bims-metlip Biomed News
on Methods and protocols in metabolomics and lipidomics
Issue of 2024–11–03
twenty-six papers selected by
Sofia Costa, Matterworks
  1. Simultaneous metabolomics and lipidomics analysis based on 4in1 online analysis system reveal metabolic signatures in atherosclerotic mice.
  2. Assay for the quantification of abemaciclib, its metabolites, and olaparib in human plasma by liquid chromatography-tandem mass spectrometry.
  3. Simultaneous Quantification of Carboxylate Enantiomers in Multiple Human Matrices with the Hydrazide-Assisted Ultrahigh-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry.
  4. Quantitative Analysis of Drugs in a Mimetic Tissue Model Using Nano-DESI on a Triple Quadrupole Mass Spectrometer.
  5. 7Method for Detection of Naturally Occurring Toxins in Human Urine Using Liquid Chromatography High Resolution Mass Spectrometry.
  6. Quantification of Epoxyeicosatrienoic acids Enantiomers: The development of reliable and practical liquid chromatography mass Spectrometry assay.
  7. A novel analytical strategy based on gas chromatography-Orbitrap high-resolution mass spectrometry combined with solid-phase extraction for the monitoring of stanozolol misuse in human urine.
  8. Understanding LC/MS-Based Metabolomics: A Detailed Reference for Natural Product Analysis.
  9. Single-Cell Lipidomics: An Automated and Accessible Microfluidic Workflow Validated by Capillary Sampling.
  10. Super Resolved Single-Cell Spatial Metabolomics from Multimodal Mass Spectrometry Imaging guided by Imaging Mass Cytometry.
  11. Advances and challenges in non-targeted analysis: An insight into sample preparation and detection by liquid chromatography-mass spectrometry.
  12. Annotating full-scan MS data using tandem MS libraries.
  13. Spatially resolved metabolomics: From metabolite mapping to function visualising.
  14. Development and application of a dual isotopic labeling method for enhanced detection and quantification of stimulants in urine samples using high-resolution mass spectrometry.
  15. Rapid analysis of N-nitrosamines in urine using ultra high-pressure liquid chromatography-mass spectrometry.
  16. Development of a UHPLC-MS/MS Method for the Determination of Moxidectin in Rat Plasma and Its Application in Pharmacokinetics.
  17. Development and validation of an UPLC-ESI-MS/MS method for simultaneous quantification of antineoplastic agents and their metabolites in human plasma after unintentional exposure.
  18. Analytical Tools for HCP Detection, Identification, and Quantitation.
  19. Development of a Desorption Electrospray Ionization-Multiple-Reaction-Monitoring Mass Spectrometry (DESI-MRM) Workflow for Spatially Mapping Oxylipins in Pulmonary Tissue.
  20. A novel high-throughput quantitative method for the determination of per- and poly-fluoroalkyl substances in human plasma based on UHPLC-Q/Orbitrap HRMS coupled with isotope internal standard.
  21. A Review of Recent Advances in Chromatographic Quantification Methods for Cyanogenic Glycosides.
  22. Metabolomics of Chinese Hamster Ovary Cells.
  23. Processing Next-Generation Mass Spectrometry Imaging Data: Principal Component Analysis at Scale.
  24. UPLC-MS/MS analysis of axitinib and pharmacokinetic application in beagle dogs.
  25. A systematic evaluation of quenching, extraction and analysis procedures for metabolomics study of the mechanism of QYSLD intervention in A549 cells.
  26. Development and Validation of an Improved HPLC-MS/MS Method for Quantifying Total and Unbound Lenalidomide in Human Plasma.
  1. Talanta. 2024 Oct 28. pii: S0039-9140(24)01488-7. [Epub ahead of print]283 127109
    Simultaneous metabolomics and lipidomics analysis based on 4in1 online analysis system reveal metabolic signatures in atherosclerotic mice.
    Meng Ding, Luwei Zheng, Xiaolin Hua, Mengxuan Chen, Qisheng Zhong, Taohong Huang, Ping Li, Hua Yang.
      Developing efficient and comprehensive analysis methods for metabolomics and lipidomics in the biological tissues and body fluids is essential for understanding the disease mechanisms. Although various two-dimensional liquid chromatography-mass spectrometry (2D-LC-MS) methods have been proposed to expand metabolite coverage, achieving higher efficiency in integrated metabolomics and lipidomics studies remains a technical challenge. In this work, a novel 4in1 online analysis system with excellent reproducibility and mass accuracy was constructed for metabolomics and lipidomics study in various biological samples from atherosclerotic mice. This system enabled the simultaneous detection in both positive and negative ion modes with extensive polarity separation in a single analytical run. Using the 4in1 online analysis system, we identified distinct but complementary metabolic signatures associated with atherosclerosis in different biological samples. Specifically, a total of 230 and 170 differential metabolites or lipids were detected in mice plasma samples and aortic tissue samples, respectively, including glycerophospholipids, sphingolipids, fatty acyls, glycerolipids, carboxylic acids, and pyrimidine nucleosides. Additionally, atherosclerosis-related metabolic pathways involved in biosynthesis of unsaturated fatty acids, sphingolipid metabolism, cholesterol metabolism, glycerophospholipid metabolism, and choline metabolism further revealed. These findings demonstrate that the novel 4in1 online analysis system is a faithful, stable and powerful tool for comprehensive metabolomics and lipidomics studies in complex biological matrices.
    Keywords:  4in1; Atherosclerosis; Lipidomics; Metabolomics; Quadrupole time-of-flight mass spectrometry; Two-dimensional liquid chromatography
    DOI:  https://doi.org/10.1016/j.talanta.2024.127109
  2. J Pharm Biomed Anal. 2024 Oct 17. pii: S0731-7085(24)00573-9. [Epub ahead of print]253 116531
    Assay for the quantification of abemaciclib, its metabolites, and olaparib in human plasma by liquid chromatography-tandem mass spectrometry.
    Kasey L Hill, Nicole L Abbott, Joo Young Na, Michelle Rudek, Kathleen Moore, Eudocia Q Lee, Mitch A Phelps.
