bims-metlip Biomed News
on Methods and protocols in metabolomics and lipidomics
Issue of 2026–06–28
37 papers selected by
Sofia Costa, Matterworks
  1. A Validated LC-MS/MS Method for the Determination of Perospirone in Human Plasma and Its Pharmacokinetic Application in Healthy Volunteers.
  2. Isotope-Coded Derivatization Enables Accurate LC-MS/MS Quantification of Urinary THC-COOH without Isotope-Labeled Cannabinoid Standards.
  3. Rapid LC-MS/MS Method for Targeted Assay of Creatine Deficiency Syndromes in Morocco.
  4. Validation of a New LC-MS/MS Method for Rapid Quantification of Neferine: Application to Lipid Nanocarriers.
  5. Simultaneous Quantification of Colistin A and B in Human Plasma Using a Small Volume with a High-Throughput LC-MS/MS Method.
  6. Practical Workflow for Building Local Mass Spectral Libraries for Untargeted Metabolomics.
  7. Benchmarking Lipid Quantitation from Skin Swab Sebum-Rich Samples to Establish Mass Spectrometry-Based Diagnostic Methodology.
  8. A robust dilute-and-inject ion chromatography - Tandem mass spectrometry for urinary glyphosate quantification.
  9. First performance verification of automated cobas® i601 LC-MS/MS in a high-throughput clinical laboratory using 25-hydroxyvitamin D, 5α-dihydrotestosterone, and 17α-hydroxyprogesterone.
  10. The silica-based sorbents for the mixed-mode extraction of risdiplam and its metabolite from serum samples.
  11. Quantitation of Tucatinib, a Novel Tyrosine Kinase Inhibitor in Dried Blood Spot (DBS) With LC-ESI-MS/MS: Application to a Pharmacokinetic Study in Mice.
  12. Development of Mass Spectrometry-Based SCFA Analysis Methods in Diverse Samples for Microbiome Research.
  13. Librarian: An Open-Access Web Application for High-Resolution Mass Spectral Library Assembly.
  14. Dried Blood Spots for Doping Purpose-Two-Step Protocol for Analysis of Non-Threshold Substances and Anabolic Steroid Esters From One Spot/Pebble.
  15. Metabolomic Signatures of Commercial Ready-to-Drink Beverages by Dual-Mode Untargeted LC-MS/MS.
  16. Column coupling for chiral LC-MS separation resolves triacylglycerol enantiomers and regioisomers in complex samples without flow recycling.
  17. Development of a validated UPLC-MS3 method for trace-level azoxystrobin determination in zebrafish liver tissues.
  18. Optimized SPE-HPLC-FLD Method for the Simultaneous Determination of Olaparib, Propranolol, and Furosemide in Human Urine.
  19. Regions of interest multivariate curve resolution for liquid chromatography with ion mobility high-resolution tandem mass spectrometry: analysis of plastic additives.
  20. Single-Laboratory Validation of Simultaneous Determination of Aflatoxins in Nutraceuticals following immune-affinity Column Cleanup and Liquid Chromatography Tandem Mass Spectrometry Analysis.
  21. Development and validation of an LC-MS method for quantification of sex steroid hormones in skeletal muscle.
  22. Improving Speed and Efficiency of DESI Imaging with the Xevo MRT Mass Spectrometer for Analyte Mapping.
  23. Plasma or Serum? A Pilot Evaluation of Matrix Selection for Integrated Metabolomics and Exposomics of Clinical Samples.
  24. Analytical Techniques for Baloxavir Marboxil: A Critical Review of Chromatographic, Mass Spectrometric, and Emerging Methods (2018-2025).
  25. LC-MS/MS Strategy Using Substructure Searching for the Annotation of Cyanopeptide Classes: Implications for New Compound Discovery and Environmental Monitoring.
  26. Mass Spectrometry-Based Lipidomics in Coffee: Linking Lipid Transformation to Flavor Formation and Quality Control.
  27. Rapid analysis of narrow therapeutic index drugs by sonic spray ionization mass spectrometry.
  28. Preliminary pharmacokinetic investigation of two discoidin domain receptor 2 inhibitors in murine plasma and brain using liquid chromatography-tandem mass spectrometry analysis.
  29. Comprehensive Pharmacokinetics of the Marine-Derived PDE4 Inhibitor LY104 and Its Major Metabolite M1 in Rats: A Validated LC-MS/MS Method with Sex Comparison, Multiple-Dose, Protein Binding, Metabolic Stability, and Excretion Studies.
  30. Validated analytical workflow for quantifying proline metabolism related amino acids in HaCaT cells with LC-Q-TOF-MS.
  31. LC-MS assay for quantifying ornithine decarboxylase activity in the biological matrix and cultured cells using stable isotope‑labeled ornithine.
  32. Evaluation of TIMS-derived peptide mobility measurements: calibration strategy and MS1/MS2 discrepancies.
  33. Matrix fidelity in microsampling: Plasma-first LC-MS/MS quantification of mycophenolic acid and MPAG.
  34. HPLC-MS/MS quantification of genomic deoxyuridine via differential protonation.
  35. A method for quantifying non-esterified cis-9,trans-11-conjugated linoleic acid in finger-prick plasma by LC/ESI-MS/MS with isotope-coded derivatization.
  36. Accelerating drug discovery with mass spectrometry-driven bioanalytical innovations and beyond.
  37. Simultaneous quantification of D-penicillamine, D-penicillamine disulfide, and L-cysteine-D-penicillamine disulfide in human plasma: optimization of sample preparation and mass spectrometry procedures to support bioequivalence studies.
  1. ACS Omega. 2026 Jun 16. 11(23): 34621-34635
    A Validated LC-MS/MS Method for the Determination of Perospirone in Human Plasma and Its Pharmacokinetic Application in Healthy Volunteers.
    Benkai Qin, Huina Zhang, Hangyu Zhao, Yuchen Su, Yifu Kong, Shengshuo Zhang, Luwei Wang, Yonghua Yu, Jie Gu, Tingting Lou.
      Perospirone hydrochloride hydrate is an atypical antipsychotic widely used in the management of schizophrenia. Reliable quantification of its plasma concentrations remains essential for pharmacokinetic characterization and dose optimization. A rapid, specific, and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and fully validated for the quantification of perospirone in human EDTA-K2 plasma using perospirone-d 8 as the internal standard (IS). Chromatographic separation was achieved on a Kinetex C18 analytical column (100 Å, 50 mm × 2.1 mm, 5 μm, 100 Å). The mobile phase consisted of an aqueous solution of 0.1% formic acid and 1 mM ammonium formate (MPA) and a solution of 95% acetonitrile containing 0.1% formic acid and 1 mM ammonium formate (MPB), delivered at a flow rate of 0.300 mL/min. The injection volume was 10 μL. Mass spectrometric detection was performed using electrospray ionization in positive-ion mode with multiple reaction monitoring (MRM). The method exhibited linearity over a concentration range of 0.0200-12.0 ng/mL. The validated method was successfully applied to fasting and postprandial pharmacokinetic studies following oral administration of perospirone hydrochloride tablets. The method demonstrates strong performance characteristics and practical utility, supporting its application in pharmacokinetic investigations, bioequivalence assessment, and clinical drug monitoring.
    DOI:  https://doi.org/10.1021/acsomega.6c02906
  2. J Am Soc Mass Spectrom. 2026 Jun 24.
    Isotope-Coded Derivatization Enables Accurate LC-MS/MS Quantification of Urinary THC-COOH without Isotope-Labeled Cannabinoid Standards.
    Maiko Kusano, Shimba Kawasue, Yu-Ki Sakamoto, Mahiro Yoshida, Taka-Aki Matsuyama.
      Quantitative determination of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in urine is the internationally accepted approach for verifying cannabis use in forensic toxicology. Accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis typically relies on stable isotope-labeled internal standards; however, isotope-labeled cannabinoid reference materials are costly, tightly regulated, and not uniformly accessible across laboratories. We report an isotope-coded derivatization (ICD) strategy that enables isotope-dilution-equivalent quantification of urinary THC-COOH without reliance on commercially available isotope-labeled cannabinoid standards. THC-COOH was derivatized with isopropyl-piperidine carboxylic acid hydrazide (IPPAH) and its deuterated analogue (IPPAH-d6), generating a chemically matched analyte/internal standard pair through parallel derivatization. The derivatives exhibited stable chromatographic coelution and closely matched ionization behavior under positive electrospray ionization. Quantitative LC-MS/MS analysis demonstrated linearity over 1-500 ng/mL (r2 > 0.999), with intra- and interday accuracy of 81.8-108.6% and precision below 10%. Internal standard-normalized matrix factors ranged from 97-109% with minimal variability, indicating effective compensation for matrix effects. This compensation enabled reliable quantification without extensive sample cleanup, allowing a simplified sample preparation workflow compared with conventional methods. Derivatized samples remained stable for at least 72 h in the autosampler without detectable isotopic exchange. This ICD strategy provides a quantitatively reliable alternative to conventional isotope dilution while eliminating dependence on isotope-labeled cannabinoid standards, thereby expanding the accessibility of high-confidence LC-MS/MS quantification and illustrating the broader potential of isotope-coded derivatization in forensic mass spectrometry.
    Keywords:  11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC−COOH); isotope-coded derivatization; liquid chromatography-tandem mass spectrometry; Δ9-tetrahydrocannabinol (Δ9-THC)
    DOI:  https://doi.org/10.1021/jasms.6c00110
  3. Metabolites. 2026 Jun 03. pii: 388. [Epub ahead of print]16(6):
    Rapid LC-MS/MS Method for Targeted Assay of Creatine Deficiency Syndromes in Morocco.
