Simultaneous quantitation of urinary albumin and creatinine for rapid clinical albuminuria diagnostics using high-throughput paper spray mass spectrometry.

Albuminuria is a clinical condition associated with poor kidney function, diagnosed by determining the ratio of albumin to creatinine concentrations in patient urine samples. We present a high-throughput paper spray mass spectrometry (PS-MS) method for simultaneous quantitation of urinary albumin and creatinine for potential diagnosis of albuminuria. Minimal (urine dilution) or no sample preparation is required. The analytical performance of the method was evaluated, achieving linear calibration curves (R2 > 0.99) with little inter-day variability in the slope (N = 5 days), exhibiting coefficient of variation (CV) of 8% and 3% for albumin and creatinine, respectively. LOD and LOQ for albumin were 2.1 and 7.0 mg L-1, and for creatinine were 0.01 and 0.03 mmol L-1, respectively. Intra- and inter-day (N = 5) precisions (%CV) and accuracies (%bias) were <10% and ±11%, respectively, for both analytes. The method was applied to determine albumin-to-creatinine ratios in anonymous human patient urine samples (N = 56), and a correlation of R2 = 0.9744 was achieved between the PS-MS results and validated clinical method results. This work demonstrates the utility of PS-MS to simultaneously quantify a large (albumin) and a small (creatinine) molecule directly in patient urine samples, and its potential as a tool for clinical albuminuria diagnostics.

Paper spray mass spectrometry combined with machine learning as a rapid diagnostic for chronic kidney disease.

A new analytical method for chronic kidney disease (CKD) detection utilizing paper spray mass spectrometry (PS-MS) combined with machine learning is presented. The analytical protocol is rapid and simple, based on metabolic profile alterations in urine. Anonymized raw urine samples were deposited (10 µL each) onto pointed PS-MS sample strips. Without waiting for the sample to dry, 75 µL of acetonitrile and high voltage were applied to the strips, using high resolution mass spectrometry measurement (15 s per sample) with polarity switching to detect a wide range of metabolites. Random forest machine learning was used to classify the resulting data. The diagnostic performance for the potential diagnosis of CKD was evaluated for accuracy, sensitivity, and specificity, achieving results >96% for the training data and >91% for validation and test data sets. Metabolites selected by the classification model as up- or down-regulated in healthy or CKD samples were tentatively identified and in agreement with previously reported literature. The potential utilization of this approach to discriminate albuminuria categories (normo, micro, and macroalbuminuria) was also demonstrated. This study indicates that PS-MS combined with machine learning has the potential to be used as a rapid and simple diagnostic tool for CKD.

PAMAM-Functionalized Paper as a New Substrate for the Paper Spray Mass Spectrometry Measurement of Proteins.

Paper surface functionalization with polyamidoamine (PAMAM) dendrimers has been developed for increased sensitivity analysis of proteins by paper spray mass spectrometry (PS-MS). PAMAM is a branched polymeric compound with an ethylenediamine core linked to repeating PAMAM units that generates an outer surface rich in primary amines. These positively charged amine groups can interact electrostatically with negatively charged residues (e.g., aspartate, glutamate) on the protein surface. PAMAM inner amide moieties can also promote hydrogen bonding with protein surface oxygens, making PAMAM a useful material for protein extraction. PAMAM-functionalized PS-MS paper strips were used to extract proteins from biofluids, dipped in acetonitrile to remove unbound constituents, dried, and then measured with PS-MS. The use of this strategy was optimized and compared with unmodified paper strips. PAMAM-functionalized paper substrates provided sixfold greater sensitivity for albumin, 11-fold for hemoglobin, sevenfold for insulin, and twofold for lysozyme. The analytical performance of the functionalized paper substrate was evaluated through the analysis of albumin in urine, achieving linearity with R2 > 0.99, LOD of 1.1 µg mL-1, LOQ of 3.8 µg mL-1, precision better than 10%, and relative recovery 70-83%. The method was applied to quantify urinary albumin from nine anonymous patient samples (concentrations ranged from 6.5 to 77.4 µg mL-1), illustrating its potential for the diagnosis of microalbuminuria. These data demonstrate the utility of paper modification with the PAMAM dendrimer for sensitive PS-MS analysis of proteins, opening a path for further applications in clinical diagnosis through the analysis of disease-related proteins.

