Laser ablation inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-TOFMS) enables the quantitative imaging of metal-based therapeutics with cellular resolution. Despite its analytical power, systematic screening of metal-containing anticancer agents remains limited by the substantial time, cost, and labor required for cell handling, staining, and analysis. Here, we present a barcoding strategy that allows us to pool multiple experiments together and analyze them simultaneously. Our labeling approach combines wheat germ agglutinin (WGA) with lanthanide-labeled anti-WGA barcodes, enabling robust discrimination of experimental conditions. Following LA-ICP-TOFMS measurement, pooled data sets can be processed using MeXpose, a data processing pipeline for single cells, to accurately assign each cell to its original barcode, i.e., experiment. In this study, we applied this novel strategy to investigate the uptake of BOLD-100 and oxaliplatin in HCT116 wild-type (WT) and oxaliplatin-resistant (OxR) colorectal cancer cells using different drug concentrations. Pooling 10 experiments into a single analytical run allowed us to reduce consumable use (mainly argon for the ICP and antibody usage if stained), measurement time, and downstream processing while increasing data consistency. Barcoding-enabled single-cell analysis confirmed a substantial reduction in oxaliplatin uptake in oxaliplatin-resistant HCT116 cells compared with the WT cell line, whereas BOLD-100 uptake was affected to a much lesser extent. These results demonstrate the utility of this strategy for the efficient and scalable assessment of metal-based therapeutics.

Single-particle inductively coupled plasma mass spectrometry (sp-ICP-MS) enables the characterization of individual nanoparticles (NPs) on a particle-by-particle basis and represents a major advance in analytical chemistry. Time-of-Flight mass analyzers (TOFMS) have emerged as a powerful platform for single-particle analysis thanks to their fast acquisition rates and simultaneous multielement detection capability. Despite its growing relevance and use in environmental and material sciences, sp-ICP-MS is rarely included in graduate teaching laboratories. This advanced lab course introduces students to the principles, operation, and data analysis of sp-ICP-MS and ICP-TOFMS through a combination of theory and hands-on experiments. First, commercially available monodisperse spherical Au and Ag NPs are analyzed so that students become familiar with the fundamental principles of sp-ICP-TOFMS, including transient signal acquisition, calibration, and particle event interpretation. Then the students apply the same workflow to consumer products using sunscreens and cosmetics to detect and quantify titanium dioxide (TiO2) NPs. Through these experiments, students strengthen their understanding of plasma-based mass spectrometry, quantitative data evaluation, and NPs characterization in complex matrices. This laboratory class bridges classical ICP-MS education with modern single-particle analysis, providing students with practical experience in advanced mass spectrometry.

Flow imaging analysis (FIA), provides composition-agnostic morphological characterization. These measurements of particle size and shape are valuable to mass-based analysis, such as single particle inductively coupled plasma time-of-flight mass spectrometry (sp-ICP-TOFMS), which provides quantitative data on elements within particles. Using these two methods together enables informed use of geometric assumptions required by sp-ICP-TOFMS, as particle mass is typically converted to a particle diameter using assumed-spherical geometry. To validate this concept, parallel measurements to determine particle diameters were performed by FIA and sp-ICP-TOFMS on four particle suspensions: 300 nm polystyrene Eu-doped nanoparticles, 1 μm Fe-rich beads, 3 μm four element calibration polystyrene beads and 5 μm polystyrene beads. The Fe-particles obtained the highest percent difference from the manufacturer's nominal diameter, as the mean diameter obtained by FIA was overestimated by 21% and sp-ICP-TOFMS underestimated the mean diameter by 20.7%. Two types of particles were selected to test the effect of varying the particle number concentrations (PNC) on sizing accuracy, and both methods accurately sized each particle population at the PNC expected. Single particle analysis of carbon has continued to be a popular research topic, with direct applications to environmental pollutants in terms of nano- and micro- plastics. Real-world plastic particles were studied, and FIA's measured circularity values demonstrated that the particles deviated from spherical geometries, therefore sp-ICP-TOFMS data should be interpreted as mass-based rather than size-based. Combining these techniques enables improved interpretation of particle populations and evaluation of particle sizes.

