bims-metlip Biomed News
on Methods and protocols in metabolomics and lipidomics
Issue of 2026–08–09
37 papers selected by
Sofia Costa, Matterworks
  1. [Determination of perchlorate in human urine by cold-induced liquid-liquid extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry].
  2. Systematic Optimization of an SLE-DAPTAD LC-MS/MS Workflow for Simultaneous Quantification of Low-Abundance Vitamin D3 Metabolites in Plasma.
  3. Evaluation of a Targeted LC-MS/MS Assay for Clinical Quantification of Urinary Organic Acids.
  4. Ultra-trace multi-matrix LC-MS/MS determination of organophosphate flame retardants and metabolites in marine waters, sediments, and algae.
  5. Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of twenty-five steroids in equine serum.
  6. Uncovering PFAS and halogenated chemicals using nontargeted analysis and the PFAS IMplementer for mass spectrometry (PIMMS).
  7. Regions of interest three-way multivariate curve resolution analysis of lipid LC-IMS-MS/MS data.
  8. An isotope dilution-liquid chromatography-tandem mass spectrometry-based candidate reference measurement procedure for the quantification of meropenem in human serum and plasma.
  9. Development and validation of an LC-MS method based on derivatization for quantification of kynurenine pathway metabolites in serum and application in cancer patients.
  10. Extending singlet oxygen lifetime to enhance sensitivity of isomer-selective lipid imaging using nanospray desorption electrospray ionization (nano-DESI).
  11. Pyridyl ethyl diazoacetate derivatization enables LC-MS annotation of cholesterol and its metabolites.
  12. Development and validation of a sensitive HPLC-MS/MS method for the analysis of semaglutide in human plasma.
  13. [Determination of 32 pesticide residues in human urine by amino multi-walled carbon nanotubes-based dispersive solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry].
  14. Quantifying the aromatic amino acid metabolome: UPLC-MS/MS analysis of aromatic amino acids and their host and co-metabolites in plasma.
  15. Optimization and Validation of Plasma-Based Assay for Detection and Quantification of Monoamine Neurotransmitter Metabolites Using LC-MS/MS.
  16. An Integrated Quality by Design RP-UPLC Method for Lumacaftor and Ivacaftor in Human Plasma.
  17. In-source freezing of Trifolium repens leaf tissue to increase spatial resolution and quench stress responses during IR-MALDESI mass spectrometry imaging.
  18. HILIC-MS/MS-based strategy for quantitative amino acid profiling in multiple biological matrices and its application in kidney disease samples.
  19. Comparative profiling of amino acids in plasma and serum across children and adults using LC-MS/MS and its implications for precision nutrition.
  20. Interactive Software for Interpreting and Curating High-Resolution Electron Ionization Mass Spectra.
  21. LC-MS/MS outperforms ELISA and UPLC-PDA for targeted quantification of 25(OH)D2 and 25(OH)D3 in human milk: A practical analytical strategy.
  22. Integrated 1D-LC-HRMS and Heart-Cutting 2D-LC-MS for Impurity Profiling and Chiral Separation of Mitiglinide Drug Substance.
  23. Tenfold Expansion Submicrometer MALDI Mass Spectrometry Imaging of Tissues and Cultured Cells.
  24. Development and validation of a LC-MS/MS method for the quantification of the E-type prostanoid receptor 4 antagonist HTL0039732 in human plasma, for a Phase I/IIa clinical trial.
  25. Profiling O-Acetylation in Sialoglycans Using MALDI-TOF and LC-MS/MS with Methylamidation and Permethylation Derivatization.
  26. Validation of a Dried Blood Spot (DBS) Liquid Chromatography - High-Resolution Mass Spectrometry (LC-HRMS) workflow for simultaneous identification of emerging opioids.
  27. Plug-and-play coaxial nested-tube supported liquid membrane extraction for the determination of nonsteroidal anti-inflammatory drugs in human plasma.
  28. A simple and sensitive QuEChERS-based LC-MS/MS method for quantifying 30 per- and polyfluoroalkyl substances (PFAS) in human serum: application to firefighter biomonitoring.
  29. Development and validation of LC-MS-based bioanalytical methods for oligonucleotide therapeutics: points to consider.
  30. A reference-guided large language model workflow for mobile phase selection in thin-layer chromatography enabled by a polarity-space tetrahedron strategy.
  31. Position-specific isotope fragmentation differential quantification (PIF-DQ) of phenylalanine positional isotopomers: A rapid UPLC-MS/MS approach for urine analysis in the IAAO method.
  32. Extraction of Short-chain Fatty Acids from Mouse Feces Using A Two-phase Acid-Solvent Method.
  33. Development, validation, and application of liquid chromatography mass spectrometry methods for the determination of juvenile hormone analogues in mealworms.
  34. O- and N-Phosphoproteomic Analysis Using Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry.
  35. A targeted lipidomics workflow for high-throughput phenotyping and mechanism-based in vitro hepatotoxicity studies.
  36. Salt-induced matrix effects on PFAS quantification in online SPE-LC-MS/MS analysis.
  37. Fit-for-purpose analytics for cannabis: identification, quantification, and standardization of cannabinoids, flavonoids and terpenes.
  1. Wei Sheng Yan Jiu. 2026 Jul;55(4): 669-675
    [Determination of perchlorate in human urine by cold-induced liquid-liquid extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry].
    Siyu Yang, Xinyu Ge, Chi Zhang, Jingguang Li, Lei Zhang, Jianping Li.
       OBJECTIVE: To develop a rapid analytical method based on cold-induced liquid-liquid extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry (CI-LLE-UPLC-MS/MS) for the determination of perchlorate in human urine.
    METHODS: After spiking urine samples with stable isotope-labeled internal standards, the samples were centrifuged with a mixture of acetonitrile and water (60∶40, V/V) and then kept at -40 ℃ for 30 min. The supernatant was diluted tenfold with acetonitrile and separated on a Poroshell 120 PFP column (50 mm×2.1 mm, 1.9 μm) with a gradient elution of methanol and 1% acetic acid aqueous solution. Perchlorate was quantified by UPLC-MS/MS using the internal standard method.
    RESULTS: The method exhibited good linearity (coefficient of determination R2>0.999), with a detection limit of 0.3 μg/L and a quantification limit of 1.0 μg/L. The average recoveries at low, medium, and high spiked levels were 101.2%-106.7%, and the intra-day and inter-day relative standard deviations were both less than 5%(n=6). The determination result of perchlorate in the human urine standard reference material (SRM 3668) were (2.75±0.11) and (13.67±0.34) μg/L, both within the reference ranges. Analysis of 243 real urine samples from China revealed a 100% detection rate for perchlorate, with concentrations ranging from 1.91 to 87.41 μg/L and a median value of 18.74 μg/L.
    CONCLUSION: The method features simple pretreatment, rapid detection, and high sensitivity, precision, and accuracy, making it suitable for large-scale detection of perchlorate in human urine.
    Keywords:  cold-induced liquid-liquid extraction (CI-LLE); perchlorate; ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS); urine
    DOI:  https://doi.org/10.19813/j.cnki.weishengyanjiu.2026.04.021
  2. Rapid Commun Mass Spectrom. 2026 Oct 30. 40(20): e70158
    Systematic Optimization of an SLE-DAPTAD LC-MS/MS Workflow for Simultaneous Quantification of Low-Abundance Vitamin D3 Metabolites in Plasma.
    Marzieh Amirmostofian, Michael D Wiese, Paul H Anderson, Peter Hoffmann.
       RATIONALE: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of vitamin D3 metabolites is analytically challenging because these compounds are structurally similar, strongly matrix-associated, and have poor ionization efficiency. These limitations are particularly important for low-abundance metabolites such as 1,25(OH)2D3. This study systematically compared extraction and derivatization strategies to develop a simplified workflow for simultaneous quantification of 25(OH)D3, 3-epi-25(OH)D3, 1,25(OH)2D3, and 24,25(OH)2D3 in plasma.
    METHODS: Five sample-preparation approaches-simple protein precipitation, liquid-liquid extraction, salting-out assisted liquid-liquid extraction, solid-phase extraction, and supported liquid extraction-were evaluated under comparable conditions. Two derivatization reagents, 4-(4'-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione (DAPTAD) and 2-fluoro-1-methylpyridinium-p-toluenesulfonate (FMP-TS), were compared. The selected workflow was validated according to bioanalytical method validation criteria.
    RESULTS: Supported liquid extraction using ethyl acetate/methyl tert-butyl ether (1:1, v/v) provided the best overall balance of signal intensity, reproducibility, simplicity, and suitability for high-throughput processing. DAPTAD derivatization produced a substantially higher MS response than FMP-TS and was selected for the final method. The optimized SLE-DAPTAD LC-MS/MS workflow showed good linearity (r2 > 0.99), acceptable accuracy and precision, recovery of 72.5%-104%, and limits of quantification of 0.08 ng/mL for 3-epi-25(OH)D3, 1,25(OH)2D3, and 24,25(OH)2D3, and 0.88 ng/mL for 25(OH)D3.
    CONCLUSIONS: The optimized SLE-DAPTAD LC-MS/MS workflow provides a simple, sensitive, and automation-compatible approach for simultaneous quantification of four vitamin D3 metabolites in plasma. The method is positioned as an analytical workflow for method development and low-volume plasma analysis, rather than as a fully standardized routine diagnostic assay for endogenous 1,25(OH)2D3 measurement.
