bims-metlip 2026-01-25 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2026–01–25
24 papers selected by
Sofia Costa, Matterworks

HPLC-MS/MS Method for the Simultaneous Quantification of SGLT2 Inhibitor YWS22026 and Its M3 Metabolite in Rat Plasma, With Application to Toxicokinetic Study.
Development and Validation of a Rapid LC-MS/MS Method for Quantification of 20 Bile Acids in Serum and Application to Murine Hepatotoxicity Models.
Rapid and selective characterization of antibody-drug conjugates in complex sample matrices by native affinity liquid chromatography-mass spectrometry.
An isotope dilution-liquid chromatography tandem mass spectrometry-based candidate reference measurement procedure for the quantification of dehydroepiandrosterone in human serum and plasma.
A Buffered LC-MS Method for Resolving and Quantifying Albiflorin and Paeoniflorin.
Quantitative Food Compounds Enable Dietary Ontology Referencing across 500 Foods and Human Plasma.
Analysis of Vitamin A Stable Isotope Tracers using LC-MS/MS.
Insect Lipidomics: Advances, Applications, and Physiological Insights.
Determination of 315 hazardous organic substances in waste textile raw materials and products by high-performance liquid chromatography-tandem mass spectrometry.
Development and application of SPE-UHPLC-MS/MS methods for quantification of 29 steroids in rat serum.
Integrated GC-MS/MS Metabolomics in Cardiovascular Disease: Targeted Nitro-Oleic Acid Quantification Meets Untargeted Profiling.
Comparison of six serum analyte workflows using routine liquid chromatography-tandem mass spectrometry methods at multiple laboratories with the Cobas i 601 analyzer, an automated mass spectrometry system.
Ultrasonic-assisted magnetic sorbent-wrapped stick dip extraction enables batch-scale and green LC-MS/MS analysis of tyrosine kinase inhibitors in plasma.
Simultaneous determination of short-chain fatty acids and tryptophan metabolites by a propyl chloroformate- derivatized GC-MS approach.
Different chromatographic techniques and recent advancements for biomedical and pharmaceutical applications.
Direct measurement of free glucocorticoids in small volumes of mouse and rat serum using ultrafiltration and liquid chromatography-tandem mass spectrometry.
Development and validation of the LC-MS/MS method based on self-made purification column for simultaneous determination of novel organophosphate esters in rice-based products.
Analytical Approaches for the Determination of Lidocaine and Its Metabolites: A Comprehensive Insight.
Multi-Solvent Suppression Ultrafast 2D COSY for High-Throughput Wine Screening.
Analytical Methods for Identification and Quantification of Quinoa Saponins: A Review.
Simultaneous determination of free and total metabolite concentrations in proteinaceous specimens by 1D 1H CPMG NMR.
Rapid, non-invasive diagnosis of tuberculosis using desorption separation ionization mass spectrometry (DSI-MS) and urinary carbonyl metabolite fingerprints.
Simultaneous Determination of Glyphosate, Aminomethylphosphonic Acid, and Glufosinate in Green Coffee Beans by LC-MS/MS: Optimization, Validation, and Field Study.
Microalgae-derived quality control material for reproducible and scalable LC-MS system suitability testing.

Biomed Chromatogr. 2026 Mar;40(3): e70366

HPLC-MS/MS Method for the Simultaneous Quantification of SGLT2 Inhibitor YWS22026 and Its M3 Metabolite in Rat Plasma, With Application to Toxicokinetic Study.

Zhiyuan Li, Qing Shao, Hongqun Qiao.

  A sensitive and robust HPLC-MS/MS method was developed and validated for the simultaneous determination of YWS22026 and its major metabolite M3 in rat plasma using protein precipitation. Chromatographic separation was achieved on a Shim-pack Scepter C18 column (4.6 × 50 mm, 3 μm) with a gradient elution program (mobile phase A: 0.2% formic acid in water; mobile phase B: methanol) at a flow rate of 0.9 mL/min. Detection was performed via positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. The method demonstrated excellent specificity, accuracy (relative error, %RE < 9.1%), precision (relative standard deviation, %RSD < 9.9%), and linearity (5-2000 ng/mL for both analytes). The lower limit of quantification (LLOQ) was 5 ng/mL. Stability studies confirmed that plasma samples remained stable under various storage conditions (at room temperature for 10 h, -20°C for 22 days, three freeze-thaw cycles, and 4°C autosampler storage for 73 h). This validated method was successfully applied to a 26-week concomitant toxicokinetic study in Sprague-Dawley rats, which revealed significant sex-related differences in exposure for both YWS22026 and M3. These findings provide valuable insights for informing the safety assessment of YWS22026 in subsequent clinical trials.

Keywords:  HPLC‐MS/MS; YWS22026; metabolite; toxicokinetics

DOI:  https://doi.org/10.1002/bmc.70366
Biomed Chromatogr. 2026 Mar;40(3): e70368

Development and Validation of a Rapid LC-MS/MS Method for Quantification of 20 Bile Acids in Serum and Application to Murine Hepatotoxicity Models.

Nianchang Zheng, Yaqin Niu, Juanna Wei, Lingjie Mo, Junyi Lin, Jingjing Cui, Liliang Li, Yan Jiang.

  Bile acids function both as facilitators of biliary lipid transport and as signaling mediators that maintain metabolic homeostasis. Perturbations in bile acid metabolism accompany diverse forms of liver injury, yet conventional clinical assays lack the sensitivity and specificity required for comprehensive bile acid profiling. To address this, we aimed to develop and validate a rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of 20 bile acids using minimal serum volume and shortened analysis time. The developed method requires only 20 μL of serum and completes analysis within 13 min. Following protein precipitation, analytes were separated on a C18 column with gradient elution. The method demonstrated excellent sensitivity with limits of detection of 1-2 ng/mL and a uniform limit of quantification of 5 ng/mL for all analytes. Validation results showed satisfactory performance: accuracy within ± 14.8%, intra-assay and interassay precision below 16.5%, extraction recovery of 72.6%-108.4%, and acceptable matrix effects. Application to murine hepatotoxicity models induced by α-amanitin (α-AMA), ethanol, and acetaminophen (APAP) revealed distinct, compound-specific bile acid alterations. This work establishes a rapid, miniaturized, and validated LC-MS/MS approach, demonstrating its utility in revealing toxin-specific bile acid signatures, which provides a valuable tool for preclinical hepatotoxicity research and potential translational applications.

