction to mass spectrometry for drugs of abuse analysis in plasma.

The direct coupling of solid-phase microextraction (SPME) to mass spectrometry (MS) (SPME-MS) has proven to be an effective method for the fast screening and quantitative analysis of compounds in complex matrices such as blood and plasma. In recent years, our lab has developed three novel SPME-MS techniques: SPME-microfluidic open interface-MS (SPME-MOI-MS), coated blade spray-MS (CBS-MS), and SPME-probe electrospray ionization-MS (SPME-PESI-MS). The fast and high-throughput nature of these SPME-MS technologies makes them attractive options for point-of-care analysis and anti-doping testing. However, all these three techniques utilize different SPME geometries and were tested with different MS instruments. Lack of comparative data makes it difficult to determine which of these methodologies is the best option for any given application. This work fills this gap by making a comprehensive comparison of these three technologies with different SPME devices including SPME fibers, CBS blades, and SPME-PESI probes and SPME-liquid chromatography-MS (SPME-LC-MS) for the analysis of drugs of abuse using the same MS instrument. Furthermore, for the first time, we developed different desorption chambers for MOI-MS for coupling with SPME fibers, CBS blades, and SPME-PESI probes, thus illustrating the universality of this approach. In total, eight analytical methods were developed, with the experimental data showing that all the SPME-based methods provided good analytical performance with R 2 of linearities larger than 0.9925, accuracies between 81% and 118%, and good precision with an RSD% ≤ 13%.

Solid-phase microextraction of endogenous metabolites from intact tissue validated using a Biocrates standard reference method kit.

Improved analytical methods for the metabolomic profiling of tissue samples are constantly needed. Currently, conventional sample preparation methods often involve tissue biopsy and/or homogenization, which disrupts the endogenous metabolome. In this study, solid-phase microextraction (SPME) fibers were used to monitor changes in endogenous compounds in homogenized and intact ovine lung tissue. Following SPME, a Biocrates AbsoluteIDQ assay was applied to make a downstream targeted metabolomics analysis and confirm the advantages of in vivo SPME metabolomics. The AbsoluteIDQ kit enabled the targeted analysis of over 100 metabolites via solid-liquid extraction and SPME. Statistical analysis revealed significant differences between conventional liquid extractions from homogenized tissue and SPME results for both homogenized and intact tissue samples. In addition, principal component analysis revealed separated clustering among all the three sample groups, indicating changes in the metabolome due to tissue homogenization and the chosen sample preparation method. Furthermore, clear differences in free metabolites were observed when extractions were performed on the intact and homogenized tissue using identical SPME procedures. Specifically, a direct comparison showed that 47 statistically distinct metabolites were detected between the homogenized and intact lung tissue samples (P < 0.05) using mixed-mode SPME fibers. These changes were probably due to the disruptive homogenization of the tissue. This study's findings highlight both the importance of sample preparation in tissue-based metabolomics studies and SPME's unique ability to perform minimally invasive extractions without tissue biopsy or homogenization while providing broad metabolite coverage.