Non-proximate Sampling and Photoionization for Damage-Free Mass Spectrometric Analysis of Intact Native American Baskets.

Selected examples of Native American woven woodsplint basketry created between 1870 and 1983 are studied to recover traditional knowledge about their manufacture by identifying dyes or colorants. An ambient mass spectrometry system is designed to sample from intact objects with minimal invasiveness, neither cutting solids from the whole, exposing objects to liquid, nor leaving a mark on a surface. Baskets up to 60 cm wide in one dimension are placed on height-adjusted mounts. A timed jet of inert nitrogen from a finely positioned probe thermally desorbs neutral material from a mounted item, and a heated transport tube carries the analyte 2 m away at 4.9 L/min. Gas phase analyte is mixed with anisole dopant from an in-line permeation tube and photoionized in a reaction tee immediately before entering the mass spectrometer, identifying dye molecules in real time. Extensive optimization and exposure tests with flat and near-flat splints of dyed wood ensure that the analysis produces no discoloration on the curved and contoured basket splints.

Flow-Optimized Model for Gas Jet Desorption Sampling Mass Spectrometry.

Thermal gas jet probes, including post-plasma desorption/ionization sources, have not been studied using computational fluid dynamics (CFD) models, as have other ambient mass spectrometry sampling techniques. Two systems were constructed: a heated nitrogen jet probe to establish practical bounds for a sampling/transmission experiment and a CFD model to study trajectories of particles desorbed from a surface through optimization of streamlines and temperatures. The physical model configuration as tested using CFD revealed large losses, transmitting less than 10% of desorbed particles. Different distances between the desorption probe and the transport tube and from the sample surface were studied. The transmission improved when the system was very close to the sample, because the gas jet otherwise creates a region of low pressure that guides the streamlines below the inlet. A baffle positioned to increase pressure in the sample region improves collection efficiency. A Lagrangian particle tracking approach confirms the optimal design leading to a transmission of almost 100%.

High-Resolution Mass Spectrometry and Nontraditional Mass Defect Analysis of Brominated Historical Pigments.

The implementation of high-resolution mass spectrometry systems offers new possibilities for the analysis of complex art samples such as historical oil paintings. However, these multicomponent systems generate large and complex data sets that require advanced visualization tools to aid interpretation, especially when no chromatographic separation is performed. In the context of this research, it was crucial to propose a data analysis tool to identify the products generated during the synthesis, drying, and aging of historical pigments. This study reports for the first time a nontraditional mass defect analysis of oil paint samples containing a fugitive brominated-organic pigment, eosin or geranium lake, by using direct infusion electrospray ionization in combination with a high-resolution Orbitrap mass spectrometer. The use of nontraditional Kendrick mass defect plots is presented in this study as a processing and visualization tool to recognize brominated species based on their specific mass defect and isotope pattern. The results demonstrate that this approach could provide valuable molecular compositional information on the degradation pathways of this pigment. We anticipate that mass defect analysis will become highly relevant in future degradation studies of many more historical organic pigments.

In-Line Dopant Generation for Atmospheric Pressure Ionization Mass Spectrometry.

A concentric trace gas permeation tube that diffuses chemical reagents to a central carrier gas stream is used to drive chemical reaction pathways and influence gas-phase chemistry for a variety of atmospheric pressure ionization sources for mass spectrometry. Tunable permeation through the reservoir-jacketed polymer membrane is triggered by the heated gas moving through the tube, evaporating the dopant into a sheath dry gas or into a sample stream in room air without diluting the analyte concentration. The permeator is used to add dopants to an electrospray plume for analyte ion charge reduction and to perform hydrogen-deuterium exchange on biomolecules in different spray conditions. Dopants are also added to atmospheric pressure chemical ionization to favor the ionization of select components of diesel fuel. Atmospheric pressure photoionization is performed with the permeation tube in line with tubing transporting sample headspace to an enclosed discharge lamp. Toluene dopant from the permeator increases the proton transfer and charge exchange signal from clove oil and mothballs many times without exposing the laboratory to reagent fumes. Water permeation is also used to humidify the sample gas stream.

Interface for Reproducible, Multishot Direct Analysis of Solid-Phase Microextraction Samples.

