Slug-Flow Microextraction Mass Spectrometry for Enhanced Detection of Analytes in Human Tear Fluids using Noninvasive Microsampling and Nanoelectrospray Ionization via a Capillary.

In vivo noninvasive sampling and sensitive analysis of human tear fluids at the microliter level is an important but challenging task in investigating eye health. In this work, capillary microsampling coupled with slug-flow microextraction mass spectrometry (SFME-MS) was developed for enhanced detection of analytes in human tear fluids. As low as 1.0 µL of human tear fluid could be directly sampled using a capillary, and extraction/spray solvent was then loaded into the capillary to perform slug-flow microextraction and direct nanoelectrospray ionization (nESI) of analytes. All analytical procedures, including tear microsampling, microextraction, and ionization of analytes, were performed using a capillary. Enhanced detection of therapeutic drugs and disease biomarkers in human tear fluids was successfully demonstrated. Acceptable analytical performances including sensitivity, reproducibility, and quantitation were obtained. It is found that the use of SFME could improve the nESI-MS detection of trace analytes over 100-fold that depends on the chemical properties of analytes. Overall, this study showed that SFME-nESI-MS is a highly effective method for enhanced detection of trace analytes in tear fluids and is expected to be a potentially powerful tool in significant biological and clinical applications.

Onsite Identification and Spatial Distribution of Air Pollutants Using a Drone-Based Solid-Phase Microextraction Array Coupled with Portable Gas Chromatography-Mass Spectrometry via Continuous-Airflow Sampling.

Hazardous air pollutants can be unintentionally and intentionally released in many cases, such as industrial emissions, accidental events, and pesticide application. Under such events, the onsite operation is highly dependent on the molecular composition and spatial distribution of air pollutants in ambient air. However, it is usually difficult for people to reach hazardous and upper sites rapidly. In this work, we designed a new drone-based microextraction sampler array in which a solid-phase microextraction (SPME) fiber was mounted on drones for remote-control sampling at different spaces and was then coupled with a portable gas chromatography-mass spectrometry (PGC-MS) approach for quickly identifying hazardous air pollutants and their spatial distribution in ambient air within minutes. Acceptable analytical performances, including good sensitivity (detection limit at nanogram per liter level), reproducibility (relative standard deviation < 20%, n = 6), analytical speed (single sample within minutes), and excellent linear dynamic response (3 orders of magnitude) were obtained for direct measurement of air samples. The drone-SPME sampling mechanism of air pollutants involving an airflow adsorptive microextraction process was proposed. Overall, this drone-SPME sampling array can access hard-to-reach and dangerous environmental sites and provide air pollution distribution in different spaces, showing versatile potential applications in environmental analysis.

Paper-in-Facemask Device for Direct Mass Spectrometry Analysis of Human Respiratory Aerosols and Environmental Exposures via Wearable Continuous-Flow Adsorptive Sampling: A Proof-of-Concept Study.

Facemasks are considered safe and wearable devices that cover the human mouth and nose for filtering exhaled aerosols and inhaled environmental exposures; various chemical and environmental residues thus can remain in facemasks. Therefore, direct analysis of residues in facemasks can be used to investigate the wearer's health and behavior. Here, we developed a simple paper-in-facemask sampling method for adsorbing a wearer's respiratory aerosol and environmental exposures by fixing paper strips at the outside and inside surfaces of facemasks, and the paper strips were then analyzed by paper spray mass spectrometry (PSMS) for directly detecting adsorbed analytes without any sample pretreatment. The applicability of this device was demonstrated by directly analyzing exhaled aerosolized saliva, breath metabolites, and inhalable environmental exposures. The technical aspects, including sampling time, sampling position, paper property, and spray solvent, were investigated. The sampling process was revealed to involve a continuous-flow adsorptive mechanism. These findings motivated us to extend this work and build a wearable sampling device that is capable of simultaneously monitoring both exhaled and inhaled biomarkers in situ to investigate human health and environmental exposure. This work highlights that facemasks are promising platforms for aerosol collection and direct MS analysis, which is expected to be a promising method for monitoring human health, diseases, and behaviors.

Schirmer Paper Noninvasive Microsampling for Direct Mass Spectrometry Analysis of Human Tears.

Rapid and sensitive detection of metabolites and chemical residues in human tears is highly beneficial for understanding eye health. In this study, Schirmer paper was used for noninvasive microsampling of human tears, and then paper spray mass spectrometry (PSMS) was performed for direct analysis of human tears. Schirmer PSMS was successfully used for rapid diagnosis of dry-eye syndrome by detecting the volume and metabolites of human tears. Drugs of abuse, therapeutic drugs, and pharmacodynamics in human tears were also investigated by Schirmer PSMS. Furthermore, specific markers of environmental exposures in the air to human eyes, including volatile organic compounds, aerosol, and smoke, were unambiguously sampled and detected in human tears using Schirmer PSMS. Excellent analytical performances were achieved, including single-use, low-sample consumption (1.0 µL), rapid analysis (the whole analytical procedure completed within 3 min), high sensitivity (absolute limit of detection less than or equal to 0.5 pg, signal-to-noise ratio greater than or equal to 3), good reproducibility (relative standard deviation less than 10%, n = 3), and accurate quantitation (average deviation less than 3%, n = 3). Overall, our results showed that Schirmer PSMS is a highly effective method for direct tear analysis and is expected to be a convenient tool for human tear analysis in significant clinical applications.

