bims-metlip 2024-07-21 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2024–07–21
fiveteen papers selected by
Sofia Costa, Matterworks

Polar metabolomics using trichloroacetic acid extraction and porous graphitic carbon stationary phase.
Development and validation of an LC-MS/MS method for ruxolitinib quantification: advancing personalized therapy in hematologic malignancies.
Hyphenation of supercritical fluid chromatography and trapped ion mobility-mass spectrometry for quantitative lipidomics.
PeakQC: A Software Tool for Omics-Agnostic Automated Quality Control of Mass Spectrometry Data.
MetaboLink: A web application for Streamlined Processing and Analysis of Large-Scale Untargeted Metabolomics Data.
Analysis and Quantification of the Mitochondrial-ER Lipidome.
Metabolomics-based profiling of five Salvia L. (Lamiaceae) species using untargeted data analysis workflow.
Wide-energy programmable microwave plasma-ionization for high-coverage mass spectrometry analysis.
Application of untargeted liquid chromatography-mass spectrometry to routine analysis of food using three-dimensional bucketing and machine learning.
Development and validation of a non-invasive method for quantifying amino acids in human saliva.
Multi-modal mass spectrometry imaging of a single tissue section.
Clinical metabolomics: Useful insights, perspectives and challenges.
Development and application of a multi-sugar assay to assess intestinal permeability.
Sensitive quantification of mevalonate pathway intermediates and prediction of relative novel analogs by chemical derivatization-based LC-MS/MS.
Development and Validation of A GC-MS Method Based on Solid Phase Extraction and Derivatization for Analysis of Free and Glucuronide-Conjugated Bisphenol F in Biological Samples.

Metabolomics. 2024 Jul 16. 20(4): 77

Polar metabolomics using trichloroacetic acid extraction and porous graphitic carbon stationary phase.

Francesca Day, Justin O'Sullivan, Farha Ramzan, Chris Pook.

   INTRODUCTION: Accurately identifying and quantifying polar metabolites using untargeted metabolomics has proven challenging in comparison to mid to non-polar metabolites. Hydrophilic interaction chromatography and gas chromatography-mass spectrometry are predominantly used to target polar metabolites.
OBJECTIVES: This study aims to demonstrate a simple one-step extraction combined with liquid chromatography-mass spectrometry (LC-MS) that reliably retains polar metabolites.
METHODS: The method involves a MilliQ + 10% trichloroacetic acid extraction from 6 healthy individuals serum, combined with porous graphitic carbon liquid chromatography-mass spectrometry (LC-MS). The coefficient of variation (CV) assessed retention reliability of polar metabolites with logP as low as - 9. QreSS (Quantification, Retention, and System Suitability) internal standards determined the method's consistency and recovery efficiency.
RESULTS: The method demonstrated reliable retention (CV < 0.30) of polar metabolites within a logP range of - 9.1 to 5.6. QreSS internal standards confirmed consistent performance (CV < 0.16) and effective recovery (70-130%) of polar to mid-polar metabolites. Quality control dilution series demonstrated that ~ 80% of annotated metabolites could be accurately quantified (Pearson's correlation coefficient > 0.80) within their concentration range. Repeatability was demonstrated through clustering of repeated extractions from a single sample.
CONCLUSION: This LC-MS method is better suited to covering the polar segment of the metabolome than current methods, offering a reliable and efficient approach for accurate quantification of polar metabolites in untargeted metabolomics.

Keywords:  HILIC; LC–MS; Polar metabolites; Porous graphitic carbon; Trichloroacetic acid; Untargeted metabolomics

DOI:  https://doi.org/10.1007/s11306-024-02146-7
J Pharm Pharm Sci. 2024 ;27 12905

Development and validation of an LC-MS/MS method for ruxolitinib quantification: advancing personalized therapy in hematologic malignancies.

Na Li, Huiying Zhang, Haochen Bai, Kaizhi Lu.