      An isotope-dilution bioanalytical assay for abemaciclib and its metabolites in combination with olaparib was developed and validated in human plasma K2 EDTA. For the quantitative assay, human plasma samples (or human plasma QC samples) were spiked with internal standard solution before a simple protein precipitation with methanol. The extract was injected onto a liquid chromatography-tandem mass spectrometry (LC-MS/MS) instrument where it was chromatographically separated by a polar end-capped reversed phase column and guard using gradient elution with water and methanol both modified with 0.2 % formic acid (v/v) as the mobile phases. The analytes and internal standards were measured by heated electrospray ionization (HESI) in positive polarity using selected reaction monitoring (SRM) on a triple quadrupole mass spectrometer. The assay was validated for linear ranges as follows: 0.4 - 1000 nM abemaciclib, 0.35 - 1000 nM M2 and M18, 0.5 - 1000 nM M20, and 0.75 - 1000 nM olaparib. The inter-day or between day precision for the quality controls (n = 18) was < 13 % and the accuracy was ± 12 %, for all analytes, including the lower limit of quantification (LLOQ). The intra-day or within day precision for the quality controls (n = 6) was ≤ 11 % and the accuracy was ± 12 % for low, mid, and high and < 19 % at LLOQ. The recovery in human plasma was determined to be between 92 % and 102 % for all analytes spanning the linear range. The validated, bioanalytical quantitative assay was designed to measure abemaciclib, its metabolites, and olaparib for pharmacokinetic evaluation of patients in clinical trials for breast, brain, and ovarian cancers.
    Keywords:  Abemaciclib; Hydroxy-N-desethylabemaciclib (M18); Hydroxyabemaciclib (M20); N-desethylabemaciclib (M2); Olaparib; Pharmacokinetic studies
    DOI:  https://doi.org/10.1016/j.jpba.2024.116531
  3. Anal Chem. 2024 Oct 30.
    Simultaneous Quantification of Carboxylate Enantiomers in Multiple Human Matrices with the Hydrazide-Assisted Ultrahigh-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry.
    Yuting Sun, Qingyu Hu, Jiali Zuo, He Wang, Zhendong Guo, Yulan Wang, Huiru Tang.
      Many chiral carboxylic acids with α-amino, α-hydroxyl, and α-methyl groups are concurrently present in mammals establishing unique molecular phenotypes and multiple biological functions, especially host-microbiota symbiotic interactions. Their chirality-resolved simultaneous quantification is essential to reveal the biochemical details of physiology and pathophysiology, though challenging with their low abundances in some biological matrices and difficulty in enantiomer resolution. Here, we developed a method of the chirality-resolved metabolomics with sensitivity-enhanced quantitation via probe-promotion (Met-SeqPro) for analyzing these chiral carboxylic acids. We designed and synthesized a hydrazide-based novel chiral probe, (S)-benzoyl-proline-hydrazide (SBPH), to convert carboxylic acids into amide diastereomers to enhance their retention and chiral resolution on common C18 columns. Using the d5-SBPH-labeled enantiomers as internal standards, we then developed an optimized ultrahigh-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous quantification of 60 enantiomers of 30 chiral carboxylic acids in one run. This enantiomer-resolved method showed excellent sensitivity (LOD < 4 fmol-on-column), linearity (R2 > 0.992), precision (CV < 15%), accuracy (|RE| < 20%), and recovery (80-120%) in multiple biological matrices. With the method, we then quantified 60 chiral carboxylic acids in human urine, plasma, feces, and A549 cells to define their metabolomic phenotypes. This provides basic data for human phenomics and a promising tool for investigating the mammal-microbiome symbiotic interactions.
    DOI:  https://doi.org/10.1021/acs.analchem.4c04187
  4. J Am Soc Mass Spectrom. 2024 Nov 01.
    Quantitative Analysis of Drugs in a Mimetic Tissue Model Using Nano-DESI on a Triple Quadrupole Mass Spectrometer.
    Alyssa M Moore, Andrew Bowman, Syeda Nazifa Wali, Miranda R Weigand, David Wagner, Junhai Yang, Julia Laskin.
      Mass spectrometry is a powerful analytical technique used at every stage of the pharmaceutical research process. A specialized branch of this method, mass spectrometry imaging (MSI), has emerged as an important tool for determining the spatial distribution of drugs in biological samples. Despite the importance of MSI, its quantitative capabilities are still limited due to the complexity of biological samples and the lack of separation prior to analysis. This makes the simultaneous quantification and visualization of analytes challenging. Several techniques have been developed to address this challenge and enable quantitative MSI. One such approach is the mimetic tissue model, which involves the incorporation of an analyte of interest into tissue homogenates at several concentrations. A calibration curve that accounts for signal suppression by the complex biological matrix is then created by measuring the signal of the analyte in the series of tissue homogenates. Herein, we use the mimetic tissue model on a triple quadrupole mass spectrometer (QqQ) in multiple reaction monitoring mode to demonstrate the quantitative abilities of nanospray desorption electrospray ionization (nano-DESI) and compare these results with those obtained using atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI). For the tested compounds, our findings indicate that nano-DESI achieves lower standard deviations than AP-MALDI, resulting in superior limits of detection for the studied analytes. Additionally, we discuss the limitations of the mimetic tissue model in the quantification of certain analytes and the challenges involved with the implementation of the model.
    DOI:  https://doi.org/10.1021/jasms.4c00345
  5. J Anal Toxicol. 2024 Nov 01. pii: bkae086. [Epub ahead of print]
    7Method for Detection of Naturally Occurring Toxins in Human Urine Using Liquid Chromatography High Resolution Mass Spectrometry.
    Bryan E Hettick, Anisha Saddy, Logan C Krajewski, Rudolph C Johnson, Elizabeth I Hamelin.