Keywords:  LC‐MS/MS; bile acids; hepatotoxicity; method validation; α‐amanitin

DOI:  https://doi.org/10.1002/bmc.70368
MAbs. 2026 Dec 31. 18(1): 2618314

Rapid and selective characterization of antibody-drug conjugates in complex sample matrices by native affinity liquid chromatography-mass spectrometry.

Dan Bach Kristensen, Nanna Sofie Eskesen, Clara Coll-Satue, Alexandre Nicolas, Jan Kirkeby Simonsen, Lykke Rasmussen, Trine Meiborg Sloth, Martin Ørgaard, Elizabeta Madzharova, Simon Krabbe, Katrine Zinck Leth, Pernille Foged Jensen, Alain Beck.

  Antibody-drug conjugates (ADCs) and other biopharmaceuticals require robust analytical methods to assess biotransformation in biological matrices. Current approaches often require off-line enrichment and extensive chromatographic separation, limiting throughput and complicating data processing. We developed a native affinity liquid chromatography-mass spectrometry (aLC-MS) method using POROS CaptureSelect FcXL columns combined with optimized solvents and MS parameters for direct analysis (1D aLC-MS) of ADCs and other antibody-derived formats in complex sample matrices, such as serum. The method was evaluated using stability studies and concentration series in mouse serum. Direct analysis enabled accurate determination of drug-antibody ratio (DAR), drug-load distribution (DLD) and relative drug abundance across samples without chromatographic peak integration. Stability studies revealed distinct ADC biotransformation profiles in serum versus PBS, including maleimide hydrolysis and disulfide exchange at under-conjugated cysteine sites. The aLC-MS method achieved excellent linearity (R2 = 0.99) over 125-2000 µg/mL in serum and demonstrated sensitivity to 31.25 µg/mL. This rapid, selective aLC-MS method enables high-throughput monitoring of ADC quality attributes in complex matrices with minimal sample preparation, supporting biopharmaceutical product development and bioanalysis applications. The method is exclusively based on MS results, which makes data processing and reporting fast and easy to automate.

Keywords:  Antibody-drug conjugate; drug-load distribution; drug-to-antibody ratio; native affinity LC-MS; serum stability

DOI:  https://doi.org/10.1080/19420862.2026.2618314
Clin Chem Lab Med. 2026 Jan 22.

An isotope dilution-liquid chromatography tandem mass spectrometry-based candidate reference measurement procedure for the quantification of dehydroepiandrosterone in human serum and plasma.

Lea Lewin, Michael Stadlmeier, Neeraj Singh, Friederike Bauland, Daniel Köppl, Alexander Gaudl, Andrea Geistanger, Uta Ceglarek, Manfred Rauh, Judith Taibon.

   OBJECTIVES: Dehydroepiandrosterone (DHEA) is a steroid prohormone and important precursor for estrogen and testosterone biosynthesis. For differential diagnosis of sexual development disorders (e.g., polycystic ovary syndrome, congenital adrenal hyperplasia), accurate measurement of DHEA in clinical testing is essential. To address that need for high-quality, reproducible assays, an isotope dilution-liquid chromatography (LC)-tandem mass spectrometry (MS/MS)-based candidate reference measurement procedure (RMP) for the quantification of DHEA was developed and validated.
METHODS: A certified primary reference material from the National Measurement Institute of Australia (NMIA), was used for DHEA assay calibration and to ensure traceability to the International System of Units (SI). Two-dimensional heart-cut chromatography was employed for LC-MS/MS in combination with a solid phase extraction sample preparation protocol, to mitigate matrix effects and prevent interference coelution. Selectivity was evaluated by spiking either the analyte or internal standard with potential interferents, such as dehydroepiandrosterone-sulfate, testosterone, and similar steroid compounds. For the evaluation of matrix effects, standard line slopes of various matrices were compared. Precision and accuracy were assessed via a multi-day validation experiment, and variability components were estimated using variance component analysis. Measurement uncertainty was evaluated in compliance with current guidelines.
RESULTS: This candidate RMP proved suitable for the analysis of DHEA in the measurement range of 0.0800-36.0 ng/mL (0.277-125 nmol/L). Predefined requirements for sensitivity and selectivity were fully met, and independence from matrix effects was demonstrated successfully. Across all tested concentration levels, intermediate precision was ≤1.5 % and repeatability was ≤1.0 %, while the relative mean bias ranged from -1.1 to 0.1 %. Regardless of DHEA concentration or sample type, the expanded measurement uncertainty for reference value assignment (n=6) was ≤1.7 %.
CONCLUSIONS: This isotope dilution-LC-MS/MS-based candidate RMP for DHEA in human serum and plasma met pre-defined analytical performance requirements such as precision, specificity and measurement uncertainty, and showed superior selectivity towards several potential interferents tested. It is suitable for application in clinical sample evaluation and routine assay standardization.

Keywords:  SI units; dehydroepiandrosterone; isotope dilution-liquid chromatography-tandem mass spectrometry; quantitative nuclear magnetic resonance spectroscopy; reference measurement procedure; traceability

DOI:  https://doi.org/10.1515/cclm-2025-1156
Biomed Chromatogr. 2026 Mar;40(3): e70353

A Buffered LC-MS Method for Resolving and Quantifying Albiflorin and Paeoniflorin.

Alina Gazizova, Ela Hümay Altincubuk, Christina Oppermann.