Single particle-inductively coupled plasma- mass spectrometry (SP-ICP-MS) is a powerful technique for characterization of the elemental and isotopic composition of individual particles. In this work, the capabilities of the newest generation of MC-ICP-MS with acquisition rates down to 50 ms were evaluated for single particle analysis, with a focus on isotopic precision achievable on a single-particle level. Nd (NdVO4) nanoparticles (similar to 120 nm in diameter) were used as case study and were first characterized in terms of mass (respective size) and particle number concentration by SP-ICP-TOF-MS and then by SP-MC-ICP-MS for isotopic precision. For the isotopic ratio measurements, the MC-ICP-MS performance was compared to the ICP-TOF-MS and it was found that the isotope ratio precision was increased (R-2 between 0.98 and 0.99) compared to ICP-TOF-MS (R-2 between 0.88 and 0.97). The accuracy attained on a single particle level, was compared to bulk digestion followed by MCICP-MS analysis, and the SP-MC-ICP-MS technique was able to determine the particle population average to be <4 %, percent relative differences for the (142) Nd/(144) Nd, (143) Nd/(144) Nd, (145) Nd/(144) Nd, (146) Nd/(144) Nd, and (148) Nd/(144) Nd ratios The detection limit for the SP-MC-ICP-MS approach was also assessed. When utilizing an all Faraday-cup based detection scheme the determined LOD for the measurements was 0.2 fg for Nd, per particle. Based on these results, the newest generation of MC-ICP-MS has demonstrated its utility for performing SP measurement, particularly when high precision isotopic determination is warranted.

Plastic pollution in marine environments poses ecological risks, in part because plastic debris can release hazardous substances, such as metal-based additives. While microplastics have received considerable attention as vectors of contaminants, less is known about larger macroplastics and their role in the spatial and temporal redistribution of substances. In this study, pristine, store-bought plastic items and macroplastics recovered from the North Pacific Subtropical Gyre (NPSG) were analysed using Fourier-Transform Infrared Spectroscopy (FTIR) to identify polymer types, and bulk acid digestion followed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) for total metal quantification. These techniques were complemented by high resolution elemental mapping by Laser Ablation Inductively Coupled Plasma Time-of-Flight Mass Spectrometry (LA-ICP-TOFMS). Detailed elemental maps revealed native metal distribution in pristine plastics, and evidence of both sorption and intrinsic metal depletion in weathered plastics. In particular, weathered plastics showed surface depletion of intrinsic metals, and enrichment of seawater-derived elements (e.g., Na, Mg, I). Linear regressions were used to quantify spatial distribution trends across cross sections, providing statistical support for directional gradients. Since pristine and weathered plastics were opportunistically collected, variability in product type, polymer chemistry, and weathering time limited direct comparisons. Instead, this study demonstrates the utility of LA-ICP-TOFMS for mapping elemental distribution in plastics, offering a novel analytical approach for investigating spatial metal distribution in plastics and laying the groundwork for future studies on weathering processes in marine environments.

This study provides an overview of a newly developed open source program written in Python, TOFHunter, which permits the rapid and untargeted screening of inductively coupled plasma (ICP)-time-of-flight (TOF)-mass spectrometry (MS) datasets. ICP-TOF-MS is an analytical tool capable of providing quasi simultaneous detection of all nuclides from Li to Pu. This capability has triggered an increase in studies investigating single-particle analysis in which the TOF-MS provides correlated elemental/isotopic signatures on a particle basis in time. Similarly, laser ablation mapping has seen rapid growth owing to ICP-TOF-MS's capacity to handle fast washout times (<10 ms) while providing a broad nuclide coverage. The caveat to this broad mass coverage and high time resolution comes in the form of large, overwhelming datasets. With datasets typically on the scale of gigabytes, it is easy for a user to only focus on very targeted analytes; however, this focus diminishes the opportunity offered by the TOF-MS detector. TOFHunter applies chemometric methods, principal component analysis (PCA), and interesting features finder (IFF) on ICP-TOF-MS data, allowing for investigation of correlations, major and minor variance sources, and sample screening. The unique spectra identified by the (IFF) are used to generate a list of mass peaks, which are then matched with both nuclides and potential interferences before being exported for the user to investigate. Several case studies are discussed herein, demonstrating TOFHunter's ability to screen aqueous injections, single-particle/single-cell analysis, and probe laser ablation mapping files for unique regions of interest.