      Natural toxins present an ongoing risk for human exposure that requires a rapid, accurate diagnosis for proper response. In this study, a qualitative liquid chromatography high resolution mass spectrometry (LC-HRMS) method was developed and validated for the detection of a large, diverse selection of natural toxins. Data-dependent acquisition was performed to identify compounds with an in-house mass spectral library of 129 hazardous toxins that originate from plants, animals, and fungi. All 129 compounds were spiked into human urine, extracted, and evaluated for spectral library matching. Of these, 92 toxins met the quality criteria and underwent validation in urine matrix based on American National Standards Institute (ANSI) guidelines. A generalized workflow for method expansion was developed and enables the rapid addition of relevant compounds to the established method. This LC-HRMS method achieves efficient detection of natural toxins in urine, and the created workflow can rapidly increase compound coverage via method expansion.
    Keywords:  Alkaloid; cardiac glycoside; high-resolution mass spectrometry; lowest reportable limit; matrix effects; method development; toxins; trichothecene
    DOI:  https://doi.org/10.1093/jat/bkae086
  6. J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 20. pii: S1570-0232(24)00355-6. [Epub ahead of print]1247 124346
    Quantification of Epoxyeicosatrienoic acids Enantiomers: The development of reliable and practical liquid chromatography mass Spectrometry assay.
    Fadumo A Isse, Sara Helal, Ahmed A El-Sherbeni, Dion R Brocks, Ayman O S El-Kadi.
      Epoxyeicosatrienoic acids (EETs) are increasingly recognized as key metabolites in the arachidonic acid (AA) metabolic pathway. EETs are epoxy derivatives of AA with two chiral centers formed by cytochrome P450 (CYP) enzymes. EETs have reported biological activities as racemates; however, knowledge on specific optical isomers of EET is lacking. A main reason is the absence of practical assay to quantify EETs isomers associated with specific pathological conditions and enzymes. The reported underivatized chiral LC-MS/MS assays utilize different mobile phases and flow rates or required long run times to achieve separation of EET stereoisomers. Others incorporated a derivatization step before the separation of EETs in their assays. Therefore, the objective of this study was to develop and validate a stereoselective assay for the simultaneous quantitation of underivatized EET enantiomers using Liquid Chromatography Mass Spectrometry (LC-MS/MS) with an optimum baseline separation using binary mobile phase and gradient elution. Herein, we report the development and validation of an LC-MS/MS assay, and its application to quantify the formation of EET enantiomers mediated by human liver microsomes. Assay linearity extends over 10-600 ng/mL with r2 > 0.99 for all EETs enantiomers. The inter-run accuracy was within ± 15 %, and precision was ≤ 15 %, and < 20 % for the LLOQ. The matrix effect for the current assay was within ≤ ±20 %, and the mean recovery for quantitative methods was 70-125 %. The assay proved to be reliable and practical for chiral analysis.
    Keywords:  Arachidonic acid; Assay; EET; Enantiomer; LC-MS/MS; Microsome matrix; Underived
    DOI:  https://doi.org/10.1016/j.jchromb.2024.124346
  7. Rapid Commun Mass Spectrom. 2025 Jan 15. 39(1): e9935
    A novel analytical strategy based on gas chromatography-Orbitrap high-resolution mass spectrometry combined with solid-phase extraction for the monitoring of stanozolol misuse in human urine.
    Siying Zheng, Ziyi Ji, Yuqi Ge, Xian Fang, Mengpan Liu, Haoyi Sun, Xiaojun Deng, Lei Liao.
       RATIONALE: Stanozolol, an anabolic androgenic steroid listed in Part S1 of the World Anti-Doping Agency Prohibited List, exhibits a low response and significant matrix interference in urine samples when using liquid-liquid extraction-gas chromatography-mass spectrometry (GC-MS). Enhancing sample preparation techniques remains essential for the effective detection of stanozolol and its metabolites.
    METHODS: A method for determining stanozolol and its metabolites (3'-OH-stanozolol, 4β-OH-stanozolol, and 16β-OH-stanozolol) in human urine was developed and validated using GC-Orbitrap high-resolution MS combined with optimized mixed-mode solid-phase extraction (SPE). This method was applied to urine samples from two volunteers who orally administered a single dose of stanozolol, with samples collected over a 30-day period post-administration.
    RESULTS: The optimized mixed-mode SPE method reduced matrix interference and achieved satisfactory extraction efficiency and high sensitivity, enabling confident identification of all targets in human urine. Validation showed extraction recovery of 74% to 81% and limits of detection from 0.1 to 0.25 ng mL-1. The method was successfully applied to detect urinary excretion profiles of stanozolol and its metabolites in positive volunteer samples.
    CONCLUSION: This study presents a novel detection protocol for stanozolol and its metabolites, enhancing the monitoring of stanozolol abuse and contributing to the integrity of sports competitions. This protocol offers a robust tool for anti-doping laboratories, aiding in the accurate detection of stanozolol misuse and supporting the enforcement of fair play in athletics.
    DOI:  https://doi.org/10.1002/rcm.9935
  8. Proteomics Clin Appl. 2024 Oct 30. e202400048
    Understanding LC/MS-Based Metabolomics: A Detailed Reference for Natural Product Analysis.
    Jyotirmay Sarkar, Rajveer Singh, Shivani Chandel.
      Liquid chromatography, when used in conjunction with mass spectrometry (LC/MS), is a powerful tool for conducting accurate and reproducible investigations of numerous metabolites in natural products (NPs). LC/MS has gained prominence in metabolomic research due to its high throughput, the availability of multiple ionization techniques and its ability to provide comprehensive metabolite coverage. This unique method can significantly influence various scientific domains. This review offers a comprehensive overview of the current state of LC/MS-based metabolomics in the investigation of NPs. This review provides a thorough overview of the state of the art in LC/MS-based metabolomics for the investigation of NPs. It covers the principles of LC/MS, various aspects of LC/MS-based metabolomics such as sample preparation, LC modes, method development, ionization techniques and data pre-processing. Moreover, it presents the applications of LC/MS-based metabolomics in numerous fields of NPs research such as including biomarker discovery, the agricultural research, food analysis, the study of marine NPs and microbiological research. Additionally, this review discusses the challenges and limitations of LC/MS-based metabolomics, as well as emerging trends and developments in this field.