A passive membrane system for on-line mass spectrometry reagent addition.

RATIONALE: Post-separation addition of chemical modifiers in liquid chromatography-mass spectrometry is widely used for improving ionization sensitivity and selectivity. This is typically accomplished using a post-column T-junction, which can result in sample dilution and imperfect mixing. We present a passive semi-permeable hollow fiber membrane approach for the addition of chemical modifiers that avoids these issues. METHODS: Model compounds were directly infused by flow injection to an electrospray ionization triple quadrupole mass spectrometer after passing through a polydimethylsiloxane hollow fiber membrane. Ionization enhancement reagents were introduced into the flowing stream by membrane permeation from aqueous solutions. Ionization enhancement from volatile acids and bases in positive and negative electrospray ionization was evaluated to assess the feasibility of this approach. RESULTS: The membrane-based apparatus resulted in relative ionization enhancement factors of up to 14×, depending upon the analyte, reagent, and ionization mode used. Ionization enhancement signal stability is reasonable (relative standard deviation of 5-7%) for extended periods from the same reagent solution, and minimal analyte dilution is observed. A proof-of-concept demonstration of the chromatographic "trifluoroacetic acid fix" strategy is presented. CONCLUSIONS: An on-line mass spectrometry ionization reagent addition method with potential post-chromatography reagent addition applications was developed using a hollow fiber polydimethylsiloxane membrane. This approach offers a promising alternative to traditional methods requiring additional hardware such as pumps and T-junctions that can result in sample dilution and imperfect reagent mixing.

A direct mass spectrometry method for cannabinoid quantitation in urine and oral fluid utilizing reactive paper spray ionization.

A direct mass spectrometry method utilizing reactive paper spray ionization was developed for sensitive cannabinoid quantitation in biofluid matrices. The ca. 2-minute sample measurements used on-paper derivatization to significantly increase paper spray mass spectrometry (PS-MS) positive ion mode sensitivity while minimizing sample preparation steps. Calibrations demonstrate high linearity, with R2 > 0.99 for (-)-trans-Δ9-tetrahydrocannabinol (THC) in oral fluid and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in urine. The limit of detection and lower limit of quantitation were 0.78 and 10 ng mL-1 for THC in oral fluid and 1.3 and 10 ng mL-1 for THC-COOH in urine, respectively. THC-COOH levels measured by reactive PS-MS in seven spiked human urine samples showed bias of -9.4 to 5.9%, and percent difference values of -16.8 to 9.8% in comparison with a reference LC-MS method. Based upon the method simplicity, validation experiments, sensitivity, and rapidity, we conclude that reactive PS-MS has potential applications for rapid cannabinoid drug testing in urine and oral fluid.

Membrane Sampling Separates Naphthenic Acids from Biogenic Dissolved Organic Matter for Direct Analysis by Mass Spectrometry.

Oil sands process waters can release toxic naphthenic acids (NAs) into aquatic environments. Analytical techniques for NAs are challenged by sample complexity and interference from naturally occurring dissolved organic matter (DOM). Herein, we report the use of a poly(dimethylsiloxane) (PDMS) polymer membrane for the on-line separation of NAs from DOM and use direct infusion electrospray ionization mass spectrometry to yield meaningful qualitative and quantitative information with minimal sample cleanup. We compare the composition of membrane-permeable species from natural waters fortified with a commercial NA mixture to those derived from weak anion exchange solid-phase extraction (SPE) using high-resolution mass spectrometry. The results show that SPE retains a wide range of carboxylic acids, including biogenic DOM, while permeation through PDMS was selective for petrogenic classically defined NAs (CnH2n+zO2). A series of model compounds (log Kow ∼1-7) were used to characterize the perm-selectivity and reveal the separation is based on hydrophobicity. This convenient sample cleanup method is selective for the O2 class of NAs and can be used prior to conventional analysis or as an on-line analytical strategy when coupled directly to mass spectrometry.

MASS SPECTROMETRY ANALYSIS OF DRUGS OF ABUSE: CHALLENGES AND EMERGING STRATEGIES.

