Analytical and Bioanalytical Chemistry最新文献

{"title":"Topographic imaging with automatic z-axis correction of Brassica oleracea var. viridis leaves by IR-MALDESI mass spectrometry imaging.","authors":"Quinn Mills, Sarah M Ashbacher, Alexandria L Sohn, David C Muddiman","doi":"10.1007/s00216-025-05820-4","DOIUrl":"10.1007/s00216-025-05820-4","url":null,"abstract":"<p><p>Mass spectrometry (MS) is a versatile technique for elucidating the chemical composition of biological samples. Beyond analysis of crude extracts, MS can be further applied to spatially resolve compounds across the area of a sample with a technique called mass spectrometry imaging (MSI). The infrared matrix-assisted laser desorption ionization (IR-MALDESI) platform combines elements of matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) to enable MSI of mammalian tissue using endogenous water in the sample as a matrix. For laser-based techniques such as IR-MALDESI, changes in topography across the sample surface cause inconsistent ablation as the sample surface moves above and below the focal plane of the laser. The localization of chemical species in plants reveals crucial information about metabolic processes as reported by Nemes and Vertes (Anal. Chem. 79 (21), 8098-8106, 2007) and biosynthetic pathways by Zou et al. (Trends in Plant Science, 2024) and can even inform selective breeding of crops as discussed by Sakurai (Breed Sci 72 (1), 56-65, 2022); however, leaf topography raises a unique challenge. Features such as veins and trichomes exhibit unique topography, but flattening risks delocalization of analytes and activation of unwanted signaling pathways, and transferring metabolites to a membrane for indirect analysis may incur delocalization and limit metabolomic coverage. To overcome these challenges, a chromatic confocal sensor probe (CA probe) was incorporated for IR-MALDESI-MSI of sections of a collard (Brassica oleracea var. viridis) leaf. The CA probe measures the height at all points of the sample, and automatic z-axis corrections (AzC) are generated from height differences to continuously raise and lower the stage. These stage height corrections keep the sample surface in focus of the laser for the duration of analysis. This method has been applied to relatively homogenous samples, but has not yet been characterized on heterogeneous leaf tissue with considerable topography. Herein, data quality is compared between MSI analyses with and without AzC applied, focusing on the localization of analytes known to be concentrated in different layers of collard leaves.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2321-2332"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11996940/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Accuracy, linearity, and statistical differences in comparative quantification in untargeted plant metabolomics using LC-ESI-Orbitrap-MS.","authors":"Christina Maisl, Rainer Schuhmacher, Christoph Bueschl","doi":"10.1007/s00216-025-05818-y","DOIUrl":"10.1007/s00216-025-05818-y","url":null,"abstract":"<p><p>High-resolution mass spectrometers, particularly when paired with liquid chromatography, are the instrument of choice for untargeted metabolomics approaches. Instruments, such as the Orbitrap, offer high sensitivity, selectivity, and exceptional mass accuracy, though they pose certain technical challenges, complicating absolute and comparative quantification. Consequently, method validation is crucial to ensure reliable results, as untargeted metabolomics approaches require the detection and quantification of a large number of metabolites in a broad dynamic range. Methods can be assessed using performance characteristics like accuracy and linearity to ensure analytical reliability. This study evaluates the suitability of untargeted metabolomics methods for discovery-based investigations. A stable isotope-assisted strategy was used with wheat extracts analyzed by a Q Exactive HF Orbitrap. Results showed that 70% of all detected 1327 metabolites displayed non-linear effects in at least one of the nine dilution levels employed. However, when considering fewer levels, 47% of all metabolites demonstrated linear behavior in at least four levels (i.e., a difference factor of 8). Moreover, the analysis further suggests that the observed abundances in less concentrated samples and those outside the linear range were mostly overestimated compared to expected abundances, but hardly ever underestimated. Consequently, during statistical analysis, which is an important step in prioritizing detected metabolites and correlating them with the biological hypothesis, the number of false-positives was not inflated, but the number of false-negatives might be increased. Generally, (non-)linear behavior did not correlate with specific compound classes or polarity, suggesting non-linearity is not easily predictable based on chemical structures.