    Keywords:  DAPTAD derivatization; LC–MS/MS; method optimization; sample preparation; supported liquid extraction; vitamin D3 metabolites
    DOI:  https://doi.org/10.1002/rcm.70158
  3. J Mass Spectrom. 2026 Aug;61(8): e70099
    Evaluation of a Targeted LC-MS/MS Assay for Clinical Quantification of Urinary Organic Acids.
    Earnest J P Daniel, Emily Garnett, Deepthi Rajapakshe, Sridevi Devaraj.
      Gas chromatography-mass spectrometry (GC-MS) is the reference method for urine organic acid analysis but requires complex sample preparation and derivatization, limiting routine clinical use. We developed and validated a targeted Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying urinary organic acids relevant to the evaluation of inborn errors of metabolism. The assay demonstrated excellent linearity, precision, accuracy, analytical sensitivity, and good agreement with GC-MS. Organic acidurias and other metabolic disorders were accurately identified through characteristic metabolite elevations. The simplified workflow eliminates derivatization and enables rapid, high-throughput implementation for newborn screening confirmation, metabolic screening, and patient monitoring in pediatric clinical laboratories.
    DOI:  https://doi.org/10.1002/jms.70099
  4. Anal Chim Acta. 2026 Oct 08. pii: S0003-2670(26)00834-2. [Epub ahead of print]1418 345884
    Ultra-trace multi-matrix LC-MS/MS determination of organophosphate flame retardants and metabolites in marine waters, sediments, and algae.
    Florentina Laura Chiriac, Iuliana Paun, Florinela Pirvu, Vasile Ion Iancu, Victor Constantin Cojocaru.
       BACKGROUND: Organophosphate flame retardants (OPFRs) are emerging contaminants increasingly detected in marine environments due to their extensive industrial use and replacement of brominated flame retardants. Their occurrence in different environmental compartments, combined with the formation of transformation products, raises concerns regarding persistence, transport, and ecological risks. However, the simultaneous determination of OPFRs and their metabolites in complex marine matrices remains analytically challenging due to their diverse physicochemical properties and low environmental concentrations.
    RESULTS: A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of eight OPFRs and three transformation products in seawater, sediments, and marine algae. The method combines solid-phase extraction for aqueous samples (200 mL) with ultrasound-assisted extraction for solid matrices, followed by chromatographic separation on a C18 column and multiple reaction monitoring (MRM) detection using electrospray ionization. Optimization of chromatographic and ionization parameters enabled the reliable separation and detection of compounds with diverse physicochemical characteristics within a single analytical run. The method demonstrated excellent linearity (R2 ≥ 0.99), satisfactory recoveries (74-105%), and good precision (relative standard deviation ≤ 10%) for all target analytes. Matrix effects, assessed by post-extraction addition, ranged from 77% to 108%, indicating only moderate signal suppression or enhancement depending on matrix type. Method detection limits ranged from 0.19 to 0.40 ng/L in seawater and remained below 0.5 ng/g in sediments and algae, confirming the suitability of the method for ultra-trace analysis.
    SIGNIFICANCE AND NOVELTY: The proposed analytical approach enables the simultaneous assessment of OPFRs and transformation products in both abiotic and biotic marine compartments using a single harmonized workflow. The inclusion of algae alongside seawater and sediments provides a more comprehensive understanding of contaminant distribution and potential bioaccumulation pathways in marine ecosystems. The method represents a robust, sensitive, and reliable analytical tool suitable for environmental assessment, contamination surveillance, and large-scale marine monitoring programs.
    Keywords:  Algae; LC–MS/MS; Monitoring; OPFRs; Seawater; Sediment; Ultra-trace
    DOI:  https://doi.org/10.1016/j.aca.2026.345884
  5. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jul 24. pii: S1570-0232(26)00311-9. [Epub ahead of print]1282 125222
    Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of twenty-five steroids in equine serum.
    Joy Ledeck, Thomas Dubrowski, Stéphanie Peeters, Caroline Le Goff, Stéfan Deleuze, Jérôme Ponthier, Etienne Cavalier.
      Steroids are potential biomarkers for monitoring equine pregnancy. However, immunoassays currently used for their quantification suffer from cross-reactivity and limited specificity, thus requiring more accurate methods. This study reports the development and validation of a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of 25 steroids covering the main biosynthetic pathways of progestogens, corticosteroids, androgens, and estrogens. Steroids were extracted by protein precipitation followed by evaporation, derivatization, and reconstitution before LC-MS/MS analysis. A surrogate matrix was used for calibration and validation to avoid endogenous interference. Validation was performed according to and partly adapted from Clinical and Laboratory Standards Institute guidelines (CLSI), including linearity, trueness, precision, limits of detection and quantification, measurement uncertainty, recovery, matrix effects, carryover, selectivity, and stability. Calibration curves were fitted using the best-performing weighted linear or quadratic regression model, yielding excellent linearity (R2 > 0.990), trueness between -9.0% and 2.3%, and intra- and inter-day precision <6.3%. Lower limits of quantification ranged from 2.07 to 2250 pg/mL depending on physiological analytes concentration. Extraction recovery averaged 24.3-114.9%, matrix effects were acceptable, and accuracy ranged from 94.4% to 98.9%. No carryover or interferences were detected. Measurement uncertainty remained <15%. This study presents the first LC-MS/MS method partially validated per CLSI criteria for the quantification of 24 steroids in equine serum. The method offers a sensitive and specific alternative to immunoassays and provides a robust tool for equine steroid profiling with potential applications in pregnancy monitoring, placentitis diagnosis, and fetal sex determination.
    Keywords:  CLSI guidelines; Equine; Liquid chromatography-tandem mass spectrometry; Reproductive monitoring; Serum; Stability; Steroid profiling
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125222
  6. Sci Adv. 2026 Aug 07. 12(32): eaeg0781
    Uncovering PFAS and halogenated chemicals using nontargeted analysis and the PFAS IMplementer for mass spectrometry (PIMMS).
    Gregory P Kudzin, James N Dodds, Erin S Baker.
      Per- and polyfluoroalkyl substances (PFASs) are synthetic chemicals of considerable epidemiological concern. While targeted liquid chromatography (LC)-tandem mass spectrometry (MS/MS) methods are established for legacy PFAS, emerging replacement compounds require nontargeted analysis (NTA) for comprehensive characterization. However, NTA of complex biological matrices like human serum is analytically challenging because of the presence of many endogenous interferences and unknown PFAS molecules. To address this, ion mobility spectrometry has been coupled with LC and MS (LC-IMS-MS). This approach provides orthogonal separation via collision cross section to resolve isobars and filter noise. Despite its utility, current software lacks efficient PFAS feature prioritization, requiring extensive manual curation. We developed the PFAS IMplementer for Mass Spectrometry (PIMMS), a vendor-neutral, open-source tool that uses advanced filtering and scoring algorithms for rapid LC-IMS-MS data analysis. The potential utility of PIMMS was demonstrated using serum from bovine and human populations. PIMMS could identify both legacy and novel PFAS in addition to eight halogenoalkane xenobiotics (containing bromine, chlorine, and iodine). Of note, these halogenoalkane species were often only singly or doubly halogenated and were still effectively prioritized by PIMMS demonstrating the potential to identify a potentially growing number of novel compounds with only a single degree of halogenation.
    DOI:  https://doi.org/10.1126/sciadv.aeg0781
  7. Anal Chim Acta. 2026 Oct 08. pii: S0003-2670(26)00800-7. [Epub ahead of print]1418 345850
    Regions of interest three-way multivariate curve resolution analysis of lipid LC-IMS-MS/MS data.
    Chenxi Peng, Anna de Juan, Romà Tauler.
       BACKGROUND: Liquid chromatography-ion mobility-tandem mass spectrometry (LC-IMS-MS/MS) enables multidimensional characterization of complex lipid mixtures. However, the resulting high-dimensional datasets pose significant challenges for reliable component resolution and interpretation. In this work, the Regions-Of-Interest-based Multivariate Curve Resolution strategy (ROIMCR) is proposed for the integrated analysis of LC-IMS-MS/MS data. It explicitly preserves ion mobility information in its native profile form rather than reducing it to a single collision cross section (CCS) value. The proposed approach simultaneously resolves chromatographic elution profiles, ion mobility profiles, and mass spectra profiles (MS1 and MS2). Bilinear or trilinear models are used to enforce chemically consistent solutions across the elution time, m/z, and ion mobility dimensions, yielding interpretable component profiles while maintaining satisfactory model performance.
    RESULTS: This study has proven the applicability and chemical interpretability of the trilinear model in resolving complex lipid mixtures using LC-IMS-MS/MS three-way data. Compound annotation and identification were achieved through direct comparison of the ROIMCR-resolved retention time, arrival time, and tandem mass spectra with those of authentic standards or reference data libraries. The applicability of the method was demonstrated using two data examples. The first one is used to validate the proposed approach using a simulated mixture of four analytes. The second example shows the application of the proposed method in the LC-IMS-MS/MS experimental analysis of a complex mixture of lipids. Overall, the proposed three-way MCR framework provides an accurate, interpretable, and scalable solution for high-dimensional LC-IMS-MS/MS data analysis.
    DOI:  https://doi.org/10.1016/j.aca.2026.345850
  8. Clin Chem Lab Med. 2026 Aug 06.
    An isotope dilution-liquid chromatography-tandem mass spectrometry-based candidate reference measurement procedure for the quantification of meropenem in human serum and plasma.