Albiflorin and paeoniflorin are bioactive isomers found in Paeoniae Radix Alba, a key component of Traditional Chinese Medicine. Their accurate and simultaneous quantification is essential for pharmacological research. Standard separation methods often rely on LC-MS using a mobile phase containing 0.1% formic acid. This study demonstrates that the use of 0.1% formic acid generates a secondary peak for albiflorin that exhibits an identical mass-to-charge ratio and similar fragmentation pattern as the main peak. To address this, an LC-MS method was developed using a Kinetex phenyl-hexyl column with a gradient elution using a buffered mobile phase of 10 mM ammonium acetate and acetic acid (pH 4.4) in water and methanol. The method was validated for linearity, precision, accuracy, and sensitivity. It successfully separated paeoniflorin and albiflorin preventing the formation of the albiflorin artifact. The method demonstrated good linearity over a concentration range of 1-50 μg/mL. Applicability was tested through the analysis of a Paeoniae Radix Alba extract. The developed LC-MS method enables accurate and simultaneous quantification of albiflorin and paeoniflorin by eliminating the formation of a second albiflorin peak. This makes the method potentially suitable for pharmacological studies of Paeoniae Radix Alba and other plants containing albiflorin and paeoniflorin.

DOI: https://doi.org/10.1002/bmc.70353
Anal Chem. 2026 Jan 18.

Quantitative Food Compounds Enable Dietary Ontology Referencing across 500 Foods and Human Plasma.

Julius Agongo, Harsha Gouda, Aubreyana McMaugh, Celeste Allaband, Seyedeh Mehrnaz Aghili, Marta Sala-Climent, Shipei Xing, Jennifer Cao, Jasmine Zemlin, Mingxun Wang, Kathleen Dorrestein, Tiffany Holt, Rob Knight, Brigid S Boland, Monica Guma, Pieter C Dorrestein.

Accurate and reproducible dietary assessment remains a persistent challenge in the clinical and nutritional sciences. We present a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the absolute quantification of 200 literature-curated food compounds across 500 food item extracts and human plasma samples. The assay uses external matrix-matched calibration and robust quality control, including postextraction recovery, matrix effect assessment, and intra- and interday precision validation, demonstrating high linearity (R2 > 0.99), low coefficients of variation (<15%), and recovery within 100 ± 15% for >75% of analytes. Using 200 authentic food standards against 500 complex food items resulted in reproducible detection and quantification of 102 food compounds across diverse food classes. Their specificity and distribution were evaluated at multiple levels of dietary ontology. Supervised multivariate analysis (PLS-DA) identified discriminative compound panels that classified foods such as citrus, dairy, and vegetables with high accuracy. Key compounds such as hesperidin, hypaphorine, and piperine demonstrated strong source specificity and were applied to human plasma samples from an inflammatory bowel disease (IBD) cohort following a Mediterranean diet. Food compound concentrations tracked with dietary intake, confirming hypaphorine's association with hummus and piperine's correlation with black-pepper-containing meals. This study demonstrates the utility of a rigorously validated targeted metabolomics workflow for both food chemistry and translational dietary intake research. The framework enables quantitative mapping of food molecules to dietary exposures and supports the development of more objective, chemistry-based dietary assessment strategies in clinical contexts.

DOI: https://doi.org/10.1021/acs.analchem.5c06769
J Vis Exp. 2025 Dec 30.

Analysis of Vitamin A Stable Isotope Tracers using LC-MS/MS.

Georg Lietz, Dan Astley, Alice Goddard, Anthony Oxley.

The retinol isotope dilution (RID) method estimates the total body stores (TBS) of vitamin A in humans through stable isotope dilution with the body's vitamin A pool. For this, it is critical to accurately and efficiently determine the plasma isotopic ratio of labeled to non-labeled retinol through mass spectrometry. To avoid extensive and time-consuming extraction and/or purification procedures, such as preparative HPLC and derivatization, LC-MS/MS is employed to conduct fast, sensitive, and simultaneous analysis of labeled and non-labeled retinoids. The method utilizes two levels of detection: (i) an initial mass/charge (m/z) separation of parent (precursor) ions, followed by (ii) detection of fragmented daughter (product) ions. This results in high sensitivity, with retinol detection limits as low as 6 fmol on-column. Despite the advantages of tandem mass spectrometry, a liquid chromatographic separation is required to separate retinol from retinyl esters since the terminal functional groups are lost during ionization, resulting in similar parent (m/z of 269) and daughter ion fragmentation patterns. The article describes the isolation of endogenous and labeled (13C or 2H) retinol from plasma by solvent extraction, followed by quantification by LC-MS/MS under atmospheric pressure chemical ionization (APCI) in positive ion mode.

DOI: https://doi.org/10.3791/69558
Adv Exp Med Biol. 2026 ;1494 261-291

Insect Lipidomics: Advances, Applications, and Physiological Insights.

Laura Palanker Musselman, Doga Cedden, Gözde Güney, Umut Toprak.

  Lipidomics, a specialized branch of metabolomics, investigates the diversity and functionality of lipids in biological systems. Lipids serve crucial roles in energy storage, membrane composition, and environmental acclimation in insects, underpinning processes such as development and stress responses. Advances in analytical technologies, such as liquid chromatography-mass spectrometry (LC-MS), have enabled precise identification and quantification of lipid species, providing unprecedented insights into lipid metabolism and dynamics. Key lipid classes, including triacylglycerols and phospholipids, exhibit structural and functional versatility, adapting to environmental pressures through mechanisms like homeoviscous adaptation. These dynamic lipid responses are essential for maintaining cellular and cuticular integrity and functionality under stress. By exploring lipid diversity and adaptations, lipidomics offers valuable perspectives on insect physiology, survival strategies, and evolutionary ecology. This chapter summarizes methods used to study insect lipidomes and highlights comparative lipidomic studies that have advanced our understanding of insect biology.

Keywords:  Cuticular hydrocarbons; Diapause; Ecdysteroids; Insect lipidomics; Mass spectrometry

DOI:  https://doi.org/10.1007/978-3-032-04842-4_878
Anal Bioanal Chem. 2026 Jan 19.

Determination of 315 hazardous organic substances in waste textile raw materials and products by high-performance liquid chromatography-tandem mass spectrometry.