An enclosed interface that joins a direct analysis in real time (DART) probe, solid-phase microextraction (SPME) fiber, and the inlet of a high-resolution mass spectrometer is described. Unlike other systems to couple SPME sampling to ambient mass spectrometry, the interface is able to perform discrete analyses on different areas of a single SPME fiber device for up to three technical replicate measurements of one sampling event. Inlet flow speed and desorption temperature are optimized, and reproducibility is demonstrated between replicate analyses on the same derivatized SPME fiber and with sequential fiber sampling events, yielding analyte measurement center of variance (CV) from 3 to 6%. Conditioning is also performed with the enclosed DART. The interface is a straightforward addition to commercially available technologies, and machine diagrams for custom components operated with SPME/DART/MS equipment are included.

Characterization of Zinc Carboxylates in an Oil Paint Test Panel.

Zinc (carboxylate) soaps, formed by reactions between zinc oxide (ZnO) and fatty acids in a drying oil, are known to cause deterioration in the paint layers of modern and contemporary oil paintings. This study investigates zinc carboxylates that developed in an oil painting test panel designed to mimic the aging and degradation encountered in actual works of art. Following accelerated and natural aging, protrusions were noted on the surface of the test panel. A large protrusion with erupted gel features was extracted from the test panel, mounted in top view, and then cut to reveal the sample's cross section. The gel features, which resulted from the unreacted oil binder's separation from the paint matrix, facilitated zinc carboxylate formation. Using reflectance µ-FTIR and SEM-EDX analysis, the morphologies and spatial distributions of zinc carboxylates within the gel regions of the protrusion were studied. A concentration gradient of zinc within the gel material was observed in the cross-sectional view, indicating patterns of zinc carboxylate formation and migration.

Rapid Evaluation of the Debromination Mechanism of Eosin in Oil Paint by Direct Analysis in Real Time and Direct Infusion-Electrospray Ionization Mass Spectrometry.

Eosin is a synthetic organic colorant prone to fading under the influence of light. On the basis of the growing interest in the understanding of the discoloration mechanism of eosin-based lakes, this study compares the ability of two ultrafast and ultrasensitive mass spectrometry techniques to detect eosin derivatives in complex matrices, such as oil media without the use of conventional separation columns or additional sample preparation protocols. Direct analysis in real time mass spectrometry (DART-MS) and direct infusion electrospray ionization mass spectrometry (DI-ESI-MS) were used to characterize the degradation pathway of eosin in oil media. The analysis protocols developed in this study are applied to discern the degradation mechanism of the lake pigment eosin (comprising the molecule per se complexed to an inorganic substrate) dispersed in linseed oil to create an oil paint. The analysis of oil paints by high resolution MS without an extraction methodology that modifies the system chemistry allowed us to identify the degradation forms without causing any additional fragmentation. Both techniques revealed the primary photodegradation pathway of eosin in linseed oil, and DI-ESI-MS provided additional information on the native conformation of the lake.

Proteomic and direct analysis in real time mass spectrometry analysis of a Native American ceremonial hat.

Complementary mass spectrometry analyses were performed to study a broken ceremonial hat of the Tlingit in the collection of the Smithsonian Institution National Museum of Natural History. The hat base and an associated cylinder are carved from wood and show multiple signs of age and breakage, as well as remnants of animal materials used for construction, decoration, and repair. Samples of animal tissues embedded in and attached to the wood were prepared for liquid chromatography-tandem mass spectrometry (LC-MS/MS), which identified proteins from five clades native to the object's area of origin. Surfaces on the hat and cylinder were analyzed using a direct analysis in real time (DART) MS system modified to accommodate the intact items. The presence of nicotine from tobacco smoke on the exterior and the relative absence of nicotine from the underside and formerly covered surfaces indicated that the cylinder was originally connected to the top of the hat. The characterization of the original object will be used to make informed decisions about reproduction of the intact hat for use by the Tlingit Kiks.ádi clan.

Humidity affects relative ion abundance in direct analysis in real time mass spectrometry of hexamethylene triperoxide diamine.

Unstable explosive hexamethylene triperoxide diamine (HMTD) is dangerous in quantity and benefits from the minimal sampling handling associated with atmospheric pressure chemical ionization for mass spectral analysis. Seasonal variation observed in HMTD mass spectra suggested a humidity dependence. Therefore, direct analysis in real time (DART) ionization mass spectra were acquired at a range of humidity values. An enclosure was designed to fit around the ion source and mass spectrometer inlet at atmospheric pressure. The enclosure was supplied with controlled amounts of humidified air from a test atmosphere generator to create programmable conditions for ambient analysis. The relative abundance and fragmentation of analyte ions were observed to change reliably with changing humidity values and, to a lesser degree, temperature. Humidity at such plasma-based ion sources should be regulated to avoid ∼90% shifts in relative ion abundance and provide stability and reproducibility of HMTD analysis.