    Rupert Schmid, Judith Taibon, Neeraj Singh, Carina Schuster, Arber Rexhaj, Michael Vogeser, Friederike Bauland, Andrea Geistanger, Elie Fux.
       OBJECTIVES: Therapeutic drug monitoring (TDM) of meropenem is critical for optimizing clinical efficacy and minimizing toxicity. We describe the validation of an isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) candidate reference measurement procedure (RMP) for the quantification of meropenem in human serum and plasma.
    METHODS: A high-purity meropenem standard was characterized using an in-house quantitative nuclear magnetic resonance protocol. This established the absolute content and ensured an unbroken chain of metrological traceability to the International System of Units. Samples were prepared via incubation with a stable isotope-labeled internal standard followed by protein precipitation. Analysis was performed using LC-MS/MS with electrospray ionization. The method was validated to assess selectivity, matrix effects, precision, accuracy, and measurement uncertainty according to international metrological guidelines.
    RESULTS: The candidate RMP demonstrated high selectivity across a linear range of 0.400-120 μg/mL and was unaffected by matrix interferences. Intermediate precision was <3.8 % for spiked serum and <4.3 % for native patient pools; repeatability ranged from 1.9 to 3.1 %. Mean bias ranged from -5.1 to 2.4 % across all serum and plasma levels, including dilution integrity assessments. Expanded measurement uncertainty (k=2) for target value assignment (n=6) was 2.6-4.3 %, while uncertainty for single measurements ranged from 6.0 to 8.8 %.
    CONCLUSIONS: The proposed method fulfills all analytical performance specifications required for a candidate RMP. By establishing a direct link from individual patient samples to primary reference materials, this approach provides a robust foundation for the global standardization of routine meropenem assays.
    Keywords:  SI units; isotope dilution-liquid chromatography-tandem mass spectrometry; meropenem; quantitative nuclear magnetic resonance characterization; reference measurement procedure
    DOI:  https://doi.org/10.1515/cclm-2026-0428
  9. J Chromatogr A. 2026 Jul 26. pii: S0021-9673(26)00629-1. [Epub ahead of print]1785 467301
    Development and validation of an LC-MS method based on derivatization for quantification of kynurenine pathway metabolites in serum and application in cancer patients.
    Kinga Ożga-Wybranowska, Paweł Stepuch, Inga Łopuszyńska, Marek Durlik, Elżbieta Radzikowska-Büchner, Ryszard Maciejewski, Ilona Sadok.
      Simultaneous determination of a broad panel of kynurenine pathway metabolites remains analytically challenging due to their diverse polarity, poor chromatographic retention, and wide concentration range in biological matrices. In this work, a liquid chromatography-mass spectrometry method, which includes a derivatization step to form ester derivatives of tryptophan metabolites, was developed to measure serum levels of kynurenine, kynurenic acid, 3-hydroxykynurenine, picolinic acid, nicotinic acid, quinolinic acid, and xanthurenic acid. Sample preparation procedure was thoroughly optimized. As part of method development, different strategies for sample purification were tested. Ultimately, serum samples after derivatization were subjected to clean-up using solid-phase extraction. This approach enables the separation of isomeric pairs (picolinic and nicotinic acids) and improved the detection of quinolinic acid. Data were normalized to a single internal standard (3-nitro-l-tyrosine), which undergoes derivatization in the same manner as the target analytes. The method demonstrated excellent linearity (R² > 0.99) over wide concentration ranges, typically from low nanomolar to several tens of micromolar levels. Low limits of quantification were determined in a surrogate matrix: 0.35 µM for picolinic acid, 0.11 µM for 3-hydroxykynurenine, and <80 nM for the other metabolites.Inter- and intraday accuracy ranged between 90-110%, with recoveries >70% for all analytes. The method was validated and successfully applied to the quantification of kynurenine pathway metabolites in serum from healthy donors and patients with gastric or pancreatic cancer.
    Keywords:  3-hydroxykynurenine; Derivatization; Gastric cancer; Kynurenine; Pancreatic cancer; Picolinic acid; Quinolinic acid
    DOI:  https://doi.org/10.1016/j.chroma.2026.467301
  10. Anal Chim Acta. 2026 Oct 08. pii: S0003-2670(26)00817-2. [Epub ahead of print]1418 345867
    Extending singlet oxygen lifetime to enhance sensitivity of isomer-selective lipid imaging using nanospray desorption electrospray ionization (nano-DESI).
    Sara Amer, Daisy Unsihuay, Emerson Hernly, Alessandra Latorre, Julia Laskin.
      Identifying and localizing lipid CC positional isomers in biological tissues remains a major analytical challenge for isomer-selective mass spectrometry imaging (iMSI). Herein, we report a straightforward yet powerful strategy to enhance the efficiency of singlet oxygen (1O2) reactions used for iMSI of lipids by extending the 1O2 lifetime using deuterated solvents. Replacing the conventional 9:1 (v/v) MeOH/H2O nanospray desorption electrospray ionization (nano-DESI) solvent with its deuterated counterpart, CD3OD/D2O, enhances the efficiency of lipid hydroperoxide (LOOH) formation by at least a factor of two. The resulting increase in signals of diagnostic fragments of LOOHs enhances sensitivity of nano-DESI iMSI for fatty acids and both mono- and polyunsaturated lipid species. Using this optimized solvent system, we achieved a deeper molecular coverage of isomeric lipids in nano-DESI iMSI of mouse brain tissue, enabling the detection and spatial mapping of lipid isomers previously undetectable with MeOH/H2O. Overall, the use of deuterated solvents is a practical and broadly applicable strategy for enhancing the sensitivity and coverage of photochemical derivatization-based nano-DESI iMSI, which will offer new insights into lipid structure and spatial distribution in complex biological systems.
    Keywords:  Double-bond localization; Lipid hydroperoxide; Lipid isomers; Mass spectrometry imaging; Photochemical derivatization; Singlet oxygen
    DOI:  https://doi.org/10.1016/j.aca.2026.345867
  11. Anal Chim Acta. 2026 Oct 08. pii: S0003-2670(26)00805-6. [Epub ahead of print]1418 345855
    Pyridyl ethyl diazoacetate derivatization enables LC-MS annotation of cholesterol and its metabolites.
    Wenhao Ke, Yikang Wang, Yang Chen, Hanchu Ye, Wen-Di Duan, Feifei Lin, Yingying Wang, Zhengjun Zhou, Zhiyong Yu, Youhong Hu, Jia Liu.
      Cholesterol and its metabolites are fundamental to membrane organization and cellular signaling, but their poor ionization efficiency and structural similarity complicate mass-spectrometric (MS) analysis. In this work, we developed pyridyl ethyl diazoacetate (PED) as a stable, efficient derivatization reagent that introduces a permanent positive charge on hydroxyl-containing sterols, thereby enhancing ionization and detection sensitivity. Systematic MS2 characterization established diagnostic fragmentation motifs of PED-tagged sterols, which we encoded into a query-based spectral-mining strategy for reproducible, rule-based annotation of sterol features in complex matrices. Application of the PED workflow to human liver tissues enabled sterol profiling across cirrhotic, hepatocellular carcinoma, and intrahepatic cholangiocarcinoma specimens, suggesting disease-context-dependent differences in sterol profiles. The workflow effectively integrates chemical derivatization and programmable data interrogation, providing a robust analytical framework for sterol profiling in biological samples and a potential basis for broader lipidomic applications.
    Keywords:  Cholesterol derivatization; Liver disease metabolomics; Mass spectrometry; Pyridyl ethyl diazoacetate; Spectral mining; Sterol profiling
    DOI:  https://doi.org/10.1016/j.aca.2026.345855
  12. Bioanalysis. 2026 Aug 03. 1-10
    Development and validation of a sensitive HPLC-MS/MS method for the analysis of semaglutide in human plasma.
    Christopher Tudan, Joel Stickling, Emily Northwick Darden, Lauren Devine, Shane Needham.
       BACKGROUND: Semaglutide is a human glucagon-like peptide-1 (GLP-1) peptide analog. To date, the literature does not report a validated HPLC-MS/MS assay using a triple-quadrupole mass spectrometer for the quantitation of semaglutide in human plasma in support of pharmacokinetics. A robust, sensitive HPLC--MS/MS assay supporting this class of compounds, is provided and full validation demonstrated.
    METHODS AND RESULTS: A triple-quadrupole HPLC-MS/MS method has been developed and fully validated for the quantitation of semaglutide in human plasma, using a stable isotope labeled internal standard and an effective, efficient protein precipitation sample extraction. The method has a sensitive analytical range of 5 ng/mL to 200 ng/mL (and to 1000 ng/mL with a dilution QC) and exhibits excellent accuracy (92.7% to 109.4%) and precision of <10.6%. This highly selective method provided high recovery of the analyte, with minimal matrix effects.
    CONCLUSION: A robust and simple HPLC-MS/MS method to quantify semaglutide in human plasma has been fully validated in accordance with the 2022 FDA M10 guidance. This method is suitable for implementation in clinical studies within the validated analytical range and stability conditions and can be extended to the bioanalysis of other GLP-1 peptide-based therapeutics with similar structural and physiochemical properties.
    Keywords:  GLP-1; HPLC-MS/MS; ICH M10; method validation; peptides; semaglutide
    DOI:  https://doi.org/10.1080/17576180.2026.2712160
  13. Wei Sheng Yan Jiu. 2026 Jul;55(4): 676-684
    [Determination of 32 pesticide residues in human urine by amino multi-walled carbon nanotubes-based dispersive solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry].