    Faïza Meiouet, François Boemer.
      Background: Creatine deficiency syndromes (CDS) are rare neurometabolic disorders caused by defects in creatine biosynthesis (AGAT and GAMT deficiencies) or creatine transport (SLC6A8 deficiency). Early biochemical recognition is crucial for timely treatment of AGAT and GAMT deficiencies and for improving neurodevelopmental outcomes. In Morocco, expanding the liquid chromatography-tandem mass spectrometry (LC-MS/MS) biomarker panel for inherited metabolic disorders is a priority to strengthen diagnostic capacity and reduce diagnostic delay. Methods: We developed and validated a rapid LC-MS/MS method for the simultaneous quantification of creatine (Cr), guanidinoacetate (GAA), and creatinine (Crn) in plasma and urine using isotopically labelled internal standards and a standardized sample preparation procedure. Analytical performance, including linearity, precision, accuracy, sensitivity, matrix effects, carryover, inter-sample contamination, stability, and measurement uncertainty, was assessed in accordance with ISO 15189:2022 requirements. Results: The assay showed excellent linearity across the analytical range (r2 > 0.99), with robust intra- and inter-day precision (CV < 10%). Limits of detection (LOD) were 0.05 µmol/L for Cr and 0.03 µmol/L for GAA in urine, and 0.05 µmol/L for Cr and GAA in plasma. The total run time was 1.1 min per sample, supporting high-throughput implementation. Method performance was further supported by satisfactory results in ERNDIM external quality assessment schemes. Preliminary internal reference ranges and expanded measurement uncertainty were calculated from the available anonymized dataset. Conclusions: This rapid LC-MS/MS method enables the measurement of key CDS biomarkers and contributes to expanding the LC-MS/MS biomarker panel for inherited metabolic disorders in Morocco.
    Keywords:  LC-MS/MS; creatine; creatine deficiency syndromes; guanidinoacetate
    DOI:  https://doi.org/10.3390/metabo16060388
  4. ACS Omega. 2026 Jun 16. 11(23): 34010-34016
    Validation of a New LC-MS/MS Method for Rapid Quantification of Neferine: Application to Lipid Nanocarriers.
    Yash Patidar, Himanshu Kathuria, Murali Monohar Pandey.
      Neferine, a natural alkaloid, is known for its multifaceted properties, including antioxidant, neuroprotective, anti-inflammatory, and autophagy-regulating effects. A simple, robust, specific, and sensitive analytical method is a key requirement for quantifying an analyte. Although a few methods have been reported for simultaneous estimation, none are available for the quantification of neferine alone in pharmaceutical or nutraceutical formulations. Whereas, the quantification of drugs alone is considered as a valuable requirement in the preformulation and formulation studies. In this research work, we report the development and validation of a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the accurate quantification of neferine in bulk and formulations. The chromatographic separation was achieved using a UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm) under an isocratic elution method. The mobile phase consisted of acetonitrile (80%) and 10 mM ammonium acetate with 0.1% formic acid (20%). The flow rate of the mobile phase was set at 0.3 mL/min. The method exhibited good linearity (R 2 = 0.9944) over a range of 25-600 ng/mL. High sensitivity of the method was demonstrated by the limit of detection (LOD) and limit of quantification (LOQ) values of 1.29 and 3.90 ng/mL, respectively. Results of the accuracy and precision study were in complete alignment with the ICH guidelines. The analytical method was found to be robust under varied analytical conditions. Furthermore, the method was successfully applied for the quantification of neferine in nanostructured lipid carriers in the diffusion studies. In conclusion, the newly developed and validated LC-MS/MS method is specific, sensitive, rapid, accurate, and precise for the quantification of neferine in bulk and formulated products.
    DOI:  https://doi.org/10.1021/acsomega.6c00933
  5. Pharmaceuticals (Basel). 2026 Jun 12. pii: 924. [Epub ahead of print]19(6):
    Simultaneous Quantification of Colistin A and B in Human Plasma Using a Small Volume with a High-Throughput LC-MS/MS Method.
    Jimin Yoon, Won Gun Kwack, Kyung-Tae Lee, Eunseo Song, Hyeon Su Kim, Ki-Ho Park, Eun Kyoung Chung.
      Background/Objectives: Colistin is a complex polymyxin antibiotic with a narrow therapeutic window and significant interindividual pharmacokinetic variability, necessitating precise concentration monitoring. Current analytical methods often utilize colistin mixtures or require large sample volumes, potentially limiting the precision and resolution of individual component quantification. This study aimed to develop a sensitive and component-specific bioanalytical assay for the simultaneous quantification of colistin A and colistin B in human plasma. Methods: A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed using pure, component-specific reference standards to ensure rigorous independent quantification of each component. Analytes were efficiently extracted from a small volume of plasma (50 µL) using solid-phase extraction. Chromatographic separation was achieved on a C18 column with a total runtime of 4 min, and detection was performed using negative-ion multiple reaction monitoring (MRM). Results: Calibration curves showed excellent linearity over a range of 0.1-20 µg/mL for both colistin A and B (R2 > 0.99). The precision (%CV ≤ 8.8%) and accuracy (86.4-105.7%) for both components met the predefined regulatory criteria. This method was clinically validated using 60 plasma samples from 15 patients, demonstrating its applicability for capturing individual concentration-time profiles within the clinically relevant range (0.323-19.579 µg/mL for colistin A and 0.065-6.132 µg/mL for colistin B). Conclusions: This validated bioanalytical assay enables precise clinical pharmacokinetic assessments in a high-throughput workflow using a small plasma volume. Therefore, it serves as a practical tool for therapeutic drug monitoring (TDM)-guided dose optimization and further clinical investigations of colistin therapy.
    Keywords:  LC–MS/MS; colistin; colistin A; colistin B; pharmacokinetics; therapeutic drug monitoring
    DOI:  https://doi.org/10.3390/ph19060924
  6. Metabolites. 2026 Jun 12. pii: 412. [Epub ahead of print]16(6):
    Practical Workflow for Building Local Mass Spectral Libraries for Untargeted Metabolomics.
    Torbjørn Norberg Myhre, Terkel Hansen, Tetiana Lutchyn, Marie Mardal, Terje Vasskog.
      Background: Metabolite identification and annotation remain major bottlenecks in untargeted metabolomics because mass spectral features often lack sufficient specificity. High-confidence annotation requires experimental validation using authentic standards analyzed under matched chromatographic and ionization conditions, providing greater reliability than in silico predictions or database matching alone. This study aimed to develop a practical and scalable workflow for constructing a high-quality mass spectral library using a commercially available analytical standards kit. Methods: A total of 603 metabolites from the MSMLS kit were organized into 42 mixtures, each containing approximately 15 compounds. Mixture design was based on molecular mass and distribution coefficient values, specifically logD at pH 3.1, with a minimum logD spacing of 0.15 to improve chromatographic separation and reduce co-elution. This strategy was used to minimize the total number of injections while maintaining spectral quality. The resulting spectra were evaluated against online spectral resources and in silico fragmentation predictions. A preliminary proof-of-concept analysis was also performed using human serum samples. Results: Using this workflow, 471 metabolites, corresponding to approximately 78% of the standards, were successfully detected and incorporated into the spectral library. Comparison with online resources and in silico fragmentation predictions demonstrated improved spectral quality and reliability. The proof-of-concept serum analysis enabled identification of endogenous metabolites using the constructed library. In addition, the robustness and applicability of the workflow were further supported by a method validation study using metabolites derived from this library. Conclusions: This workflow provides a scalable strategy for constructing mass spectral libraries that balances spectral quality with analytical throughput. By using rational mixture design and authentic standards analyzed under matched experimental conditions, the approach enables substantial metabolite coverage while maintaining data reliability and minimizing experimental effort.
    Keywords:  LC-MS/MS; MSMLS; compound discoverer; fragmentation spectra; mass spectral library; mzVault
    DOI:  https://doi.org/10.3390/metabo16060412
  7. Anal Chem. 2026 Jun 24.
    Benchmarking Lipid Quantitation from Skin Swab Sebum-Rich Samples to Establish Mass Spectrometry-Based Diagnostic Methodology.
    Minhui Zhu, Caitlin Walton-Doyle, Eleanor Sinclair, Katherine Hollywood, Monty Silverdale, Michael Morris, Perdita Barran.
      Skin swab sampling provides an accessible, noninvasive method for collecting lipid-rich material from the skin surface, with growing potential for biomarker discovery and diagnostic testing. However, quantitative sebum analysis remains challenging because of variability in the amount collected and the complexity of the lipid matrix. This study evaluates the robustness of skin swab sampling for lipid analysis and the feasibility of quantifying skin-surface components using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Multiple reaction monitoring was used to assess analytical linearity, matrix effects, and recovery following established protocols for swab collection, storage, and extraction. Robust linearity was observed for both spiked isotope-labeled internal standards (IS) and endogenous lipids across concentrations of 0.15-550 ng/mL (R2 > 0.97). IS recovery varied between lipid classes, with consistent under-recovery. Although IS corrected for analytical variability during extraction and analysis, it could not fully account for differences in the amount of material collected. Endogenous lipids were, therefore, evaluated as normalization markers. Ratios between selected triacylglycerol (TG) species were highly conserved across samples, with coefficients of variation below 15% for TG 45:1/TG 47:1, TG 46:1/TG 48:1, and TG 47:1/TG 48:1. These ratios were stable across individuals and were independent of disease status. The reproducibility of TG recovery and stability of TG ratios suggest that endogenous lipid normalization can mitigate sampling and extraction variability. This approach provides a framework for improving quantitative reliability in skin swab lipidomics and represents a big step toward the development of reproducible mass spectrometry workflows for skin-swat-based biomarker studies.