Mass spectrometry has been the "gold standard" for drugs of abuse (DoA) analysis for many decades because of the selectivity and sensitivity it affords. Recent progress in all aspects of mass spectrometry has seen significant developments in the field of DoA analysis. Mass spectrometry is particularly well suited to address the rapidly proliferating number of very high potency, novel psychoactive substances that are causing an alarming number of fatalities worldwide. This review surveys advancements in the areas of sample preparation, gas and liquid chromatography-mass spectrometry, as well as the rapidly emerging field of ambient ionization mass spectrometry. We have predominantly targeted literature progress over the past ten years and present our outlook for the future. © 2020 Periodicals, Inc. Mass Spec Rev.

Direct, Isomer-Specific Quantitation of Polycyclic Aromatic Hydrocarbons in Soils Using Membrane Introduction Mass Spectrometry and Chemical Ionization.

Polycyclic aromatic hydrocarbons (PAHs) are routinely screened for in soils, where quantitation of structural isomers is critical due to varying toxicity within PAH isomer classes. While chromatographic methods provide isomer resolution, such strategies are cost and time intensive. To address these challenges, we present condensed phase membrane introduction mass spectrometry using liquid electron ionization/chemical ionization (CP-MIMS-LEI/CI) as a direct mass spectrometry technique that provides rapid, quantitative results for PAH isomer measurements in soil samples. A methanol acceptor phase is flowed through a probe-mounted polydimethylsiloxane hollow fiber membrane directly immersed into a dichloromethane/soil slurry. PAHs and dichloromethane co-permeate the membrane into the acceptor solvent, whereas particulates and charged matrix components remain in the sample. A nanoflow of the membrane permeate is then directly infused into a LEI/CI interfaced triple quadrupole mass spectrometer. Diagnostic PAH adduct ions were formed at either M + 45 ([M + CH2Cl + CH3OH-HCl]+) or M + 47 ([M + CHCl2-HCl]+). This allowed the development of specific MS/MS transitions for individual PAH isomers. These transitions were subsequently used for the direct analyses of PAHs in real soils where CP-MIMS-LEI/CI was shown to be rapid (15 soil samples/h) and sensitive (ng/g level detection limits). CP-MIMS-LEI/CI results compared well to those obtained using GC-MS (average percent difference of -9% across 9 PAHs in 8 soil samples), presenting a compelling argument for direct, quantitative screening of PAHs in soils by CP-MIMS-LEI/CI, particularly given the simple workflow and short analytical duty cycle.

Direct Analysis of Polyaromatic Hydrocarbons in Soil and Aqueous Samples Using Condensed Phase Membrane Introduction Tandem Mass Spectrometry with Low-Energy Liquid Electron Ionization.

Polyaromatic hydrocarbons (PAHs) are listed as priority pollutants by the United States Environmental Protection Agency (U.S. EPA). PAH-contaminated samples often require extensive sample cleanup before analysis, with the method used dependent upon the sample matrix. We present condensed phase membrane introduction mass spectrometry with liquid electron ionization (CP-MIMS-LEI) as a sensitive and universal technique that can directly analyze both aqueous and soil samples for PAHs without the need for sample clean up or instrumental modifications for different matrixes. The method uses a semipermeable hollow fiber membrane immersion probe to transfer analytes from complex samples into a solvent acceptor phase that is directly entrained at nanoliter/min flows to an LEI-interfaced mass spectrometer. The resulting aerosol is desolvated under vacuum leading to analyte vaporization and subsequent electron ionization. Electron energy and LEI vaporization capillary position were examined and optimized for PAHs. The CP-MIMS probe was directly immersed in complex aqueous matrixes, demonstrating low nanogram per liter PAH detection limits and response times of ≤1.6 min. For soil sample analysis, 2-propanol was found to be the optimal PAH sampling solvent. Soil samples were briefly sonicated in 2-propanol, followed by direct CP-MIMS measurement. Soil sample throughput was ca. 15 samples/h, with PAH quantitation successful at microgram per kilogram levels. The workflow is remarkably simple, fast, green, and leads to reproducible results that enable high-throughput screening of heterogeneous soil samples.