   Background: Hematologic malignancies such as leukemia and lymphoma present treatment challenges due to their genetic and molecular heterogeneity. Ruxolitinib, a Janus kinase (JAK) inhibitor, has demonstrated efficacy in managing these cancers. However, optimal therapeutic outcomes are contingent upon maintaining drug levels within a therapeutic window, highlighting the necessity for precise drug monitoring.
Methods: We developed a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify ruxolitinib in human plasma, improving upon traditional methods in specificity, sensitivity, and efficiency. The process involved the use of advanced chromatographic techniques and robust mass spectrometric conditions to ensure high accuracy and minimal matrix effects. The study was conducted using samples from 20 patients undergoing treatment, with calibration standards ranging from 10 to 2000 ng/mL.
Results: The method displayed linearity (R 2 > 0.99) across the studied range and proved highly selective with no significant interference observed. The method's precision and accuracy met FDA guidelines, with recovery rates consistently exceeding 85%. Clinical application demonstrated significant variability in ruxolitinib plasma levels among patients, reinforcing the need for individualized dosing schedules.
Conclusion: The validated LC-MS/MS method offers a reliable and efficient tool for the therapeutic drug monitoring of ruxolitinib, facilitating personalized treatment approaches in hematologic malignancies. This approach promises to enhance patient outcomes by optimizing dosing to reduce toxicity and improve efficacy.

Keywords:  LC-MS/MS; hematologic malignancies; precision medicine; ruxolitinib; therapeutic drug monitoring

DOI:  https://doi.org/10.3389/jpps.2024.12905
Anal Chim Acta. 2024 Aug 15. pii: S0003-2670(24)00714-1. [Epub ahead of print]1317 342913

Hyphenation of supercritical fluid chromatography and trapped ion mobility-mass spectrometry for quantitative lipidomics.

Johannes Scholz, Edward Rudt, Anna Gremme, Carina M Gaßmöller Née Wienken, Julia Bornhorst, Heiko Hayen.

   BACKGROUND: Lipidomics studies require rapid separations with accurate and reliable quantification results to further elucidate the role of lipids in biological processes and their biological functions. Supercritical fluid chromatography (SFC), in particular, can provide this rapid and high-resolution separation. The combination with trapped ion mobility spectrometry (TIMS) has not yet been applied, although the post-ionization separation method in combination with liquid chromatography or imaging techniques has already proven itself in resolving isomeric and isobaric lipids and preventing false identifications. However, a multidimensional separation method should not only allow confident identification but also provide quantitative results to substantiate studies with absolute concentrations.
RESULTS: A SFC method was developed and the hyphenation of SFC and TIMS was further explored towards the separation of different isobaric overlaps. Furthermore, lipid identification was performed using mass spectrometry (MS) and parallel accumulation serial fragmentation (PASEF) MS/MS experiments in addition to retention time and collision cross section (CCS). Quantification was further investigated with short TIMS ramps and performed based on the ion mobility signal of lipids, since TIMS increases the sensitivity by noise filtering. The final method was, as an exemplary study, applied to investigate the function of different ceramide synthases (CerS) in the nematode and model organism Caenorhabditis elegans (C. elegans). Loss of three known CerS hyl-1, hyl-2 and lagr-1 demonstrated different influences on and alterations in the sphingolipidome.
SIGNIFICANCE: This method describes for the first time the combination of SFC and TIMS-MS/MS, which enables a fast and sensitive quantification of lipids. The results of the application to C. elegans samples prove the functionality of the method and support research on the metabolism of sphingolipids in nematodes.

Keywords:  Caenorhabditis elegans; Ceramides; Lipidomics; SFC-TIMS-MS; Sphingolipids

DOI:  https://doi.org/10.1016/j.aca.2024.342913
J Am Soc Mass Spectrom. 2024 Jul 16.

PeakQC: A Software Tool for Omics-Agnostic Automated Quality Control of Mass Spectrometry Data.

Andrea Harrison, Josie G Eder, Priscila M Lalli, Nathalie Munoz, Yuqian Gao, Chaevien S Clendinen, Daniel J Orton, Xueyun Zheng, Sarah M Williams, Sneha P Couvillion, Rosalie K Chu, Vimal K Balasubramanian, Arunima Bhattacharjee, Christopher R Anderton, Kyle R Pomraning, Kristin E Burnum-Johnson, Tao Liu, Jennifer E Kyle, Aivett Bilbao.

