MS Inlet Research Articles

Long term online desalting analysis of MS/LC-MS using thermal assisted recrystallization ionization

Mass spectrometric analysis of non-volatile salts containing samples remains challenging due to salt-induced ion suppression and contamination. This challenge is even more pronounced for a liquid chromatography-mass spectrometry analysis, where the accumulation of salts in the transmission system poses an ongoing problem. In this study, a novel thermal assisted recrystallization ionization mass spectrometry (TARI-MS) device was developed to achieve efficient on-line desalting and prolonged analysis of saline samples. The core component of this device was a heated plate positioned between the electrospray unit and the MS inlet. The desalting mechanism was demonstrated as the spontaneous separation of target molecules from salts during the “crystallization” process. After optimization, the angle between the nebulizer and the heated plate was 45°; the distance between the front end of the heated plate and the MS inlet was 2 mm; the distance between the front edge of the heated plate and the center of the sample spray projected onto the heating plate was 3 mm; the distance between the emitter of nebulizer and the heated plate was 3 mm. TARI-MS realized direct analysis of eight drugs dissolved in eight commonly used non-volatile salts solutions (up to 0.5 mol/L). The high sensitivity, repeatability, linearity, accuracy, and intra- and inter-day precision of TARI-MS confirm its reliability as a robust tool for the analysis of saline samples. Furthermore, TARI-MS allowed continuous analysis of salty eluates of LC for up to nearly 1 h without maintenance and verified the feasibility of LC-MS analysis through detecting a five-drug mixture and a crude aripiprazole product. Finally, six impurities in the crude aripiprazole product were successfully detected by LC-TARI-MS. The established method holds promise for applications across academic and pharmaceutical domains.

Rapid and selective detection of AFB1 by direct mass spectrometry using immunoaffinity paper substrate

An antibody-modified Teslin paper was prepared in combination with direct mass spectrometry for the rapid and sensitive analysis of trace aflatoxin B1 (AFB1) in grain and its related products. Antibodies were covalently immobilized to Teslin surface by glutaraldehyde method, and an immunoaffinity paper substrate (IAP) was obtained. IAP can not only be used as a carrier for enrichment of analytes, but also coupled with the MS inlet for desorption/ionization detection. Compared to antibody-free Teslin, IAP could have excellent selectivity for the enrichment of AFB1. It took less than 1 min for detection. Among the matrices (wheat flour, peanut oil and corn), the proposed method showed satisfactory selectivity and linearity. The LOD and LOQ were 0.40–0.80 μg kg−1 and 1.20 μg kg−1, respectively. In conclusion, the IAP can provide a new powerful tool for rapid and sensitive analysis of mycotoxins.

Quantitative Electrochemistry-Mass Spectrometry: Real-Time Detection of Volatile Products for Electrocatalysis and Batteries

A major challenge both within electrocatalysis and battery research concerns the real-time (i) identification and (ii) quantification of volatile reaction products as a function of the applied electrochemical parameters and the reaction conditions. Analysis of both steadily evolving gases and of fast- and transient reaction phenomena can reveal key insights into reaction mechanisms. To this end, methods like Differential Electrochemical Mass Spectrometry (DEMS)1 and On-line Electrochemical Mass Spectrometry (OLEMS)2 have been used successfully to study reaction mechanisms to an ever-increasing degree in recent years, primarily in aqueous electrolytes. Similarly, Online Electrochemical Mass Spectrometers (OEMS)3 have been essential for the study of gas evolution in batteries.However, to gain an in-depth understanding of these mechanisms, the ability to accurately relate the electrochemical charge transferred in the reaction to the amount of evolved product is essential. To this end, accurate calibration of MS signals is paramount. Calibration procedures available for DEMS systems are by themselves cumbersome and unreliable and full quantification is not possible for OLEMS.Herein, we show how a simple gas-based calibration procedure using chip-based Electrochemistry-Mass Spectrometry (EC-MS) can be used for reliable, fully quantitative real-time analysis of volatile electrochemical reaction products. We validate this calibration with electrochemistry-based calibration methods described previously.4,5 The new procedure allows to extend the quantitative analysis to systems with no a priori knowledge of faradaic efficiencies. We demonstrate calibration of important gases like H2, O2 and C2H4 in aqueous and non-aqueous electrolytes and exemplify the calibration methodology’s usefulness on standard electrochemical reactions. References H. Baltruschat, J Am Soc Mass Spectrom, 15, 1693–1706 (2004).A. H. Wonders, T. H. M. Housmans, V. Rosca, and M. T. M. Koper, J Appl Electrochem, 36, 1215–1221 (2006).N. Tsiouvaras, S. Meini, I. Buchberger, and H. A. Gasteiger, J Electrochem Soc, 160, A471–A477 (2013).D. B. Trimarco et al., Electrochim Acta, 268 (2018).S. B. Scott, PhD thesis, Technical University of Denmark (2019). Figure 1: a) Schematic of the MS inlet chip’s working principle. b) Uncalibrated M2 and M32 signals (dotted lines) and the same signals after calibration for H2 and O2 (full lines) in a cyclic voltammogram of PtPoly in 0.1 M HClO4 obtained at room temperature at 20 mV/s. Figure 1