    Keywords:  data pre‐processing; liquid chromatography; mass spectrometry; metabolomics; natural products method development
    DOI:  https://doi.org/10.1002/prca.202400048
  9. Anal Chem. 2024 Oct 26.
    Single-Cell Lipidomics: An Automated and Accessible Microfluidic Workflow Validated by Capillary Sampling.
    Anastasia Kontiza, Johanna von Gerichten, Kyle D G Saunders, Matt Spick, Anthony D Whetton, Carla F Newman, Melanie J Bailey.
      We report the first demonstration of a microfluidics-based approach to measure lipids in single living cells using widely available liquid chromatography mass spectrometry (LC-MS) instrumentation. The method enables the rapid sorting of live cells into liquid chambers formed on standard Petri dishes and their subsequent dispensing into vials for analysis using LC-MS. This approach facilitates automated sampling, data acquisition, and analysis and carries the additional advantage of chromatographic separation, aimed at reducing matrix effects present in shotgun lipidomics approaches. We demonstrate that our method detects comparable numbers of features at around 200 lipids in populations of single cells versus established live single-cell capillary sampling methods and with greater throughput, albeit with the loss of spatial resolution. We also show the importance of optimization steps in addressing challenges from lipid contamination, especially in blanks, and demonstrate a 75% increase in the number of lipids identified. This work opens up a novel, accessible, and high-throughput way to obtain single-cell lipid profiles and also serves as an important validation of single-cell lipidomics through the use of different sampling methods.
    DOI:  https://doi.org/10.1021/acs.analchem.4c03435
  10. bioRxiv. 2024 Oct 24. pii: 2024.10.21.619323. [Epub ahead of print]
    Super Resolved Single-Cell Spatial Metabolomics from Multimodal Mass Spectrometry Imaging guided by Imaging Mass Cytometry.
    Efe Ozturk, Abhijeet Venkataraman, Felix G Rivera Moctezuma, Ahmet F Coskun.
      Mass spectrometry imaging (MSI) is a powerful technique for spatially resolved analysis of metabolites and other biomolecules within biological tissues. However, the inherent low spatial resolution of MSI often limits its ability to provide detailed cellular-level information. To address this limitation, we propose a guided super-resolution (GSR) approach that leverages high-resolution Imaging Mass Cytometry (IMC) images to enhance the spatial resolution of low-resolution MSI data. By using these detailed IMC images as guides, we improve the resolution of MSI images, creating high-resolution metabolite maps. This enhancement facilitates more precise analysis of cellular structures and tissue architectures, providing deeper insights into super-resolved spatial metabolomics at the single-cell level.
    DOI:  https://doi.org/10.1101/2024.10.21.619323
  11. J Chromatogr A. 2024 Oct 19. pii: S0021-9673(24)00833-1. [Epub ahead of print]1737 465459
    Advances and challenges in non-targeted analysis: An insight into sample preparation and detection by liquid chromatography-mass spectrometry.
    Vivek Mandal, Jinal Ajabiya, Nasir Khan, Rakesh K Tekade, Pinaki Sengupta.
      Unknown impurities, metabolites and harmful pollutants present in pharmaceutical products, biological and environmental samples, respectively are of high concern in terms of their detection and quantification. The targeted analysis aims to quantify known chemical entities, but it lacks the ability to identify unknown components present in a sample. Non-targeted analysis is an analytical approach that can be made applicable to various disciplines of science to effectively search for unknown chemical, biological, or environmental entities that can answer various baffling mysteries of research. It employs various high-end analytical techniques that can specifically screen out multiple unknown compounds from complex mixtures. Non-targeted analysis is also applicable for complex studies such as metabolomics to search unidentified metabolites of new chemical entities. This review critically discusses the current advancements in non-targeted analysis related to the analysis of pharmaceutical, biological, and environmental samples. Various steps like sample collection, handling, preparation, extraction, its analysis using advanced techniques like high-resolution mass spectrometry, liquid chromatography mass spectrometry, and lastly interpretation of the huge amounts of complex data obtained upon analysis of complex matrices have been discussed broadly in this article. Besides the advantages of non-targeted analysis over targeted analysis, limitations, bioinformatics tools, sources of error, and research gaps have been critically analyzed.
    Keywords:  Bio-informatics tools; Bioanalysis; LC-MS/MS; Non-targeted analysis; Sample preparation
    DOI:  https://doi.org/10.1016/j.chroma.2024.465459
  12. bioRxiv. 2024 Oct 17. pii: 2024.10.14.618269. [Epub ahead of print]
    Annotating full-scan MS data using tandem MS libraries.
    Shipei Xing, Vincent Charron-Lamoureux, Yasin El Abiead, Pieter C Dorrestein.
      Full-scan mass spectrometry (MS) data from both liquid chromatography (LC) and MS imaging capture multiple ion forms, including their in-source fragments. Here we leverage such fragments to structurally annotate full-scan data from LC-MS or MS imaging by matching against peak intensity scaled tandem MS spectral libraries using precursor-tolerant reverse match scoring. Applied to inflammatory bowel disease and imaging datasets, we show the approach facilitates re-analyses of data in public repositories.
    DOI:  https://doi.org/10.1101/2024.10.14.618269
  13. Clin Transl Med. 2024 Nov;14(11): e70031
    Spatially resolved metabolomics: From metabolite mapping to function visualising.
    Xinyue Min, Yiran Zhao, Meng Yu, Wenchao Zhang, Xinyi Jiang, Kaijing Guo, Xiangyi Wang, Jianpeng Huang, Tong Li, Lixin Sun, Jiuming He.
      Mass spectrometry imaging (MSI)-based spatially resolved metabolomics addresses the limitations inherent in traditional liquid chromatography-tandem mass spectrometry (LC-MS)-based metabolomics, particularly the loss of spatial context within heterogeneous tissues. MSI not only enhances our understanding of disease aetiology but also aids in the identification of biomarkers and the assessment of drug toxicity and therapeutic efficacy by converting invisible metabolites and biological networks into visually rendered image data. In this comprehensive review, we illuminate the key advancements in MSI-driven spatially resolved metabolomics over the past few years. We first outline recent innovations in preprocessing methodologies and MSI instrumentation that improve the sensitivity and comprehensiveness of metabolite detection. We then delve into the progress made in functional visualization techniques, which enhance the precision of metabolite identification and annotation. Ultimately, we discuss the significant potential applications of spatially resolved metabolomics technology in translational medicine and drug development, offering new perspectives for future research and clinical translation. HIGHLIGHTS: MSI-driven spatial metabolomics preserves metabolite spatial information, enhancing disease analysis and biomarker discovery. Advances in MSI technology improve detection sensitivity and accuracy, expanding bioanalytical applications. Enhanced visualization techniques refine metabolite identification and spatial distribution analysis. Integration of MSI with AI promises to advance precision medicine and accelerate drug development.