  Mass spectrometry is broadly employed to study complex molecular mechanisms in various biological and environmental fields, enabling 'omics' research such as proteomics, metabolomics, and lipidomics. As study cohorts grow larger and more complex with dozens to hundreds of samples, the need for robust quality control (QC) measures through automated software tools becomes paramount to ensure the integrity, high quality, and validity of scientific conclusions from downstream analyses and minimize the waste of resources. Since existing QC tools are mostly dedicated to proteomics, automated solutions supporting metabolomics are needed. To address this need, we developed the software PeakQC, a tool for automated QC of MS data that is independent of omics molecular types (i.e., omics-agnostic). It allows automated extraction and inspection of peak metrics of precursor ions (e.g., errors in mass, retention time, arrival time) and supports various instrumentations and acquisition types, from infusion experiments or using liquid chromatography and/or ion mobility spectrometry front-end separations and with/without fragmentation spectra from data-dependent or independent acquisition analyses. Diagnostic plots for fragmentation spectra are also generated. Here, we describe and illustrate PeakQC's functionalities using different representative data sets, demonstrating its utility as a valuable tool for enhancing the quality and reliability of omics mass spectrometry analyses.

Keywords:  computational tool; data processing; data-dependent acquisition; data-independent acquisition, data quality control, proteomics; ion mobility spectrometry; lipidomics, algorithm; liquid chromatography; metabolomics

DOI:  https://doi.org/10.1021/jasms.4c00146
Bioinformatics. 2024 Jul 17. pii: btae459. [Epub ahead of print]

MetaboLink: A web application for Streamlined Processing and Analysis of Large-Scale Untargeted Metabolomics Data.

Ana Mendes, Jesper Foged Havelund, Jonas Lemvig, Veit Schwämmle, Nils J Færgeman.

MOTIVATION: The post-processing and analysis of large-scale untargeted metabolomics data face significant challenges due to the intricate nature of correction, filtration, imputation, and normalization steps. Manual execution across various applications often leads to inefficiencies, human-induced errors, and inconsistencies within the workflow.
RESULTS: Addressing these issues, we introduce MetaboLink, a novel web application designed to process LC-MS metabolomics datasets combining established methodologies and offering flexibility and ease of implementation. It offers visualization options for data interpretation, an interface for statistical testing, and integration with PolySTest for further tests and with VSClust for clustering analysis.
AVAILABILITY: Fully functional tool is publicly available at https://computproteomics.bmb.sdu.dk/Metabolomics/. The source code is available at https://github.com/anitamnd/MetaboLink and a detailed description of the app can be found at https://github.com/anitamnd/MetaboLink/wiki. A tutorial video can be found at https://youtu.be/ZM6j10S6Z8Q.
SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

DOI: https://doi.org/10.1093/bioinformatics/btae459
Bio Protoc. 2024 Jul 05. 14(13): e5028

Analysis and Quantification of the Mitochondrial-ER Lipidome.

Alexis R Diaz-Vegas, Anthony S Don, James G Burchfield.

  Mitochondria are vital organelles essential for cellular functions, but their lipid composition and response to stressors are not fully understood. Recent advancements in lipidomics reveal insights into lipid functions, especially their roles in metabolic perturbations and diseases. Previous methods have focused on the protein composition of mitochondria and mitochondrial-associated membranes. The advantage of our technique is that it combines organelle isolation with targeted lipidomics, offering new insights into the composition and dynamics of these organelles in pathological conditions. We developed a mitochondria isolation protocol for L6 myotubes, enabling lipidomics analysis of specific organelles without interference from other cellular compartments. This approach offers a unique opportunity to dissect lipid dynamics within mitochondria and their associated ER compartments under cellular stress. Key features • Analysis and quantification of lipids in mitochondria-ER fraction through liquid chromatography-tandem mass spectrometry-based lipidomics (LC-MS/MS lipidomics). • LC-MS/MS lipidomics provide precise and unbiased information on the lipid composition in in vitro systems. • LC-MS/MS lipidomics facilitates the identification of lipid signatures in mammalian cells.

Keywords:  Cardiolipin; Ceramides; Endoplasmic reticulum; Lipidomics; Mitochondria; Subcellular fractionation

DOI:  https://doi.org/10.21769/BioProtoc.5028
Phytochem Anal. 2024 Jul 14.