Development of a Simple and Stable NanoESI Spray System Using Suction Wind from the MS Inlet.

Improving the reproducibility of proteome analysis systems presents a challenge; therefore, in this study, we developed a new insertion spray ionization (InSpIon) system wherein an InSpIon tube was designed with a spray tip inserted into the tube. This system stabilized the spray and subsequently improved the reproducibility of the analysis.

Development of a dual-electrospray ionization source with in-line absorbance-based voltage control.

Emitter tip arrays for electrospray ionization have been used for a variety of MS sample introduction purposes, including detection of multiple sample eluent streams and improved accuracy through parallel infusion of an internal standard. User control is typically required for targeted application of high voltage to specific channels to maximize analyte signal and minimize other background signals. In this communication, an automated approach to applying electrospray voltage only when a detectable analyte is present is described. An in-line absorbance detector is used to identify the presence of an analyte in the fluidic path between the sample introduction valve and the mass spectrometer. A Raspberry Pi-controlled system is then used to apply high voltage to a downstream emitter tip at the MS inlet following a delay volume between the detectors. Demonstration of this technique on two parallel sample channels is reported, including a pulsed voltage application to maximize signal when analytes elute on each channel simultaneously.

Thin layer chromatography/desorption flame-induced atmospheric pressure chemical ionization/mass spectrometry for the analysis of volatile and semi-volatile mixtures.

A micro-sized oxyacetylene flame was generated using a small concentric tube system. Hot gas flow and primary reactive species from the micro-flame were directed toward a developed TLC gel plate to thermally desorb and ionize analytes on the gel surface. The resulting analyte ions subsequently entered the MS inlet for detection. A 1-1.5-mm-wide light-brown line was observed on the TLC plate after the desorption FAPCI/MS (DFAPCI/MS) analysis, revealing that the gel surface withstood a high temperature from the impact of the micro-flame. Volatile and semi-volatile chemical compounds, including amine and amide standards, drugs, and aromatherapy oils, were successfully desorbed, ionized, and detected using this TLC/DFAPCI/MS. The limit of detection of TLC-DFAPCI/MS was determined to be 5ng/spot for dibenzylamine and ethenzamide. TLC/DFAPCI/MS is one of the simplest TLC-MS interfaces showing the advantages such as low costs and an easy set up. The technique is useful for characterizing thermally stable volatile and semi-volatile compounds in a mixture.

Rapid determination of aldehydes in food by high-throughput reactive paper spray ionization mass spectrometry

Due to endogenous existence and hazardous properties, the quantification of fatty and aromatic aldehydes in food matrices is indispensable to food safety. Current analytical methodologies for aldehydes in food suffer from several drawbacks (typically complicated pretreatment, long analysis time and low analytical throughput, low sensitivity, etc.). In this work, a highly sensitive, high-throughput and sample pretreatment-free method for aldehyde analysis was developed to address the above problems, where in situ extraction and derivatization of aldehydes with hydrazines to hydrazones were integrated into reactive paper spray ionization mass spectrometry (reactive PSI-MS) considering weakly ionizable properties of aldehydes. Four aldehydes, formaldehyde, acetaldehyde, propionaldehyde and benzaldehyde were reacted with dansyl hydrazine in 1.5 % v/v HAc acidified methanol on the qualitative fast-speed filter paper for 5 min, and then a voltage (+4 kV) was applied to initiate paper spray for mass spectrometry detection with internal calibration by dansyl amide. Once the completion of MS analysis, the glass plate was moved to introduce the second triangular tip towards the MS inlet (5 mm apart) for next trial, achieving high-throughput reactive PSI-MS analysis (60 h−1). The four aldehydes showed good linearity in the concentration range from 2 to 150 μM with low detection limits of 0.03–0.15 μM. The method was applied to direct quantification of four aldehydes in 15 kinds of foods such as vegetables, fruits and meat products. The proposed method has the advantages of simple operation, high efficiency and high sensitivity, making it attractive to routine analysis of aldehydes in food, environmental and biological samples.