    Keywords:  clinical translation; drug discovery; mass spectrometry imaging; spatially resolved metabolomics; tumour metabolism
    DOI:  https://doi.org/10.1002/ctm2.70031
  14. Anal Bioanal Chem. 2024 Oct 25.
    Development and application of a dual isotopic labeling method for enhanced detection and quantification of stimulants in urine samples using high-resolution mass spectrometry.
    Yang Wang, Wanli Li, Xiaojun Deng.
      Given the critical nature of anti-doping efforts, the detection of stimulant substances is shifting from accurate qualitative analysis to precise quantitative analysis. Additionally, the use of liquid chromatography-high-resolution mass spectrometry (LC-HRMS) in detecting stimulants is becoming more widespread. However, the lack of isotope-labeled internal standards is causing increasing issues with quantitative accuracy. Furthermore, challenges such as the mass spectrometric response of small molecules and the separation of isomers present additional difficulties. We have developed a quantitative method for stimulant substances containing amine or phenol hydroxyl groups, using a dual-label derivatization system. This method offers a new perspective for analyzing and detecting low molecular weight substances, isomers, or those with poor LC-MS response, and proposes a solution to the problem of missing isotope-labeled internal standards. Methodological validation has shown that this approach has promising application potential.
    Keywords:  Dual isotopic labeling; High-resolution mass spectrometry (HRMS); LC–MS response enhancement; Quantitative analysis; Stimulant detection
    DOI:  https://doi.org/10.1007/s00216-024-05612-2
  15. Anal Methods. 2024 Oct 29.
    Rapid analysis of N-nitrosamines in urine using ultra high-pressure liquid chromatography-mass spectrometry.
    S Shinde, K D Croft, J M Hodgson, C P Bondonno.
      N-Nitrosamines, carcinogenic compounds present in dietary and environmental sources and formed endogenously, are believed to be linked with the presence of nitrate and nitrite, both within dietary sources and after intake. To fully evaluate this potential threat to human health, an accurate analytical method to measure N-nitrosamines in biological matrices is necessary. We report a simple, fast, selective mass spectrometry method to detect N-nitrosamines in human urine. Analysis of seven N-nitrosamines, N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosopiperdine (NPIP), N-nitrosopyrrolidine (NPYR), N-nitrosodi-N-propylamine (NDPA) and N-nitrosodi-N-butylamine (NDBA) in urine was quantitated using Ultra High-Pressure Liquid Chromatography-tandem Mass spectrometry (UHPLC-MS/MS). A Sorbent supported Liquid Extraction (SLE) method was employed to extract N-nitrosamines from 24 hour collected human urine samples. The percent recovery varied between 74.3 to 110 and the limit of detection and limit of quantification ranged from 0.1 to 0.85 ng mL-1 and 0.22 to 2.06 ng mL-1 respectively. Precision for inter-day and intra-day assay yielded a % coefficient of variation between 4-10% for all measured compounds in urine. Linear regression analysis of calibration curves for N-nitrosamines measured in urine in the concentration range 0.4-12.8 ng mL-1 gave correlation coefficients, R2 0.9874-0.9962. Urinary excretion of N-nitrosamines measured in ten healthy subjects resulted in detection of most of the N-nitrosamines including NDMA, NDEA, NPYR, NDPA and NDBA by this method.
    DOI:  https://doi.org/10.1039/d4ay01870b
  16. Molecules. 2024 Oct 10. pii: 4786. [Epub ahead of print]29(20):
    Development of a UHPLC-MS/MS Method for the Determination of Moxidectin in Rat Plasma and Its Application in Pharmacokinetics.
    Hongjuan Zhang, Zhen Yang, Baocheng Hao, Di Wu, Dan Shao, Yu Liu, Wanxia Pu, Shouli Yi, Ruofeng Shang, Shengyi Wang.
      The aim of the present study was to establish a simple and reliable ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method and apply it for the determination of pharmacokinetics of moxidectin-loaded microspheres (MOX-MS) in rats. Plasma samples were processed using a simplified liquid-liquid extraction method and were separated using an Agilent Zorbax Eclipse Plus C18 column (50 mm × 2.1 mm, 1.8 μm) with a mobile phase consisting of a 10 mM ammonium formate solution with 0.1% formic acid (A) and acetonitrile (B) at a flow rate of 0.4 mL/min for 5 min. Avermectin B1a was used as an internal standard (IS). The sample was injected at a volume of 10 μL with a column temperature of 35 °C and detected in a positive ion mode. A good linear response across the concentration range of 1.00-200 ng/mL (r2 > 0.99) and a lower limit of quantification (LLOQ) of 1.00 ng/mL were achieved. The extraction recovery of moxidectin exceeded 94.1%, the matrix effect was between 91.2% and 96.2%, the accuracy ranged from 100.1 to 103.6%, and the relative standard deviation (RSD) did not exceed 15% for the intra- and inter-day accuracy and precision. The pharmacokinetic results showed that MOX-MS significantly decreased Cmax, prolonged T1/2, and improved bioavailability. The developed method significantly reduced the assay volume, shortened detection time, simplified sample processing methods and saved assay costs, which may contribute to the development of the new antiparasitic drug.
    Keywords:  UPLC-MS/MS; microspheres; moxidectin; pharmacokinetics; rat
    DOI:  https://doi.org/10.3390/molecules29204786
  17. Arch Toxicol. 2024 Oct 31.
    Development and validation of an UPLC-ESI-MS/MS method for simultaneous quantification of antineoplastic agents and their metabolites in human plasma after unintentional exposure.