    Lingzhen Xu, Ye Wang, You Xia.
       OBJECTIVE: An analytical method for the determination of 32 pesticide residues in human urine was established using amino multi-walled carbon nanotubes (NH2-MWCNTs)-based dispersive solid-phase extraction combined with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).
    METHODS: 25% saturated sodium chloride solution was added to precipitate protein in the urine. The pH was adjusted to 5 using 20 mmol/L ammonium acetate solution. The target analytes were adsorbed onto amino multi-walled carbon nanotubes, followed by desorption with acetonitrile. Finally, the extract was analyzed by UPLC-MS/MS.
    RESULTS: Good linearity for the 32 pesticides was observed within the concentration range of 1-100 μg/L, with correlation coefficients (r) all above 0.999. Fortified at three concentration levels (4, 40, and 200 μg/L, n=6), average recoveries were obtained in the range of 91.2%-110.0%, with relative standard deviations ranging from 1.1%-4.6%. The limits of detection and quantification were determined to be 0.003-0.03 μg/L and 0.01-0.1 μg/L, respectively. The adsorption process of the 32 pesticides onto the NH2-MWCNTs was found to be well described by the pseudo-second-order kinetic model and the Langmuir isotherm model.
    CONCLUSION: This method is simple and rapid to operate, accurate, and highly sensitive, enabling the rapid detection of 32 pesticide residues in human urine.
    Keywords:  amino multi-walled carbon nanotubes; dispersive solid-phase extraction; human urine; pesticide residues; ultra-performance liquid chromatography-tandem mass spectrometry
    DOI:  https://doi.org/10.19813/j.cnki.weishengyanjiu.2026.04.022
  14. J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Aug 03. pii: S1570-0232(26)00331-4. [Epub ahead of print]1282 125242
    Quantifying the aromatic amino acid metabolome: UPLC-MS/MS analysis of aromatic amino acids and their host and co-metabolites in plasma.
    Marlene Thaitumu, Domniki Gallou, Olga Begou, Georgios Theodoridis, Helen Gika.
      Aromatic amino acids (AAAs), tryptophan, phenylalanine, and tyrosine along with their pathway metabolites have been implicated in the pathogenesis of diseases ranging from cardiovascular, neurological, inflammatory, and cancer diseases, among others. As such, the measurement of the primary AAAs, their host pathway metabolites, and microbiome derived co-metabolites in blood can provide a sensitive reflection of systemic health. The aim of the study was to develop a method for the quantification of 17 metabolites, the three AAAs and various of their metabolites in plasma using a high-throughput ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. The method demonstrated a dynamic range (1 to 16,700 ng/mL), with detection limits (LOD) as low as 0.05 ng/mL. Quantification limits ranged from 3 to 5019 ng/mL (LLOQ) and up to 16,700 ng/mL (ULOQ). Recovery at LQC, MQC, and HQC was satisfactory and consistent across most metabolites, with significant matrix effects observed only for 4-ethylphenol sulfate. Furthermore, intra and inter-day accuracy and precision met all acceptance criteria at all quality control concentrations for most of the metabolites. Measurement of NIST SRM 1950 showcased the method's accuracy for most of the metabolites. Finally, the method was applied on the analysis of plasma samples from 55 individuals (13 males and 42 females) providing information on AAAs and their pathway metabolites relevant concentrations in human plasma.
    Keywords:  Aromatic amino acids pathways; LC–MS/MS; Metabolites; Microbial-host; Phenylalanine; Tryptophan; Tyrosine
    DOI:  https://doi.org/10.1016/j.jchromb.2026.125242
  15. ACS Omega. 2026 Jul 28. 11(29): 43625-43637
    Optimization and Validation of Plasma-Based Assay for Detection and Quantification of Monoamine Neurotransmitter Metabolites Using LC-MS/MS.
    Lloyd Tauro, Kusuma C G, Nirpendra Singh, Vykuntaraju K Gowda, Rakesh K Mishra, Runa Hamid.
      One of the essential subclasses of biogenic amines is monoamine neurotransmitters, which include dopamine, serotonin, norepinephrine, and epinephrine neurotransmitters. The biosynthesis pathways of these neurotransmitters share several common substrates, metabolites, cofactors, and enzymes. Genetic mutations in these components lead to a group of rare inherited conditions termed monoamine neurotransmitter metabolic disorders (mNMDs). Existing studies report methods for measurement of a particular set of metabolites belonging to this pathway for diagnosis, frequently across different biological matrices such as cerebrospinal fluid (CSF), plasma, serum, or urine. Hence, the available assays and data are methodologically heterogeneous. In a biochemical pathway, disorder-specific metabolic patterns arise from paired alterations in multiple upstream and downstream metabolites, and these patterns can be captured only when these metabolites are measured simultaneously from the same biological matrix under similar analytical conditions. The lack of concurrent measurement limits the use of metabolite ratios, which can be of diagnostic value and pattern-based interpretation in the case of mNMDs. In this study, we report the development, optimization, and validation of a plasma-based targeted liquid chromatography-mass spectrometry (LC-MS/MS) method for the simultaneous detection and quantification of 11 key metabolites of the monoamine biosynthesis pathway. The assay was optimized for sample preparation, chromatographic separation, MS detection, and validated according to the M10 Bioanalytical Method Validation and Study Sample Analysis, US -FDA guidelines (November 2022). The method demonstrates good linearity across a broad range, high sensitivity and specificity, acceptable interday- and intraday-precision and accuracy, acceptable coefficient of variation, and lower limit of detection and quantification. The applicability of the method was further validated using plasma samples from healthy participants. As mNMDs are potentially treatable if identified early, this validated plasma-based method can provide a quick and predictive alternative to currently used different biological matrices or invasive CSF sampling for diagnosis with additional utility for longitudinal therapeutic monitoring of these treatable conditions.
    DOI:  https://doi.org/10.1021/acsomega.6c02312
  16. Biomed Chromatogr. 2026 Sep;40(9): e70587
    An Integrated Quality by Design RP-UPLC Method for Lumacaftor and Ivacaftor in Human Plasma.
    Subbarao Jhampani, Rayini Venkata Sai Mounica, Suryadevara Vidhyadhara.
      Cystic fibrosis, a genetic disorder caused by mutations in the CFTR gene, is commonly treated using combination therapy with IVA and LUMA. Accurate quantification of these drugs in human plasma is essential for therapeutic monitoring; however, conventional analytical methods often lack sufficient sensitivity and robustness. In this study, a QbD-based RP-UPLC method was developed and validated for the simultaneous estimation of IVA and LUMA in human plasma. The primary objective was to achieve efficient separation of analytes from plasma interferences with reduced run time and consistent analytical performance. Chromatographic separation was carried out using a Waters Acquity UPLC system equipped with a CSH C18 column (100 mm × 2.1 mm, 1.8 μm). Method optimization was performed using a Central Composite Design model. The mobile phase is 0.01 N ammonium acetate and acetonitrile (60:40, v/v) at a flow rate of 0.3 mL/min, with the column maintained at 30°C. EMT was used as the internal standard, and detection was conducted at 260 nm. The retention times were 1.45, 1.77, and 1.99 min for EMT, IVA, and LUMA, respectively. The method is reliable, rapid and selective, exhibiting excellent linearity, precision, and accuracy, complying with USFDA guidelines.
    Keywords:  IVA and LUMA; bioanalysis; internal standard; quality by design; reverse‐phase ultra performance liquid chromatography
    DOI:  https://doi.org/10.1002/bmc.70587
  17. Anal Bioanal Chem. 2026 Aug 03.
    In-source freezing of Trifolium repens leaf tissue to increase spatial resolution and quench stress responses during IR-MALDESI mass spectrometry imaging.
    Quinn Mills, David C Muddiman.
      Mass spectrometry imaging (MSI) is a robust analytical tool for resolving the spatial localization of diverse metabolites in a tissue sample simultaneously. Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is particularly advantageous for analyzing leaf tissue, since it bypasses the cuticle and uses endogenous water in the sample as a matrix, avoiding delocalization and reducing costs associated with sample preparation. While direct analysis of fresh tissue offers substantial benefits, it also presents a less-discussed challenge: plants initiate a cascade of metabolic responses to various forms of stress (e.g., mechanical wounding, heat stress), so freshly-harvested tissue can undergo major metabolic changes before and during analysis. Herein, a method is introduced which retains the benefits of IR-MALDESI MSI but increases spatial resolution and quenches metabolic processes by quickly freezing leaf tissue within the source enclosure immediately after harvest. Leaf samples were mounted on a glass slide with carboxymethylcellulose gel immediately after harvest and moved into the source enclosure, humidity was purged, and the sample was frozen using the Peltier-cooled stage. Camera images demonstrated minimal ice deposition on the sample surface, and evaluation of the phenylpropanoid pathway suggested that freezing the sample inhibited post-harvest stress responses during and prior to analysis. Frozen tissue analysis significantly decreased ablation spot diameter from 65.4 to 43.9 µm, enabling approximately a 1.5-fold increase in spatial resolution with a conventional 2.97-µm IR laser.
    Keywords:  IR-MALDESI; Mass spectrometry imaging; Plant imaging; Plant metabolomics; Plant stress response
    DOI:  https://doi.org/10.1007/s00216-026-06710-z
  18. Anal Bioanal Chem. 2026 Aug 04.
    HILIC-MS/MS-based strategy for quantitative amino acid profiling in multiple biological matrices and its application in kidney disease samples.