    DOI:  https://doi.org/10.1021/acs.analchem.5c08056
  8. MethodsX. 2026 Dec;17 104001
    A robust dilute-and-inject ion chromatography - Tandem mass spectrometry for urinary glyphosate quantification.
    Mari-Vorgan Louyer, Cécile Jabalot, Arthur David, Fabien Mercier.
      Glyphosate is one of the most widely used herbicides in the world, whose potential effects on human health are still the subject of debate today. To assess the associated risks, reliable biomonitoring data are required using validated robust and sensitive analytical methods. In this context, an ion chromatography coupled with tandem mass spectrometry (IC-MS/MS) method was developed to quantify glyphosate in human urine. The method is based on direct injection of diluted urine samples. A labeled internal standard (13C2 15N1 glyphosate) is added prior to injection to correct for losses or matrix effects during the analysis. The method limit of quantification (LOQ) is 0.1 ng/mL in urine. The working range was validated between 0.1 and 20 ng/mL. Method trueness was assessed at three concentration levels (LOQ, 2 and 20 ng/mL), and a full validation of the method was performed to characterize the analytical performance in terms of precision and measurement uncertainty.•Direct injection ion chromatography coupled with tandem mass spectrometry•Validation of the method according to international standards.
    Keywords:  Biomarker of exposure; Environmental health; Herbicide; Human biomonitoring; IC-MS/MS
    DOI:  https://doi.org/10.1016/j.mex.2026.104001
  9. Clin Chem Lab Med. 2026 Jun 23.
    First performance verification of automated cobas® i601 LC-MS/MS in a high-throughput clinical laboratory using 25-hydroxyvitamin D, 5α-dihydrotestosterone, and 17α-hydroxyprogesterone.
    Nathalie Eckel, Karsten Stemmerich, Detlef Lutz, Karina Morch, Tanja Westenberger, Anja Blödel, Beate Schmidt, Annika Werle, Sebastian Wildt, Sarah Filbrecht, Eberhard Wieland, Oliver Harzer, Maria Shipkova.
       OBJECTIVES: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the analytical gold standard for measuring many parameters in clinical laboratories but remains labor-intensive, time-consuming, and insufficiently standardized. Automating LC-MS analytics is one way to integrate the advantages of the methodology into laboratory routine. This study aimed to evaluate the performance of the fully automated LC-MS/MS analyzer cobas® i601 for 25-hydroxyvitamin D (25(OH)D), 5α-dihydrotestosterone (DHT), and 17α-hydroxyprogesterone (17-OHP), and to assess its suitability for routine clinical use.
    METHODS: Analytical performance of the cobas® i601 was verified for 25(OH)D, DHT, and 17-OHP according to established quality standards by assessing precision, accuracy, linearity, limit of quantitation (LOQ), and measurement uncertainty. Comparability with existing certified or validated routine methods as well as consistency of two chromatographic lines within the instrument were assessed using Passing-Bablok regression and Bland-Altman analysis.
    RESULTS: Satisfactory analytical performance was observed for all analytes on both chromatographic lines. Imprecision and bias relative to target values fulfilled the acceptance criteria, with values ≤4.2 % and ≤6.0 %, respectively. Linearity and LOQ claimed by the manufacturer were confirmed. Strong agreement with our conventional LC-MS/MS methods was observed for 25(OH)D and with LC-MS/MS data from external quality assessment schemes for 17-OHP. In contrast, 17-OHP patient sample concentrations were considerably lower and DHT concentrations were slightly lower compared to the radioimmunoassay (DIAsource) and the laboratory-developed LC-MS/MS method we used as references, respectively.
    CONCLUSIONS: The fully automated cobas® i601 LC-MS/MS analyzer provides robust analytical performance and its results are both precise and accurate as well as comparable to established LC-MS/MS methods. It enables, for the first time, an efficient high-throughput vitamin D and steroid testing by random-access mass spectrometry in routine clinical laboratories, without having to rely on specially trained personnel.
    Keywords:  17-hydroxyprogesterone; 25-hydroxyvitamin D; 5α-dihydrotestosterone; analytical performance; automation; liquid chromatography-tandem mass spectrometry
    DOI:  https://doi.org/10.1515/cclm-2026-0421
  10. J Chromatogr A. 2026 Jun 23. pii: S0021-9673(26)00539-X. [Epub ahead of print]1783 467210
    The silica-based sorbents for the mixed-mode extraction of risdiplam and its metabolite from serum samples.
    Natalia Balińska, Anna Lemska, Maria Mazurkiewicz-Bełdzińska, Sylwia Studzińska.
      Analysis of risdiplam and its main metabolite is critical for pharmacokinetic studies and therapeutic drug monitoring in spinal muscular atrophy. However, current analytical methods face challenges due to low analyte stability, limited selectivity, and pronounced matrix effects, with only four studies published to date. In this work, a comprehensive analytical strategy was developed for the first time, combining optimized chromatographic separation with a novel sample preparation approach. The effects of methanol content, salt concentration, and pH on retention, peak symmetry, and resolution were systematically investigated. Chromatographic conditions were optimized using a central composite design, enabling rapid selection of mobile phase and achieving efficient separation of both compounds within 3.5 min. General trends in mobile phase composition were identified, indicating optimal performance at either low salt concentration and high pH or high salt concentration and low pH. For sample preparation, dispersive solid-phase extraction was introduced using newly synthesized hydrophobic and mixed-mode sorbents designed to provide hydrophobic, π-π, electrostatic, and hydrogen-bonding interactions. The extraction procedure was extensively optimized with respect to sorbent type and elution conditions (solvent composition and the addition of salt, formic acid, or ammonia). Compared with conventional non-polar sorbents, mixed-mode materials containing alkylamide, benzoic acid, cholesterol, and residual aminopropyl groups demonstrated superior recoveries and reproducibility. Finally, an alkylamide sorbent with acidified methanol-water elution was selected. The method enabled direct extraction from serum without protein precipitation, yielding recoveries of 86% and 53% for risdiplam and its metabolite, respectively. The proposed strategy provides a robust, selective, and efficient approach for risdiplam bioanalysis.
    Keywords:  Dispersive solid phase extraction (dSPE); High performance liquid chromatography; Hydrophobic-hydrophilic sorbents; Metabolite; Risdiplam; Serum
    DOI:  https://doi.org/10.1016/j.chroma.2026.467210
  11. Biomed Chromatogr. 2026 Aug;40(8): e70527
    Quantitation of Tucatinib, a Novel Tyrosine Kinase Inhibitor in Dried Blood Spot (DBS) With LC-ESI-MS/MS: Application to a Pharmacokinetic Study in Mice.
    Abhishek Dixit, Arunava Das.
      Tucatinib (Irbinitinib, ARRY-380), a potent oral, selective HER2 kinase inhibitor that the FDA, has approved for the treatment of HER2-positive metastatic breast cancer and colorectal cancer. Tucatinib provides a powerful, targeted therapy for HER2-positive cancers, showing substantial benefits in survival and disease control, especially for patients with difficult-to-treat brain metastases, extending options beyond standard chemotherapy. This research aimed to create and validate a new rapid, sensitive, and specific LC-MS/MS technique for measuring tucatinib in dried blood spots (DBS) from mice, with afatinib serving as an internal standard (IS) following regulatory guidelines in the linearity range from 0.178 to 1009 ng/mL (r > 0.990). The chromatographic separation of tucatinib and IS was achieved using the Acquity BEH C18 column with an isocratic mobile phase at a flow rate of 0.6 mL/min within 1.15 min total run time and 2 μL injection volume. Detection and quantification of tucatinib and the IS was done by using a triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode. Tucatinib remained stable under various storage conditions. Comparison of DBS versus plasma samples concentrations showed a strong correlation, suggesting that DBS can serve as a valid alternative to plasma for pharmacokinetic evaluation.
    Keywords:  DBS; LC–MS/MS; method validation; pharmacokinetics; tucatinib
    DOI:  https://doi.org/10.1002/bmc.70527
  12. Life (Basel). 2026 Jun 09. pii: 974. [Epub ahead of print]16(6):
    Development of Mass Spectrometry-Based SCFA Analysis Methods in Diverse Samples for Microbiome Research.
    Chaeeun Park, Md Abdur Rahim, Indrajeet Barman, Hanieh Tajdozian, Youjin Yoon, Sukyung Kim, Mijung Kim, Hoonhee Seo, Ho-Yeon Song.
      With the growing interest in the microbiome, short-chain fatty acids (SCFAs) have emerged as key metabolites due to their critical roles in host physiology, including immune regulation, energy homeostasis, and inflammatory control. As a result, the accurate quantification of SCFAs in various biological samples has become increasingly important. However, reliable and standardized methods for measuring SCFAs across different sample types remain underdeveloped, highlighting the need for methodological refinement. To address this need, we optimized two analytical methods, headspace GC-MS and GC-MS/MS, for SCFA quantification. These techniques were applied to a range of biological matrices, including pure microbial cultures, low-abundance animal liver, animal feces, and standardized simulated human fecal samples. The headspace GC-MS approach enables direct analysis with minimal sample preparation, thereby enhancing throughput and ease of use. In contrast, the GC-MS/MS method, involving methanol extraction, alkaline treatment, and derivatization with MTBSTFA, offers superior sensitivity and precision, making it particularly suitable for small-volume and low-abundance samples. Together, these optimized protocols provide robust, sensitive platforms for profiling SCFAs across diverse biological matrices, facilitating a deeper understanding of microbiome-host interactions and supporting future translational applications.