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2293-2309"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11996957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143595961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enzymatic bioelectrodes based on ferrocene-modified metal-organic layers for electrochemical glucose detection.","authors":"Lingling Dong, Xuefu Zeng, Yu Xiong, Xinxin Xiao, Dongping Zhan, Shizhen Wang","doi":"10.1007/s00216-025-05808-0","DOIUrl":"10.1007/s00216-025-05808-0","url":null,"abstract":"<p><p>Metal-organic frameworks (MOFs) are often applied for enzyme immobilization, while they are limited for bioelectrochemical applications due to poor electronic conductivity. Two-dimensional (2D) metal-organic layers (MOLs) with an ultra-thin lamellar structure can effectively shorten the electron transport path and improve the electron transfer rate. In this study, ferrocene as an electron mediator is covalently bound to a 2D-MOL (Fc-NH₂-Hf-BTB-MOL) to accelerate electron transfer between the electrode surface and enzyme. Glucose oxidase (GOx) is immobilized on the electrode modified with Fc-NH₂-Hf-BTB-MOL with the addition of chitosan and carboxylated carbon nanotubes. Electrochemical tests such as cyclic voltammetry are carried out on the glucose biosensor, which shows linear detection ranges of 5 ~ 400 μM and 3 ~ 9 mM, with a detection limit of 3.9 μM (S/N = 3). Therefore, this strategy of construction of an enzyme electrode based on 2D-MOLs with enhanced electron transfer results in a biosensor with excellent specificity and activity for practical glucose detection.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2217-2224"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143514159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation of bovine milk-derived extracellular vesicles via a capillary-channeled polymer (C-CP) fiber stationary phase.","authors":"Carolina Mata, Jerisa M Pimentel, Kun Huang, Alexis Stamatikos, R Kenneth Marcus","doi":"10.1007/s00216-025-05824-0","DOIUrl":"10.1007/s00216-025-05824-0","url":null,"abstract":"<p><p>Extracellular vesicles (EVs) are released by all cell types into the extracellular environment. A subset of EVs, known as exosomes, range in size from 30 to 200 nm and are of biochemical interest due to their function as vehicles of intercellular communication. Their ability to transport proteinaceous species and genetic material at the cellular level makes them prime candidates as vectors in gene therapies. Focusing on biotherapeutics, bovine milk-derived extracellular vesicles (MDEVs) hold particular promise as an alternative to other exosome sources for therapeutics delivery. Bovine milk poses unique challenges due to the complex colloidal matrix, composed predominantly of fats and proteins like casein, which form micelles that exhibit exosome-like characteristics, specifically size and density. When faced with complex matrices like milk, conventional size/density-based isolation methods including ultracentrifugation and size exclusion chromatography struggle to provide high purity yields on practical time and cost scales. When paired with a stepwise hydrophobic interaction chromatography (HIC) gradient, polyester (PET) capillary-channeled polymer (C-CP) fibers in column and spin-down tips formats have been used effectively to isolate exosomes from highly diverse sources. Here, PET C-CP fiber columns are demonstrated to isolate MDEVs from pre-treated raw milk, yielding concentrations of 1.5 × 10¹⁰ particles mL⁻¹ with purities of ~2 × 10¹⁰ EVs µg^-1 protein in less than 20 min. The efficacy of the isolation process is verified by a suite of characterization methods. Implementing PET C-CP fiber columns for MDEV isolation addresses the challenges associated with conventional isolation methods, holding promise for scale-up towards therapeutic applications.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2345-2359"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11996999/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143623014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of separation and configuration identification of the four tetrabenazine stereoisomers in determining their pharmacokinetics.","authors":"Jianxin Li, Haoran Li, Kai Liu, Alan Kueichieh Chang, Ying Pei, Wenbao Liu, Jiao Ai, Nan Wang, Yuhui Liu, Zhen Jiang, Lijiang Chen, Xiao Liang","doi":"10.1007/s00216-025-05813-3","DOIUrl":"10.1007/s00216-025-05813-3","url":null,"abstract":"<p><p>Tetrabenazine (TBZ) is used in the treatment of psychiatric diseases, and it works by inhibiting vesicular monoamine transporter 2 (VMAT2) protein to exert a curative effect. TBZ is administered as the mixture of stereoisomers in clinical treatment. TBZ has two chiral centers, and therefore, it has four stereoisomers, and this greatly makes it difficult to separate the stereoisomers and to identify their configurations because of their high susceptibility to structural transformation. This study aims to develop a method to resolve TBZ into four individual peaks, corresponding to the four stereoisomers (1-4). Based on the different binding affinities between TBZ stereoisomers and VMAT2, the UF-UHPLC-QQQ/MS method is used to determine the absolute configuration of TBZ stereoisomers 1 and 2. Molecular docking simulations are used to verify the accuracy of UF-UHPLC-QQQ/MS. The configurations of stable stereoisomers 3 and 4 were confirmed by electron circular dichroism (ECD). The established analytical method was applied to determine the pharmacokinetics of each TBZ stereoisomer in vivo. It was found that the stereoisomer 1 (3R,11bR-TBZ) showed better bioavailability and more excretion than the other stereoisomers. The results of tissue distribution experiments indicated a much higher content of 3R,11bR-TBZ in the brain, suggesting that it may better penetrate the blood-brain barrier and exert its therapeutic effects there. The paper addresses the complex problem of separating and identifying stereoisomers with multiple chiral centers, which is a significant challenge in pharmaceutical chemistry. And this work could provide a basis for the preparation of TBZ stereoisomers and a reference for the method of separating drugs with multichiral centers and identifying unstable drugs based on their configurations.