    Xiongwei Yin, Jean-Paul Decuypere, Djalila Mekahli, Bert Bammens, Rudi Vennekens, Erwin Adams, Ann Van Schepdael.
      Amino acid metabolism disorders have strong links to various kidney diseases, and the kidney is essential for maintaining systemic amino acid homeostasis. Given the lack of obvious clinical symptoms in the early stages of kidney disease, searching for reliable metabolic biomarkers is essential for early diagnosis. Endogenous AAs are widely present in human biological matrices, making it difficult to obtain analyte-free blank matrices for method validation. This poses challenges in the preparation of calibration standards, leading to uncertainty and reduced consistency in the results. This study aimed to develop a rapid hydrophilic interaction liquid chromatographic method coupled with tandem mass spectrometry. It could analyze 47 amino acids and related compounds, present in various biological matrices (human plasma, urine, and cyst fluids). The calibration curves were constructed by stripping each matrix to obtain a surrogate matrix. The slopes of the different calibration curves were systematically evaluated and the analytical results were compared with results obtained through the method of standard addition. The calibration curves established using both the standard addition method and the matrix stripping method were parallel, hence matrix stripping effectively mitigates matrix effects under a variety of matrix conditions, thereby assuring the accuracy and reliability of quantitative analysis results. To further investigate the applicability of this method to large-scale sample analysis, the analysis of plasma samples and urine samples obtained from patients with two types of kidney disease was performed.
    Keywords:  Amino acids; Cyst fluid; LC-MS/MS; Matrix effect; Plasma; Urine
    DOI:  https://doi.org/10.1007/s00216-026-06687-9
  19. Front Nutr. 2026 ;13 1850080
    Comparative profiling of amino acids in plasma and serum across children and adults using LC-MS/MS and its implications for precision nutrition.
    Yanhui Ma, Yi Liu, Yuchan Huangfu, Yunlan Zhou, Yingxia Zheng.
       Objective: Circulating amino acids (AAs) are important biomarkers of nutritional status, metabolic homeostasis, and disease related physiological changes. However, the influence of specimen type and age on AA measurements remains incompletely understood. This study aimed to establish a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying 29 amino acids (AAs), compare serum and plasma matrices, and investigate age related variations in healthy populations.
    Methods: In a cross-sectional study, a total of 231 healthy adults (≥18 years) and 140 children (<18 years) were recruited from the Shanghai area with a subgroup of 26 adults providing paired serum and plasma samples. Twenty-nine AA quantification was performed using LC-MS/MS with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatization. Method validation followed Clinical and Laboratory Standards Institute (CLSI) guidelines for linearity, lower limit of quantitation (LLOQ), precision, accuracy, carryover, and stability.
    Results: The assay demonstrated excellent linearity (R 2 > 0.99), precision (CV < 15%), and accuracy (bias < 10%) with minimal carryover and acceptable LLOQs for all 29 AAs. Matrix differences and correlations were evaluated using the paired samples, while age related AA profiles were assessed in largely independent serum and plasma cohorts of pediatric and adult. Serum and plasma concentrations correlated well for most AA. However, significant matrix dependent differences were observed for several amino acids, including GABA, Ser, Tau, Sar, and Asp (R 2 < 0.5). Age related analysis revealed that most essential AAs (Leu, Ile, Trp, Phe, His, Tyr) exhibited higher median concentrations in children compared to adults in serum. Conversely, lysine and threonine were significantly higher in adult serum. In plasma, age related differences were less pronounced for essential AAs but consistent for specific non-essential AAs. Bubble plot showed Cit, Arg, and Gln were positively associated with age, while GABA was negatively associated with age.
    Conclusion: The validated LC-MS/MS platform provides a reliable platform for nutritional biomarker research. Matrix selection and age are critical factors in AA interpretation, offering valuable insights for clinical diagnostics and translational metabolomics research.
    Keywords:  LC–MS/MS; age; amino acids; fasting serum; plasma
    DOI:  https://doi.org/10.3389/fnut.2026.1850080
  20. J Mass Spectrom. 2026 Aug;61(8): e70096
    Interactive Software for Interpreting and Curating High-Resolution Electron Ionization Mass Spectra.
    Dmitriy D Matyushin, Anastasia Yu Sholokhova.
      Gas chromatography coupled with electron ionization and high-resolution mass spectrometry (GC-EI-HRMS) has been increasingly used in analytical chemistry in recent years. Databases (libraries) of such mass spectra are emerging, and various studies require processing batches of such data. This necessitates the development of user-friendly software for curation and batch processing of such data sets. In this work, we propose the "gchrmsexplain" software specifically developed for this purpose. It includes both a graphical user interface and a command-line interface. The software receives a peak list as input and finds a formula for each peak using in silico fragmentation or combinatorial enumeration. The isotopic pattern is checked. The fraction of the ion current due to the annotated peaks is calculated. The absolute error in determining m/z can be specified in either millidaltons or parts per million. Batch processing is possible. The software was applied to six publicly available GC-EI-HRMS data sets: RECETOX (exposome and metabolome libraries), HREI-MSDB, library from Castro's work (2022), and two subsets of MassBank EU. In total, these data sets contain 1811 mass spectra. Impurity peaks, systematic errors in m/z, and completely incorrect mass spectra were detected. It was shown that some of the data sets contain low-quality mass spectra. It was demonstrated that the presented software can be used for the curation of such data. The software is free and available online: https://github.com/mtshn/gchrmsexplain.
    Keywords:  electron ionization; gas chromatography–mass spectrometry; high resolution mass spectrometry; isotopic pattern; molecular formula
    DOI:  https://doi.org/10.1002/jms.70096
  21. Food Res Int. 2026 Oct 01. pii: S0963-9969(26)01470-5. [Epub ahead of print]241 119787
    LC-MS/MS outperforms ELISA and UPLC-PDA for targeted quantification of 25(OH)D2 and 25(OH)D3 in human milk: A practical analytical strategy.
    Yuanyuan Zhao, Yongmao Lu, Tian Yang, Shaohua Hu, Yuanyuan Yan, Xingguo Wang, Wei Wei.
      Accurate analysis of vitamin D in human milk is essential for infant nutrition but remains analytically challenging due to low levels and a complex matrix. This study evaluates a practical multi-method strategy using LC-MS/MS, ELISA, and UPLC-PDA for vitamin D analysis in human milk. LC-MS/MS precisely quantified 25(OH)D₂ and 25(OH)D₃ with assay LOQ of 0.045 ng/mL and 0.100 ng/mL, respectively. ELISA showed a 11.58 ng/mL positive bias of vitamin D which limits use for accurate quantification. UPLC-PDA profiled vitamins A and E effectively but lacked sensitivity for vitamin D. Longitudinally, LC-MS/MS results show 25(OH)D₃ and total 25(OH)D concentrations remained relatively stable, with medians ranging from 0.141 to 0.281 ng/mL and 0.263 to 0.315 ng/mL, respectively, across the study period, while 25(OH)D₂ peaked at 8 weeks before declining. This study establishes LC-MS/MS as the preferred analytical strategy for quantifying 25(OH)D2 and 25(OH)D3 in human milk, providing critical data to assess infant nutritional intake and guide clinical interventions.
    Keywords:  Bioactive lipids; Clinical screening; ELISA; Human milk; LC-MS/MS; Nutritional epidemiology; Vitamin D
    DOI:  https://doi.org/10.1016/j.foodres.2026.119787
  22. Rapid Commun Mass Spectrom. 2026 Oct 30. 40(20): e70144
    Integrated 1D-LC-HRMS and Heart-Cutting 2D-LC-MS for Impurity Profiling and Chiral Separation of Mitiglinide Drug Substance.
    Chenxi Wang, Shiwen Zhou, Wanhao Sun, Yuanjiang Pan, Hongru Feng.
       RATIONALE: The comprehensive quality control of chiral pharmaceuticals like mitiglinide necessitates simultaneous assessment of chemical impurities and enantiomeric purity, yet conventional workflows address these separately, leading to inefficiency. This study develops a comprehensive analytical strategy to overcome this challenge for the anti-diabetic drug mitiglinide.
    METHODS: For chiral analysis, an online heart-cutting two-dimensional liquid chromatography-high-resolution mass spectrometry (2D-LC-HRMS) method was developed. First, impurity profiling of mitiglinide was accomplished using a one-dimensional reversed-phase LC-HRMS (1D-LC-HRMS) method. Subsequently, the 2D-LC-HRMS system achieved enantiomer separation by online coupling of a C18 column (first dimension) with a polysaccharide-based chiral column (second dimension), with the separated analytes detected by an Orbitrap mass spectrometer.
    RESULTS: 1D-LC-HRMS identified five major impurities, structurally characterizing four, with the main component accounting for only 49.12% of the total integrated peak area (relative abundance by EIC peak area normalization, not absolute purity). The 2D-LC-HRMS method achieved effective enantiomer separation. A consistent third minor chromatographic peak was observed across six replicate analyses, which is tentatively assigned as a potential diastereomeric impurity based on stereochemical interpretation of the chromatographic behavior; confirmatory evidence is required for definitive identification.
    CONCLUSIONS: This work successfully establishes a comprehensive strategy that efficiently consolidates impurity profiling and chiral purity assessment for mitiglinide. It provides a reliable, more informative approach for the quality control of complex chiral pharmaceuticals.