    Eline Verscheure, Ilana Struys, Matteo Creta, Katrien Poels, Jeroen Vanoirbeek, Liesbeth Lenaerts, Frédéric Amant, Manosij Ghosh, Lode Godderis.
      Cyclophosphamide, daunorubicin, epirubicin, doxorubicin and paclitaxel are commonly used drugs in cancer treatment. However, there are no methods available enabling simultaneous measurement of these compounds and their metabolites in human plasma. Our aim was to develop and validate a sensitive method for simultaneous quantification of multiple antineoplastic drugs and their major metabolites in plasma. Solid phase extraction with Oasis PRiME HLB cartridges was used for sample clean-up. The samples were separated on an Acquity UPLC BEH C18 column, ionised by electrospray ionisation and detected with tandem mass spectrometry. The method was validated based on selectivity, extraction efficiency, matrix effect, process efficiency, linearity, sensitivity, precision and accuracy. The established LLOQs were 0.05 ng/mL (cyclophosphamide), 30 ng/mL (4-oxo-cyclophosphamide), 0.3 ng/mL (doxorubicin, daunorubicinol), 0.7 ng/mL (epirubicin, epirubicinol, doxorubicinol), 1 ng/mL (daunorubicin and paclitaxel) and 5 ng/mL (6-alpha-hydroxypaclitaxel). Afterwards, the method was tested in a real-life, unintentional exposure setting. Twenty-two plasma samples of matched maternal and cord blood pairs from pregnant cancer patients treated with chemotherapy were analysed. This resulted in two positive samples, with cyclophosphamide concentrations up to 0.37 ng/mL. The validated method is now ready to be applied in the field.
    Keywords:  Antineoplastic drug; Biomonitoring; Liquid chromatography-mass spectrometry; Method validation
    DOI:  https://doi.org/10.1007/s00204-024-03900-5
  18. Methods Mol Biol. 2025 ;2853 235-248
    Analytical Tools for HCP Detection, Identification, and Quantitation.
    Faiza Kanwal, Michael Henry, Paula Meleady.
      Identifying and quantifying host cell proteins (HCPs) are crucial in developing and manufacturing biopharmaceutical products. ELISA has been used widely for identifying and quantifying HCPs in biopharmaceutical products. Liquid chromatography-mass spectrometry (LC-MS) has recently emerged as an orthogonal tool for HCP analysis, providing comprehensive and quantitative information on individual HCPs. This chapter explores the advancements in LC-MS/MS methodologies for HCP analysis, including sample preparation, data acquisition modes, and data analysis strategies. It also discusses the challenges of HCP identification and quantitation, and recent innovations to overcome these limitations. The application of LC-MS/MS methods in HCP profiling holds significant promise for improving process control and ensuring product safety in biopharmaceutical manufacturing.
    Keywords:  Biopharmaceutical manufacturing; Data acquisition modes; Data analysis strategies; Host cell proteins; LC-MS/MS; Sample preparation
    DOI:  https://doi.org/10.1007/978-1-0716-4104-0_15
  19. Anal Chem. 2024 Oct 26.
    Development of a Desorption Electrospray Ionization-Multiple-Reaction-Monitoring Mass Spectrometry (DESI-MRM) Workflow for Spatially Mapping Oxylipins in Pulmonary Tissue.
    Matthew J Smith, Mu Nie, Mikael Adner, Jesper Säfholm, Craig E Wheelock.
      Oxylipins are a class of low-abundance lipids formed via oxygenation of fatty acids. These compounds include potent signaling molecules (e.g., octadecanoids, eicosanoids) that can exert essential functions in the pathophysiology of inflammatory diseases including asthma. While some oxylipin signaling cascades have been unraveled using LC-MS/MS-based methods, measurements in homogenate samples do not represent the spatial heterogeneity of lipid metabolism. Mass spectrometry imaging (MSI) directly detects analytes from a surface, which enables spatial mapping of oxylipin biosynthesis and migration within the tissue. MSI has lacked the sensitivity to routinely detect low-abundance oxylipins; however, new multiple-reaction-monitoring (MRM)-based MSI technologies show increased sensitivity. In this study, we developed a workflow to apply desorption electrospray ionization coupled to a triple quadrupole mass spectrometer (DESI-MRM) to spatially map oxylipins in pulmonary tissue. The targeted MSI workflow screened guinea pig lung extracts using LC-MS/MS to filter oxylipin targets based on their detectability by DESI-MRM. A panel of 5 oxylipins was then selected for DESI-MRM imaging derived from arachidonic acid (TXB2, 11-HETE, 12-HETE), linoleic acid (12,13-DiHOME), and α-linolenic acid (16-HOTrE). To parse this new data type, a custom-built R package (quantMSImageR) was developed with functionality to label regions of interest as well as quantify and analyze lipid distributions. The spatial distributions quantified by DESI-MRM were supported by LC-MS/MS analysis, with both indicating that 16-HOTrE and 12-HETE were associated with airways, while 12,13-DiHOME and arachidonic acid were mapped to parenchyma. This study realizes the potential of targeted MSI to routinely map low-abundance oxylipins with high specificity at scale.
    DOI:  https://doi.org/10.1021/acs.analchem.4c02350
  20. J Hazard Mater. 2024 Oct 13. pii: S0304-3894(24)02717-1. [Epub ahead of print]480 136138
    A novel high-throughput quantitative method for the determination of per- and poly-fluoroalkyl substances in human plasma based on UHPLC-Q/Orbitrap HRMS coupled with isotope internal standard.
    Wenting Zhang, Qiuyan Lu, Huafeng Chen, Yuxiang Li, Yongyou Hua, Jing Wang, Fa Chen, Renjin Zheng.