This work describes an analytical procedure, single particle-inductively coupled plasma-time-of-flight-mass spectrometry (SP-ICP-TOF-MS), that was developed to determine the platinum binding efficiency of protein-coated magnetic microparticles.

Nanoplastics, solid polymer particles smaller than 1 μm, are suspected to be widely present in the environment, food and air, and may pose a potential threat to human health. Detecting nanoplastics in and associated with individual cells is crucial to understand their mechanisms of toxicity and potential harm. In this context, we developed a single-cell inductively coupled plasma time-of-flight mass spectrometry (sc-ICP-TOFMS) method for the sensitive and rapid quantification of metal-doped model nanoplastics in human cells. By providing multi-elemental fingerprints of both the nanoplastics and the cells, this approach can be advantageous in laboratory toxicological studies as it allows for the simultaneous acquisition of a full mass spectrum with high time resolution. As a proof-of-concept study, we exposed two different human cell lines relevant to inhalation exposures (A549 alveolar epithelial cells and THP-1 monocytes) to Pd-doped nanoplastics. The sc-ICP-TOFMS analysis revealed a similar dose-dependent endocytotic capacity of THP-1 and A549 cells for nanoplastics uptake, and particle internalization was confirmed by transmission electron microscopy. Moreover, single-cell quantification showed that a considerable proportion of the exposed cells (72% of THP-1; 67% of A549) did not associate with any nanoplastics after exposure to 50 μg L-1 for 24 h. This highlights the importance to include single-cell analysis in the future safety assessment of nanoplastics in order to account for heterogeneous uptake within cell populations and to identify the origins and response trajectories of nanoplastics in biological tissues. In this regard, sc-ICP-TOFMS can be a powerful approach to provide quantitative data on nanoplastics-cell associations at single cell level for a large number of individual cells.

This study describes an interlaboratory comparison (ILC) among nine (9) laboratories to evaluate and validate the standard operation procedure (SOP) for single-particle (sp) ICP-TOFMS developed within the context of the Horizon 2020 project ACEnano. The ILC was based on the characterization of two different Pt nanoparticle (NP) suspensions in terms of particle mass, particle number concentration, and isotopic composition. The two Pt NP suspensions were measured using icpTOF instruments (TOFWERK AG, Switzerland). Two Pt NP samples were characterized and mass equivalent spherical sizes (MESSs) of 40.4 ± 7 nm and 58.8 ± 8 nm were obtained, respectively. MESSs showed <16% relative standard deviation (RSD) among all participating labs and <4% RSD after exclusion of the two outliers. A good agreement was achieved between the different participating laboratories regarding particle mass, but the particle number concentration results were more scattered, with <53% RSD among all laboratories, which is consistent with results from previous ILC studies conducted using ICP-MS instrumentation equipped with a sequential mass spectrometer. Additionally, the capabilities of sp-ICP-TOFMS to determine masses on a particle basis are discussed with respect to the potential for particle density determination. Finally, because quasi-simultaneous multi-isotope and multi-element determinations are a strength of ICP-TOFMS instrumentation, the precision and trueness of isotope ratio determinations were assessed. The average of 1000 measured particles yielded a precision of below ±1% for intensity ratios of the most abundant Pt isotopes, i.e.194Pt and 195Pt, while the accuracy of isotope ratios with the lower abundant isotopes was limited by counting statistics.