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2253-2266"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Towards quality-assured measurements of microplastics in soil using fluorescence microscopy.","authors":"Quynh Nhu Phan Le, Crispin Halsall, Stoyana Peneva, Olivia Wrigley, Melanie Braun, Wulf Amelung, Lorna Ashton, Ben W J Surridge, John Quinton","doi":"10.1007/s00216-025-05810-6","DOIUrl":"10.1007/s00216-025-05810-6","url":null,"abstract":"<p><p>Fluorescence microscopy is increasingly seen as a fast, user-friendly, and high-throughput method for detecting microplastics (MPs) in soil; however, its effectiveness across diverse MP types and soil properties remains underexplored. This study tested a fluorescence microscopy-Nile red (NR) staining approach on eight MP types, covering both biodegradable and non-biodegradable plastics, in three size ranges (≤ 150 µm, 100-250 µm, 500-1000 µm) across loamy, clayey, and sandy soils. Each sample, processed in triplicate, underwent a relatively quick and straightforward extraction procedure involving density separation, organic digestion, and NR staining, followed by fluorescence and bright-field microscopy. A new digital image analysis pipeline using Image J was developed to expedite and (semi)automate MP quantification. Recoveries ranged from 80% to 90% for MPs with a Feret diameter of 500-1000 µm, regardless of soil type. In contrast, the recovery of smaller MPs (Feret dia. ≤ 250 µm) varied depending on the soils and plastic types: recoveries for low-density polyethylene (LDPE) reached 85% in sandy soil and 90% in loamy soil, whereas those for biodegradable polybutylene adipate terephthalate/polylactic acid (PBAT/PLA) were only 60% and 10%, respectively. The lowest recovery rate was observed in clayey soil and for biodegradable plastics. The method was tested on non-agricultural soil samples, yielding a MP mean number concentration of 20.7 ± 9.0 MPs/g for MPs sized from dia. ≥ 25 µm, comparable to Fourier transform infrared (FPA-µ-FTIR) results of 13.1 ± 7.3 MPs/g (p > 0.05). We conclude that fluorescence microscopy with NR staining and automated particle quantification offers a time-efficient, reproducible, and accurate method for MP detection in light-textured soils, whereas limitations remain for reliable MP analysis in clay-dominated soils.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2225-2238"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11996956/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"UHPLC-QTOF-MS-based metabolomics joint high-throughput RNA sequencing transcriptomics approach for the analysis of fecal and liver biological samples and application in a case study for the mechanism of Qing-Kai-Ling oral liquid in treating MASLD.","authors":"Kaiwei Cai, Zihao Chen, Jingyun Wu, Jingtao Yu, Jingheng Chen, Xiaoqin Zhou, Biyan Pan, Zhiyong Xie, Qiuyun Wang, Pei Li, Qiongfeng Liao","doi":"10.1007/s00216-025-05892-2","DOIUrl":"https://doi.org/10.1007/s00216-025-05892-2","url":null,"abstract":"<p><p>Qing-Kai-Ling (QKL) oral liquid has been increasingly used in metabolic dysfunction-associated steatotic liver disease (MASLD). However, the specific metabolic differentials and metabolic pathway mechanisms that affect the MASLD regulated by QKL remained unclear. In this study, serum biochemical analyses and hematoxylin-eosin staining of the liver revealed QKL reduced liver injury and enhanced lipid metabolism ability, respectively. To clarify the therapeutic mechanism of the QKL in the treatment of MASLD, UHPLC-QTOF-MS non-target metabolomics and RNA-Seq high-throughput sequencing analysis were used to explore the mechanism of the QKL in the treatment of MASLD from the perspective of metabolic-gene interactions. UHPLC-QTOF-MS-based untargeted metabolomics further revealed that there were 196 common differentially expressed metabolites identified among 3 groups; QKL significantly up-regulated 44 metabolites, while 11 metabolites (including N-phenylacetylglutamic acid and glycocholic acid) were downregulated significantly. Moreover, the main metabolic pathways regulated by QKL included amino acids, peptides, bile acids, carbohydrates, linoleic acids, etc. Additionally, the result of the RNA sequencing-based transcriptomics showed that a total of 984 differential genes (DEGs) were identified and 9 important DEGs were obtained. The result of the Kyoto Encyclopedia of Genes and Genomes (KEGG) demonstrated that the 984 DEGs were linked to bile acid metabolism, glycerophospholipid metabolism, insulin resistance, AMPK signaling pathway, etc. Overall, this work was the first to show that QKL regulated metabolites and genes to alleviate MASLD by the UHPLC-QTOF-MS-based untargeted metabolomics joint high-throughput RNA sequencing-based transcriptomics analysis, providing the basis and research method for the treatment of metabolic diseases by QKL and other drugs.