Quantitative detection of caffeine in beverages using flowing atmospheric-pressure afterglow (FAPA) ionization high-resolution mass spectrometry imaging and performance evaluation of different thin-layer chromatography plates as sample substrates

Ambient desorption/ionization mass spectrometry (ADI-MS) is widely used as a rapid screening tool of samples in their native state without sample preparation. While analysis times are much less than 1 min per sample, one challenge of ADI-MS is the possibility to perform quantitative analysis of analytes in complex matrices. Typically, the goal is to probe a variety of different analytes in a complex matrix from a solid, liquid, or otherwise uncharacterized surface in the open air in front of the MS inlet. In this study, it is demonstrated that a carefully selected surface for analyte spot sampling and co-deposited isotopically labeled standards both significantly improve the capabilities of flowing atmospheric-pressure afterglow (FAPA) high-resolution (HR) MS for direct quantitative analysis. Specifically, a systematic study of different surfaces (glass, steel mesh, high-performance thin-layer chromatography (HPTLC) stationary phases including silica, reversed-phase (RP)-modified silica, and cyano (CN)-modified silica) and their suitability for spot sampling with FAPA-MS was performed. A set of different caffeine-containing standards and beverages (Red Bull, Coca-Cola, coffee, and black tea) was deposited on the surfaces and direct FAPA-HR-MS analysis of caffeine was performed using internal calibration with co-deposited 13C3-caffeine. For TLC surfaces, it was demonstrated that quantitative results could be achieved with the matrix and concomitants present and that a preceding chromatographic separation was not mandatory for this application. In addition, the use of a CN-HPTLC surface resulted in a significantly more intense caffeine signal in the beverage samples compared to the other surfaces studied, with the highest increase compared to the silica (200-fold higher) and the lowest increase compared to the steel mesh (30-fold higher). The utilization of TLC-based surfaces as sample carriers is considered an attractive tool in the ADI-MS toolbox for fast and efficient mass spectrometric investigations of complex samples without time-consuming sample preparation.Graphical

Tee-Shaped Sample Introduction Device Coupled with Direct Analysis in Real-Time Mass Spectrometry for Gaseous Analytes.

Ambient ionization mass spectrometry (AIMS) is simple to operate for analytes adsorbed on the surface of various shaped probes. However, gaseous substances or liquids that are easy to evaporate, diffuse, and escape in the atmosphere are harder to capture. In this work, a Tee-shaped sample introduction device coupled with direct analysis in real time mass spectrometry (DART-MS) is developed. The Tee-shaped device is placed between the DART ion source and the MS inlet with a heated sample transfer tube. Gaseous samples from either a Tedlar sampling bag or liquids evaporated from a graduated syringe were tested. The Tee-shaped device was used for several volatile organic compounds with a wide range of boiling points, and detection limits of ng/mL to fg/mL were obtained. To test the device for real-life samples, puff-by-puff analysis of a complex gaseous mainstream smoke was performed. Individual puffs can be analyzed rapidly, and there is no cross contamination between consecutive puffs. The dynamic changes of chemical components among different puffs for different types of cigarettes can be observed. This work provides a universal Tee-shaped sampling device to enhance AIMS for the analysis of volatile compounds and gases, which is adapted to different sampling modules applicable for various forms of samples. The device enables direct exploration of chemical components in complex gaseous samples without tedious sample preparation and time-consuming LC or GC separation.