The increasing release of engineered nanoparticles (NPs) into aquatic ecosystems makes it crucial to understand the interactions of NPs with aquatic organisms, such as algae. In this study, the association of CeO2 NPs with unicellular algae (Raphidocelis subcapitata) and changes to the cellular elemental profile were investigated using three exposure concentrations (1, 50, and 1000 µg CeO2/L) at two different algal growth conditions—exponential and inhibited growth (1% glutaraldehyde). After a 24 h-exposure, algal suspensions were settled by gravity and CeO2-NP/algae association was analyzed by single-cell inductively coupled plasma quadrupole mass spectrometry (sc-ICP-QMS) and ICP time-of-flight MS (sc-ICP-TOFMS). Concurrent detection of the cellular fingerprint with cerium indicated NP association with algae (adsorption/uptake) and changes in the cellular elemental profiles. Less than 5% of cells were associated with NPs when exposed to 1 µg/L. For 50 µg/L exposures in growing and inhibited cell treatments, 4% and 16% of cells were associated with CeO2 NPs, respectively. ICP-TOFMS analysis made it possible to exclude cellular exudates associated with CeO2 NPs due to the cellular fingerprint. Growing and inhibited cells had different elemental profile changes following exposure to CeO2 NPs—e.g., growing cells had higher Mg and lower P contents independent of CeO2 concentration compared to inhibited cells.

Multiple analytical techniques to measure microplastics (MPs) in complex environmental matrices are currently under development, and which is most suited often depends on the aim(s) of the research question and the experimental design. Here, we further broaden the suite of possible techniques which can directly detect MPs in suspension while differentiating the carbon contained in MPs from other natural particles and dissolved organic carbon (DOC). Single particle inductively coupled plasma mass spectrometry (sp-ICP-MS) is well suited to measuring particles at trace concentrations, and the use of ICP time-of-flight-MS (ICP-TOFMS) allows one to simultaneously monitor the entire elemental spectrum to assess the full elemental composition of individual particles through developing elemental fingerprints. Because carbon is not detected in a standard operation mode with icp TOF, a dedicated optimization was necessary. Subsequently, to assess the feasibility of monitoring 12C particle pulses for the detection of MPs in more complex natural waters, two proof-of-principle studies were performed to measure MPs in waters with environmentally relevant DOC backgrounds (≤20 mg/L) and in the presence of other carbon containing particles, here, algae. Elevated DOC concentrations did not impact the enumeration of particles in suspension, and individual MPs, single algae, and aggregates of MPs and algae were clearly distinguished. The simultaneous identification of different analytes of interest allows for multiplexed sp-ICP-TOFMS experiments utilizing elemental fingerprinting of particles and is a step forward in quantifying MPs in aqueous environmental samples.

The work described herein assesses the ability to characterize gold nanoparticles (Au NPs) of 50 and 100 nm, as well as 60 nm silver shelled gold core nanospheres (Au/Ag NPs), for their mass, respective size, and isotopic composition in an automated and unattended fashion. Here, an innovative autosampler was employed to mix and transport the blanks, standards, and samples into a high-efficiency single particle (SP) introduction system for subsequent analysis by inductively coupled plasma–time of flight–mass spectrometry (ICP-TOF-MS). Optimized NP transport efficiency into the ICP-TOF-MS was determined to be >80%. This combination, SP-ICP-TOF-MS, allowed for high-throughput sample analysis. Specifically, 50 total samples (including blanks/standards) were analyzed over 8 h, to provide an accurate characterization of the NPs. This methodology was implemented over the course of 5 days to assess its long-term reproducibility. Impressively, the in-run and day-to-day variation of sample transport is assessed to be 3.54 and 9.52% relative standard deviation (%RSD), respectively. The determination of Au NP size and concentration was of <5% relative difference from the certified values over these time periods. Isotopic characterization of the 107Ag/109Ag particles (n = 132,630) over the course of the measurements was determined to be 1.0788 ± 0.0030 with high accuracy (0.23% relative difference) when compared to the multi-collector–ICP-MS determination.