    Keywords:  GC-MS/MS; biological matrices; headspace analysis; microbiome; short-chain fatty acids (SCFAs)
    DOI:  https://doi.org/10.3390/life16060974
  13. Metabolites. 2026 Jun 22. pii: 433. [Epub ahead of print]16(6):
    Librarian: An Open-Access Web Application for High-Resolution Mass Spectral Library Assembly.
    Jacob Ahlberg Weidenfors, Bénilde Bonnefille, Stefano Papazian.
      Background: Confident chemical annotation in nontarget small-molecule mass spectrometry critically depends on the availability of high-quality tandem mass spectral (MS2) reference libraries. While community efforts have driven significant expansion of open-access repositories, technical challenges in assembling standardized, metadata-rich records continue to limit broader participation, underscoring the need for improved computational tools to assist contributors. Methods: To promote the creation and sharing of standardized reference MS2 spectral records, we have developed Librarian, a free, open-access web application designed for rapid and scalable assembly of high-resolution MS2 libraries. Librarian integrates automated retrieval and harmonization of chemical identifiers and metadata from PubChem, compound mixture design for high-resolution mass spectrometry (HRMS) acquisition, and assembly of curated MS2 spectra into repository-ready records compatible with public spectral databases. Results: Through a simple in-browser interface, Librarian offers a flexible end-to-end workflow compatible with popular open-source pre-processing tools to lower technical barriers and facilitate broader community participation in library development. As a demonstration, we used Librarian to create and deposit a spectral library comprising over 1500 new MS2 records into MassBank, which was further applied in retrospective analysis of environmental datasets. Conclusions: Librarian streamlines the creation of standardized, metadata-rich and repository-ready MS2 reference records. Addressing a key bottleneck in community spectral library development and sharing, Librarian supports the continued growth of open-access resources for metabolomics, exposomics, and environmental mass spectrometry. The Librarian web application is publicly accessible via the SciLifeLab Serve platform.
    Keywords:  FAIR; HRMS; MS-DIAL; MassBank; NTA; exposomics; mass spectrometry; metabolomics; open-source; spectral database
    DOI:  https://doi.org/10.3390/metabo16060433
  14. Biomed Chromatogr. 2026 Aug;40(8): e70531
    Dried Blood Spots for Doping Purpose-Two-Step Protocol for Analysis of Non-Threshold Substances and Anabolic Steroid Esters From One Spot/Pebble.
    B Stojanovic, D Pecher, V Jeitler, S Portenschlager, T Geisendorfer, G Gmeiner.
      Analytical performance in anti-doping science continues to evolve through the identification of novel metabolites, improved detection sensitivity, and the use of alternative biological matrices. In recent years, dried blood spots (DBS) have gained increasing attention in doping control due to advantages such as improved analyte stability and simplified, cost-effective sample collection, shipment, and storage. However, DBS sampling is inherently limited by the small sample volume and the restricted number of spots available for analysis. Therefore, sensitive and comprehensive analytical strategies are required to maximize the information obtained from a single DBS or Tasso M20 devices. This study aimed to develop and validate a two-step analytical workflow enabling the detection of 91 target compounds, including selected non-threshold substances (NTS) and anabolic steroid esters (ASE), in two dried blood matrices: cellulose-based DBS cards and volumetric absorptive microsampling devices (Tasso M20 pebbles). The protocol comprises extraction with a methanol-acetonitrile mixture (4:1, v/v), followed by LC-MS/MS analysis for NTS. Subsequently, the extract is evaporated, derivatized with Girard P reagent, and analyzed by LC-MS/MS for ASE detection. The validated method proved suitable for qualitative detection of the selected prohibited substances.
    Keywords:  LC/MS/MS; anabolic steroid esters; dried blood spot; non‐threshold substances; two‐step protocol
    DOI:  https://doi.org/10.1002/bmc.70531
  15. Metabolites. 2026 Jun 10. pii: 404. [Epub ahead of print]16(6):
    Metabolomic Signatures of Commercial Ready-to-Drink Beverages by Dual-Mode Untargeted LC-MS/MS.
    Ivana Blaženović, Kara Bresnahan, Shunyang Wang.
      Background: The rapid expansion of functional ready-to-drink (RTD) beverages-formulated with prebiotic fibers, botanical extracts, and reduced sugar-has outpaced systematic characterization of their small-molecule composition. Methods: We applied dual-mode untargeted high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS), integrating hydrophilic interaction (HILIC) and reversed-phase C18 separations, to profile five commercial RTD beverages spanning distinct formulation categories: Coca-Cola®, Poppi® Orange, OLIPOP® Cream Soda, Pure Leaf® Unsweetened Black Tea, and BeePop™ Peach + Orange Blossom Honey. Results: Across all products, 478 compounds were structurally annotated at Metabolomics Standards Initiative (MSI) Levels 1 and 2, of which 42 matched compounds with reported bioactivity in a curated literature-based reference database. Seventeen compounds-including the NAD+ precursor trigonelline and multiple B vitamins-were detected across all five products. The number and diversity of compounds with reported bioactivity varied substantially by product and correlated with botanical ingredient complexity. Conclusions: This work presents a qualitative molecular survey of the RTD beverage category using standardized, dual-mode untargeted metabolomics, providing a reference dataset for future targeted quantitation studies.
    Keywords:  LC–MS/MS; bioactive compounds; functional beverages; molecular profiling; polyphenols; ready-to-drink beverages; untargeted metabolomics
    DOI:  https://doi.org/10.3390/metabo16060404
  16. Talanta. 2026 Jun 23. pii: S0039-9140(26)00858-1. [Epub ahead of print]311 130202
    Column coupling for chiral LC-MS separation resolves triacylglycerol enantiomers and regioisomers in complex samples without flow recycling.
    Jenny M Nilsson, David Balgoma, Edvin Dedering Stephansson, Sofia Landström, Curt Pettersson, David Dahlgren, Hans Lennernäs, Mikael Hedeland.
      The positional distribution of fatty acids in the glycerol of triacylglycerols (TGs) gives multiple regioisomeric and enantiomeric species with different absorption, metabolism and tissue distribution. This diversity is further enhanced by the positional isomers of double bonds in these fatty acids which present an analytical challenge. To improve the determination of this plethora of isomers, we coupled chiral liquid chromatography with electrospray ionization high-resolution mass spectrometry. For the separation of enantiomers and regioisomers, we used an amylose tris(3-chloro-4-methylphenylcarbamate) stationary phase (Chiralpak IF-3) and evaluated the key chromatographic parameters: mobile phase composition, column temperature and flow-rate. Acetonitrile/water (98:2, v/v) as mobile phase at 35 °C and 0.15 mL/min was selected for separation of enantiomers and regioisomers. In addition, coupling three columns (750 mm) improved efficiency and resolution for the separation of key enantiomers and regioisomers. In addition, our method discriminated TGs differing in double-bond position and cis/trans configuration, which are difficult to resolve using conventional reversed-phase chromatographic systems. The separation method showed stable retention and reproducible determination of enantiomeric percentages in liver matrices. The application of the method revealed the stereochemical TG diversity in rat liver extracts that is unresolved by conventional RP chromatography. Furthermore, it enabled detection of differences in enantiomeric distribution following exposure to doxorubicin. This highlights the importance of resolving TG isomers in biological samples. In conclusion, this work reports a robust MS-compatible lipidomics chiral platform for stereospecific TG analysis providing an analytical foundation for future investigation of lipid remodelling and de novo lipogenesis at the stereochemical level.
    Keywords:  Chiral LC-MS; Chiralpak IF-3; Double-bond isomers; Enantiomeric separation; Regioisomeric separation; Stereospecific analysis; Triacylglycerols
    DOI:  https://doi.org/10.1016/j.talanta.2026.130202
  17. Anal Methods. 2026 Jun 22.
    Development of a validated UPLC-MS3 method for trace-level azoxystrobin determination in zebrafish liver tissues.
    Yufeng Guo, Chunpeng Feng, Mengchuan Lai, Meichen Liu, Yalin Xi, Lei Yin, Meiyun Shi.
      Azoxystrobin is a widely used strobilurin fungicide. Its environmental persistence and potential toxicity to aquatic organisms demand accurate trace-level quantification in biological tissues. However, its sensitive and specific determination in small tissue samples using conventional methods remains challenging. In this study, a novel high-sensitivity analytical method based on ultra-performance liquid chromatography coupled with triple-stage mass spectrometry (UPLC-MS3) was established for the trace determination of azoxystrobin in zebrafish liver tissues. Following protein precipitation extraction, chromatographic separation was performed on a C18 column using a gradient of 0.1% aqueous formic acid and acetonitrile. Detection relied on an optimized MS3 transition (m/z 404.0 → 371.9 → 344.2). The method exhibited excellent linearity (r > 0.9984) from 0.1 to 20 ng mL-1, with accuracy between -3.33% and 3.67% and precision (CV) between 5.57 and 10.19%. Consistent recoveries (94.93-106.64%) and minimal matrix effects (99.45-104.79%) were achieved across all tissue matrices. Compared to conventional MRM, MS3 scanning significantly enhanced specificity by reducing endogenous interference. The validated approach was successfully applied to tissue distribution studies in zebrafish, confirming its reliability for environmental toxicology research and providing a robust platform for investigating fungicide biodistribution in aquatic organisms.
    DOI:  https://doi.org/10.1039/d6ay00445h
  18. Biomed Chromatogr. 2026 Aug;40(8): e70529
    Optimized SPE-HPLC-FLD Method for the Simultaneous Determination of Olaparib, Propranolol, and Furosemide in Human Urine.
    Georgios Kamaris, Antonia Kalagia, Catherine K Markopoulou.