Ambient ion focusing for paper spray ionisation

Paper Spray (PS) is an ambient ionisation technique that is conceptually simple, economical, direct and versatile. In its simplest form, the procedure for operation relies on the application of an electric potential to a triangular-shaped paper substrate with the addition of an extraction/spray solvent to generate charge droplets, without requiring any pneumatic assistance. Despite its promise and rapidly growing popularity, there are some practical challenges associated with PS implementation with scope to enhance its performance further. One such challenge relates to only a small fraction of the Taylor cone expansion being sampled at the MS inlet. In this work we propose a new arrangement for PS, which retains the inherent advantages of this popular technique without introducing additional variables, by using an ambient ion focusing lens that is held at the same potential as the paper substrate. A thorough investigation, consisting of visual spray inspection, electric field simulations and analytical evaluation, including analysis of paracetamol from saliva, shows that ambient ion focusing can provide up to a 50% improvement in spray stability, 60% increase in signal intensity and a 30% improvement in detection limits for routine paper spray mass spectrometry analysis.

Evaluation of a nanoflow interface based on the triple-tube coaxial sheath-flow sprayer for capillary electrophoresis-mass spectrometry coupling in metabolomics

The performance of an original CE-MS interface that allows the in-axis positioning of the electrospray with respect to the MS inlet was evaluated. The variations in the geometrical alignment of this configuration in the absence of a nebulizing gas afforded a significant reduction in the sheath-liquid flow rate from 3 µL/min to as low as 300 nL/min. The sheath liquid and BGE were respectively composed of H2O—iPrOHCH3COOH 50:50:1 (v/v/v) and 10% acetic acid (pH 2.2). A significant gain in sensitivity was obtained, and it was correlated to the effective mobility of the analytes. Compounds with low mobility values showed a greater sensitivity gain. Special attention was paid to the detection of proteinogenic amino acids. Linear response functions were obtained from 15 ng/mL to 500 ng/mL. The limits of quantification, as low as 34.3 ng/mL, were improved by a factor of up to six compared to the conventional configuration. The in-axis setup was ultimately applied to the absolute quantification of four important amino acids, alanine, tyrosine, methionine and valine, in standard reference material (NIST plasma). The accuracies ranged from 78 to 113%, thus demonstrating the potential of this configuration for metabolomics.

Electrospray ionization stability and concentration sensitivity in capillary electrophoresis‐mass spectrometry using a flow‐through microvial interface

Concentration sensitivity is a key performance indicator for analytical techniques including for capillary electrophoresis-mass spectrometry (CE-MS) with electrospray ionization (ESI). In this study, a flow-through microvial interface was used to couple CE with MS and improve the ESI stability and detection sensitivity. By infusing a peptide mixture through the interface into an MS detector at a typical flow rate for CE-MS analysis, the spatial region near the interface was mapped for MS signal intensity. When the sprayer tip was within a 6 × 6.5 × 5mm region in front of the MS inlet, the ESI was stable with no significant loss of signal intensity for ions with m/z 239. Finite element simulations showed that the average electric field strength at the emitter tip did not change significantly with minor changes in emitter tip location. Experiments were conducted with four different mass spectrometer platforms coupled to CE via the flow-through microvial interface. Key performance indicators, that is, limit of detection (LOD) and linearity of calibration curves were measured for nine amino acids and five peptides. Inter- and intraday reproducibility were also tested. The results were shown to be suitable for quantification when internal standards were used.

Nebulization dielectric barrier discharge ionization mass spectrometry: Rapid and sensitive analysis of acenaphthene

A rapid, simple and sensitive method was proposed for low-polar acenaphthene analysis by coupling nebulization with dielectric barrier discharge ionization (N-DBDI). The sample solution was nebulized followed by heating and converted to be gas-phase analyte molecules prior to DBDI. This boosts the collision efficiency of analyte molecules with reactive species and thus the sensitivity, and the high-velocity gas from nebulization guides ions directed to the MS inlet without deflection. The dependence of sensitivity on the operation parameters was systematically investigated. The LOD and LOQ of acenaphthene were determined to be 0.61 ng/L and 2.05 ng/L, respectively, which were superior approximately 30 folds compared to those obtained by other methods. Parameters, including accuracy, precision, reproducibility and utility, were tested to further evaluate the performance of N-DBDI. Real environmental samples, including river water, initial rainwater and mineral water, were analyzed with good accuracy (93.61–103.50%) and satisfactory precision (RSD ≤ 8.92%). These findings suggest that the N-DBDI allows the determination of non/low-polar species at sub-pg/mL possible, and would benefit for the non/low-polar species analysis in real environmental samples.