    Keywords:  chiral separation; heart‐cutting; high‐resolution mass spectrometry; impurity profiling; mitiglinide; two‐dimensional liquid chromatography
    DOI:  https://doi.org/10.1002/rcm.70144
  23. Angew Chem Int Ed Engl. 2026 Aug 07. e1256471
    Tenfold Expansion Submicrometer MALDI Mass Spectrometry Imaging of Tissues and Cultured Cells.
    Chengyi Xie, Jianing Wang, Lei Guo, Xin Diao, Thomas Ka-Yam Lam, Yanyan Chen, Ruxin Li, Yue Zhang, Xiaoxiao Wang, Jiacheng Fang, Zhongping Yao, Klaus Dreisewerd, Zongwei Cai.
      Achieving  submicrometer lateral resolution in matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) is essential for molecular characterization at the single-cell and subcellular levels but is fundamentally limited by optical focusing and matrix crystallization. Here, we present tenfold expansion mass spectrometry imaging (10X ExMSI), an expansion MALDI-MSI workflow that reaches an effective lateral resolution of ∼500 nm on standard commercial instruments using a 5 µm acquisition step size without pixel oversampling. The SDS-free digestion protocol minimizes lipid leaching and supports broad detection of major lipid classes, including glycerophospholipids and sphingolipids, although a reduction in primary amine-containing lipids is observed due to cross-linking. Using 10X ExMSI, we resolve subcellular structures in mouse brain tissues, such as dendritic arborizations, that are challenging to access with existing MALDI-MSI implementations. We further demonstrate expansion-based MSI on cultured A549 cells, achieving subcellular-level lipid mapping. The method is fully compatible with standard MALDI-MSI systems, providing an accessible and scalable route to high-resolution, label-free molecular imaging. These capabilities open new opportunities for studying cellular architecture and molecular heterogeneity in biological and biomedical research.
    Keywords:  MALDI; expansion; mass spectrometry imaging; subcellular lateral resolution; subcellular structure
    DOI:  https://doi.org/10.1002/anie.1256471
  24. Bioanalysis. 2026 Aug 06. 1-10
    Development and validation of a LC-MS/MS method for the quantification of the E-type prostanoid receptor 4 antagonist HTL0039732 in human plasma, for a Phase I/IIa clinical trial.
    Dan Astley, Svatopluk Svetlik, Shelby Barnett, Gareth J Veal.
       BACKGROUND: HTL0039732 (HTL) is a novel E-type prostanoid receptor 4 (EP4) antagonist for the treatment of advanced solid tumors. An analytical method was developed to facilitate the analysis of patient samples as part of an ongoing Phase I/IIa clinical trial.
    METHODS AND RESULTS: A LC-MS/MS method for quantifying HTL in human plasma was developed and validated per regulatory guidelines. The assay was selective, with a 2 ng/mL detection limit observed. It demonstrated good precision (CV ≤11.9%) and accuracy (86-101%), with linearity from 10 to 2000 ng/mL. Recovery was consistent, and no significant matrix, anticoagulant, or carryover effects were observed.
    CONCLUSIONS: A sensitive and selective method for measuring HTL in human plasma was successfully developed and applied to clinical samples, generating first‑in‑human pharmacokinetic data for HTL. The assay is now being utilized in an early‑phase clinical trial setting.
    CLINICAL TRIAL REGISTRATION: https://clinicaltrials.gov/study/NCT05944237.
    Keywords:  EP4; HTL0039732; LC-MS/MS; cancer; pharmacokinetics; prostaglandin; validation
    DOI:  https://doi.org/10.1080/17576180.2026.2712785
  25. Methods Mol Biol. 2026 ;3018 211-225
    Profiling O-Acetylation in Sialoglycans Using MALDI-TOF and LC-MS/MS with Methylamidation and Permethylation Derivatization.
    Zhaoguan Wu, Isabelle Sirois.
      O-acetylation of sialoglycans plays a critical role in modulating the structural and functional properties of glycoproteins, with implications in immune response, cell signaling, and disease pathology. Among sialic acids, N-acetylneuraminic acid (Neu5Ac) and its derivatives have garnered attention as biomarkers for various cancers and inflammatory conditions due to their terminal positioning on glycan chains and their susceptibility to biochemical modification. Despite their biological relevance, comprehensive analysis of O-acetylation patterns remains challenging due to their chemical lability and loss during conventional glycomic workflows, such as permethylation. This protocol outlines a robust workflow combining methylamidation and permethylation with porous graphitic carbon (PGC)-based purification and high-resolution MALDI-MS and LC-MS/MS to analyze O-acetylation in serum/plasma-derived N-sialoglycans. We demonstrate sensitive and reproducible profiling of sialylated glycan structures. This method enables deeper insight into glycan modification dynamics and can be broadly applied to biomarker discovery and comparative glycomics.
    Keywords:  Cancer biomarkers; Glycomics; LC–MS/MS; MALDI–MS; Methylamidation; N-acetylneuraminic acid; O-acetylation; PGC purification; Posttranslational modification; Sialoglycans
    DOI:  https://doi.org/10.1007/978-1-0716-5166-7_14
  26. Forensic Sci Int. 2026 Aug 04. pii: S0379-0738(26)00282-3. [Epub ahead of print]388 113094
    Validation of a Dried Blood Spot (DBS) Liquid Chromatography - High-Resolution Mass Spectrometry (LC-HRMS) workflow for simultaneous identification of emerging opioids.
    Serena Mestria, Sara Odoardi, Valeria Valentini, Giulia Biosa, Sabina Strano-Rossi.
      The rapid emergence of novel synthetic opioids, including fentanyl analogues and nitazene-class substances, represents a critical public health concern. Their high potency and continuous structural modification demand highly sensitive and adaptable analytical approaches for reliable forensic detection. This study evaluates dried blood spot (DBS) sampling as a robust alternative to conventional liquid matrices for the identification of synthetic opioids. A method based on liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was developed and validated. Data acquisition was performed in full-scan data-dependent acquisition (DDA) mode. The combined DBS-LC-HRMS methodology is suitable for the detection of known fentanyl analogues and nitazenes, while also allowing the identification of potentially unknown compounds through data analysis. The limits of detection (LOD) and quantification (LOQ) ranged from 0.05 to 0.2 ng/mL and 0.1-0.75 ng/mL, respectively. The method established a linear range of 0.5-3 ng/mL for all analytes, with validation results supporting the suitability of DBS as a sample storage and pre-treatment approach, with simplified collection, storage and transport compared to traditional matrices. Stability was also assessed, demonstrating that DBS is a more robust and cost-effective approach for analyte preservation than frozen whole blood over a 7 days period. Furthermore, the method was successfully applied to real samples, confirming its suitability as an effective tool for forensic toxicology screening and analysis. The proposed workflow demonstrated satisfactory analytical performance, enabling the identification and quantification of 37 synthetic opioids; Overall, this strategy provides a flexible and efficient tool for routine forensic toxicology, with the capacity to adapt to the dynamic and evolving landscape of illicit synthetic opioids.
    Keywords:  DBS; Forensic toxicology; HRMS; Synthetic opioids
    DOI:  https://doi.org/10.1016/j.forsciint.2026.113094
  27. RSC Adv. 2026 Aug 04.
    Plug-and-play coaxial nested-tube supported liquid membrane extraction for the determination of nonsteroidal anti-inflammatory drugs in human plasma.
    Huifen Huang, Mengyuan Lv, QianQian Wang, Qiwen Zhang, Yongjie Yang, Di Chen, Zifei Qin.
      A robust coaxial nested-tube supported liquid membrane extraction (CNT-SLME) method was developed for determining nonsteroidal anti-inflammatory drugs (NSAIDs) in human plasma. The device was adapted from commercial nucleic acid purification columns using industrial-grade polypropylene fabric as a stable, hydrophobic support for a dihexyl ether liquid membrane. This "plug-and-play" three-phase system (acidic donor/organic membrane/alkaline acceptor) facilitates efficient analyte enrichment and reduced matrix interference. The setup allows simultaneous processing of multiple samples using a standard multi-well shaker, overcoming the low-throughput bottlenecks of traditional liquid-phase microextraction. Under optimized conditions, the method yielded satisfactory recoveries (46.89-66.63%), negligible matrix effects (86.50-103.51%), excellent linearity (R 2 > 0.998), and detection limits of 3.2-55.3 ng mL-1. Validation in spiked plasma demonstrated high accuracy (recoveries 90.05-104.69%) and precision (RSDs ≤ 8.26%). Given its simplicity, cost-effectiveness, and batch-processing capability, CNT-SLME provides a practical platform for routine therapeutic drug monitoring and clinical pharmacokinetic studies.
    DOI:  https://doi.org/10.1039/d6ra03853k
  28. Talanta. 2026 Aug 05. pii: S0039-9140(26)01056-8. [Epub ahead of print]312(Pt A): 130400
    A simple and sensitive QuEChERS-based LC-MS/MS method for quantifying 30 per- and polyfluoroalkyl substances (PFAS) in human serum: application to firefighter biomonitoring.
    Crystal L Sweeney, Royin Suwanratanabus, Neda Seyedsadjadi, Dac Nguyen, Jason Rennert, Farzaneh Masoud.