      A novel method for the quantitative analysis of 56 per- and polyfluoroalkyl substances (PFASs) in human plasma was established on the basis of ultrahigh performance liquid chromatography tandem quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS) in combination with accurate customized mass databases and isotopic internal standards. A streamlined, high-throughput, and high-recovery (RE) sample pretreatment method was developed. The method's performance was evaluated in terms of linearity, limit of quantification, RE, repeatability, reproducibility, and matrix effect. The proposed method was applied in the simultaneous analysis of 56 PFASs in human plasma, and its results demonstrated high sensitivity, accuracy, and precision. The optimized method was implemented to analyze PFASs in 135 plasma samples, and 12 components were detected. The comparative analysis of the results from 135 plasma samples with domestic and international studies revealed elevated contents of PFOA, PFOS, PFBA, and PFTrDA, the moderate amounts of PFHxS, PFUdA, PFBS, and PFHpS, and the low concentrations of PFNA and PFDA. Notably, GenX was detected in human plasma for the first time. This finding suggests that the study region is contaminated with this substance. Correlation analysis revealed a strong relationship among PFNA, PFDA, and PFUdA, implying that these substances may have similar exposure sources.
    Keywords:  Human plasma; Isotope internal standard method; Per- and polyfluoroalkyl substance; UHPLC-Q/Orbitrap HRMS
    DOI:  https://doi.org/10.1016/j.jhazmat.2024.136138
  21. Molecules. 2024 Oct 11. pii: 4801. [Epub ahead of print]29(20):
    A Review of Recent Advances in Chromatographic Quantification Methods for Cyanogenic Glycosides.
    Yao Zhao, Shuai Wen, Yan Wang, Wenshuo Zhang, Xiangming Xu, Yi Mou.
      Cyanogenic glycosides are naturally occurring compounds found in numerous plant species, which can release toxic hydrogen cyanide upon hydrolysis. The quantification of cyanogenic glycosides is essential for assessing their potential toxicity and health risks associated with their consumption. Liquid chromatographic techniques coupled with various detectors have been widely used for the quantification of cyanogenic glycosides. In this review, we discuss recent advances in chromatographic quantification methods for cyanogenic glycosides, including the development of new stationary phases, innovative sample preparation methods, and the use of mass spectrometry. We also highlight the combination of chromatographic separation with mass spectrometric detection for the identification and quantification of specific cyanogenic glycosides and their metabolites in complex sample matrices. Lastly, we discuss the current challenges and future perspectives in the development of reliable reference standards, optimization of sample preparation methods, and establishment of robust quality control procedures. This review aims to provide an overview of recent advances in chromatographic quantification methods for cyanogenic glycosides and their applications in various matrices, including food products, biological fluids, and environmental samples.
    Keywords:  chromatography; cyanogenic glycosides; quantification
    DOI:  https://doi.org/10.3390/molecules29204801
  22. Methods Mol Biol. 2025 ;2853 205-234
    Metabolomics of Chinese Hamster Ovary Cells.
    Rita Singh, Sneh Bajpai, Amardeep Singh, Phulwanti Sharma, Yashwant Kumar, Niraj Kumar.
      Increasing demand of protein biotherapeutics produced using Chinese hamster ovary (CHO) cell lines necessitates improvement in the production yield of the bioprocess. Various cell engineering, improved media formulation and process-design based approaches utilizing the power of OMICS technologies, specifically, genomics and proteomics, have been employed; however, the potential of metabolomics largely remains unexplored. Metabolomics enables the detection, identification, and/or quantitation of small molecules, commonly known as metabolites, in and around the cells and may help to unlock the cellular molecular mechanism(s) that regulates cell growth and productivity in the bioprocess and improves cellular performance during the bioprocess. Currently, liquid chromatography (LC)/gas chromatography (CG)- coupled with mass-spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy are the most commonly used approaches for metabolomics. Therefore, in this chapter, we have discussed the standard procedures of investigating CHO metabolites using LC/GC-MS and/or NMR-based approaches.
    Keywords:  Adherent and suspension cell culture; Cell metabolism; Chinese hamster ovary (CHO) cells; Data analysis; Gas chromatography-mass spectrometry (GC-MS); Liquid chromatography-mass spectrometry (LC-MS); Metabolite extraction; Nuclear magnetic resonance (NMR) spectroscopy
    DOI:  https://doi.org/10.1007/978-1-0716-4104-0_14
  23. J Am Soc Mass Spectrom. 2024 Oct 28.
    Processing Next-Generation Mass Spectrometry Imaging Data: Principal Component Analysis at Scale.
    Kasper Krijnen, Paul Blenkinsopp, Ron M A Heeren, Ian G M Anthony.
      Mass spectrometry imaging (MSI) is constantly improving in spatial resolving power, throughput and mass resolution. Although beneficial, these improvements increase data set size and content. The larger data requires correspondingly fast computer-based analyses. However, these analyses often do not scale well with increased data size. Principal component analysis (PCA) is an important analytical tool commonly used with MSI data; however, most PCA algorithms load and process the entire data set within random access memory (RAM) which is most often insufficient for large data sets. PCA algorithms that use less RAM than the data set exist but are usually much slower or sacrifice precision and are rarely used for MSI data processing. Incremental PCA (IPCA) is an alternative algorithm that avoids large RAM allocations while also preserving speed and analytical precision. Here, we demonstrate and benchmark the use of differing implementations of IPCA, PCA, and commercial software on large and often complex MSI data sets. We show that using an already-published Python-based IPCA algorithm, IPCA can be successfully applied to MSI data sets too large to fit with RAM. Furthermore, our benchmarks demonstrate that, contrary to expectations, IPCA is faster than all other tested PCA implementations on all large data sets that can be directly compared.
    Keywords:  Python; algorithm; incremental principal component analysis; mass spectrometry imaging; principal component analysis; random access memory
    DOI:  https://doi.org/10.1021/jasms.4c00314
  24. Heliyon. 2024 Oct 30. 10(20): e39422
    UPLC-MS/MS analysis of axitinib and pharmacokinetic application in beagle dogs.
    Hengyu Bi, Yixin Wang, Xiaochen Ding, Jiahang Li, He Qi, Xiangjun Qiu.
       Objective: To develop a method for determining the concentration of axitinib in beagle dog plasma and utilize this method to investigate the pharmacokinetics of orally administered axitinib in beagle dogs.
