Development of electrospray ionization on solid substrates (solid-substrate ESI) avoids the clogging problem encountered in conventional capillary-based ESI, allows more convenient sampling and permits new applications. So far, solid-substrate ESI with various materials, e.g., metals, paper, wood, fibers and biological tissue, has been developed, and applications ranging from analysis of pure compounds to complex mixtures as well as in vivo study were demonstrated. Particularly, the capability of solid-substrate ESI in direct analysis of complex samples, e.g., biological fluids and foods, has significantly facilitated mass spectrometric analysis in real-life applications and led to increasingly important roles of these techniques nowadays. In this review, various solid-substrate ESI techniques and their applications are summarized and the prospects in this field are discussed.

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Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) can be used to visualize the distribution of biomolecules (proteins, peptides, metabolites) and drugs on tissue surfaces. In MALDI-IMS, sample preparation is crucial for successful results. A variety of conditions, such as tissue sampling methods, tissue thickness and matrix application procedure can have an impact on the results. In this review, we summarize each sample preparation step in an orderly sequence with practical examples. In addition, we discuss the importance of the organic solvent used in the matrix solution. The composition of the organic solvent used in the matrix solution is critical for achieving a high sensitivity in this procedure.

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The amino acid residues susceptible to in-source decay (ISD) in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry have been identified from both positive and negative ion ISD spectra of cytochrome c, myoglobin, thioredoxin and bovine serum albumin. Backbone cleavages at the N–C_α bonds of Xxx–Asp, Xxx–Asn, Xxx–Cys, and Gly–Xxx residues gave discontinuous intense peaks of c-ions, independent of positive and negative ion mode. The intensity values for c-ions, Int(c), were defined to allow estimation of the discontinuous intense peaks of c-ions. The identities of the high intensity value residues Asp, Asn, Cys, and Gly were compared with those identified using other measures of flexibility such as the B-factor, turn preferential factor and protection factor. The comparison indicates that Asp, Asn, and Gly residues are common to all measures. Thus, the intensity values of c-ions can be adopted as a measure of protein flexibility.

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Pressurizing the ionization source to gas pressure greater than atmospheric pressure is a new tactic aimed at further improving the performance of atmospheric pressure ionization (API) sources. In principle, all API sources, such as ESI, APCI and AP-MALDI, can be operated at pressure higher than 1 atm if suitable vacuum interface is available. The gas pressure in the ion source can have different role for different ionization. For example, in the case of ESI, stable electrospray could be sustained for high surface tension liquid (e.g., pure water) under super-atmospheric pressure, owing to the absence of electric discharge. Even for nanoESI, which is known to work well with aqueous solution, its stability and sensitivity were found to be enhanced, particularly in the negative mode when the ion source was pressurized. For the gas phase ionization like APCI, measurement of gaseous compound also showed an increase in ion intensity with the ion source pressure until an optimum pressure at around 4–5 atm. The enhancement was due to the increased collision frequency among reactant ion and analyte that promoted the ion/molecule reaction and a higher intake rate of gas to the mass spectrometer. Because the design of vacuum interface for API instrument is based on the upstream pressure of 1 atm, some coupling aspects need to be considered when connecting the high pressure ion source to the mass spectrometer. Several coupling strategies are discussed in this paper.

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A specific property of silver oxide-based nanoparticles permits the ionization of an analyte, giving rise to various applications of a smart analytical methodology. The nanoparticles (d=6.7 nm) contained an Ag₂O core. The detection of several model componds (a nucleobase and two hair growth promoters) via the use of silver oxide nanoparticles is described. Adducts were produced between the target molecules and the two silver stable isotopes (Ag¹⁰⁷ and Ag¹⁰⁹), resulting in the formation of specific signals as well as a protonated signal. Thus, it was possible to easily determine whether the given signals were correlated with the target molecule or not.

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Microfluidic chips have been used as platforms for a diversity of research purposes such as for separation and micro-reaction. One of the suitable detectors for microfluidic chip is mass spectrometry. Because microfluidic chips are generally operated in an open air condition, mass spectrometry coupled with atmospheric pressure ion sources can suit the requirement with minimum compromise. In this study, we develop a new interface to couple a microfluidic chip with mass spectrometry. A capillary tip coated with a layer of graphite, capable of absorbing energy of near-infrared (NIR) light is used to interface microfluidic chip with mass spectrometry. An NIR laser diode (λ=808 nm) is used to irradiate the capillary tip for assisting the generation of spray from the eluent of the microfluidic chip. An electrospray is provided to fuse with the spray generated from the microfluidic chip for post-ionization. Transesterification is used as the example to demonstrate the feasibility of using this interface to couple microfluidic chip with mass spectrometry.