Metabolomics-based profiling of five Salvia L. (Lamiaceae) species using untargeted data analysis workflow.

Navaz Kharazian, Farzaneh Jafari Dehkordi, Chun-Lei Xiang.

   INTRODUCTION: The genus Salvia L., a member of the family Lamiaceae, is a keystone genus with a wide range of medicinal properties. It possesses a rich metabolite source that has long been used to treat different disorders.
OBJECTIVES: Due to a deficiency of untargeted metabolomic profiling in the genus Salvia, this work attempts to investigate a comprehensive mass spectral library matching, computational data annotations, exclusive biomarkers, specific chemotypes, intraspecific metabolite profile variation, and metabolite enrichment by a case study of five medicinal species of Salvia.
MATERIAL AND METHODS: Aerial parts of each species were subjected to QTRAP liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis workflow based on untargeted metabolites. A comprehensive and multivariate analysis was acquired on the metabolite dataset utilizing MetaboAnalyst 6.0 and the Global Natural Products Social Molecular Networking (GNPS) Web Platform.
RESULTS: The untargeted approach empowered the identification of 117 metabolites by library matching and 92 nodes annotated by automated matching. A machine learning algorithm as substructural topic modeling, MS2LDA, was further implemented to explore the metabolite substructures, resulting in four Mass2Motifs. The automated library newly discovered a total of 23 metabolites. In addition, 87 verified biomarkers of library matching, 58 biomarkers of GNPS annotations, and 11 specific chemotypes were screened.
CONCLUSION: Integrative spectral library matching and automated annotation by the GNPS platform provide comprehensive metabolite profiling through a workflow. In addition, QTRAP LC-MS/MS with multivariate analysis unveiled reliable information about inter and intraspecific levels of differentiation. The rigorous investigation of metabolite profiling presents a large-scale overview and new insights for chemotaxonomy and pharmaceutical studies.

Keywords:  Lamiaceae; Salvia; metabolomics; molecular network; profiling

DOI:  https://doi.org/10.1002/pca.3423
Nat Commun. 2024 Jul 18. 15(1): 6075

Wide-energy programmable microwave plasma-ionization for high-coverage mass spectrometry analysis.

Fengjian Chu, Gaosheng Zhao, Wei Wei, Nazifi Sani Shuaibu, Hongru Feng, Yuanjiang Pan, Xiaozhi Wang.

Although numerous ambient ionization mass spectroscopy technologies have been developed over the past 20 years to address diverse analytical circumstances, a single-ion source technique that can handle all analyte types is still lacking. Here, a wide-energy programmable microwave plasma-ionization mass spectrometry (WPMPI-MS) system is presented, through which MS analysis can achieve high coverage of substances with various characteristics by digitally regulating the microwave energy. In addition, ionization energy can be rapidly scanned using programmable waveforms, enabling the simultaneous detection of biomolecules, heavy metals, non-polar molecules, etc., in seconds. WPMPI-MS performs well in analyzing real samples, rapidly analyzing nine toxicological standards in one drop of serum, and demonstrating good quantification and liquid chromatography coupling capability. The WPMPI-MS has also been used to detect soil extracts, solid pharmaceuticals, and landfill leachate, further demonstrating its robust analytical capabilities for real samples. The prospective uses of the technology in biological and chemical analysis are extensive, and it is anticipated to emerge as a viable alternative to commercially available ion sources.

DOI: https://doi.org/10.1038/s41467-024-50322-z
Sci Rep. 2024 Jul 18. 14(1): 16594

Application of untargeted liquid chromatography-mass spectrometry to routine analysis of food using three-dimensional bucketing and machine learning.

Jule Hansen, Christof Kunert, Hella Münstermann, Kurt-Peter Raezke, Stephan Seifert.