Fast-switching high-voltage porous-tip electrospray ionization mass spectrometry for rapid detection of antirheumatic drugs in adulterated herbal dietary supplements.

RATIONALE: Herbal dietary supplements (HDSs) adulterated with undeclared synthetic drugs can lead to serious health problems METHODS: A fast-switching positive/negative high-voltage (+/- HV) was developed to apply on electrospray ionization mass spectrometry (ESI-MS) with porous tips for rapid screening of five antirheumatic drugs in antirheumatic HDSs. The fast-switching (switch-time: 100 ms) negative and positive ions were alternately generated to perform full-MS and tandem-MS analysis, providing an effective method for rapid detection of analytes in whichever mode of detection was most suitable (negative or positive ion mode). The use of different tips and solvents was also optimized in this work. RESULTS: The limits of detection of the five antirheumatic drugs were found to be less than 0.1 ng/g (S/N > 3). The reproducibility of the five drugs was measured to be 10.0-23.3% (n = 5). A single sample analysis could be completed within 1 min. Rapid screening of a total of 28 real HDS samples collected from the market was examined by the fast-switching HV substrate-tip ESI-MS method, and the screening result was further validated by conventional liquid chromatography/mass spectrometry. CONCLUSIONS: Overall, our results demonstrated that fast-switching HV substrate-tip ESI-MS is a rapid, reliable, and effective method for simultaneous screening of various analytes in complex samples.

Direct mass spectrometry analysis of biological tissue for diagnosis of thyroid cancer using wooden-tip electrospray ionization.

Direct mass spectrometry (MS) analysis of human tissue at the molecular level could gain insight into biomarker discovery and disease diagnosis. Detecting metabolite profiles of tissue sample play an important role in understanding the pathological properties of disease development. Because the complex matrices in tissue samples, complicated and time-consuming sample preparation processes are usually required by conventional biological and clinical MS methods. Direct MS with ambient ionization technique is a new analytical strategy for direct sample analysis with little sample preparation, and has been proven to be a simple, rapid, and effective analytical tools for direct analysis of biological tissues. In this work, we applied a simple, low-cost, disposable wooden tip (WT) for loading tiny thyroid tissue, and then loading organic solvents to extract biomarkers under electrospray ionization (ESI) condition. Under such WT-ESI, the extract of thyroid was directly sprayed out from wooden tip to MS inlet. In this work, thyroid tissue from normal and cancer parts were analyzed by the established WT-ESI-MS, showing lipids were mainly detectable compounds in thyroid tissue. The MS data of lipids obtained from thyroid tissues were further analyzed with MS/MS experiment and multivariate variable analysis, and the biomarkers of thyroid cancer were also investigated.

Direct Detection of Lysozyme in Viscous Raw Hen Egg White Binding to Sodium Dodecyl Sulfonate by Reactive Wooden-tip Electrospray Ionization Mass Spectrometry.

Direct characterization of native protein binding to ligands in raw biological samples is a challenging task, because the ligand solution might induce proteins to aggregation, flocculation and denaturation. In this work, we developed a reactive wooden-tip electrospray ionization mass spectrometry (ESI-MS) for formation and characterization of protein-ligand complexes upon rapid mixing in electrospray droplets. Raw viscous hen egg white (HEW) was directly loaded onto a wooden tip to induce spray ionization, and sodium dodecyl sulfonate (SDS) solution was directly loaded into the HEW spray by a pipette tip, and thus lysozyme-DS complexes were then formed in the electrospray droplets and were detected subsequently by mass spectrometry. The new approach was successfully applied to investigate interaction of SDS and native lysozyme in electrospray droplets of standard solution and raw egg white. Our results showed that wooden-tip ESI-MS is a promising method to form and characterize protein-ligand complexes.

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.

A high-quality genome assembly for the endangered golden snub-nosed monkey (Rhinopithecus roxellana).

BACKGROUND: The golden snub-nosed monkey (Rhinopithecus roxellana) is an endangered colobine species endemic to China, which has several distinct traits including a unique social structure. Although a genome assembly for R. roxellana is available, it is incomplete and fragmented because it was constructed using short-read sequencing technology. Thus, important information such as genome structural variation and repeat sequences may be absent. FINDINGS: To obtain a high-quality chromosomal assembly for R. roxellana qinlingensis, we used 5 methods: Pacific Bioscience single-molecule real-time sequencing, Illumina paired-end sequencing, BioNano optical maps, 10X Genomics link-reads, and high-throughput chromosome conformation capture. The assembled genome was ∼3.04 Gb, with a contig N50 of 5.72 Mb and a scaffold N50 of 144.56 Mb. This represented a 100-fold improvement over the previously published genome. In the new genome, 22,497 protein-coding genes were predicted, of which 22,053 were functionally annotated. Gene family analysis showed that 993 and 2,745 gene families were expanded and contracted, respectively. The reconstructed phylogeny recovered a close relationship between R. rollexana and Macaca mulatta, and these 2 species diverged ∼13.4 million years ago. CONCLUSION: We constructed a high-quality genome assembly of the Qinling golden snub-nosed monkey; it had superior continuity and accuracy, which might be useful for future genetic studies in this species and as a new standard reference genome for colobine primates. In addition, the updated genome assembly might improve our understanding of this species and could assist conservation efforts.