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The development of a MALDI-TOF mass spectrometer that utilizes spiral ion trajectory, SpiralTOF, is reported. The total flight path length was 17 m, which is five times longer than that in commonly used reflectron ion optical system. The SpiralTOF reduced the dependence of the mass resolving power on the mass of the analyte, while improving the accuracy of the mass measurements. Furthermore, SpiralTOF has two advantages that can be exploited for the separation of minor abundant isobaric components in mass spectra. One is the reduction in chemical background due to the post source decay (PSD), which is achieved through PSD ion elimination by electrostatic sectors contained within the SpiralTOF. The other is that the stabilities of peak positions are improved during mass spectrum accumulation. The peak drift caused by the fine structure of matrix crystals and the small irregularities on the sample surface can be reduced by extending the flight path. In this study, these advantages are demonstrated via the analysis of a block copolymer and mass spectrometry imaging (MSI) of lipids.

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Parallel Fragmentation Monitoring (PFM), which is an analogue of selected reaction monitoring (SRM), is a recently developed method for quantification of small molecules by MALDI-TOF/TOF mass spectrometry (MS). It is well known that isobaric interference substances can be occasionally present in complex biological samples, and affect the accuracy of measurement by SRM. Unfortunately, by design it is not possible to assess whether isobaric interference happens in a SRM analysis. In contrast, the unique design of PFM should allow quick inspection for isobaric interference and subsequent correction. In this study, using arginine as an example, interference effect of isobaric structural analogs on the quantification of citrulline by PFM was evaluated. Our results showed that the presence of arginine affected the measured concentrations of citrulline standard solutions in a concentration dependent manner. Such interference could be observed readily in the MS/MS spectra, and contributed by [arginine+H–NH₃]⁺ fragment ion. Because of having highly similar mass, ¹³C-isotope of [arginine+H–NH₃]⁺ fragment ion overlapped with monoisotope of [citrulline+H–NH₃]⁺ fragment ion, which was used as the report ion for quantification. However, such interference could be mathematically eliminated or minimized through estimation of the signal intensity of the ¹³C-isotopic peak of [arginine+H–NH₃]⁺ from the intensity of the corresponding monoisotopic peak according to isotope distribution of elements. Furthermore, the presence of interfering fragment ions could be avoided by optimizing MALDI ionization condition. In conclusion, isobaric interference can happen in PFM, but can be easily identified in the mass spectra and eliminated (minimized) with simple methods.

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Chiral recognition of D- and L-amino acids is achieved by a method combining electrospray ionization (ESI) and in-source collision-induced dissociation (CID) mass spectrometry (MS). Trimeric cluster ions [Cu^II(A)(ref)₂-H]⁺ are formed by ESI of mixtures of D- or L-analyte amino acid (A), chiral reference (ref) and CuSO₄. By increasing the applied voltage in the ESI source region, the trimeric ions become unstable and dissociate progressively. Thus chiral differentiation of the analyte can be achieved by comparing the dependence of their relative intensities to a reference ion on applied voltages. The method does not need MS/MS technique, thus can be readily performed on single-stage MS instruments by turning the voltage of sampling cone.

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Ion mobility spectrometry (IMS) as a stand-alone technique has become increasingly important for applications in security, defense, and environmental monitoring, and also in biological applications such as molecular structure and -omic analysis when combined with mass spectrometry. Yet, the majority of these devices are drift cell based and limited by low duty cycles because of ion gating. Differential Mobility Analyzers (DMAs) are attractive alternatives due to their continuous ion transmission and success in analyzing aerosol particles in real time environmental tests. But, the resolution of a DMA is low due to difficulties in achieving laminar gas flow, low sample gas flow to sheath gas flow ratio, and high velocity sheath gas using small pumps, if portability is a concern. To overcome these challenges, we will introduce a new ion mobility spectrometer that increases the amount of work done on the ions during separation by introducing an electric field opposing the gas flow direction while simultaneously preserving laminar gas flow. The development of the Periodic Focusing Differential Mobility Analyzer (PFDMA) can lead to a portable device that exhibits both high resolution and sensitivity, to meet the needs of today’s expanding applications.

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