Xuwei Guo, Youzhi Su, Fang Li, Yanmei Li, Jun Liu.

  A method was established and verified for identifying 315 toxic organic compounds-such as chlorophenols, dyes, pesticides, perfluoroalkyl acids, and fungicides-in waste textile materials and finished products through ultrasound-assisted extraction coupled with high-performance liquid chromatography-tandem mass spectrometry. Acetonitrile was utilized as the extraction solvent in two sequential ultrasonic extraction steps to isolate the analytes from the samples. For liquid chromatography, a C18 column was used for separation. In electrospray ionization positive mode, mobile phase gradient elution was performed with a mixture of acetonitrile and a 0.1% formic acid solution (containing 2 mmol/L ammonium acetate) in water. A mobile phase gradient elution using acetonitrile and a 2 mmol/L aqueous ammonium acetate solution was applied under electrospray ionization in negative mode, with detection conducted via multiple reaction monitoring (MRM). The analysis revealed that all 315 target compounds maintained strong linearity across their calibration ranges, with correlation coefficients (R2) consistently above 0.9901. The detection limits (LOD) were between 0.01 and 3.8 µg/kg, whereas the quantification limits (LOQ) ranged from 0.03 to 10.90 µg/kg. Method validation through spiking at concentrations of 10, 20, and 100 µg/kg was performed on three sample types: waste cotton lint, waste wool yarn, and waste hemp yarn. The recoveries ranged from 61.3 to 119.9%, with relative standard deviations (RSDs) between 1.1 and 15.9%. Overall, the method offers high sensitivity, accuracy, and reproducibility, rendering it effective for the routine detection of residual organic contaminants in recycled textile materials.

Keywords:  HPLC‒MS/MS; Organic hazardous substances; Ultrasonic extraction; Waste textile raw materials and products

DOI:  https://doi.org/10.1007/s00216-025-06301-4
J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jan 11. pii: S1570-0232(26)00013-9. [Epub ahead of print]1271 124924

Development and application of SPE-UHPLC-MS/MS methods for quantification of 29 steroids in rat serum.

Haiyang Cui, Dongliang Yang, Yonggang Zheng, Jing Li, Hairui Yan, Xiaoli Gao.

  Steroids and their metabolites play crucial roles in reproductive health, endocrine diseases, and tumor development. Comprehensive analysis of these compounds is essential for both mechanistic investigations and clinical evaluations. Although comprehensive analytical methods have been established for steroids in human clinical samples, their direct application to rat specimens remains limited, particularly with the absence of validated methods for detecting certain hydroxylated steroid metabolites in vivo. This study developed highly sensitive solid-phase extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry methods for the quantitative analysis of 29 steroid metabolites. Through optimized derivatization of steroids and integration of two-dimensional liquid chromatography techniques, the method effectively reduces interference from non-volatile salts and overcomes trace-level detection challenges. Method validation demonstrated a wide linear range (0.002-250 ng/mL), low limits of quantification (2-100 pg/mL), high accuracy (88-114%), and precision (intra- and inter-batch CV ≤14%). The method exhibited acceptable extraction recovery, matrix effects, and stability, confirming compliance with regulatory standards. The validated method was successfully applied to characterize serum steroid metabolic profiles in both N-methyl-N-nitrosourea-induced breast cancer and ovariectomized rat models, revealing alterations in estrogen 2-hydroxylation and pregnenolone 21-hydroxylation pathways mediated by key steroidogenic enzymes, including CYP1A1/CYP1B1 and potentially CYP21A2-independent mechanisms, providing robust support for preclinical research on steroid-related diseases.

Keywords:  Breast cancer; CYP1A1; CYP1B1; CYP21A2; Derivatization; Steroids; UHPLC-MS/MS

DOI:  https://doi.org/10.1016/j.jchromb.2026.124924
Anal Chem. 2026 Jan 22.

Integrated GC-MS/MS Metabolomics in Cardiovascular Disease: Targeted Nitro-Oleic Acid Quantification Meets Untargeted Profiling.

Yasmin Elshoura, Magy Herz, Mohamed Z Gad, Rasha Hanafi.

In recent decades, interest in nitrated fatty acids (NO2-FAs) has grown due to their role as endogenous signaling molecules involved in health and disease. As a result, their metabolic profiling has gained increasing attention. For metabolite analysis, GC-MS/MS offers greater sensitivity and robustness than LC-MS/MS, with more reliable annotation libraries. This study investigates metabolic dysregulation in cardiovascular disease (CVD) patients by using advanced metabolomics. A novel GC-MS/MS method for profiling NO2-FAs was developed, showing improved precision using 17-BrHDA as an internal standard compared to previous HDA-based methods. It is also the first report of alkylation and silylation derivatization of 17-BrHDA, demonstrating superior GC-MS sensitivity for pentafluorobenzyl-alkylated fatty acids over their silylated counterparts in positive ion mode. Untargeted metabolomics was applied to plasma samples from acute myocardial infarction (AMI) patients and healthy controls using both derivatization techniques. Multivariate analysis (PCA and PLS-DA) revealed distinct metabolic profiles. Key metabolites, identified based on VIP scores, were annotated via the Human Metabolome Database and literature. Findings highlight the complementary nature of both derivatization approaches for comprehensive plasma metabolome analysis. Notably, NO2-OA levels were significantly elevated (p < 0.01) in AMI patients, indicating its possibility to be utilized as a cardiovascular biomarker. This study represents the first use of alkylation derivatization in untargeted metabolomics for AMI and introduces a highly sensitive GC-MS/MS method with an innovative internal standard and optimized derivatization for cardiovascular biomarker discovery. The method demonstrates the potential to discriminate between groups of patients and healthy subjects.

DOI: https://doi.org/10.1021/acs.analchem.5c05647
Clin Chem Lab Med. 2026 Jan 26.

Comparison of six serum analyte workflows using routine liquid chromatography-tandem mass spectrometry methods at multiple laboratories with the Cobas i 601 analyzer, an automated mass spectrometry system.