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A dual-mode RNA-splitting aptamer biosensor for sensitive HIV Tat peptide detection via colorimetry and fluorescence.","authors":"Rui Huang, Li-Kang Yin, Can Yang, Ze-Lin Wang, Rui-Min Ni, Han-Ying Zhan, Zhi-Qi Zhang","doi":"10.1007/s00216-025-05823-1","DOIUrl":"10.1007/s00216-025-05823-1","url":null,"abstract":"<p><p>Early diagnosis of human immunodeficiency virus (HIV) is critical for effective treatment; however, traditional antibody methods encounter challenges during the infection window, and nucleic acid tests require specialized equipment. In this study, a dual-mode ribonucleic acid (RNA)-splitting aptamer biosensor was developed to target the HIV trans-activator of transcription (Tat) protein, a key HIV biomarker for viral replication throughout the infection cycle. The biosensor integrates colorimetric and fluorescent detection techniques by utilizing gold nanoparticles (AuNPs) and two types of aptamers, one labeled with carboxyfluorescein (FAM). In the presence of Tat, RNA-splitting aptamers adsorb onto AuNPs, protecting them from recombination, while the fluorescence of FAM is quenched via fluorescence resonance energy transfer (FRET). Aptamers form a ternary complex with Tat, preventing adsorption and leading to thioflavin T-induced aggregation of AuNPs, accompanied by a visible color change and fluorescence signal restoration. The biosensor demonstrated excellent sensing performance, with a linear range of 0.5-60 nM and a detection limit of 0.28 nM, successfully detecting Tat in human serum. Therefore, this low-cost dual-mode detection platform offers a promising tool for early HIV diagnosis and potential applications in clinical and point-of-care fields.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2333-2343"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Predicting bone aging using spatially offset Raman spectroscopy: a longitudinal analysis on mice.","authors":"Hongmei Dou, Wendong Sun, Shuo Chen, Keren Chen","doi":"10.1007/s00216-025-05819-x","DOIUrl":"10.1007/s00216-025-05819-x","url":null,"abstract":"<p><p>Osteoporosis, a global health concern, poses an increasing challenge due to the aging population. While dual-energy X-ray absorptiometry (DXA) scans measuring bone mineral density (BMD) remain the clinical standard for osteoporosis diagnosis, this method's inability to detect changes in bone chemical composition limits its effectiveness in early diagnosis. This study applies Raman spectroscopy on examining bone aging in Senescence Accelerated Mouse Prone 6 (SAMP6) mice compared to their senescence-resistant controls (SAMR1) over an age period from 6 to 10 months. We performed Raman spectroscopic analysis on mouse tibiae both transcutaneously and on exposed bone. Leave-one-out cross-validation combined with partial least squares regression (LOOCV-PLSR) was applied to analyze Raman spectra to predict age, BMD, and maximum torque (MT) as determined by biomechanical testing. Our results revealed significant correlations between Raman spectroscopic predictions and reference values, particularly for age determination. To our knowledge, this study represents the first demonstration of transcutaneous Raman spectroscopy for accurate bone aging prediction, showing a strong correlation with established reference measurements.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2311-2320"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143571871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two-dimensional isotopic shifts for steroid isomer delineation with high-resolution cyclic ion mobility separations.","authors":"Noah D Roberts, Gabriella Sprague, Gabe Nagy","doi":"10.1007/s00216-025-05806-2","DOIUrl":"10.1007/s00216-025-05806-2","url":null,"abstract":"<p><p>Recently, the use of mass distribution-based isotopic shifts in high-resolution ion mobility spectrometry-mass spectrometry-based separations have enabled isomer delineation by measuring the relative arrival times of their heavy and light isotopologues. However, all previous efforts to induce such shifts have focused solely on the introduction of one type of isotopic substitution for a given molecule or isomer set. Herein, for the first time, we present a two-dimensional isotopic labeling strategy where two unique derivatizations are performed on various steroid isomer molecules to induce two distinct isotopic shifts and thus simultaneously measure them in a single ion mobility separations experiment. Derivatization strategies were chosen to target two specific functional groups in these steroids (i.e., hydroxyl and carbonyl), and heavy-labeled versions of the derivatizing reagents were used to induce isotopic shifts at each of these positions. We found that isotopic shifts were orthogonal to one another, diagnostic for certain steroid isomers, and that the simultaneous analysis of two different isotopic shifts was necessary for complete characterization of each steroid isomer set. We envision this multidimensional isotopic shift strategy as a new method for delineating amongst isomeric molecules, especially those with several different functional groups causing their isomerism.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2207-2216"},"PeriodicalIF":3.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143497881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}