    Methods: Plasma samples were processed using acetonitrile precipitation and analyzed by UPLC-MS/MS with sunitinib as an internal standard (IS). Chromatographic separation was achieved on a Waters Acquisition UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm) with a gradient elution of acetonitrile and 0.1 % formic acid. Mass spectrometry uses an electrospray ion source for positive ion detection in a multiple reaction monitoring mode. The monitored ion transitions for axitinib and sunitinib were m/z 387 → 355.96 and m/z 399.3 → 282.96, respectively. Six beagle dogs were administered 0.33 mg/kg of axitinib orally, and venous blood samples were collected at 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 h post-dose for pharmacokinetic analysis.
    Results: The assay demonstrated a linear range of 0.5-100 ng/mL (r2 = 0.9992), and the lower limit of quantification was up to 0.5 ng/mL. Precision, as assessed by relative standard deviation (RSD), was within 8.64 % for both intraday and interday variability. The relative error (RE) for precision from -2.77 %-1.20 %. The recovery rate of the analytes exceeded 85.28 % and the matrix effect was approximately 100 %. Plasma samples maintained stability under various conditions, including room temperature storage for 12 h, processed on an automatic sampler at 4 °C for 6 h, three freeze-thaw cycles, and long-term storage at -80 °C for 60 days. Pharmacokinetic parameters were determined using DAS 2.0 software, revealing a half-life (T1/2) of 6.05 h and an area under the curve (AUC (0 → ∞)) of 97.13 ng h/mL for axitinib.
    Conclusions: The UPLC-MS/MS method developed in this study offers high specificity, rapid analysis, high recovery, excellent linearity, and minimal plasma volume requirements, making it well-suited for pharmacokinetic and drug interaction studies in beagles dogs.
    Keywords:  Axitinib; Beagles; Pharmacokinetics; UPLC-MS/MS
    DOI:  https://doi.org/10.1016/j.heliyon.2024.e39422
  25. Anal Bioanal Chem. 2024 Oct 29.
    A systematic evaluation of quenching, extraction and analysis procedures for metabolomics study of the mechanism of QYSLD intervention in A549 cells.
    Chang Chen, Ting Zheng, Yang Chen, Zegeng Li, Huan Wu.
      The preparation of cellular metabolomics samples and how to achieve comprehensive coverage of different polar metabolites in cell samples in the analysis pose a challenge for cellular metabolomics. In this study, we optimized a metabolomics protocol based on ultra-high-performance liquid chromatography high-resolution mass spectrometry (UPLC/HRMS) for the extraction and detection of metabolites in A549 cells and exploration of the intervention effect of Qi-Yu-San-Long decoction (QYSLD) on A549 cells. The results indicate that the lowest level of ATP leakage was observed when A549 cells were quenched under liquid nitrogen. MeOH/chloroform/H2O (1:2:1) extraction yielded more chromatographic peaks and excellent reproducibility, and the relative extraction efficiency of most target metabolites was also high. And we optimized the chromatographic separation conditions in both HILIC and RPLC modes, enabling comprehensive detection and analysis of metabolites with varying polarities. Then, we applied the optimized method to UPLC-Q-TOF/MS-based metabolomics of A549 cells to study the mechanism of QYSLD intervention in non-small cell lung cancer (NSCLC). The CCK-8, EdU staining, and cell cycle assay showed that QYSLD inhibited the proliferation of A549 cells by interfering with the cell cycle and blocking them in the G1 phase. A total of 36 differential metabolites associated with the antitumor effects of QYSLD on NSCLC were identified, mainly involving nicotinate and nicotinamide metabolism, sphingolipid metabolism, and glycerophospholipid metabolism. And western blotting confirmed that the change in 1-methylnicotinamide levels after QYSLD intervention was associated with the inhibition of nicotinamide N-methyltransferase expression in A549 cells.
    Keywords:  A549 cells; Cell metabolomics; HILIC-HRMS; RPLC-HRMS; Sample preparation
    DOI:  https://doi.org/10.1007/s00216-024-05563-8
  26. Pharmaceutics. 2024 Oct 19. pii: 1340. [Epub ahead of print]16(10):
    Development and Validation of an Improved HPLC-MS/MS Method for Quantifying Total and Unbound Lenalidomide in Human Plasma.
    Suhyun Lee, Seungwon Yang, Wang-Seob Shim, Eunseo Song, Seunghoon Han, Sung-Soo Park, Suein Choi, Sung Hwan Joo, Seok Jun Park, Beomjin Shin, Donghyun Kim, Hyeonsu Kim, Yujung Jung, Kyung-Tae Lee, Eun Kyoung Chung.
       BACKGROUND/OBJECTIVES: This study aimed to develop a fully validated HPLC-MS/MS method for quantifying total and unbound lenalidomide concentrations in human plasma.
    METHODS: Unbound concentrations were measured using plasma ultrafiltrate prepared with Amicon® Centrifugal Filters. Lenalidomide and lenalidomide-d5 (internal standard) were extracted from 50 μL of human plasma using liquid-liquid extraction. Chromatography was conducted with a Halo® C18 column using 0.1% formic acid and methanol (20:80, v/v) as the mobile phase. The mass spectrometer was operated in a positive ion mode with an electrospray ionization interface and multiple reaction monitoring modes.
    RESULTS: Calibration curves were linear over the range of 5 to 1000 ng/mL (r2 > 0.996) for both the total and unbound lenalidomide. For total lenalidomide concentrations, between-run precision (coefficients of variation) and accuracy were 1.70-7.65% and 94.45-101.10%, respectively. For unbound concentrations, inter-day precision and accuracy were 1.98-10.55% and 93.95-98.48%, respectively.
    CONCLUSIONS: We developed a highly reproducible, sensitive, and efficient bioanalytical method using a smaller volume of plasma sample (50 μL) with a relatively short run time (2.5 min). The proposed analytical method was successfully applied to measure total and unbound lenalidomide concentrations at various time points in multiple myeloma patients with renal impairment.
    Keywords:  HPLC-MS/MS; human plasma; lenalidomide; protein binding; quantitative analysis; validation
    DOI:  https://doi.org/10.3390/pharmaceutics16101340
This page is being maintained by Thomas Krichel. It was last updated on 2024‒11‒22 at 4:21.