Pieter Vermeersch, Robert de Jonge, Takamaro Miyazawa, Mads Nybo, Palle Fruekilde, Barry J Toole, Philippe Metz, Eva Albrecht, Karl Slusarczyk, Michael Vogeser, Robert L Fitzgerald.

   OBJECTIVES: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers substantial analytical advantages over traditional immunoassay methods, including greater specificity and robustness. Conventional LC-MS/MS methods are labor-intensive, requiring highly trained, dedicated laboratory staff. The Cobas® i 601 analyzer (Roche Diagnostics International Ltd., Rotkreuz, Switzerland) is an automated, random-access, mass spectrometry (MS) system designed to improve workflows and reduce turnaround times. This study evaluated turnaround times for analyses with the i 601 analyzer vs. routine LC-MS/MS using batch processing.
METHODS: We performed a workflow analysis for six sets of analytes (mycophenolic acid, antibiotics, antiepileptics, voriconazole, testosterone, and 25-hydroxyvitamin D) in seven laboratories across Europe, North America, and Asia. We recorded batch size, result turnaround times, and hands-on times (including quality control) for 93 batches analyzed on routine LC-MS/MS. These were compared to equivalent measurements generated using the i 601 analyzer in batch mode as part of a prototype pilot study at Ludwig Maximilian University, Munich.
RESULTS: Turnaround times were shorter with the i 601 analyzer vs. routine LC-MS/MS for all tested workflows, independent of batch size and analyte workflow. Median time from batch start to final result availability with the i 601 analyzer was 6.8-fold shorter vs. routine LC-MS/MS (1 h, 34 min vs. 10 h, 37 min, respectively). In addition, median total hands-on time was 19-fold shorter with the i 601 analyzer vs. routine LC-MS/MS (5 vs. 93 min).
CONCLUSIONS: Compared with routine LC-MS/MS methods, the i 601 analyzer substantially reduced turnaround and hands-on times for common analytes, regardless of batch size.

Keywords:  automation; liquid chromatography-tandem mass spectrometry; steroids; therapeutic drug monitoring; workflow

DOI:  https://doi.org/10.1515/cclm-2025-0994
Anal Bioanal Chem. 2026 Jan 23.

Ultrasonic-assisted magnetic sorbent-wrapped stick dip extraction enables batch-scale and green LC-MS/MS analysis of tyrosine kinase inhibitors in plasma.

Dong Wang, Mengrong Li, Xiangyu Zhang, Shiwei Chai, Di Chen.

  A novel ultrasound-assisted magnetic sorbent-wrapped stick dip extraction (UA-MSWS-DE) method was developed for the batch-scale determination of seven tyrosine kinase inhibitors (TKIs) in human plasma. The extraction device was fabricated by immobilizing octadecylphosphonic acid-functionalized magnetic nanoparticles onto flame-sealed glass capillaries embedded with magnets. Ultrasonic-assisted dip extraction was used to extract TKIs from diluted plasma, followed by quantification via liquid chromatography-tandem mass spectrometry (LC-MS/MS). Key extraction parameters were systematically optimized, yielding the following optimal conditions: sample pH 5.0 and volume 1.0 mL, sorbent particle size 20 nm and dosage 10 mg, extraction time 20 min, and desorption with 150 µL of ethanol for 8 min. The method demonstrated excellent linearity (R2 ≥ 0.99), low limits of quantification (0.1-5.0 ng/mL), and good intra-/inter-day precision (RSDs < 14.95%). The accuracy in blinded plasma samples, tested at 0.25-25% of the studied concentration range, ranged from 87.13 to 112.33%. Compared to conventional magnetic dispersive solid-phase extraction, the UA-MSWS-DE strategy simplifies handling and is designed to facilitate potential parallel processing using only an ultrasonic bath. Five greenness metrics confirmed the method's high sustainability. Furthermore, the modular design enables adaptation to other analytes by modifying the sorbent coating, making UA-MSWS-DE a sensitive, green, and automation-ready method for therapeutic drug monitoring.

Keywords:  Dip extraction; Liquid chromatography–tandem mass spectrometry; Magnetic sorbent; Therapeutic drug monitoring; Tyrosine kinase inhibitors

DOI:  https://doi.org/10.1007/s00216-026-06335-2
J Chromatogr B Analyt Technol Biomed Life Sci. 2026 Jan 12. pii: S1570-0232(26)00014-0. [Epub ahead of print]1271 124925

Simultaneous determination of short-chain fatty acids and tryptophan metabolites by a propyl chloroformate- derivatized GC-MS approach.

Yefeng Han, Xiaofang He, Yueling Gong, Mingxiao Li, Lili Sheng, Ningning Zheng, Jianbo Wan, Houkai Li, Yuanyuan Li.

  Short-chain fatty acids (SCFAs) and tryptophan metabolites, serve as key mediators of host-microbiota crosstalk, influencing physiological and pathological processes. Their interconnected roles necessitate simultaneous quantification to fully elucidate the potential mechanism of gut microbiota in metabolic diseases. However, they are difficult to be detected simultaneously due to differences in content or different polarity which contain specific carboxylic group, amino group or phenolic hydroxyl groups. In current study, our primary goal is to establish a rapid and sensitive quantitative measurement for SCFAs and tryptophan metabolites by using propyl chloroformate-(PCF) derivatization based on gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) analysis. Then, we applied the established method in measurement of serum samples from healthy subjects and metabolic associated fatty liver disease (MAFLD) patients. First, we optimized the reaction conditions including PCF volume, reaction time, extraction reagent ratio, and alkaline reagent concentration, enabling the simultaneous detection of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, 2-methylvaleric acid, hexanoic acid, indole, succinic acid, quinolinic acid, indole acetic acid, glutamine, indole butyric acid, 3-hydroxyanthranilic acid, melatonin, kynurenine, tyrosine, and tryptophan. 2-chloro-L-phenylalanine was set as the internal standard. The optimized condition showed good linearity, and the intra/inter-day precision achieved in the range from 0.81% to 14.88%. The recovery ranged from 85.68% to 114.50% and the matrix effect ranged from 85.81% to 113.42%. In addition, the influence of different storage conditions on sample stability was also acceptable. Finally, the quantitation result of serum samples indicated that isovaleric acid, quinolineic acid, indoleacetic acid, glutamine, melatonin, tyrosine, and tryptophan had significant difference between healthy subjects and MAFLD patients. The levels of isovaleric acid, indoleacetic acid, glutamine, tyrosine, and tryptophan had significant correlations with clinical indicators such as ALT, AST, Cap, Cr and TG, supporting their potential for further translational studies. In summary, this improved method is applicable for quantitative measurement of SCFAs and tryptophan metabolites, as well as those endogenous metabolites containing carboxylic, amino and phenolic hydroxyl groups.