  For the detection of food adulteration, sensitive and reproducible analytical methods are required. Liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) is a highly sensitive method that can be used to obtain analytical fingerprints consisting of a variety of different components. Since the comparability of measurements carried out with different devices and at different times is not given, specific adulterants are usually detected in targeted analyses instead of analyzing the entire fingerprint. However, this comprehensive analysis is desirable in order to stay ahead in the race against food fraudsters, who are constantly adapting their adulterations to the latest state of the art in analytics. We have developed and optimized an approach that enables the separate processing of untargeted LC‑HRMS data obtained from different devices and at different times. We demonstrate this by the successful determination of the geographical origin of honey samples using a random forest model. We then show that this approach can be applied to develop a continuously learning classification model and our final model, based on data from 835 samples, achieves a classification accuracy of 94% for 126 test samples from 6 different countries.

Keywords:  Food profiling; Honey; Machine learning; Mass spectrometry; Routine analysis

DOI:  https://doi.org/10.1038/s41598-024-67459-y
RSC Adv. 2024 Jul 12. 14(31): 22292-22303

Development and validation of a non-invasive method for quantifying amino acids in human saliva.

Md Mehedi Hasan, Mamudul Hasan Razu, Sonia Akter, Salma Akter Mou, Minhazul Islam, Mala Khan.

As an analytical matrix, saliva has superior characteristics than blood and urine. Saliva collection is, first and foremost, non-invasive, making it convenient, painless, and secure for more susceptible people. Second, it does not need professional training for medical personnel, resulting in cost-effectiveness and suitability for extensive collection in support of research. In this study, we developed a method and used it to quantify 13 salivary-free amino acid (SFAA) profiles to support the early clinical diagnosis of diseases using LC-MS/MS. Using an Intrada Amino Acid column (100 × 3 mm, 3 μm), chromatographic separation was accomplished with a binary gradient elution, and an electrospray ionisation source running in the positive ionisation mode was chosen for data collection using the multiple reaction monitoring (MRM) modes. Amino acids were extracted from saliva using acetonitrile. In the MRM mode, LODs and LOQs for ten amino acids were in the range of 0.06-2.50 μM and 0.19-7.58 μM, respectively, and those values were in the range of 1.00-3.00 μM and 3.00-8.50 μM, respectively, for three amino acids. Matrix-matched six-point calibration curves showed a linear correlation coefficient (r 2) of ≥0.998. Recovery experiments validated the method by spiking the control sample at three different concentration levels (5, 50 and 100 μM), and the accuracy level was 85-110%. Except for Thr and Ser, intra- (n = 3) and inter-day (n = 3) precision fell between 0.02 and 7.28. Salivary amino acids can serve as possible biomarkers for various malignancies, with fluctuations in body fluids being crucial for cancer diagnosis; therefore, examining amino acid patterns in saliva can assist in early cancer detection. LC-MS offers improved selectivity and sensitivity for non-derivatised amino acid analysis, surpassing conventional methods and offering proactive quality assurance, making it suitable for complicated sample matrices. These discoveries could be significant in investigating new pathways and cancer treatments and looking for possible AA biomarkers for other malignancies and diseases.

DOI: https://doi.org/10.1039/d4ra01130a
J Mass Spectrom. 2024 Aug;59(8): e5074

Multi-modal mass spectrometry imaging of a single tissue section.

Lingpeng Zhan, Yanyi Huang, Guanbo Wang.

  Mass spectrometry imaging (MSI) was developed to visualize spatial chemical information within tissues, thereby facilitating spatial multi-omic analysis. However, due to the limited spatial information provided by individual modal MSI, correlating various chemical data within tissues remains a significant challenge. In recent years, multimodal MSI has garnered considerable attention due to its ability to visualize the spatial distributions of multiple biomolecules within tissues. Among the strategies employed in this field, multimodal imaging on a single tissue section circumvents multiple issues introduced by integration of images of consecutive tissue sections. In this minireview, we provide an overview of multimodal MSI on a single tissue section, with a particular focus on the use of Matrix-Assisted Laser Desorption/Ionization-MSI for spatial multi-omic investigations that offer a comprehensive and in-depth elucidation of the biological state and activities, aiming to inspire the development of new approaches in this field.

Keywords:  MALDI; mass spectrometry imaging; matrix selection; multimodal imaging; multi‐omics; spatial omics

DOI:  https://doi.org/10.1002/jms.5074
Metabol Open. 2024 Jun;22 100290

Clinical metabolomics: Useful insights, perspectives and challenges.