      Occupational exposure to per- and polyfluoroalkyl substances (PFAS) is under scrutiny as a potential cancer risk for firefighters. Existing methods for PFAS biomonitoring involve solid-phase extraction (SPE), either conventional or online, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). SPE requires a significant investment of time and technical skill for optimizing many variables to achieve maximum accuracy and precision. This increases analytical costs, making routine PFAS biomonitoring cost-prohibitive for a majority of fire service members. The motivation for this study was to develop an analytical method for measuring 30 PFAS in firefighters' serum (N = 84) using a QuEChERS (quick, easy, cheap, effective, rugged, and safe) protocol. For all target analytes, process efficiency (PE) and recovery efficiency (RE) were between 89 and 126% at 1 μg L-1 and between 85 and 133% at 25 μg L-1 (%RSD ≤ 13) (N = 5). At 1 and 25 μg L-1, matrix effects (ME) for all target PFAS ranged from 78 to 110% and 87 to 117%, respectively (%RSD ≤ 11). Precision criteria were met for all target analytes in both intra-day and inter-day precision assessments (%RSD ≤ 19). Limits of detection (LOD) and quantitation (LOQ) ranged from 0.006 to 0.3 μg L-1 and 0.02 to 0.9 μg L-1, respectively. This high-performance, high-throughput, and cost-effective alternative to the current gold standard for PFAS measurement in serum advances current firefighter biomonitoring capabilities by decreasing the time and cost of analysis, reducing constraints imposed by specialized laboratory equipment and expertise, and optimizing analytical batch processing and workflow.
    Keywords:  Firefighter biomonitoring; LC-MS/MS; Occupational exposures; PFAS; QuEChERS; Serum
    DOI:  https://doi.org/10.1016/j.talanta.2026.130400
  29. Drug Metab Pharmacokinet. 2026 Jun 19. pii: S1347-4367(26)00031-5. [Epub ahead of print]70 101545
    Development and validation of LC-MS-based bioanalytical methods for oligonucleotide therapeutics: points to consider.
    Yuchen Sun, Kosuke Saito, Shin-Ichiro Nitta, Ryoya Goda, Yuki Kishino, Hisashi Fujita, Masaaki Kakehi, Yoshiharu Hayashi, Satoshi Ito, Tomomi Ishida, Kiyomi Kikuchi, Kazuko Inoue, Kazutaka Shimbo, Takeru Yamaguchi, Fumihiro Jinno, Kaori Murakoshi, Chie Murata, Kentaro Takahara, Mitsuhiko Kawabata, Ruri Kikura-Hanajiri, Yoshiro Saito.
      The analysis of oligonucleotide therapeutics, including antisense oligonucleotides and siRNAs, using existing bioanalytical methods presents unique analytical challenges owing to their distinctive physicochemical properties. Although the ICH M10 guideline provides general bioanalytical validation frameworks, specific considerations for oligonucleotide therapeutics remain undefined, highlighting an urgent need for targeted points to be considered. This white paper presents comprehensive recommendations developed through expert consensus and a literature review by the Japan Agency for Medical Research and Development research groups, addressing the critical technical challenges in liquid chromatography-mass spectrometry (LC-MS)-based bioanalytical methods for oligonucleotide therapeutics. It focuses on ion-pair reversed-phase LC-MS methodologies and emerging techniques, such as hybridization extraction, addressing key issues, including mobile phase stability and degradation, metal adsorption mitigation, carryover prevention, internal standard selection, and matrix effects unique to nucleic acids. The comprehensive coverage includes sample preparation strategies, chromatographic optimization, mass spectrometric detection approaches, and validation parameters adapted from ICH M10, with fit-for-purpose modifications on a case-by-case basis when conventional acceptance criteria cannot be applied. This white paper provides practical recommendations for the development and validation of analytical methods specific to oligonucleotide therapeutics, supports regulatory submissions, and promotes the international harmonization of bioanalytical standards in this rapidly expanding therapeutic modality.
    Keywords:  Bioanalysis; ICH M10; LC–MS; Method validation; Oligonucleotide therapeutics; Regulatory submission; White paper
    DOI:  https://doi.org/10.1016/j.dmpk.2026.101545
  30. Anal Chim Acta. 2026 Oct 08. pii: S0003-2670(26)00809-3. [Epub ahead of print]1418 345859
    A reference-guided large language model workflow for mobile phase selection in thin-layer chromatography enabled by a polarity-space tetrahedron strategy.
    Wei-Song Kong, Song-Xue Shao, Li-Na Zhu, Xi Yu.
      Mobile phase selection in thin-layer chromatography (TLC) still relies heavily on empirical trial-and-error. Existing machine learning methods often exhibit limited predictive performance, as they depend on manual descriptor engineering and are sensitive to data quality. Here, a reference-guided large language model (LLM) workflow for TLC mobile phase recommendation is proposed. The LLM is used as a flexible inference interface that integrates structural similarity, polarity descriptors, and example-based analogical inference. The key methodological contribution is a polarity-space tetrahedron strategy for selecting reference compounds. Three polarity descriptors, including molecular refractivity, topological polar surface area, and n-octanol-water partition coefficient, are used to construct a three-dimensional space, and a target compound is constrained within a tetrahedron formed by four reference compounds to enable interpolation-based inference. A similarity-weighted version of this tetrahedron strategy is also developed. Using a publicly available high-throughput automated TLC dataset and DeepSeek-Reasoner as the inference engine, four reference selection strategies are compared. The similarity-weighted tetrahedron method achieves the best performance, with an availability of 81.48% and a final score of 0.8436 across 45 compound pairs evaluated in triplicate. The success rate of 92.16% confirmed by independent experimental validation supports the practical relevance of the recommendations. The framework also provides interpretable outputs, including structured rationales and experimental suggestions. This work demonstrates that combining a locally constrained reference construction strategy with an LLM offers a practical and interpretable tool for TLC mobile phase optimization, without requiring large labeled datasets or task-specific model training.
    Keywords:  DeepSeek; Large language model (LLM); Mobile phase screening; Tetrahedron strategy; Thin-layer chromatography (TLC)
    DOI:  https://doi.org/10.1016/j.aca.2026.345859
  31. J Chromatogr A. 2026 Jul 30. pii: S0021-9673(26)00639-4. [Epub ahead of print]1785 467311
    Position-specific isotope fragmentation differential quantification (PIF-DQ) of phenylalanine positional isotopomers: A rapid UPLC-MS/MS approach for urine analysis in the IAAO method.
    Ippei Suzuki, Mai Matsumoto, Keiko Komikado, Ryosei Sakai, Yoichi Hatamoto.
      The indicator amino acid oxidation (IAAO) method is a minimally invasive approach for estimating amino acid and protein requirements using a ¹³C-labeled indicator amino acid. In IAAO studies using l-[1-¹³C]phenylalanine ([1-¹³C]Phe), accurate determination of tracer-derived enrichment relative to unlabeled phenylalanine is important for calculating the tracer-to-tracee ratio (TTR). A key analytical challenge is distinguishing the administered [1-¹³C]Phe tracer from naturally occurring M + 1 phenylalanine isotopologues. In this study, we developed and validated a novel analytical method, termed Position-specific Isotope Fragmentation Differential Quantification (PIF-DQ), using ultra-high performance liquid chromatography-tandem mass spectrometry for the simultaneous quantification of l-phenylalanine (Phe) and its isotopologues in human urine. By optimizing collision energy, we established specific multiple reaction monitoring transitions to differentially quantify the species: m/z 166.1 > 120.2 for Phe; m/z 167.1 > 78.1, selective for the phenyl-group labeled Natural-(M + 1)-Phe; and m/z 167.1 > 77.1 for Total-(M + 1)-Phe (the sum of natural and tracer isotopologues). The method demonstrated good linearity (R² > 0.995), sensitivity (LLOQ ≤ 0.0004 µg/mL), accuracy (recoveries of 94.0-111.0 %), and precision (RSD < 13.1 %) over a 4.5-minute run time. Application to urine samples from an IAAO study successfully distinguished the tracer-derived elevation in the Total-(M + 1)-Phe/Phe ratio from the stable natural abundance ratio, demonstrating the robustness of the method against physiological variations like urinary concentration.
    Keywords:  IAAO method; Multiple reaction monitoring (MRM); Phenylalanine; Positional isotopomer; UPLC-MS/MS
    DOI:  https://doi.org/10.1016/j.chroma.2026.467311
  32. J Vis Exp. 2026 Jul 17.
    Extraction of Short-chain Fatty Acids from Mouse Feces Using A Two-phase Acid-Solvent Method.
    Chenyi Yang, Vincent Xu, John Wolf, Renee Frenette, Haiyun Wang, Mingrui Jasmine Zheng, Wei Qi, Wei Li, Yiying Zhang.
      This article describes a protocol for extracting short-chain fatty acids (SCFAs) from low-input mouse fecal samples using a two-phase acid-solvent extraction strategy. The method is designed for individual fecal inputs of approximately 20-50 mg and employs dilute hydrochloric acid and ethyl acetate to enable phase separation without derivatization. The protocol consists of fecal homogenization, acidification, organic solvent partitioning, and low-temperature centrifugation, and can be performed using standard laboratory equipment commonly available in research laboratories. Following extraction, SCFAs are suitable for downstream analysis by routine gas chromatography-based platforms. Successful execution of the protocol is indicated by the detection of clearly resolved SCFA peaks at characteristic retention times above background noise, as demonstrated for major fecal SCFAs including acetate, propionate, and butyrate from individual mouse samples. This protocol addresses a common procedural challenge in small-animal studies by providing a standardized workflow for fecal SCFA extraction from limited sample material while maintaining compatibility with routine analytical workflows.