Keywords:  Gas chromatography–mass spectrometry; Metabolic associated fatty liver disease; Propyl chloroformate derivatization; Short-chain fatty acids; Tryptophan metabolites

DOI:  https://doi.org/10.1016/j.jchromb.2026.124925
North Clin Istanb. 2025 ;12(6): 739-752

Different chromatographic techniques and recent advancements for biomedical and pharmaceutical applications.

Ozlem Coskun, Sama Akbarzadeh, Basak Guncer.

  Chromatography remains a cornerstone analytical technique in pharmaceutical and biomedical sciences, with recent innovations significantly expanding its capabilities. Advances such as fast chromatography, two-dimensional liquid chromatography (2D-LC), supercritical fluid chromatography (SFC), and hyphenated techniques, including liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS), have broadened the scope of its applications. These developments enhance resolution, sensitivity, and efficiency, enabling more robust analysis of complex biological and pharmaceutical samples. These methods address complex analytical challenges, improving precision, speed, and efficiency in separating and analyzing biomolecules. Emerging technologies, including miniaturized liquid chromatography, shear flow chromatography, column arrays, and microfluidic chip-based systems, present exciting opportunities for the future. These developments enhance the capability of chromatography to analyze trace compounds, optimize drug formulations, and ensure the quality control of pharmaceuticals. Chromatography is also increasingly integrated with cutting-edge techniques like metabolomics and proteomics, furthering its impact on biomarker discovery and personalized medicine. This paper reviews recent advancements in chromatographic methods and their practical applications in the pharmaceutical and biomedical fields. It highlights the critical role of chromatography in drug discovery, purification of therapeutic compounds, and metabolite profiling.

Keywords:  Biomedical analysis; biomarker discovery; chromatography; pharmaceutical applications; protein

DOI:  https://doi.org/10.14744/nci.2025.42966
PLoS One. 2026 ;21(1): e0341089

Direct measurement of free glucocorticoids in small volumes of mouse and rat serum using ultrafiltration and liquid chromatography-tandem mass spectrometry.

Anna Mazurenko, Melody Salehzadeh, Kiran K Soma.

Glucocorticoids are critical steroid hormones secreted from the adrenal glands. In mice and rats, over 90% of circulating corticosterone is bound to proteins such as corticosteroid-binding globulin and albumin, and the rest is unbound (free). Only free glucocorticoids can enter cells and bind receptors, so it is crucial to measure free glucocorticoids. Some studies have estimated free glucocorticoid levels, but such estimates might be inaccurate as they do not take temperature and protein binding by competing steroids into account. Far fewer studies have directly measured free glucocorticoid levels in serum, and current methods to do so are time-consuming and require sample volumes (200 + µl) that are difficult to obtain from mice. Here, we developed a method to directly measure free glucocorticoids in a small volume of rodent serum using ultrafiltration and liquid chromatography-tandem mass spectrometry. We validated this method to measure free 11-deoxycorticosterone (corticosterone precursor), corticosterone, and 11-dehydrocorticosterone (corticosterone metabolite) in as little as 30 µl of mouse and rat serum. Ultrafiltration produces results that are qualitatively similar to those from equilibrium dialysis, an established method that requires a much larger sample volume (200 + µl). We then applied our novel method to examine the effects of lipopolysaccharide (LPS), an immune stressor that is known to increase total corticosterone levels, free corticosterone levels, and percent free corticosterone. We administered saline vehicle or LPS to adult male and female mice, collected blood 4 hr later, and measured total and free glucocorticoid levels in serum from individual mice. As expected, LPS increased total and free corticosterone levels and percent free corticosterone. This simple, robust, and rapid method allows direct measurement of free corticosterone and other steroids in 30 µl of rodent serum.

DOI: https://doi.org/10.1371/journal.pone.0341089
Food Chem X. 2026 Jan;33 103428

Development and validation of the LC-MS/MS method based on self-made purification column for simultaneous determination of novel organophosphate esters in rice-based products.

Haiyan Zhou, Jie Cao, Fang Xiao, Mingxin Gu, Zhoupeng Li, Dan Qiao, Xiao Yang, Xianggui Chen, Bing Shao, Yukun Huang.

  Organophosphate esters (OPEs), as commonly used flame retardant and plasticizer, emerging organic pollutants, exhibit concerning contamination levels, particularly in widely consumed rice-based foods. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of 5 OPEs in rice-based products. According to recoveries and matrix effects as indicators, quality and proportion of different purification fillers were mainly optimized to fabricate a purification cartridge, simplifying and reducing the pre-treatment process and cost. The proposed method showed excellent linearity (0.08-500 μg kg-1, with all correlation coefficients exceeding 0.99), with improved method detection and quantification limit (MDL and MQL): 0.0047-0.1309 and 0.0490-0.6479 μg kg-1. Recoveries were 62.16-104.83 % with relative standard deviations (RSDs) at 0.67-15.83 % and negligible matrix effects (<20 %). Contaminants were widely detected (8.62-100 %) in real samples, with higher levels in rice bran than in rice, especially triethyl phosphate (TEP) (10.736 μg kg-1) in southern China samples.

Keywords:  Contamination monitoring; LC-MS/MS; Organophosphate esters; Rice; Rice bran

DOI:  https://doi.org/10.1016/j.fochx.2025.103428
Crit Rev Anal Chem. 2026 Jan 18. 1-37

Analytical Approaches for the Determination of Lidocaine and Its Metabolites: A Comprehensive Insight.