Mass Spectrometry Based Approach for Organic Synthesis Monitoring.

Current mass spectrometry-based methodologies for synthetic organic reaction monitoring largely use electrospray ionization (ESI), or other related atmospheric pressure ionization-based approaches. Monitoring of complex, heterogeneous systems may be problematic because of sampling hardware limitations, and many relevant analytes (neutrals) exhibit poor ESI performance. An alternative monitoring strategy addressing this significant impasse is condensed phase membrane introduction mass spectrometry using liquid electron ionization (CP-MIMS-LEI). In CP-MIMS, a semipermeable silicone membrane selects hydrophobic neutral analytes, rejecting particulates and charged chemical components. Analytes partition through the membrane, and are then transported to the LEI interface for sequential nebulization, vaporization, and ionization. CP-MIMS and LEI are both ideal for continuous monitoring applications of hydrophobic neutral molecules. We demonstrate quantitative reaction monitoring of harsh, complex reaction mixtures (alkaline, acidic, heterogeneous) in protic and aprotic organic solvents. Also presented are solvent-membrane compatibility investigations and, in situ quantitative monitoring of catalytic oxidation and alkylation reactions.

Direct quantitation and characterization of fatty acids in salmon tissue by condensed phase membrane introduction mass spectrometry (CP-MIMS) using a modified donor phase.

Existing mass spectrometric methods for the analysis of fatty acids often require derivatization, chromatographic separations, and/or extensive sample preparation. Direct mass spectrometry strategies can avoid these requirements, but may also suffer from poor quantitation and/or lack of sensitivity. Condensed phase-membrane introduction mass spectrometry (CP-MIMS) provides direct quantitative measurements of analytes in complex samples with little or no sample preparation. CP-MIMS uses a semipermeable membrane to transfer neutral, hydrophobic compounds from real-world samples to a mass spectrometer. The results presented utilize aqueous/organic sample solvent (donor) mixtures to allow for the sensitive (pptr) detection of a range of fatty acids. The relative sensitivity across a homologous series of fatty acids is observed to change, favoring short- or long-chain fatty acids, depending on the amount of miscible co-solvent added to the donor phase. Further, lithium acetate added online via the acceptor phase was used in tandem mass spectrometry experiments to determine the location of double bonds in polyunsaturated fatty acids (PUFAs). The method was applied to direct measurements and structural determinations for selected PUFAs in salmon tissue samples. Standard addition was employed to quantify the amount of PUFAs in a variety of salmon samples, yielding 0.27-0.42 and 0.40-0.84 w/w % for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), respectively, for Sockeye and Chinook salmon, in good agreement with the literature. This work presents, to our knowledge, the first use of CP-MIMS for the direct analysis of fatty acids in oily foodstuff samples. Graphical abstract ᅟ.

Rapid analysis of fentanyls and other novel psychoactive substances in substance use disorder patient urine using paper spray mass spectrometry.

RATIONALE: Drug overdose deaths due to fentanyls and other novel psychoactive substances (NPS) are on the rise. The higher potencies of fentanyl analogs compared with morphine require new technologies to identify and quantitate NPS. METHODS: Paper spray tandem mass spectrometry (MS/MS) and high-resolution mass spectrometry were used to identify and measure fentanyl analogs as well as common drugs of abuse in urine samples from substance use disorder clinics. Ten-microliter urine samples were deposited directly on paper spray cartridges previously loaded with internal standards, dried, and analyzed with no other sample treatment. Quantitative results were obtained using MS/MS. Individual drugs were identified using high-resolution accurate mass spectrometry, and confirmed by data-dependent MS/MS. RESULTS: Calibration curves in urine were linear over a range of 0.5-50 ng/mL with R2 of 0.99 or better for eight representative fentanyl analogs. Cartridges preloaded with internal standards demonstrated satisfactory quantitative results compared with LC/MS. Direct identification and confirmation of fentanyl analogs and other common drugs of abuse in urine using high-resolution accurate mass and MS/MS fragmentation were demonstrated at low picogram levels. CONCLUSIONS: Paper spray mass spectrometry can reliably identify and quantitate fentanyl analogs and other drugs of abuse in urine. Using paper spray cartridges as collection devices reduces exposure and transportation risks associated with biological fluids. Cartridges preloaded with labeled internal standards can be effective for targeted screening of fentanyl analogs and other drugs of abuse.