      Olaparib belongs to the PARP inhibitors and is widely used as an anticancer drug. Due to frequent side effects, it is usually co-administered with an antihypertensive, such as propranolol and/or an antidiuretic, such as furosemide. In this study, a highly sensitive HPLC-FLD method was developed for the simultaneous determination of the three APIs in urine. For their analysis, a Nucleosil C8 column (250 mm × 4.6 mm, 5 μm) at 50°C was used as the stationary phase and an isocratic system of methanol-acetonitrile-20 mM NaH2PO4 at pH 3.6, 12:20:68 v/v/v as the mobile phase. The calibration curves for the analytes were generated both in urine and in the pure diluent. The reliability of the method was evaluated according to ICH guidelines in terms of accuracy and repeatability. For sample purification, protein precipitation techniques involving the addition of organic solvents, acids, or salts, as well as the freezing-out technique, were studied. Liquid-liquid extraction, using various solvents, and solid-phase extraction (SPE) were also tested, where the latter was found more suitable. SPE optimization was achieved using experimental design and a D-optimal mixture design. Reliable analyte recoveries demonstrate the capability of the proposed method, making it a suitable tool for biological studies.
    Keywords:  furosemide; olaparib; propranolol; solid‐phase extraction (SPE); urine
    DOI:  https://doi.org/10.1002/bmc.70529
  19. Anal Bioanal Chem. 2026 Jun 22.
    Regions of interest multivariate curve resolution for liquid chromatography with ion mobility high-resolution tandem mass spectrometry: analysis of plastic additives.
    Ana Torres-Agullo, Roma Tauler, Silvia Lacorte, Tytus D Mak, Yamil Simón-Manso.
      Ion mobility (IM) enhances liquid chromatography-mass spectrometry identification capabilities by providing an additional dimension that improves peak capacity, mass detection, and sensitivity, being particularly valuable for untargeted analysis. However, the high dimensionality and large amount of information in the datasets obtained in this type of analysis are a big challenge for data processing, slowing its broader application and potential. The regions of interest multivariate curve resolution (ROIMCR) is a chemometrics approach based on the bilinear model intrinsic structure of the data. It represents an effective strategy for efficient feature extraction, deconvolution, and resolution of overlapping signals in complex datasets without losing relevant information and maintaining instrumental mass accuracy. In this work, the application of ROIMCR is shown for the first time for liquid chromatography ion mobility mass spectrometry (LC-IM-MS) in data-dependent acquisition (DDA) mode, including MS1 and MS2 datasets for the analysis of plastic additives. An improvement in the discrimination of isomeric species and minimization of interferences was shown by working with mobility profiles. IM-MS data from five microsphere standards and three polyethylene (PE) plastic leachates, containing complex mixtures of unknown additives, were analyzed and resolved, and collision cross-section (CCS) values were calculated for each component. Using ROIMCR, 14 different plastic additives, including phthalates, siloxanes, and phosphates, present in the original PE polymers, were identified. Our findings highlight that ROIMCR provides an effective and scalable tool for resolving complex IM datasets and enhancing the identification of potential additives associated with microplastics.
    Keywords:  Collision cross-section; Ion mobility mass spectrometry; Microplastics; Non-targeted analysis; ROIMCR
    DOI:  https://doi.org/10.1007/s00216-026-06632-w
  20. J AOAC Int. 2026 Jun 26. pii: qsag059. [Epub ahead of print]
    Single-Laboratory Validation of Simultaneous Determination of Aflatoxins in Nutraceuticals following immune-affinity Column Cleanup and Liquid Chromatography Tandem Mass Spectrometry Analysis.
    Saravanan J, Santhosh Kumar J Urumarudappa, Vijay Bommuluri, Ashutosh Kumar Mittal, Peter Chang, Gary Swanson.
       BACKGROUND: Aflatoxins B1, B2, G1, and G2 are highly carcinogenic mycotoxins frequently detected in nutraceutical products. Due to their toxicological significance and the complex composition of nutraceutical matrices, sensitive and validated analytical methods are required to ensure regulatory compliance, product safety, and routine surveillance.
    OBJECTIVE: This study aimed to develop and perform single-laboratory validation of a reliable and sensitive method for the simultaneous determination of aflatoxins B1, B2, G1, and G2 in nutraceutical products, in accordance with AOAC Appendix F.
    METHOD: Samples were extracted using an organic solvent system and purified through aflatoxin-specific immunoaffinity columns to reduce matrix interference. Quantitative analysis was performed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Method validation evaluated linearity, limits of detection (LOD) and quantitation (LOQ), precision (repeatability and intermediate precision), and accuracy across three representative nutraceutical matrices.
    RESULTS: The method showed linearity for all analytes, with coefficients of determination (R2) between 0.9984 and 0.9989. The LOD and LOQ values ranged from 0.1268 to 0.6170 µg/kg and from 0.2733 to 1.1290 µg/kg, respectively. Precision studies demonstrated acceptable repeatability and intermediate precision, with relative standard deviations ≤20%. Accuracy assessment across beverage and protein-based nutraceutical products yielded recoveries between 88% and 119% at multiple fortification levels, meeting AOAC performance requirements for mycotoxin analysis.
    CONCLUSIONS: The validated LC-MS/MS method is sensitive, accurate, and precise for the determination of aflatoxins B1, B2, G1, and G2 in complex nutraceutical products. The method is fit for purpose of routine quality control, regulatory testing, and surveillance of aflatoxins in nutraceutical matrices.
    HIGHLIGHTS: Optimized sample extraction and immunoaffinity cleanup enabled sensitive LC-MS/MS determination of aflatoxins B1, B2, G1, and G2. The method was validated for linearity, LOD, LOQ, sensitivity, precision, and accuracy, and was found to be suitable for routine aflatoxin estimation in nutraceutical products.
    Keywords:  Aflatoxins; LC-MS/MS; immune-affinity cleanup; method development; quantification
    DOI:  https://doi.org/10.1093/jaoacint/qsag059
  21. Steroids. 2026 Jun 24. pii: S0039-128X(26)00085-1. [Epub ahead of print] 109823
    Development and validation of an LC-MS method for quantification of sex steroid hormones in skeletal muscle.
    Viktor Engman, Séverine Lamon, Shaun Mason.
      Sex steroid hormones are not exclusively localised in the circulation and can be found in numerous extragonadal tissues, in concentrations unrelated to the circulating fraction. Existing methodology to measure intramuscular steroid hormone concentrations includes both immune-based assays and liquid chromatography-mass spectrometry (LC-MS), the gold standard for hormone measurements. To date, no LC-MS based methods validation has been published on the measurement of intramuscular sex steroid hormones, despite clear biological relevance. Here, we describe the development and validation of a simple, high-throughput LC-MS Orbitrap method for the measurement of 10 intramuscular sex steroid hormones, namely pregnenolone, progesterone, dehydroepiandrosterone, androstenedione, testosterone, epitestosterone, dihydrotestosterone, oestrone, oestradiol, and oestriol. In brief, isotope labelled standards were added to 5-6 mg of lyophilised muscle tissue, homogenised and extracted with ethyl acetate. The extracts were dried down and sequentially derivatised with 1-methylimidazole-2-sulfonyl chloride and hydroxylamine hydrochloride to target both the phenolic hydroxy groups and oxo groups. The limit of detection was 1.0 ± 1.0 pg/mg (range 0.36-3.26 pg/mg), with a R2 > 0.99 for all analytes. Matrix effects were 90-110% for all analytes except for dihydrotestosterone (143.6%), and precision was <10 CV% for all analytes in the presence of a muscle matrix. Our method allows for 20-40 samples to be prepared in ~4 h, with a sample data acquisition time of 13 min. Moreover, our method provides the opportunity for specific analysis of steroid hormone concentrations in skeletal muscle, allowing target tissue specificity instead of relying on proxy measures from the circulation.
    Keywords:  Hormone; Muscle; Orbitrap; Testosterone
    DOI:  https://doi.org/10.1016/j.steroids.2026.109823
  22. Metabolites. 2026 Jun 18. pii: 429. [Epub ahead of print]16(6):
    Improving Speed and Efficiency of DESI Imaging with the Xevo MRT Mass Spectrometer for Analyte Mapping.
    Mark Towers, Emmanuelle Claude, Lisa Towers, Helen Yates, Joanne Ballantyne.
      Background: Recent technology improvements have enabled desorption electrospray ionisation (DESI) mass spectrometry imaging to achieve down to 5 µm (pixel) image resolution. However, operating at this resolution introduces challenges, particularly regarding increased total analysis time and the need for sufficient instrument sensitivity to detect analytes from very small tissue areas. Methods: High mass and image resolution DESI imaging was performed on rat brain tissue using a Xevo™ MRT benchtop mass spectrometer equipped with a multi-reflecting time-of-flight mass analyser and a DESI XS source. Data acquisition was conducted at speeds of up to 100 Hz. Sensitivity was assessed using a dilution series of five Active Pharmaceutical Ingredients (APIs) spotted onto porcine liver tissue. Signal detection limits were evaluated using extracted ion chromatograms (XICs) with signal-to-noise (S/N) calculations against blank samples. Additionally, enhanced duty cycle (EDC) was applied to evaluate improvements in analyte signal intensity across specific mass ranges in both positive and negative ionisation modes. Results: At acquisition speeds of up to 100 Hz, excellent data quality was achieved, with signal intensity remaining suitable for analytical applications. All five tested APIs were detectable at concentrations of 25 pg/mm2. Three of the five compounds were further detected at concentrations as low as 2.5 pg/mm², with signal-to-noise ratios greater than 5. The application of EDC resulted in a significant increase in analyte signal intensity within the targeted mass ranges, particularly for small molecule endogenous metabolites and lipids, in both ionisation modes. Furthermore, the system demonstrated substantially improved spectral quality, achieving mass resolution up to 100,000 FWHM. This enabled the resolution of previously indistinguishable analytes with significantly improved mass accuracy compared to systems operating at approximately 30,000 FWHM. Conclusions: The Xevo™ MRT mass spectrometer with DESI XS source enables high-resolution DESI imaging at speeds up to 100 Hz without compromising data quality or sensitivity. The system demonstrates excellent detection limits for pharmaceutical compounds and improved performance through enhanced duty cycle operation. Overall, the combination of high spatial resolution, increased mass resolution, and improved spectral quality allows for more accurate analyte differentiation, representing a significant advancement over lower-resolution systems.