Hemn A H Barzani, Rebaz Anwar Omer, Nergz Bayiz Abdulrahman, Zanco Hassan Jawhar, Seerwan Hamadameen Sulaiman.

  Lidocaine (LID), an amide-type local anesthetic and antiarrhythmic agent, remains one of the most extensively used drugs in medical, dental, and surgical practice owing to its rapid onset, potent analgesic activity, and reversible nerve-blocking effects. However, its narrow therapeutic index, rapid hepatic metabolism, and typically low plasma concentrations necessitate the development of highly sensitive and selective analytical approaches for accurate quantification in biological, pharmaceutical, and environmental matrices. This review provides a comprehensive and critical evaluation of more than two decades of methodological progress in LID and metabolites analysis (monoethylglycinexylidide (MEGX) and glycinexylidide (GX)). High-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) have emerged as the gold-standard techniques due to their superior sensitivity, reproducibility, and specificity at trace levels. Complementary spectrophotometric, electroanalytical, and capillary electrophoresis methods also demonstrate distinct advantages in cost, simplicity, and eco-sustainability. Recent advancements highlight the integration of Artificial Intelligence (AI) and Machine Intelligence (MI) algorithms to enhance data processing, optimize chromatographic conditions, and improve pattern recognition in complex matrices, thereby enabling automated, real-time analysis with higher precision. Persistent analytical challenges such as matrix interference, compound instability, and ultra-trace detection underscore the need for continued innovation. Future perspectives emphasize the combination of AI-driven modeling, nanomaterial-based sensors, and green analytical platforms to establish smart, miniaturized, and environmentally sustainable systems for LID determination, facilitating reliable pharmacokinetic monitoring, clinical diagnostics, and pharmaceutical quality assurance.

Keywords:  HPLC; Lidocaine; electroanalytical techniques; pharmaceutical quality control; spectroscopy

DOI:  https://doi.org/10.1080/10408347.2026.2615681
Magn Reson Chem. 2026 Jan 18.

Multi-Solvent Suppression Ultrafast 2D COSY for High-Throughput Wine Screening.

Pia S Mayer, Jérémy Marchand, Marine P M Letertre, Jean-Nicolas Dumez, Søren B Engelsen, Patrick Giraudeau.

  Nuclear magnetic resonance (NMR) is a powerful analytical tool for wine analysis to identify and quantify a metabolite composition. However, a limiting factor of 1D 1H NMR spectroscopy is the overlap of signals in complex mixtures. While conventional 2D NMR methods disperse the signals over two dimensions, they are associated with long experiment times. In the case of wine, interesting metabolites are also often masked by the large water and ethanol peaks. To improve wine analysis by NMR, a method that uses the advantages of 2D NMR while suppressing solvent signals and being within the timeframe of 1D NMR is highly desirable. Interleaved ultrafast COSY (iuf-COSY) offers a possibility for fast acquisition of a 2D spectrum and has been demonstrated as a powerful tool in metabolomics studies, as a complement to 1D NMR methods. Here, the iuf-COSY experiment has been adapted to suppress water and ethanol signals by using a shaped pulse and a NOESY block. This approach efficiently suppresses solvent signals and gives a 2D COSY spectrum of wine in approximately 20 min. Important metabolites that originally were covered by solvent signals could be annotated, while minimal interleaving artefacts were observed. This is an efficient method to acquire a COSY spectrum of a wine sample, which can aid with the identification and discrimination of metabolites in future wine studies through additional cross peaks, while working within a high-throughput time scale. This might be particularly interesting in the field of wine metabolomics, quality control, authenticity and fraud.

Keywords:  1H; 2D; COSY; NMR; high‐throughput analysis; solvent suppression; ultrafast; wine

DOI:  https://doi.org/10.1002/mrc.70078
ACS Omega. 2026 Jan 13. 11(1): 46-69

Analytical Methods for Identification and Quantification of Quinoa Saponins: A Review.

Rodrigo Villagomez, Maribel Lozano, Yonny R Flores, Yasufumi Kobayashi, Yasunari Fujita, Giovanna R Almanza.

Quinoa saponins (SAPs) are key secondary metabolites occurring as complex mixtures mainly in the seed coat of Chenopodium quinoa Willd. Although traditionally removed due to their bitter taste and potential toxicity, quinoa SAPs display diverse biological activities, including anti-inflammatory, hypocholesterolemic, antifungal, molluscicidal, hemolytic, and cytotoxic effects, that support their potential applications in pharmaceuticals, functional foods, cosmetics, and biopesticides. Their amphiphilic nature also enables their use as natural emulsifiers. This review (1981-2024) summarizes advances in analytical methodologies for quinoa SAPs, emphasizing that while GC-MS and LC-MS/MS are widely applied for profiling, full structural elucidation still requires isolation and analysis by NMR and MS. We discuss key considerations for quinoa SAPs identification using GC-MS, LC-MS/MS, and NMR. Quantification remains challenging and is often based on relative estimations, with afrosymmetric, UV-vis, and GC-MS methods being the most frequently employed, while HPLC-DAD, LC-MS, and GC-MS/MS offer greater sensitivity. Ultimately, the selection of the analytical method and standard critically determines accuracy.

DOI: https://doi.org/10.1021/acsomega.5c01812
Cell Rep Methods. 2026 Jan 16. pii: S2667-2375(25)00327-3. [Epub ahead of print] 101291

Simultaneous determination of free and total metabolite concentrations in proteinaceous specimens by 1D 1H CPMG NMR.

Alexander Reindl, Claudia Samol, Silke Haerteis, Helena U Zacharias, Katja Dettmer, Peter J Oefner, Wolfram Gronwald.