Online measurement of phthalate-particulate matter interactions by membrane introduction mass spectrometry (MIMS).

To enable further study and assessment of indoor inhalation exposure risk, an online apparatus enabling measurement of semi-volatile compound partitioning on household particulates was developed. An example for use of the apparatus is described using dimethyl phthalate (DMP). The system employs direct measurement by membrane introduction mass spectrometry (MIMS). The MIMS system was calibrated using known gas phase DMP concentrations produced by gravimetrically calibrated permeation devices. The quantity of DMP sorbed by particles is described first using a model particle type, a reverse-phase liquid chromatography packing material, and then with a household dust sample. In addition, the desorption of semi-volatile compounds from a household dust sample was monitored using the apparatus, and characteristic fragment ion signals for phthalate compounds were observed.

Polymer Inclusion Membranes with Condensed Phase Membrane Introduction Mass Spectrometry (CP-MIMS): Improved Analytical Response Time and Sensitivity.

Condensed phase membrane introduction mass spectrometry (CP-MIMS) is an online, in situ analysis technique for low volatility analytes. Analytes diffuse through a hollow fiber membrane, where they are then dissolved by a liquid (condensed) acceptor phase flowing through the membrane lumen. Permeating analytes are entrained to an atmospheric pressure ionization source for subsequent measurement by a mass spectrometer. Larger analytes, with inherently lower diffusivities, suffer from lengthy response times and lower sensitivity, limiting the use of CP-MIMS for their online, real-time measurement. We present the use of a heptane cosolvent in a methanol acceptor phase in combination with a polydimethylsiloxane (PDMS) membrane. The heptane generates an in situ polymer inclusion membrane (PIM) with the PDMS. We report improved measurement response times and greater sensitivity across a suite of analytes studied (gemfibrozil, nonylphenol, triclosan, 2,4,6-trichlorophenol, and naphthenic acids), with detection limits in the low parts per trillion (ppt) range. These improvements are attributed to increasing analyte diffusivities, as well as increased analyte partitioning across the PIM. Response times are ∼3× faster for the larger analytes studied, and calibration sensitivity is improved by up to ∼3.5× using 0.046 mole fraction heptane in the methanol acceptor. We report the use of short sample exposure times and the use of non-steady-state signals to reduce the analytical duty cycle, and illustrate that the use of a PIM provides a simple and robust variant of CP-MIMS amenable to rapid screening of analytes in complex samples.

A membrane introduction mass spectrometer utilizing ion-molecule reactions for the on-line speciation and quantitation of volatile organic molecules.

RATIONALE: The ability of membrane introduction mass spectrometry to quantitatively resolve low molecular weight volatile organic compounds (VOCs) such as benzene, toluene, ethylbenzene and xylene (BTEX) using electron ionization (EI) can be compromised by isobaric interferences. This work focuses on reducing isobaric interferences with ion-molecule reactions in a portable quadrupole ion trap mass spectrometer for the analysis of VOCs. METHODS: EI was used to produce reagent ions from precursors (chloroform, methyl iodide, trichloroethylene or chlorobenzene) that were continually infused into the helium acceptor phase upstream of the membrane introduction mass spectrometry (MIMS) sampling interface. The reagent ions were selectively stored in the ion trap, and then allowed to react with target VOC analytes in air samples via ion-molecule reactions within the trap storage volume. A variety of reaction times were examined (50-5000 ms), and the resulting product ions were analyzed in positive ion mode. RESULTS: The detection limits achieved were comparable with those obtained using EI (low ppbv), and in some cases better than for EI coupled with tandem mass spectrometry (MS/MS). For the VOCs studied, isobaric interferences were greatly reduced or eliminated using chloroform as a reagent gas. The predominant ionization mechanism was via adduct formation, although charge transfer and hydride abstractions were also observed. An internal standard was shown to be effective at correcting for signal changes due to consumption of reagent ions when complex mixtures were sampled. CONCLUSIONS: Ion-molecule reactions were exploited to eliminate isobaric interferences that are often encountered in direct, real-time analysis strategies for atmospheric VOC mixtures. The use of a continuously infused internal standard will improve quantitative results in field applications where analyte concentration and sample complexity may be wide ranging.