    Keywords:  DESI; Xevo MRT MS; imaging; lipidomics; metabolites
    DOI:  https://doi.org/10.3390/metabo16060429
  23. Toxics. 2026 Jun 06. pii: 494. [Epub ahead of print]14(6):
    Plasma or Serum? A Pilot Evaluation of Matrix Selection for Integrated Metabolomics and Exposomics of Clinical Samples.
    Xiaowen Ji, Julian Edwards, Miaomiao Wang, Juan C Irwin, Binya Liu, Amanda M Gutierrez, Lin Li, Jeannette C Lager, Camran R Nezhat, David K Stevenson, Tomiko T Oskotsky, Marina Sirota, Dimitri Abrahamsson, Linda C Giudice, Tracey J Woodruff, Joshua F Robinson, June-Soo Park.
      Serum and plasma are the most widely used matrices in metabolomics and human biomonitoring studies; however, the optimal matrix for integrated non-targeted analysis (NTA) workflows combining metabolomics and exposomics has not been systematically evaluated. This pilot study applied parallel NTA workflows to paired serum and plasma samples from five individuals to characterize matrix-dependent differences and provide an empirical basis for matrix selection in integrated studies. Three analytical methods were employed: one metabolomic method (Method 1) using Hydrophilic Interaction Liquid Chromatography (HILIC) and Reversed-Phase Liquid Chromatography (RPLC) columns and one exposomics (Method 2) method using an RPLC column, each analyzed in both electrospray ionization (ESI) positive and negative modes. Overall, serum and plasma showed broad similarity, with substantial overlap in detected features and strong linear correlations between paired samples (R2 = 0.70-0.87). However, PCA revealed systematic differences between the two matrices along PC1 and PC2, likely attributable to matrix effects arising from coagulation-related compositional changes in serum. For metabolomics, glycerophospholipids, sphingolipids, and acylcarnitines were consistently enriched in serum, attributable to platelet activation and phospholipase release during blood coagulation, consistent with prior reports. In contrast, oxidized fatty acid species were predominantly elevated in plasma, warranting caution in oxylipin-focused studies using serum. For exogenous chemical profiling, the two matrices performed comparably, with 32 out of 36 annotated features showing no significant matrix-dependent differences (p > 0.05), including PFAS, pharmaceuticals, and diverse xenobiotics. These findings support the interchangeability of serum and plasma for broad exposomics studies. Overall, while both matrices provided broadly comparable global coverage, plasma may represent a more appropriate matrix for integrated NTA workflows, as it better preserves in vivo metabolite composition and minimizes coagulation-induced confounding, though validation in larger cohorts is needed.
    Keywords:  exposomics; matrix selection; metabolomics; non-targeted analysis; plasma; serum
    DOI:  https://doi.org/10.3390/toxics14060494
  24. Crit Rev Anal Chem. 2026 Jun 23. 1-24
    Analytical Techniques for Baloxavir Marboxil: A Critical Review of Chromatographic, Mass Spectrometric, and Emerging Methods (2018-2025).
    Samar M Mahgoub, A B M Sharif Hossain, Aya Shaban, Ahmed A Allam, Rehab Mahmoud.
       PURPOSE: Baloxavir Marboxil, a first-in-class cap-dependent endonuclease inhibitor, is a prodrug used in influenza therapy. Reliable analytical methods are essential for its quantification in pharmaceutical formulations and biological matrices to support drug development, quality control, and pharmacokinetic studies. This review evaluates current analytical approaches, and future directions.
    METHODS: A systematic literature search identified relevant analytical studies, which were critically compared based on principles, validation performance, and applicability.
    RESULTS: UHPLC-MS/MS is considered the gold standard, offering high sensitivity (LLOQ as low as 0.1 ng/mL), strong selectivity, and simultaneous determination of baloxavir marboxil and its active metabolite. Sample preparation techniques, particularly protein precipitation and liquid-liquid extraction, significantly influence method performance, while isotopically labeled internal standards effectively minimize matrix effects. The compound shows pronounced instability under alkaline conditions, whereas oxidative, thermal, and photolytic degradation are relatively limited. Emerging spectrofluorimetric methods provide greener alternatives but with reduced sensitivity and limited bioanalytical applicability.
    CONCLUSION: This review compares major analytical platforms, highlights regulatory validation requirements, and discusses clinical applications including therapeutic drug monitoring. It underscores the need for standardized, high-throughput, and sustainable analytical methods to support future research and clinical implementation.
    Keywords:  Analytical methods; Baloxavir Marboxil; UHPLC-MS/MS; green analytical chemistry; sample preparation; stability-indicating; therapeutic drug monitoring
    DOI:  https://doi.org/10.1080/10408347.2026.2687138
  25. Environ Sci Technol. 2026 Jun 20.
    LC-MS/MS Strategy Using Substructure Searching for the Annotation of Cyanopeptide Classes: Implications for New Compound Discovery and Environmental Monitoring.
    Runjie Xia, Lindsey Ahn, Michaela Burkhauser, Ross Youngs, Matthew J Bertin.
      Cyanobacterial harmful algal blooms (cyanoHABs) are a major ecological and public health concern, commonly monitored for hepatotoxic microcystins and cylindrospermopsins and neurotoxic anatoxins and saxitoxins. However, the broader suite of bioactive metabolites produced during blooms remains undercharacterized. Here, we interrogated a chromatography fraction library generated from a cyanoHAB in Muskegon, Michigan. From this library, we isolated two new micropeptins (1 and 2), including an analog bearing a bishomologated tyrosine residue, and we confirmed the structure of ferintoic acid C (3). Structures were established using complementary spectrometric and spectroscopic methods. To expand chemical space coverage beyond isolated compounds, we analyzed liquid chromatography-tandem mass spectrometry (LC-MS/MS) data using the Global Natural Products Social Molecular Networking 2 (GNPS2) Analysis Hub query language for product ion searching, enabling annotation of cyanopeptide classes and class-specific modifications across the fraction set, which provided a practical and user-friendly strategy for identifying cyanopeptide classes. One of the new micropeptins (1) exhibited moderate inhibition of neutrophil elastase, consistent with roles in ecological interactions and potential relevance to human exposure. Analysis of field samples from ongoing Lake Erie blooms showed recurring micropeptins but no evidence of microcystins. Together, these results challenge microcystin-centric assessments of bloom hazard and support expanded monitoring of nonmicrocystin cyanopeptides.
    Keywords:  LC-MS/MS; cyanoHABs; cyanopeptides; micropeptins; protease inhibition
    DOI:  https://doi.org/10.1021/acs.est.6c05607
  26. Foods. 2026 Jun 18. pii: 2196. [Epub ahead of print]15(12):
    Mass Spectrometry-Based Lipidomics in Coffee: Linking Lipid Transformation to Flavor Formation and Quality Control.
    Yanbing Wang, Xiaoyuan Wang, Ping Du, Xiaogang Liu.
      Mass spectrometry-based lipidomics has created new opportunities to investigate the role of lipids in coffee quality formation and stability across the production chain. Coffee lipids contribute to flavor precursor formation, aroma release, mouthfeel, and storage behavior, but their molecular remodeling during maturation, processing, roasting, and storage remains insufficiently integrated. This review summarizes recent progress in lipidomics methodologies relevant to coffee research, with emphasis on sample preparation, mass spectrometry platforms, data analysis, and the strengths and limitations of current lipid annotation strategies. It further examines how lipid profiles change during bean maturation, how they differ among coffee species and varieties, and how they are reshaped by postharvest processing, roasting, and storage. However, it is important to note that most of these associations are currently correlational rather than causal; direct evidence linking specific lipid species to particular sensory attributes remains limited. Existing studies suggest that lipid composition, rather than total lipid content alone, is more informative for understanding coffee quality differences and for identifying candidate markers associated with origin, processing method, roasting degree, and storage conditions. In particular, alterations in glycerolipids, glycerophospholipids, fatty acids, diterpenes, and other minor lipid constituents are increasingly associated with lipid oxidation, thermal degradation, and flavor-related transformations in coffee. However, current evidence is still limited by incomplete structural annotation, isomeric ambiguity, platform dependence, and the frequent gap between statistical discrimination and mechanistic validation. Future work integrating high-resolution mass spectrometry, ion mobility, targeted quantification, stable isotope tracing, sensory analysis, and multi-omics approaches will be essential to improve marker reliability and to clarify the functional roles of coffee lipids. Overall, lipidomics provides a promising framework for linking molecular composition with coffee quality control, traceability, and process optimization, although substantial work is still needed to establish mechanistic links to flavor formation.
    Keywords:  aroma formation; coffee; flavor formation; lipidomics; roasting; storage stability
    DOI:  https://doi.org/10.3390/foods15122196
  27. Anal Sci. 2026 Jun 23.
    Rapid analysis of narrow therapeutic index drugs by sonic spray ionization mass spectrometry.