  Nuclear magnetic resonance (NMR) spectroscopy is often used for the analysis of metabolites in proteinaceous biological specimens. However, the binding of metabolites to proteins impedes accurate quantitation of total metabolite concentrations by NMR, unless protein binding is disrupted by organic solvent precipitation, which increases variance and may result in the loss of volatile metabolites during post-extraction drying. Here, we present an approach for the inference of total metabolite concentrations from Carr-Purcell-Meiboom-Gill NMR spectra via computation of metabolite and sample-specific factors derived from the individual broadening of spectral peaks due to protein-metabolite binding. The method was validated on both synthetic proteinaceous samples and plasma and urine specimens including a certified reference plasma. Furthermore, results were compared with those obtained for methanol extracts of plasma specimens. In summary, our approach obviates the need for protein precipitation, is easy to use, and allows precise and reliable determination of total metabolite concentrations.

Keywords:  CP: metabolism; CPMG; NMR; metabolites; protein binding; quantification

DOI:  https://doi.org/10.1016/j.crmeth.2025.101291
Talanta. 2026 Jan 13. pii: S0039-9140(25)01831-4. [Epub ahead of print]302 129340

Rapid, non-invasive diagnosis of tuberculosis using desorption separation ionization mass spectrometry (DSI-MS) and urinary carbonyl metabolite fingerprints.

Yile Yu, Xin Liu, Jiameng Sun, Ruiyue Wang, Huihui Liu, Xiaoyan Zhong, Lixia Jiang, Yuze Li, Zongxiu Nie.

  Tuberculosis (TB) remains a leading global infectious disease that demands rapid, non-invasive diagnostic solutions. Here, we present a rapid urine-based diagnostic platform that integrates desorption separation ionization mass spectrometry (DSI-MS) with Amine reagents N-(2-Aminoethyl) piperidine (AED) carbonyl derivatization to enhance detection of urinary carbonyl metabolites. Minimal sample pretreatment was required: 100 μL urine samples were mixed with 2 μL AED and an 10 μL internal standard (9-phenylacridine), heated (70 °C, 5 min), applied to a sand-core filter and analyzed on an LTQ Orbitrap Velos Pro coupled to the DSI device (heater 230 °C; positive-ion acquisition m/z 50-500; acquisition time 3 min). Using this approach, we identified 65 metabolites (32 derivatized and 33 underivatized) in urine samples from a cohort of 151 tuberculosis patients and 151 healthy controls, with samples collected at the First Affiliated Hospital of Gannan Medical University and Beijing Hospital. Derivatization substantially increased ionization efficiency for carbonyl-containing metabolites and expanded coverage of low-abundance species. Pathway analysis highlighted perturbation of energy- and nitrogen-related metabolism, notably arginine and proline metabolism with elevated ornithine in TB patients. A neural-network classifier trained on the combined metabolite fingerprint achieved high discriminative performance (AUC = 0.927 on the training set and AUC = 0.922 on the test set). These results demonstrate that DSI-MS with AED derivatization, combined with machine learning, provides a rapid, non-invasive approach for TB screening and yields metabolic insights warranting further validation.

Keywords:  Carbonyl derivatization; Desorption separation ionization mass spectrometry (DSI-MS); Rapid diagnosis; Tuberculosis (TB); Urinary metabolic fingerprints

DOI:  https://doi.org/10.1016/j.talanta.2025.129340
J Agric Food Chem. 2026 Jan 20.

Simultaneous Determination of Glyphosate, Aminomethylphosphonic Acid, and Glufosinate in Green Coffee Beans by LC-MS/MS: Optimization, Validation, and Field Study.

Júlio César R M da Silva, Millena Christie F Avelar, Márcia C M Ribeiro, Mariana de O Almeida, Vanessa H F de Faria, Vanessa M Osório, Adriana F Faria.

  Despite the widespread use of glyphosate in coffee crops, there is a lack of simple, low-cost, validated methods for detecting glyphosate and related compounds in green coffee beans, the primary export form subject to international regulation. This study optimized and validated a liquid chromatography with tandem mass spectrometry method for the determination of glyphosate, aminomethylphosphonic acid (AMPA), and glufosinate in green coffee, using a modified QuPPe-PO extraction followed by a cleanup combining liquid-liquid extraction and dispersive solid-phase extraction. Validation followed SANTE/11312/2021 guidelines, confirming the method's linearity, precision, accuracy, and quantification limits. Limits of quantification were 0.02 mg kg-1 (glyphosate), 0.04 mg kg-1 (AMPA), and 0.01 mg kg-1 (glufosinate). The method was applied to a field study with Arabica coffee and Conilon Coffee, revealing that plant height was the primary factor influencing residue levels. The proposed method enables reliable monitoring of polar herbicides in coffee without the need for derivatization.

Keywords:  QuPPe-PO; dispersive solid-phase extraction; green coffee; polar herbicides; validation

DOI:  https://doi.org/10.1021/acs.jafc.5c09979
Anal Methods. 2026 Jan 22.

Microalgae-derived quality control material for reproducible and scalable LC-MS system suitability testing.

Wonseok Bang, Hyejeong Jung, Yejin Jeon, Minjung Ju, Seong-Joo Hong, Jong-Moon Park, Hookeun Lee.

Ensuring the accuracy and reliability of liquid chromatography-mass spectrometry (LC-MS/MS)-based proteomic analysis requires robust quality control (QC) strategies. However, widely used cell line-based QC materials present challenges in reproducibility, scalability, and ethical compliance. In this study, we developed a microalgae-derived QC material based on Synechocystis sp. PCC6803 and evaluated its suitability for LC-MS/MS system performance monitoring. The Synechocystis-based QC samples demonstrated high identification reproducibility, with 76% of the protein groups being consistently detected across five replicates and maintaining a median coefficient of variation (CV) of 6.7% in quantitative precision. Furthermore, QC peptides exhibited excellent linearity (R2 ≥ 0.98) and reproducibility (CV ≤ 12.06%) across varying injection amounts. These results indicate that Synechocystis-derived QC materials provide a reproducible and scalable approach that complements conventional QC workflows. They are particularly well suited for routine LC-MS/MS system performance monitoring in proteomics workflows involving non-human or mixed biological samples.

DOI: https://doi.org/10.1039/d5ay01931a