Mass Spectrometry

A team of researchers at the University of Brighton in England used a combination of geochemical and statistical approaches to determine where Stonehenge’s sarsen (silcrete) megalith stones came from.

Gas chromatography–mass spectrometry (GC–MS) with cold electron ionization (EI) is based on interfacing the GC and MS instruments with supersonic molecular beams (SMB) along with electron ionization of vibrationally cold sample compounds in SMB in a fly-through ion source (hence the name cold EI). GC–MS with cold EI improves all the central performance aspects of GC–MS. These aspects include enhanced molecular ions, improved sample identification, an extended range of compounds amenable for analysis, uniform response to all analytes, faster analysis, greater selectivity, and lower detection limits. In GC–MS with cold EI, the GC elution temperatures can be significantly lowered by reducing the column length and increasing the carrier gas flow rate. Furthermore, the injector temperature can be reduced using a high column flow rate, and sample degradation at the cold EI fly-through ion source is eliminated. Thus, a greater range of thermally labile and low volatility compounds can be analyzed. The extension of the range of compounds and applications amenable for analysis is the most important benefit of cold EI that bridges the gap with LC–MS. Several examples of GC–MS with cold EI applications are discussed including cannabinoids analysis, synthetic organic compounds analysis, and lipids in blood analysis for medical diagnostics.

Per- and poly-fluoroalkyl substances (PFAS) are a family of potentially thousands of synthetic compounds that have long been used in the manufacture of a variety of common products with stain-repellent and nonstick properties. Their signature strong fluorine and carbon bonds make them difficult to break down and, as a result, they are among the most persistent of today’s environmental pollutants. Alarmingly, PFAS can be found in drinking water and have been shown to accumulate in the body with the potential to cause multiple health problems, such as hormone disruption and cancer. Advances in mass spectrometry have facilitated the detection of known PFAS contaminants as well as the identification of poorly studied and novel compounds in watersheds. This article explores the detection of known and novel PFAS contaminants in aqueous film-forming foams and raw drinking water sources in North Carolina, using new advances in mass spectrometry and data acquisition to improve identification and quantitation.

Over the last decade, matrix-assisted laser desorption–ionization (MALDI) imaging has become an indispensable tool for a broad range of applications, from studying plant metabolomics to discovering biomarkers of disease to developing new therapies. As such, MALDI imaging is revolutionizing preclinical drug discovery pipelines by providing direct distribution monitoring of therapeutic compounds and their metabolites along with untargeted pharmacodynamic information. A key application of MALDI imaging is tissue analysis for oncology, and recent developments in MALDI technology promise greater benefits to cancer research. The combination of MALDI with laser-induced post-ionization (PI) enhances the detection and imaging of pharmaceutical compounds and other classes of compounds, allowing for significant advances in the use of MALDI imaging for studying drug metabolism and pharmacodynamics in tumor tissues. This article describes the value of MALDI Imaging for oncology applications and examines the potential for laser-induced PI, including the ability to achieve up to three orders of magnitude higher sensitivity and to image metabolite classes previously undetectable with traditional MALDI.

Long chain fatty acids (LCFAs) function as a source of metabolic energy, substrates for membrane biogenesis, and storage of metabolic energy. Oxylipins, oxygenated derivatives of LCFAs, regulate the activity of many cellular processes. Existing methods for the analysis of LCFAs and oxylipins have limited compound coverage and sensitivity that, therefore, prevent their application in biological studies. In this work, we developed a high-throughput LC–MS method for analysis of 51 LCFAs and oxylipins. LCFAs and oxylipins were first extracted from biological samples via solid-phase extraction. The extracted molecules were analyzed by targeted comparative metabolomics. Saturated and monounsaturated LCFAs were analyzed in single ion reaction mode, while polyunsaturated LCFAs and oxylipins were analyzed in multiple reaction monitoring mode. Using this method, we successfully quantified 31 LCFAs and oxylipins from mouse livers.

Liquid chromatography coupled with high-resolution mass spectrometry (LC–HRMS) is used in combination with a comprehensive data analysis workflow to screen water samples for potentially hazardous transformation products from organic micropollutants to determine the efficacy of different water treatment methods.

When explosives are encountered on the battlefield, the use of portable GC–MS is valuable for the detection and confirmatory identification of pre- and post-detonation threats. In addition, this technique provides information about the source of explosives based on the detection and identification of trace-level chemicals in the sample. The data presented here confirm this capability.

Size-exclusion chromatography (SEC), with the use of ammonium acetate buffer, can be coupled on-line to electrospray ionization MS for the characterization of size variants of therapeutic monoclonal antibodies (mAbs). A quadrupole time-of-flight (QTOF) MS system was employed, and the MS method was optimized to achieve favorable sensitivity for high-mass detection, while maintaining the structural integrity of the aggregates (or high molecular weight species) and fragments (or low molecular weight species).

Food quality differences are dependent on botanical and geographical origins of primary food ingredients as well as storage and handling. Quality assessment for food materials, including cocoa and olive oil, is demonstrated by applying two-dimensional gas chromatography (GC×GC) combined with time-of-flight mass spectrometry (TOF-MS) and pattern recognition.

Drug discovery using high-throughput screening of discreet compounds, and the discovery of natural products with pharmacological mechanisms of action, rely on bioassay-guided fractionation analysis. Recent applications of affinity selection–mass spectrometry (AS-MS) are useful for exploring the discovery of ligands to membrane-bound proteins and RNA targets.

Per- and polyfluoroalkyl substances (PFAS) are found in firefighting foams and consumer products. They are ubiquitous in the environment and are an emerging human health concern. This work compares the 2009 and 2018 revised US Environmental Protection Agency (EPA) LC–MS/MS methods of analysis for PFAS in drinking water.