    Bingxin Yu, Shaojun Qian, Chongyang Yao, Qingli Shen, Lei Wang.
      Narrow therapeutic index drugs (NTIDs) require precise monitoring to ensure efficacy and prevent toxicity, yet their analysis in complex biological matrices remains challenging. This study presents a novel sonic-spray ionization mass spectrometry (SSI-MS) method for the rapid and sensitive analysis of NTIDs, using tacrolimus as a model compound. The custom-built SSI source operates without high voltage or heating, relying solely on a coaxial nitrogen flow for gentle and efficient ionization. Under optimized conditions, the method achieved a low limit of quantitation (5 ng/mL) and excellent linearity (R² = 0.999) across the clinically relevant therapeutic window (5-200 ng/mL). Compared to electrospray ionization, SSI provided a cleaner background and a 1.88-fold improvement in the signal-to-noise ratio. The accuracy of this method was validated in mice serum, with recoveries ranging from 99.8% to 106.8% and relative standard deviations below 8.4%. Furthermore, the platform successfully detected tacrolimus directly in a saline matrix. These results demonstrate that SSI-MS is a powerful, high-performance tool for the rapid and reliable quantification of NTIDs, offering significant potential for advancing therapeutic drug monitoring in clinical practice.
    Keywords:  Mass spectrometry; Narrow therapeutic index drugs; Sonic spray ionization; Tacrolimus
    DOI:  https://doi.org/10.1007/s44211-026-00941-x
  28. Sci Rep. 2026 Jun 24.
    Preliminary pharmacokinetic investigation of two discoidin domain receptor 2 inhibitors in murine plasma and brain using liquid chromatography-tandem mass spectrometry analysis.
    Paul Gueroue, Audrey Burban, Ahmad Sharanek, Guillaume Bougueon, Stéphane Bouchet, Dominique Ducint, Mathieu Molimard, Sarah Djabarouti, Joris Guyon.
      Owing to its involvement in extracellular matrix-glial cell interactions, discoidin domain receptor 2 (DDR2) has emerged as a promising pharmacological target in various human diseases, including cancers and neurodegenerative disorders. Two allosteric inhibitors, WRG-28 and DDR2-IN-1, selectively target DDR2. Their therapeutic efficacy is likely to depend on blood-brain barrier (BBB) penetration; however, a lack of analytical methods has so far left their pharmacokinetic properties and BBB permeability profiles largely unexplored. In this study, a liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of WRG-28 and DDR2-IN-1 in plasma and brain tissue. The extraction procedure was simple and based on protein precipitation followed by lipid removal. This analytical method met the acceptance criteria of the European Medicines Agency guidelines. The calibration range was linear from 1 to 1000 ng/mL for both compounds. In addition, five prediction software tools were used to estimate pharmacokinetic parameters relevant to the BBB penetration of the two compounds. Finally, the method was applied in preclinical pharmacokinetic studies using elacridar, an efflux transporter inhibitor.
    Keywords:  Brain; Discoidin domain receptor 2 (DDR2); Elacridar; LC-MS/MS; Pharmacokinetic; Tyrosine kinase inhibitor (TKI)
    DOI:  https://doi.org/10.1038/s41598-026-59287-z
  29. Mar Drugs. 2026 Jun 15. pii: 215. [Epub ahead of print]24(6):
    Comprehensive Pharmacokinetics of the Marine-Derived PDE4 Inhibitor LY104 and Its Major Metabolite M1 in Rats: A Validated LC-MS/MS Method with Sex Comparison, Multiple-Dose, Protein Binding, Metabolic Stability, and Excretion Studies.
    Xiaochen Niu, Jun Zhao, Deqi Ding, Wei He, Guanhua Du, Jiejie Hao, Jianchun Zhao.
      LY104 (previously designated as B7) is a selective phosphodiesterase 4 inhibitor with promising activity against chronic obstructive pulmonary disease. We previously reported its single-dose pharmacokinetics and tissue distribution in rats. In the present study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous quantification of LY104 and its major metabolite M1 in rat plasma following ICH M10 guidelines. The method showed excellent linearity over 20-1200 ng/mL for both analytes, with retention times of 2.85 min (LY104) and 3.22 min (M1). Using this method, we extended our previous work in several directions. Re-analysis of previously published single-dose pharmacokinetic and tissue distribution data revealed no significant sex differences for LY104. Newly generated multiple-dose studies (1 mg/kg daily for 7 days) demonstrated no accumulation of LY104 or M1. The pharmacokinetic profile of M1 was quantified for the first time. Comprehensive in vitro investigations included plasma and liver microsomal stability, plasma protein binding, and excretion studies. This systematic preclinical pharmacokinetic characterization of LY104 and M1, incorporating re-analysis of existing data with sex stratification, newly generated multiple-dose and metabolite data, excretion studies, and comprehensive in vitro investigations, provides useful information to support further drug development and clinical trial design.
    Keywords:  LC-MS/MS; LY104; excretion; metabolic stability; pharmacokinetics; phosphodiesterase 4 inhibitor; sex difference
    DOI:  https://doi.org/10.3390/md24060215
  30. Sci Rep. 2026 Jun 22.
    Validated analytical workflow for quantifying proline metabolism related amino acids in HaCaT cells with LC-Q-TOF-MS.
    Szymon Plewa, Magdalena Nizioł, Katarzyna Bielawska, Agnieszka Klupczynska-Gabryszak, Jan Matysiak, Wojciech Miltyk.
      It is known that altered amino acid metabolism can influence immunological responses, thus studying the molecular mechanisms underlying these changes requires analytical methods for reliably determining the quantities of individual analytes. To address the ongoing need for quantitative assessment of amino acid composition, a validated method for determining their quantities was proposed. This study presents the development and validation of a novel method for the quantitative analysis of selected amino acids using high-performance liquid chromatography coupled with a quadrupole time-of-flight (Q-TOF) mass spectrometer equipped with an electrospray ionization source. The method has been satisfactorily validated in terms of linearity, range, precision, accuracy, and stability. Its applicability to biological matrices was demonstrated by the successful quantification of amino acids in HaCaT cell lysates. Owing to the use of a robust, state-of-the-art analytical platform based on Q-TOF technology, which enables high-resolution mass detection, the method is readily adaptable for the inclusion of additional analytes and can be extended to other complex biological samples. The applicability of the method for the simultaneous determination of selected amino acids was demonstrated, and the method successfully applied to the HaCaT cell lysates.
    Keywords:  Amino acids; Cell culture; Liquid chromatography-mass spectrometry; Metabolomics; Proline
    DOI:  https://doi.org/10.1038/s41598-026-58884-2
  31. Amino Acids. 2026 Jun 21.
    LC-MS assay for quantifying ornithine decarboxylase activity in the biological matrix and cultured cells using stable isotope‑labeled ornithine.
    Yuma Shiomi, Naoya Ogawa, Kentaro Takahama, Atsushi Murai, Kyohei Furukawa.
      Polyamines (putrescine, spermidine, and spermine) are essential for animal health and development, and their intracellular levels must be tightly regulated to maintain normal cellular functions. Ornithine decarboxylase (ODC) catalyzes the rate-limiting decarboxylation step in polyamine biosynthesis, and thus accurate assessment of its activity is vital for studies of polyamine metabolism. However, conventional ODC assays rely on radiolabeled substrates and require specialized facilities for radioactive handling. To overcome these limitations, we developed a sensitive and non-radioactive ODC assay using stable isotope-labeled ornithine in combination with liquid chromatography-mass spectrometry (LC-MS). In this protocol, animal tissues (e.g., 0.5 g) or cultured cells (e.g., 2.0 × 106 cells) are homogenized, and cytosolic fractions are prepared by centrifugation. These fractions are incubated with d7-ornithine under 37 °C conditions, and the produced d7-putrescine is derivatized with dansyl chloride and quantified by LC-MS analysis. Reaction samples showed a distinct peak corresponding to d7-putrescine, whereas negative control displayed negligible signals. Moreover, d7-spermidine and d7-spermine were not detectable under these conditions, indicating d7-putrescine production directly reflects ODC activity. We optimized reaction time and substrate concentrations to ensure linearity and precision, and confirmed that the assay responds appropriately to pharmacological inhibition of ODC. Collectively, this protocol provides a practical, sensitive, and non-radioactive method for quantifying ODC activity in both animal tissues and cultured cells, and it offers an accessible tool for polyamine metabolism research.
    Keywords:  DFMO; Dansyl chloride; LC–MS; Ornithine decarboxylase; Polyamines; Stable isotope
    DOI:  https://doi.org/10.1007/s00726-026-03539-9
  32. Anal Bioanal Chem. 2026 Jun 26.
    Evaluation of TIMS-derived peptide mobility measurements: calibration strategy and MS1/MS2 discrepancies.
    Anh Tran, Tytus Mak, Meghan Burke Harris.
      The widespread adoption of trapped ion mobility coupled to mass spectrometry for proteome analysis necessitates high-quality data measurement of both ion mobility and mass spectra. In this study, the accuracy of ion mobility measurements in the context of mass spectral (MS) library building and searching is evaluated. Key factors influencing measurement accuracy, such as calibration strategy and the timing of acquisition relative to the chromatographic apex, are systematically investigated. A calibration system based on a synthetic peptide mixture is shown to improve calibration performance. Further, reproducibility of MS1- and MS2-associated mobility measurements in data-dependent acquisition (DDA) is analyzed. Overall, this work identifies methods to improve ion mobility measurement accuracy for proteomics analyses using a standardized synthetic peptide mixture.