A High-Throughput Screening Method for Determining the Optimized Synthesis Conditions of Quinoxaline Derivatives Using Microdroplet Reaction

Quinoxaline derivatives demonstrate many distinguished chemical, biological, and physical properties and have a wide application in dyes, electroluminescent material, organic semiconductors, biological agents, etc. However, the synthesis of quinoxaline still suffers from several drawbacks, for instance, longer reaction time, unsatisfactory yields, and use of metal catalysts. Here, utilizing microdroplet-assisted reaction, we demonstrate the successive synthesis of several important quinoxaline derivatives. For case studies of 1H-indeno [1, 2-b] quinoxaline and 3,5-dimethyl-2-phenylquinoxaline, the present microdroplet approach can complete in milliseconds and the conversion rate reached 90% without adding any catalyst, which is considerably quicker and higher than conversional bulk-phase reactions. When combined with MS detection, high-throughput screening of the optimal reaction conditions can be achieved. Several impacts of droplet volume, reaction flow rate, distance from the MS inlet, spray voltage, and flow rate of the auxiliary gas can be screened on-site quickly for enhanced reaction speed and yields. More importantly, this platform is capable to be used for the scaled-up microdroplet synthesis of quinoxaline diversities. Considering the facile, economic, and environmentally friendly features of the microdroplet approach, we sincerely hope that the current strategy can effectively promote the academic research and industrial fabrications of functional quinoxaline substances for chemical, biological, and pharmaceutical application developments.

Contactless electrospray ionization mass spectrometry for direct detection of analytes in living organisms.

In this study, we developed contactless electrospray ionization mass spectrometry (ESI-MS) for in vivo analysis of living organisms in different applications. The in vivo sampling and direct analysis processess of living organisms were integrated into an operation that only requires the organism close to MS inlet that was applied to a high voltage. Living plants and animals were directly induced to generate spray ionization. Direct detection and in vivo monitoring of metabolites and chemical residues in various living organisms were successfully demonstrated. Analysis of a single sample could be completed within 30 s. Overall, contactless ESI-MS provides an attractive in vivo method to straightforward investigation of living organisms.

Vibrating tip spray ionization mass spectrometry for direct sample analysis.

In this work, a vibrating tip spray ionization source was developed for direct mass spectrometric analysis of raw samples under voltage-free condition. A solid tip was mounted on a vibrator, and the solid tip was placed on the front of MS inlet. Liquid, viscous, and bulk solid samples could be directly loaded on the tip-end surface, and then a drop of solvent at microliter level was subsequently loaded on the tip for dissolution and extraction of analytes, and a vibrator was then started to atomize and ionize the analytes under ambient condition. We demonstrated vibrating tip spray mass spectrometry in various applications, including food safety, pharmaceutical analysis, and forensic science. Furthermore, in situ analysis of biological tissues and in vivo analysis of living plants were conveniently performed, due to voltage-free. Different vibration frequencies and solvent compositions were investigated. The analytical performances, including sensitivity, reproducibility, and linear range, were investigated. The ionization process and mechanism were also discussed in this work.

Design Characteristics to Eliminate the Need for Parameter Optimization in Nanoflow ESI-MS.

The sampling efficiency in electrospray ionization-mass spectrometry (ESI-MS) can be improved by decreasing the liquid flow rate to the nanoflow regime, where it is possible to draw a large fraction of the ESI plume into the mass spectrometer. This mode of operation is typically more difficult than ESI-MS at higher flow rates because it requires careful optimization of a number of parameters to achieve optimal sampling efficiency. In this work, we screened the relative impact on signal intensity and spray stability of factors that included sprayer position, spray electrode protrusion, sprayer tip shape, spray angle relative to the MS inlet, nebulizer gas flow rate, ESI potential, and means for generating the electric field to initiate electrospray. Based on the screening results, we explore the possibility of providing fixed optimal values for many of the key source parameters to eliminate much of the tuning that is required for conventional nanoflow sources. This approach has potential to greatly simplify nanoflow ESI-MS, while providing optimized sensitivity, stability, and robustness, with decreased variability between analyses.