    DOI:  https://doi.org/10.3791/70082
  33. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2026 Aug 07. 1-12
    Development, validation, and application of liquid chromatography mass spectrometry methods for the determination of juvenile hormone analogues in mealworms.
    Sjors Rasker, Rita Boerrigter-Eenling, Helmoed Cammin, Ricky Potjes, Lucas Agterberg, Marco H Blokland, Ane Arrizabalaga-Larrañaga, Saskia S Sterk.
      Juvenile hormone analogues (JHAs) are known to affect insect growth and can potentially be used in edible insect rearing. In this study, a reversed-phase liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method was developed for the detection of five common juvenile hormone analogues (fenoxycarb, pyriproxyfen, methoprene, kinoprene, and hydroprene) in mealworms. Sample clean-up was performed using reversed-phase solid-phase extraction cartridges. To enhance ionisation, methoprene, kinoprene, and hydroprene are detected as their 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) derivatives. The method was fully validated according to Commission Implementing Regulation (EU) 2021/808 and is capable of both quantitative and qualitative confirmation of fenoxycarb, pyriproxyfen, methoprene, and kinoprene (CCα values of 6.0-29 µg kg-1), and qualitative screening of hydroprene (with a CCβ of 7.7 µg kg-1). Additionally, it was demonstrated that liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) can also be used for the screening of JHA residues at relevant levels. The LC-MS/MS method was used to demonstrate the presence of methoprene in an incurred sample of mealworms. Finally, 14 mealworm samples originating from the Dutch national residue control plan were analysed, and none of the target analytes were found.
    Keywords:  Juvenile hormone analogue; food safety; mealworm; residue; veterinary drug
    DOI:  https://doi.org/10.1080/19440049.2026.2710625
  34. Methods Mol Biol. 2026 ;3018 111-122
    O- and N-Phosphoproteomic Analysis Using Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry.
    Yuqiu Wang, Minchu Tang, Wenfan Xie.
      Protein phosphorylation (O-linked and N-linked) is associated with a wide range of biological processes and cell signaling pathways. Aberrant phosphorylation is often observed as a hallmark of diseases, including cancer, neurodegenerative disorders, and metabolic syndromes. Mass spectrometry (MS) is an indispensable tool for studying protein phosphorylation. This is primarily due to its high sensitivity and throughput, which allows for comprehensive identification and quantification of phosphorylation events in complex biological samples. In this chapter, we describe a detailed protocol for phosphoproteomic analysis using one-dimensional online alkaline-pH reversed-phase nanoelectrospray-tandem MS (alkaline-pH-MS/MS). It complements traditional online low-pH reversed-phase nanoelectrospray-tandem MS (low-pH-MS/MS) by modulating the charge state distribution of phosphopeptides, which may facilitate the characterization of phosphoproteins with significant biological functions.
    Keywords:  Alkaline-pH-MS/MS; Mass spectrometry; Phosphoproteomics; Phosphorylation; Posttranslational modification
    DOI:  https://doi.org/10.1007/978-1-0716-5166-7_7
  35. Toxicol In Vitro. 2026 Aug 01. pii: S0887-2333(26)00098-6. [Epub ahead of print]117 106290
    A targeted lipidomics workflow for high-throughput phenotyping and mechanism-based in vitro hepatotoxicity studies.
    Tzu-Hsin Yen, Chander K Negi, Courtney Sakolish, Erin S Baker, Ivan Rusyn.
      Animal-based toxicity testing is limited in throughput and mechanistic characterization, underscoring the need for cell-based approaches. Incorporating molecular readouts, such as lipidomics, can enhance the translational relevance of cell-based assays. Because untargeted lipidomic analyses are time-consuming and can require large sample volumes, a targeted workflow was used in this study to characterize drug-induced liver injury-related lipid responses in primary hepatocytes from multiple species. Multiple reaction monitoring was applied in a targeted lipidomics workflow by coupling liquid-chromatography with triple quadrupole mass spectrometry (LC-MS/MS) for 148 lipid targets across 3 categories and 11 classes. We used primary hepatocytes from human, rat, monkey, and dog cultured in 96-well plates and exposed to chlorpromazine (1-30 μM), bosentan (1-200 μM), and fialuridine (1-100 μM) from culture day 4 to 8. Chlorpromazine induced the most pronounced lipid alterations in human, monkey, and rat hepatocytes, while responses in dog hepatocytes were minimal, with human cells showing a dose-dependent trend up to 10 μM. Bosentan induced dose-dependent lipid changes across species, strongest in human hepatocytes, whereas fialuridine caused limited lipid alterations. Overall, lipidomic effects closely aligned with conventional toxicity markers, reflecting compound and species-specific hepatotoxic sensitivity. The targeted lipidomics workflow provided mechanism-informed phenotyping of chemical-induced lipid responses in cultured hepatocytes. Moreover, it provided a framework for mechanistic evaluation, supported cross-species comparison, and complemented in vitro hepatotoxicity assessments.
    Keywords:  Cross species; DILI; In vitro Lipidomics; NAMS
    DOI:  https://doi.org/10.1016/j.tiv.2026.106290
  36. Anal Bioanal Chem. 2026 Aug 03.
    Salt-induced matrix effects on PFAS quantification in online SPE-LC-MS/MS analysis.
    Aleum Lee, Soo Min Song, Hee Sun Moon, Sung Pil Hyun.
      Residual inorganic ions in automated online solid-phase extraction (SPE) workflows can alter electrospray ionization and bias LC-MS/MS quantification of perfluoroalkyl substances (PFAS) in environmental waters. We evaluated the influence of modest salt concentration (10-30 mg/L NaCl or CaCl2) on the PFAS analytical response using an online SPE-LC-MS/MS system in negative ESI-MRM (Electrospray Ionization-Multiple Reaction Monitoring) mode. PFBA (50 ng/L) was non-detectable under salt-free conditions but produced well-defined peaks after salt addition, with detection limits improving to 0.17-0.22 ng/L (LOD) and 0.57-0.74 ng/L (LOQ) at 20-30 mg/L of NaCl and CaCl2. The isotopically labeled internal standard remained stable across conditions, but the PFBA/13C4-PFBA area ratio increased from 1.67 to 1.89, demonstrating incomplete compensation for salt-dependent response changes by isotope dilution. A matrix effect assessment across the full PFAS target list revealed a wide range from - 74% to + 1005%, with short-chain compounds exhibiting the strongest enhancements (up to tenfold for PFPeA) and longer-chain PFAS showing weaker or negative effects. NaCl amplified chain length-dependent contrast more strongly than CaCl2 did, producing divergent responses, particularly for C8-C9 compounds. These findings demonstrate that the environmentally relevant salt concentrations substantially alter PFAS response patterns in automated online SPE-LC-MS/MS systems, with implications for method validation and data interpretation. The ionic composition should be explicitly considered when quantifying PFAS in water matrices with varying ionic composition, particularly for short-chain compounds where internal standardization provides incomplete correction.
    Keywords:  Extraction; Matrix effects; Online SPE-LC–MS/MS; Per- and polyfluoroalkyl substances (PFAS); Quality assurance/control
    DOI:  https://doi.org/10.1007/s00216-026-06699-5
  37. Crit Rev Anal Chem. 2026 Aug 07. 1-22
    Fit-for-purpose analytics for cannabis: identification, quantification, and standardization of cannabinoids, flavonoids and terpenes.
    Eman Shawky, Lutfun Nahar, Satyajit D Sarker, Dina A Selim.
      Cannabis-based products used in medicine and wellness are chemically complex mixtures of cannabinoids, terpenes, flavonoids (including cannflavins), and degradation products whose profiles can shift substantially with cultivar genetics, cultivation conditions, extraction workflows, formulation choices, and storage history. This field-to-field and lab-to-lab variability, compounded by inconsistent analytical practices and uneven validation standards, can undermine potency labeling, cross-product comparability, and the interpretation of clinical and pharmacological findings. This review critically summarizes current quantitative and fingerprint-based strategies for measuring cannabinoids and related constituents across major product types (flowers/biomass, oils, edibles, and vape products) and proposes a practical "field-to-result" framework spanning sampling, method selection, validation, and lifecycle quality control. Drawing on peer-reviewed literature and metrological guidance, we evaluate LC-UV/LC-MS/MS, GC-FID/GC-MS, quantitative and structural NMR, and selected spectroscopic and aerosol approaches, with emphasis on isotope-dilution concepts, calibration design, matrix effects, validation criteria, and chemometric tools for multivariate fingerprints. The evidence is synthesized to map analyte classes and matrices to fit-for-purpose platforms and sample preparation schemes, highlight best practices for internal standards and matrix-matched calibration, and summarize commonly applied acceptance expectations for accuracy and precision. Strategies to reduce or correct matrix effects are detailed, and the role of high-dimensional fingerprints is explained as a complement to targeted potency panels for identity confirmation, process drift monitoring, lot release decisions, and traceability. Overall, combining validated potency assays with statistically governed fingerprinting offers a pragmatic route to harmonize cannabinoid testing, improve data integrity, and better support future phytomedicine research, clinical translation, and regulatory oversight.
    Keywords:  Cannabinoids; Cannabis sativa; chemometrics; flavonoids; terpenes
    DOI:  https://doi.org/10.1080/10408347.2026.2714459
This page is being maintained by Thomas Krichel. It was last updated on 2026‒08‒09 at 13:46.