We describe the outcome of a large international interlaboratory study of the measurement of particle number concentration of colloidal nanoparticles, project 10 of the technical working area 34, "Nanoparticle Populations" of the Versailles Project on Advanced Materials and Standards (VAMAS). A total of 50 laboratories delivered results for the number concentration of 30 nm gold colloidal nanoparticles measured using particle tracking analysis (PTA), single particle inductively coupled plasma mass spectrometry (spICP-MS), ultraviolet-visible (UV-Vis) light spectroscopy, centrifugal liquid sedimentation (CLS) and small angle X-ray scattering (SAXS). The study provides quantitative data to evaluate the repeatability of these methods and their reproducibility in the measurement of number concentration of model nanoparticle systems following a common measurement protocol. We find that the population-averaging methods of SAXS, CLS and UV-Vis have high measurement repeatability and reproducibility, with between-labs variability of 2.6%, 11% and 1.4% respectively. However, results may be significantly biased for reasons including inaccurate material properties whose values are used to compute the number concentration. Particle-counting method results are less reproducibile than population-averaging methods, with measured between-labs variability of 68% and 46% for PTA and spICP-MS respectively. This study provides the stakeholder community with important comparative data to underpin measurement reproducibility and method validation for number concentration of nanoparticles.

Single-cell analysis has recently become a growing research field, and has found many applications in a variety of disciplines such as biochemistry, toxicology, metallomics, and even medical diagnosis and pharmaceutics for drug and cancer research. Although several analytical methods exist for the analysis of individual cells and their cellular content, this article focuses on highlighting the latest applications and trends in which inductively coupled plasmatime-of-flight mass spectrometry (ICP-TOF- MS) is applied to singlecell analysis. The novelty of this method relies on the determination of the elemental fingerprint through the detection of elements intrinsically present in cells. By always measuring a full mass spectrum, there is no need for compromising which analytes are measured, and nothing is missed. This is particularly beneficial for nanotoxicology studies, where a direct and complete picture of the nanoparticlecell association is provided.

Characterization of dissolved background signals as compound Poisson distributed enables more accurate and robust nanoparticle detection thresholds in single-particle ICP-TOFMS.

Microdroplets with known concentrations of element standards are introduced concomitant with NP-containing solutions into the ICP to provide online matrix-matched calibration of analyte NPs for single-particle-ICP-TOFMS.

1. ETH Zurich, Department of Chemistry and Applied Biosciences, Laboratory of Inorganic Chemistry, Vladimir-Prelog-Weg 1, 8093 Zurich, Switzerland 2. BAM Federal Institute for Materials Research and Testing, Richard-Willstätter Str. 11, 12489, Berlin, Germany 3. ETH Zurich, Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, VladimirPrelog-Weg 1, 8093 Zurich, Switzerland 4. Rauschenbuschstr. 31, 44319 Dortmund, Germany # These authors contributed equally.

Due to the rapid development of nanotechnologies, engineered nanomaterials (ENMs) and nanoparticles (ENPs) are becoming a part of everyday life: nanotechnologies are quickly migrating from laboratory benches to store shelves and industrial processes. As the use of ENPs continues to expand, their release into the environment is unavoidable; however, understanding the mechanisms and degree of ENP release is only possible through direct detection of these nanospecies in relevant matrices and at realistic concentrations. Key analytical requirements for quantitative detection of ENPs include high sensitivity to detect small particles at low total mass concentrations and the need to separate signals of ENPs from a background of dissolved elemental species and natural nanoparticles (NNPs). To this end, an emerging method called single-particle inductively coupled plasma mass spectrometry (sp-ICPMS) has demonstrated great potential for the characterization of inorganic nanoparticles (NPs) at environmentally relevant concentrations. Here, we comment on the capabilities of modern sp-ICPMS analysis with particular focus on the measurement possibilities offered by ICP-time-of-flight mass spectrometry (ICP-TOFMS). ICP-TOFMS delivers complete elemental mass spectra for individual NPs, which allows for high-throughput, untargeted quantitative analysis of dispersed NPs in natural matrices. Moreover, the multi-element detection capabilities of ICP-TOFMS enable new NP-analysis strategies, including online calibration via microdroplets for accurate NP mass quantification and matrix compensation.

High time resolution of TOFMS allows to study variations in ion transit times caused by collisions within the pressurized cell.