Photosensitized degradation kinetics of trace halogenated contaminants in natural waters using membrane introduction mass spectrometry as an in situ reaction monitor.

The photochemically mediated dechlorination of polyhalogenated compounds represents a potential decontamination strategy and a relevant environmental process in chemically reducing media. We report the UV irradiation of natural and artificial waters containing natural dissolved organic matter to effect the photo-sensitized degradation of chlorinated organic compounds, including tetrachloromethane, 1,1,1-tricloroethane, perchloroethene, 1,2-dibromo-3-chloropropane and chlorobenzene at trace (ppb) levels in aqueous solution. The degradation kinetics are followed in situ using membrane introduction mass spectrometry. By re-circulating the reaction mixture in a closed loop configuration over a semi-permeable hollow fiber polydimethylsiloxane membrane in a flow cell interface, volatile and semi-volatile compounds are continuously monitored using a quadrupole ion trap mass spectrometer. The time resolved quantitative information provides useful mechanistic insights, including kinetic data. Pseudo first-order rate constants for the degradation of contaminant mixtures in natural waters are reported.

Delicate polydimethylsiloxane hollow fibre membrane interfaces for condensed phase membrane introduction mass spectrometry (CP-MIMS).

RATIONALE: On-line analytical techniques such as condensed phase membrane introduction mass spectrometry (CP-MIMS) permit direct and rapid analyte measurements in complex samples. Direct, rapid analytical methods are desirable because they eliminate potential contamination and/or dilution from sample workup steps, facilitate rapid sample screening and allow 'real-time' monitoring applications. METHODS: PDMS hollow fibre membrane (HFM) flow cell interfaces (215 µm, 35 µm, and 0.5 µm thick composite) were coupled with an electrospray ionization (ESI) triple quadrupole mass spectrometer. A simultaneous push/pull methanol acceptor phase delivery system and membrane mounting via epoxy potting ensured that the delicate membranes were not ruptured during construction or sample measurements. Both flow cell and direct insertion 'J-Probe' interfaces using the 0.5 µm thick composite PDMS HFM were utilized for direct naphthenic acid measurements. RESULTS: Delicate HFM CP-MIMS interfaces were used for the rapid screening and continuous, on-line monitoring of carboxylic acids and hydroxylated compounds directly in complex sample matrices under ambient conditions at pptr - ppb detection limits. Push/pull acceptor phase (methanol) delivery maintained ambient hydrostatic pressures within the HFMs, improving ESI stability and analytical sensitivity, especially with stopped acceptor flow operation. Signal response times less than 2 min were achieved for thin, composite PDMS HFMs at 30°C. The continuous monitoring of naphthenic acid degradation was demonstrated. CONCLUSIONS: Delicate PDMS HFM CP-MIMS interfaces were developed and used for the direct, on-line detection of low volatility, polar analytes in complex aqueous samples. Composite PDMS HFM interfaces yielded the best overall analytical performance improvements, and were used to demonstrate the direct measurement of naphthenic acids in complex aqueous samples.

Monitoring microplastic-contaminant sorption processes in real-time using membrane introduction mass spectrometry.

Microplastics are environmentally ubiquitous and their role in the fate and distribution of trace contaminants is of emerging concern. We report the first use of membrane introduction mass spectrometry to directly monitor the rate and extent of microplastic-contaminant sorption. Target contaminant (naphthalene, anthracene, pyrene, and nonylphenol) sorption behaviours were examined at nanomolar concentrations with four plastic types: low density polyethylene (LDPE), high density polyethylene (HDPE), polypropylene (PP), and polystyrene (PS). Under the conditions employed here, short-term sorption kinetics were assessed using on-line mass spectrometry for up to one hour. Subsequent sorption was followed by periodically measuring contaminant concentrations for up to three weeks. Short-term sorption followed first order kinetics with rate constants that scaled with hydrophobicity for the homologous series of polycyclic aromatic hydrocarbons (PAHs). Sorption rate constants on LDPE for equimolar solutions of naphthalene, anthracene, and pyrene were 0.5, 2.0, and 2.2 h-1, respectively, while nonylphenol did not sorb to pristine plastics over this time period. Similar trends among contaminants were observed for other pristine plastics with 4- to 10-fold faster sorption rates associated with LDPE when compared to PS and PP. Sorption was largely complete after three weeks, with the percent analyte sorbed ranging from 40-100% across various microplastic-contaminant combinations. Photo-oxidative ageing of LDPE had little effect on PAH sorption. However, a marked increase in nonylphenol sorption was consistent with increased hydrogen-bonding interactions. This work provides kinetic insights into surface interactions and describes a powerful experimental platform to directly observe contaminant sorption behaviours in complex samples under a variety of environmentally relevant conditions.