In the human food supply, public confidence is affected by contaminants and misreporting of nutritional information. This article highlights three events that required development of new mass spectrometry methods, including the detection of pesticides (such as fipronil and glyphosate), and the detection and quantification of fat-soluble vitamins.

In late-stage pharmaceutical development a new generation of high-resolution mass spectrometers and ion mobility mass spectrometers operate as orthogonal separation techniques and have greatly increased the ability to resolve impurities and increase the level of analytical information gained from a single analysis.

In this study, atmospheric pressure photoionization (APPI) is compared to the default ionization method, electrospray ionization (ESI), for solution-phase samples. These mass spectrometry methods are compared and optimized relative to artificial wastewater for the detection and quantitation of pharmaceuticals frequently found as environmental contaminants.

An arsenic speciation method using LA–ICP-MS was developed to provide a more accurate procedure for the determination of arsenic species in marine oils. It was validated for the analysis of five arsenic species in krill oil, and should also prove useful when quantitating inorganic arsenic species in other marine oils.

Method setup and optimization steps are explored to illustrate how an ICP-MS/MS method can be defined and tested to ensure consistent performance. Users can benefit from improved interference removal performance without the complex method development inherent in the use of ion-molecule reaction chemistry

A simple analytical method, requiring no sample pretreatment, was developed for determination of chromium, iron, nickel, and zinc in mouthwash by inductively coupled plasma‒optical emission spectrometry (ICP-OES). This method allowed the study of potential migration by iron, chromium, and nickel from stainless steel containers.

Our annual review of products introduced at Pittcon or during the previous year

Complex isobaric and polyatomic spectral interferences can be mitigated using triple quadrupole ICP-MS (ICP-MS/MS) with a collision–reaction cell (CRC). This configuration allows for the multielement characterization and detection of smaller nanoparticle sizes.

The structural complexity of monoclonal antibodies (mAbs) challenges the capabilities of even the most advanced chromatography and mass spectrometry techniques. This study examines the use of micro-pillar array columns in combination with mass spectrometry for peptide mapping of both mAbs and antibody–drug conjugates (ADCs).

Under a suitable thermal oxidation regime, vegetable oils yield a mixture of volatile and semivolatile organics that exhibit very high antimicrobial activities against a variety of microbial species. Volatile and semivolatile products were characterized with GC–MS using electron ionization and chemical ionization. The thermal oxidation of vegetable oils resulted in the formation of an array of shortand medium-chain acids, aldehydes, and ketones that act synergistically to yield a potent antimicrobial disinfectant.

Since glycans are responsible for bioactivity, solubility, immunogenicity, and clearance rate from circulation, it is vital to have a detailed map of glycans in therapeutic glycoproteins. Detailed glycoprotein structural analysis must be able to identify the peptide sequence where the glycans are attached as well as the structure of the glycan portion, including oligosaccharide sequence and glycosyl linkages. This article details methods for mass spectrometry experiments on both released glycans (“glycomics”), as well as on intact glycopeptides (“glycoproteomics”) using electron transfer dissociation, high-energy collision dissociation, and collisioninduced dissociation fragmentation pathways, which are needed to fully elucidate the structure of glycoproteins.

In this study, general extract screening of food storage materials was done with nontargeted analytical methods to understand what analytes could potentially leach into food or beverages. GC and mass spectral deconvolution effectively separated analytes within the complex mixture and TOF-MS provided full mass range spectral data for identification. This workflow can be used for confident characterization of components present as extractables from food packaging materials.

There is growing interest in the determination of endogenous proteins in biological samples for diagnostic purposes, because a concentration increase or decrease of such proteins can allows us to monitor the state of a pathological condition such as cancer. Immunocapture LC–MS/MS analysis combines the workflow of conventional immunological assays with LC–MS analysis. This article describes typical challenges, such as cross reactivity and the mass spectrometer’s dynamic range, as well as the advantages of isoform differentiation and multiplexing.

Interest in connecting ion mobility spectrometry (IMS) to GC and especially to LC is now growing. One favorable property of IMS is that it can work with ambient pressure and can be easily connected to a gas or liquid chromatograph. Analytical applications of GC–MS and LC–MS are very different and encompass investigations into food, medical science, environment, drugs of abuse, chemical warfare agents, and explosives.

In drug development, quantitative determination of a candidate drug and its metabolites in biofluids is an important step. The standard technique for quantitative metabolite profiling is radiolabeling followed by HPLC with radiodetection, but there are disadvantages to this approach, including cost and time, as well as safety and ethical concerns related to administering radiolabeled compounds to humans. Frank Vanhaecke and his research group at Ghent University have been developing an alternative technique, and he recently spoke to us about this work.

We present a brief review of this year’s ASMS conference, which took place June 4–8 in Indianapolis, Indiana.

Simultaneous, enantiomer-specific identification of chiral molecules in multi-component mixtures is extremely challenging. Many established techniques for single-component analysis fail to provide selectivity in multi-component mixtures and lack sensitivity for dilute samples. Mass spectrometry is chirally blind, and so cannot directly distinguish the two enantiomers of chiral molecules. Here we discuss how enantiomers may be differentiated by Mass Spectrometry correlated with PhotoElectron Circular Dichroism (MS-PECD) using an electron–ion coincidence imaging spectrometer. Following an ionizing circular polarized laser pulse, ions and electrons are detected in coincidence on their respective time- and position sensitive detectors. The MS-PECD asymmetry measured on electrons tagged by the mass of their corresponding parent ion directly reveals that the compound with identified mass is chiral without the need for any prior enantiomeric separation or enantiomer-selective complexation. MS-PECD enables direct enantiomeric excess measurement of multi-component chiral samples in a table-top mass spectrometer.

Our annual review of new spectroscopy products introduced at Pittcon, or during the previous year.