NanoBlow: A Simple Device To Limit Contaminants during NanoLC-MS.

Limiting contamination of liquid chromatography/tandem mass spectrometry (LC-MS) systems and reducing the downtime associated with maintenance and cleaning is essential for productivity. We developed a simple device that creates a gas curtain barrier to prevent ions from entering the MS inlet. The gas can be quickly and easily applied when certain contaminant ions are known to elute. We show that the device can prevent the buildup of contaminants on the heated transfer capillary following >100 injections of a crude tissue lysate and improves peptide identifications. The device may provide a promising approach toward improving instrument robustness.

Addition of Serine Enhances Protein Analysis by DESI-MS.

Previous studies have suggested that the loss in sensitivity of DESI-MS for large molecules such as proteins is due to the poor dissolution during the short time scale of desorption and ionization. An investigation into the effect of serine as a solvent additive leads to the interesting observation that there is a concentration-dependent improvement in protein signal intensity when micromolar to low millimolar concentrations of serine is combined with a suitable co-additive in DESI spray. This effect, however, was not observed during similar ESI-MS experiments, where the same solvents and proteins were sprayed directly into the MS inlet. This suggests that the mechanism of signal improvement in DESI is associated with the desorption step of proteins, possibly by facilitating dissolution or improving solubility of proteins on the surface in the solvent micro-layer formed during DESI. Other than poor dissolution, cation adduction such as by sodium ions is also a major contributing factor to the mass-dependent loss in sensitivity in both ESI and DESI, leading to an increase in limits of detection for larger proteins. The adduction becomes a more pressing issue in native-state studies of proteins, as lower charge states are more susceptible to adduction. Previous studies have shown that addition of amino acids to the working spray solution during ESI-MS reduces sodium adduction and can help in stabilization of native-state proteins. Similar to the observed reduction in sodium adducts during native-state ESI-MS, when serine is added to the desorbing spray in DESI-MS, the removal of up to 10mM NaCl is shown. A selection of proteins with high and low pI and molecular weights was analyzed to investigate the effects of serine on signal intensity by improvements in protein solubility and adduct removal. Graphical Abstract.

Reversed-Phase Functionalised Multi-lumen Capillary as Combined Concentrator, Separation Column, and ESI Emitter in Capillary-LC–MS

For the first time, a multi-lumen capillary (MLC) (126 parallel channels of 4.2 µm i.d) has been modified to produce a C18-functionalised silica porous layer open tubular (PLOT) capillary column for both on-capillary preconcentration and separation. The modified multi-lumen capillary used in this dual mode provided significant advantages over typical nano/capillary-LC–MS systems, in that it facilitated both higher sample loading capacity, the use of elevated flow rates, and simplified equipment requirements. Following modification, 100% of the channels displayed a homogenous porous silica layer, 257 ± 36 nm thick. The PLOT-MLC was first evaluated for on-capillary solid-phase extraction. Extraction of caffeine, ofloxacin, atrazine, and diuron was carried out offline using an 8-cm-long PLOT-MLC, with quantification achieved using HPLC coupled to a quadrupole-time of flight (QTOF) mass spectrometer. The results confirmed reversed-phase selectivity and average recoveries obtained were around 70%. Subsequently, a 65-cm-long PLOT-MLC was evaluated as a separator column using a capillary liquid chromatography (Cap-LC) system equipped with a nano-injector and coupled to the mass spectrometer. The short PLOT-MLC provided a baseline separation in isocratic mode [water:acetonitrile (each with 0.1% formic acid) = 70:30, v/v] of ofloxacin, atrazine, and diuron. Finally, direct coupling of the PLOT-MLC with the QTOF via a capillary electrosprayer facilitated the simultaneous use of the modified capillary as a solid-phase concentrator, separator column (carrying out concentration-focusing-separation on the PLOT-MLC) and electrospray emitter. This configuration greatly simplifies the traditional capillary-LC–MS equipment requirements, via the removal of all connectors and additional capillary between injector and MS inlet, and is demonstrated herein with large volume sample loading and step-gradient elution/separation with sensitive MS detection.