Selective Sampling to and Desorption from Single Solid-Phase Microextraction Arrow Fiber for Replicative and Quantitative Analysis.

New analytical functionality is demonstrated with an enclosed interface that joins a solid phase microextraction (SPME) device, a direct analysis in real time (DART) probe, and a high-resolution mass spectrometer. With a single 20 mm long SPME Arrow, the interface is able to perform five discrete DART analyses on different areas of the same fiber in 1 min of practical operation time. Three-fiber replicates for 15 runs total produce 15% or better center of variance (CV) values for both volatile headspace sampling and direct immersion sampling of a solvated analyte. Chemometric analysis of rapidly acquired headspace data is able to distinguish volatile profiles. Selective desorption within the interface also confers the ability to selectively sample to discrete areas of a fiber, and three different headspace samples or five different liquid samples can be acquired and differentiated with one Arrow. A five-point standard addition curve is constructed to measure the concentration of the solvated analyte. Unmodified commercial components of the analysis system include the fiber itself, the Orbitrap and AccuTOF mass spectrometer platforms, and the conditioning gas chromatograph inlet. Machine diagrams for the SPME-DART interface and Arrow fiber holder are included.

Minimally Invasive Liquid Microjunction Sampling to Identify Dye on a 17th Century Spanish Illuminated Manuscript.

Common sampling methods for mass spectrometry such as sectioning are undesirably damaging to cultural heritage objects. A liquid microjunction sampling technique is developed that uses minimal solvent volume for analysis. Painted illustrations on a 17th century parchment manuscript from Spain were analyzed to identify the organic red pigment throughout its pages. By extracting with 0.1 µL solvent, the pigment was supplied for direct infusion electrospray MS, and the resulting disruption to the object surface was practically invisible to the naked eye.

Humidity Effects on Fragmentation in Plasma-Based Ambient Ionization Sources.

Post-plasma ambient desorption/ionization (ADI) sources are fundamentally dependent on surrounding water vapor to produce protonated analyte ions. There are two reports of humidity effects on ADI spectra. However, it is unclear whether humidity will affect all ADI sources and analytes, and by what mechanism humidity affects spectra. Flowing atmospheric pressure afterglow (FAPA) ionization and direct analysis in real time (DART) mass spectra of various surface-deposited and gas-phase analytes were acquired at ambient temperature and pressure across a range of observed humidity values. A controlled humidity enclosure around the ion source and mass spectrometer inlet was used to create programmed humidity and temperatures. The relative abundance and fragmentation of molecular adduct ions for several compounds consistently varied with changing ambient humidity and also were controlled with the humidity enclosure. For several compounds, increasing humidity decreased protonated molecule and other molecular adduct ion fragmentation in both FAPA and DART spectra. For others, humidity increased fragment ion ratios. The effects of humidity on molecular adduct ion fragmentation were caused by changes in the relative abundances of different reagent protonated water clusters and, thus, a change in the average difference in proton affinity between an analyte and the population of water clusters. Control of humidity in ambient post-plasma ion sources is needed to create spectral stability and reproducibility.

Isobutane Made Practical as a Reagent Gas for Chemical Ionization Mass Spectrometry.

As a reagent gas for positive- and negative-mode chemical ionization mass spectrometry (CI-MS), isobutane (i-C4H10) produces superior analyte signal abundance to methane. Isobutane has never been widely adopted for CI-MS because it fouls the ion source more rapidly and produces positive CI spectra that are more strongly dependent on reagent gas pressure compared with methane. Isobutane was diluted to various concentrations in argon for use as a reagent gas with an unmodified commercial gas chromatograph-mass spectrometer. Analyte spectra were directly compared using methane, isobutane, and isobutane/argon mixtures. A mixture of 10% i-C4H10 in argon produced twice the positive-mode analyte signal of methane, equal to pure isobutane, and reduced spectral dependence on reagent gas pressure. Electron capture negative chemical ionization using 1% i-C4H10 in argon tripled analyte signal compared with methane and was reproducible, unlike pure isobutane. The operative lifetime of the ion source using isobutane/argon mixtures was extended exponentially compared with pure isobutane, producing stable and reproducible CI signal throughout. By diluting the reagent gas in an inert buffer gas, isobutane CI-MS experiments were made as practical to use as methane CI-MS experiments but with superior analytical performance. Graphical Abstract ᅟ.