    Keywords:  Ion mobility; Mass spectrometry; Proteomics
    DOI:  https://doi.org/10.1007/s00216-026-06633-9
  33. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 23. pii: S1570-0232(26)00286-2. [Epub ahead of print]1281 125197
    Matrix fidelity in microsampling: Plasma-first LC-MS/MS quantification of mycophenolic acid and MPAG.
    Arkadiusz Kocur, Agnieszka Czajkowska, Jacek Rubik.
      Quantitative microsampling is increasingly used in bioanalysis; however, for plasma-referenced analytes, whole-blood microsampling may introduce matrix-dependent bias related to hematocrit (HCT) and blood-to-plasma analyte distribution. This study developed and validated a rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method incorporating miniaturized micro-QuEChERS extraction for the simultaneous quantification of mycophenolic acid (MPA) and mycophenolic acid glucuronide (MPAG) in plasma, quantitative dried plasma spots (qDPS; Capitainer SEP10®), and quantitative dried blood spots (qDBS; Capitainer B®). Method validation, performed according to International Council for Harmonization (ICH) M10 and International Association of Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT) recommendations, demonstrated satisfactory selectivity, linearity, accuracy, precision, recovery, matrix-effect performance, stability, and incurred sample reanalysis across all matrices. In paired samples from pediatric kidney transplant recipients (n = 50), Passing-Bablok regression and Bland-Altman analysis showed close agreement between qDPS and conventional plasma, with low systematic deviation. For MPA, a small statistically significant proportional bias was observed, although most results remained within predefined clinical acceptance limits. In contrast, qDBS showed marked proportional bias and systematic underestimation compared with plasma. HCT- and regression-based correction strategies improved qDBS agreement but did not fully eliminate matrix-related discrepancies and require further external validation. These findings demonstrate that plasma-first microfluidic microsampling improves comparability with conventional plasma for MPA and MPAG by controlling matrix composition at the sampling stage. The proposed qDPS-based LC-MS/MS workflow provides a low-volume, matrix-aligned approach for plasma-referenced therapeutic drug monitoring and may support decentralized sampling in pediatric transplant recipients.
    Keywords:  LC–MS/MS; Mycophenolic acid; Mycophenolic acid glucuronide; Quantitative dried blood spot; Quantitative dried plasma spot; Therapeutic drug monitoring
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125197
  34. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 16. pii: S1570-0232(26)00277-1. [Epub ahead of print]1281 125188
    HPLC-MS/MS quantification of genomic deoxyuridine via differential protonation.
    Tianjiao Fang, Yifan Zhang, Hailin Wang.
      Precise quantification of trace genomic deoxyuridine (dU) is critical for elucidating DNA damage mechanisms. This analytical process is often hampered by severe isobaric mass spectrometric interference originating from the M + 1 isotope of highly abundant deoxycytidine (dC). Herein, we established a highly sensitive HPLC-MS/MS analytical strategy for the quantitative analysis of dU. By utilizing an acidic mobile phase system on a Zorbax SB-C18 column, dC and dU exhibited distinct protonation states, enabling their successful baseline separation. Methodological validation demonstrated a low limit of detection of 0.5 nM (2.5 fmol on-column with 5 μL injection), with excellent linearity, precision, and accuracy. Analysis of biological samples revealed significant dU basal heterogeneity across seven cell lines, with the highest accumulation observed in A549 cells, and precisely captured the dynamic increase in dU within 293T cells following 5-fluorouracil (5-FU) exposure. This study provides a reliable quantitative tool for investigating nucleotide metabolic imbalances and DNA damage repair.
    Keywords:  DNA damage; Deoxyuridine; Differential protonation; HPLC-MS/MS; Isobaric interference
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125188
  35. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jun 18. pii: S1570-0232(26)00283-7. [Epub ahead of print]1281 125194
    A method for quantifying non-esterified cis-9,trans-11-conjugated linoleic acid in finger-prick plasma by LC/ESI-MS/MS with isotope-coded derivatization.
    Kodai Maeda, Hayato Ichimura, Shoujiro Ogawa, Eiko Ito, Shinya Sugimoto, Takahisa Sasahara, Shunji Kawamura, Koji Matsunaga, Tatsuya Higashi.
      cis-9,trans-11-Conjugated linoleic acid (c9,t11-CLA) is the major CLA consumed from ruminant-derived foods and exhibits anti-carcinogenic, anti-atherogenic and anti-Alzheimer's disease effects for humans. Determination of the circulating non-esterified c9,t11-CLA helps provide a better understanding of its health benefits, but very few methods are available for this purpose. In this study, a liquid chromatography/electrospray ionization-tandem mass spectrometry-based method was developed and validated for quantifying non-esterified c9,t11-CLA in plasma. The DEMECAL device enabled not only the minimally invasive collection of finger-prick blood, but also an easy and rapid preparation of the plasma sample. The developed method employed the derivatization with 4-(4-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione (DAPTAD), which achieved the complete separation of c9,t11-CLA from its positional and geometric isomers on an octadecylsilyl silica column (resolution, ≥ 1.92), elimination of the negative influence from linoleic acid, and a higher sensitivity (limit of detection, 210 amol) than any methods currently available. In addition, high precision (relative standard deviations in five repetitive measurements, ≤ 6.1%) and accuracy (analytical recovery rates, 99.4-103.7%) were obtained by the isotope-coded derivatization using deuterium-labeled DAPTAD. This method also had a sufficient applicability to real sample analysis; we successfully determined the plasma non-esterified c9,t11-CLA concentrations of Japanese healthy subjects and detected their changes due to the CLA supplementation.
    Keywords:  Finger-prick plasma; Isotope-coded derivatization; LC/ESI-MS/MS; Non-esterified form; cis-9,trans-11-conjugated linoleic acid
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125194
  36. Bioanalysis. 2026 Jun 21. 1-20
    Accelerating drug discovery with mass spectrometry-driven bioanalytical innovations and beyond.
    Joseph N Mwangi, Ryan Hill, Avery Runnebohm, Gregg Czerwieniec, Michael Berna.
      The emergence of structurally complex therapeutic modalities, including bispecific antibodies, antibody-drug conjugates, fusion proteins, incretins, radioligand therapeutics, antibody-oligonucleotide conjugates, and small interfering RNAs, demands advanced mass spectrometry workflows across the ADME pipeline. This forward-looking perspective examines the bioanalytical challenges these next-generation therapeutics present, from absorption and distribution through metabolism and elimination. Current MS platforms, LC-MS/MS, high-resolution MS, quantitative mass spectrometry imaging, and ICP-MS, while highly capable, present opportunities for sensitivity enhancement to fully characterize low-dose therapeutics through the elimination phase. To quantify this need, we developed a pharmacokinetic-driven prediction framework and applied it to FDA-approved complex therapeutics, demonstrating that enhanced sensitivity would enable comprehensive metabolite characterization, particularly during the elimination phase. We identify five convergent innovation priorities: (1) addressing modality complexity through integrated ADME workflows; (2) continued advancement of instrument sensitivity for low-dose therapeutics; (3) resolving biotransformation challenges through enhanced analytical resolution; (4) improving qMSI capabilities for therapeutic-level tissue detection; and (5) implementing AI/ML-driven automation for data complexity management. Rather than advocating for MS-only solutions, we recommend integration of mass spectrometric structural specificity with complementary high-throughput technologies; immunoassays, hybridization ELISA, qPCR, and element-specific detection, to enable comprehensive bioanalysis across all modalities, accelerating the path from discovery to development.
    Keywords:  ADME; Mass spectrometry; antibody–drug conjugates; biotransformation; oligonucleotide therapeutics; pharmacokinetics; quantitative mass spectrometry imaging; radioligand therapeutics
    DOI:  https://doi.org/10.1080/17576180.2026.2687591
  37. Bioanalysis. 2026 Jun 25. 1-11
    Simultaneous quantification of D-penicillamine, D-penicillamine disulfide, and L-cysteine-D-penicillamine disulfide in human plasma: optimization of sample preparation and mass spectrometry procedures to support bioequivalence studies.
    Jianghong Gu, Sabrina Li, Guozhang Zou, Nilufer Tampal, Li Xia, Manjinder Kaur, Daniel Seungtaek Oh, Xiaoming Xu, Patrick J Faustino, David A Keire, Diaa Shakleya.
       AIM: D-penicillamine (PSH) is an active pharmaceutical ingredient used for the treatment of various diseases, such as Wilson's disease, rheumatoid arthritis, cystinuria, and heavy metal poisoning. However, evaluating PSH in plasma poses substantial analytical challenges due to drug instability, thiol-disulfide exchange reactions with endogenous thiols, and the potential formation of multiple chemical forms of PSH. This study describes the development and validation of an analytical method to ensure the stability of PSH during analysis and enable accurate bioavailability measurements and bioequivalence assessments in human plasma.
    METHODS: An ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated to simultaneously determine PSH and its two major metabolites, D-penicillamine disulfide and L-cysteine-D-penicillamine disulfide, in human plasma. PSH stability was investigated under various experimental conditions to identify the optimal sample preparation procedure. The method was then applied to conduct a 90-day stability study of PSH in human plasma stored at -80°C.
    RESULTS AND CONCLUSION: A sensitive and specific analytical method was developed and validated in accordance with the US FDA M10 guidance. PSH remained stable under optimized conditions for at least 90 days. This method provides pharmaceutical researchers with a standardized approach for PSH pharmacokinetic analysis and bioequivalence evaluations.
    Keywords:  D-penicillamine; D-penicillamine disulfide; L-cysteine-D-penicillamine disulfide; UHPLC-MS/MS; bioequivalence; plasma; stability; validation
    DOI:  https://doi.org/10.1080/17576180.2026.2690263
This page is being maintained by Thomas Krichel. It was last updated on 2026‒06‒28 at 12:44.