Carfentanil structural analogs found in street drugs by paper spray mass spectrometry and their characterization by high-resolution mass spectrometry.

Carfentanil is one of the most potent synthetic opioids ever developed, with an estimated analgesic potency approximately 20-100 times that of fentanyl and 10,000 times that of morphine. Carfentanil has been appearing in the illicit drug supply in many regions and has been linked to fatal overdose events. A subset of 59 street drug samples obtained in Victoria, B.C., that were confirmed to contain carfentanil were analyzed by mass spectrometry for this study. Carfentanil quantitation by paper spray mass spectrometry ranged from 0.05 to 2.95 w/w% (median = 0.32%) in the original drug sample. Paper spray mass spectrometry analysis also detected two unknown peaks at m/z 380.2 and 381.2 in 31 of these 59 samples (53%). Initial tandem mass spectrometry experiments revealed structural similarities between these unknown compounds and carfentanil, suggesting they were potential structural analogs, possibly arising from incomplete purification during synthesis. High-resolution mass spectrometry determined the chemical formulas of these compounds as C23 H29 N3 O2 (m/z 380.2333) and C23 H29 N2 O3 (m/z 381.2137). Literature and tandem mass spectrometry results were used to determine the identity of these potential new psychoactive substances, C23 H29 N3 O2 as desmethylcarfentanil amide and C23 H29 N2 O3 as desmethylcarfentanil acid. µ-Opioid receptor binding modeling determined that the binding poses of these analogs were nearly identical to that of carfentanil with relative binding energy calculations of 0.544 kJ/mol (desmethylcarfentanil amide) and -0.171 kJ/mol (desmethylcarfentanil acid); these data suggest they may share the toxic effects of carfentanil and have similar potencies.

Automated, High-Throughput Analysis of Tire-Derived p-Phenylenediamine Quinones (PPDQs) in Water by Online Membrane Sampling Coupled to MS/MS.

The tire-derived contaminant N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine quinone (6-PPDQ) was recently identified as a potent toxin to coho salmon (Oncorhynchus kisutch). Studies investigating 6-PPDQ have employed solid-phase extraction (SPE) or liquid-liquid extraction (LLE) with liquid chromatography-mass spectrometry (LC-MS), providing excellent sensitivity and selectivity. However, cleanup and pre-enrichment steps (SPE/LLE) followed by chromatographic separation can be time- and cost-intensive, limiting sample throughput. The ubiquitous distribution of 6-PPDQ necessitates numerous measurements to identify hotspots for targeted mitigation. We recently developed condensed phase membrane introduction mass spectrometry (CP-MIMS) for rapid 6-PPDQ analysis (2.5 min/sample), with a simple workflow and low limit of detection (8 ng/L). Here, we describe improved quantitation using isotopically labeled internal standards and inclusion of a suite of PPDQ analogues. A low-cost autosampler and data processing software were developed from a three-dimensional (3D) printer and Matlab to fully realize the high-throughput capabilities of CP-MIMS. Cross-validation with a commercial LC-MS method for 10 surface waters provides excellent agreement (slope: 1.01; R2 = 0.992). We employ this analytical approach to probe fundamental questions regarding sample stability and sorption of 6-PPDQ under lab-controlled conditions. Further, the results for 192 surface water samples provide the first spatiotemporal characterization of PPDQs on Vancouver Island and the lower mainland of British Columbia.