Minimizing thermal degradation in gas chromatographic quantitation of pentaerythritol tetranitrate.

An analytical method for establishing calibration curves for the quantitation of pentaerythriol tetranitrate (PETN) from sorbent-filled thermal desorption tubes by gas chromatography with electron capture detection (TDS-GC-ECD) was developed. As PETN has been demonstrated to thermally degrade under typical GC instrument conditions, peaks corresponding to both PETN degradants and molecular PETN are observed. The retention time corresponding to intact PETN was verified by high-resolution mass spectrometry with a flowing atmospheric pressure afterglow (FAPA) ionization source, which enabled soft ionization of intact PETN eluting the GC and subsequent accurate-mass identification. The GC separation parameters were transferred to a conventional GC-ECD instrument where analytical method-induced PETN degradation was further characterized and minimized. A method calibration curve was established by direct liquid deposition of PETN standard solutions onto the glass frit at the head of sorbent-filled thermal desorption tubes. Two local, linear relationships between detector response and PETN concentration were observed, with a total dynamic range of 0.25-25ng.

Quantification of allergenic bovine milk α(S1)-casein in baked goods using an intact ¹5N-labeled protein internal standard.

Intact bovine ¹5N-α(S1)-casein was used as an internal standard in a selected reaction monitoring (SRM) assay for milk protein in baked food samples containing fats, sugar, and gums. Effects on SRM results of sample matrix composition in two biscuit recipes containing nonfat dry milk (NFDM) were studied, including samples from a milk allergen ELISA proficiency trial. Following extraction of defatted samples with carbohydrate-degrading enzymes and acid precipitation of casein, the SRM assay exhibited an LOQ of <3 ppm NFDM with 60-80% recovery. NFDM levels measured by the SRM assay were 1.7-2.5 times greater than median levels determined by ELISA. Differences were observed in the α(S1)-casein interpeptide SRM ion abundance profile between recipes and after baking. ¹5N-α(S1)-Casein increases SRM analysis accuracy by correcting for extraction recovery but does not eliminate underestimation of allergen concentrations due to baking-related milk protein transformation (modifications).

A new approach to IRMPD using selective ion dissociation in a quadrupole ion trap.

Infrared multiphoton photodissociation (IRMPD) in a quadrupole ion trap is not selective for a parent ion. Product ions are decreased in abundance by continuous sequential dissociation and may be lost below the low mass cut-off. The IRMPD process is made selective by resonantly exciting trapped ions into an axially offset laser path. Product ions form and collisionally relax out of the laser path to accumulate in the center of the trap. The technique, termed selective broadband (SB) IRMPD, limits sequential dissociation to preserve first generation product ion abundance. The abundances of larger product ions are maximized by completely dissociating the parent ion, but continuous sequential dissociation does not form small product ions below the low mass cut-off associated with conventional IRMPD. Smaller product ions are further increased in abundance in another tandem mass spectrum by performing sequential stages of SB-IRMPD, adjusting the trapping rf amplitude to dissociate larger product ions at the same q(z) range. Thermal assistance is used to perform SB-IRMPD at higher bath gas pressures for increased sensitivity.

Improving IRMPD in a quadrupole ion trap.

A focused laser is used to make infrared multiphoton photodissociation (IRMPD) more efficient in a quadrupole ion trap mass spectrometer. Efficient (up to 100%) dissociation at the standard operating pressure of 1 x 10(-3) Torr can be achieved without any supplemental ion activation and with shorter irradiation times. The axial amplitudes of trapped ion clouds are measured using laser tomography. Laser flux on the ion cloud is increased six times by focusing the laser so that the beam waist approximates the ion cloud size. Unmodified peptide ions from 200 Da to 3 kDa are completely dissociated in 2.5-10 ms at a bath gas pressure of 3.3 x 10(-4) Torr and in 3-25 ms at 1.0 x 10(-3) Torr. Sequential dissociation of product ions is increased by focusing the laser and by operating at an increased bath gas pressure to minimize the size of the ion cloud.