Maria Dalamaga.

  Metabolomics, a cutting-edge omics technique, is a rapidly advancing field in biomedical research, concentrating on the elucidation of pathogenetic mechanisms and the discovery of novel metabolite signatures predictive of disease risk, aiding in earlier disease detection, prognosis and prediction of treatment response. The capacity of this omics approach to simultaneously quantify thousands of metabolites, i.e. small molecules less than 1500 Da in samples, positions it as a promising tool for research and clinical applications in personalized medicine. Clinical metabolomics studies have proven valuable in understanding cardiometabolic disorders, potentially uncovering diagnostic biomarkers predictive of disease risk. Liquid chromatography-mass spectrometry is the predominant analytical method used in metabolomics, particularly untargeted. Metabolomics combined with extensive genomic data, proteomics, clinical chemistry data, imaging, health records, and other pertinent health-related data may yield significant advances beneficial for both public health initiatives, clinical applications and precision medicine, particularly in rare disorders and multimorbidity. This special issue has gathered original research articles in topics related to clinical metabolomics as well as research articles, reviews, perspectives and highlights in the broader field of translational and clinical metabolic research. Additional research is necessary to identify which metabolites consistently enhance clinical risk prediction across various populations and are causally linked to disease progression.

Keywords:  Mass spectrometry; Metabolite; Metabolomics; Multi-omics; Obesity; Omics; Precision medicine

DOI:  https://doi.org/10.1016/j.metop.2024.100290
Bioanalysis. 2024 Jul 18. 1-13

Development and application of a multi-sugar assay to assess intestinal permeability.

Nicole A Schneck, Ahmed Moghieb, Claire Teague, Hermes Licea Perez.

  Aim: Bioanalytical assays to measure rhamnose, erythritol, lactulose and sucralose in human urine and plasma were developed to support an indomethacin challenge study for intestinal permeability assessment in healthy participants. Methods: The multi-sugar assays utilized 5-μl sample matrix and a simple chemical derivatization with acetic anhydride, followed by RPLC-MS/MS detection. Results: Rhamnose and erythritol quantification was established between 1.00-1,000 μg/ml in urine and 250-250,000 ng/ml in plasma. For lactulose and sucralose, dynamic ranges of 0.1-100 μg/ml (urine) and 25-25,000 ng/ml (plasma) were applied for biological measurements. Conclusion: This work helped overcome some of the common analytical challenges associated with the bioanalysis of mono- and disaccharides and achieved improved limits of quantification.

Keywords:  LC-MS/MS; ammonium adducts; bioanalysis; carbohydrates; derivatization; saccharides

DOI:  https://doi.org/10.1080/17576180.2024.2374168
J Chromatogr A. 2024 Jul 14. pii: S0021-9673(24)00537-5. [Epub ahead of print]1731 465163

Sensitive quantification of mevalonate pathway intermediates and prediction of relative novel analogs by chemical derivatization-based LC-MS/MS.

Meiyu Gao, Jiarui Sun, Qinwen Xiao, Yuanyuan Zhai, Yuan Tian, Zunjian Zhang, Fengguo Xu, Pei Zhang.

  The mevalonate (MVA) pathway plays a crucial role in the occurrence and progression of various diseases, such as osteoporosis, breast cancer, and lung cancer, etc. However, determining all the MVA pathway intermediates is still challenging due to their high polarity, low concentration, chelation effect with metal compartments, and poor mass spectrometric response. In this study, we established a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method coupled with N2, N2, N4, N4-tetramethyl-6-(4-(piperazin-1-ylsulfonyl) phenyl)-1,3,5-triazine-2,4-diamine (Tmt-PP) labeling for the simultaneous analysis of all MVA intermediates in biospecimens. Chemical derivatization significantly improved the chromatographic retention, peak shape, and detection sensitivity of the analytes. Moreover, we employed a method named mass spectrum calculation to achieve the absolute quantification of the isomers, i.e., isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). The established method was fully qualified and applied to explore the difference of these metabolites in cisplatin-resistant non-small cell lung cancer (NSCLC) cells. Additionally, several MVA intermediate analogs, including isopentenyl monophosphate or dimethylallyl monophosphate (IMP/DMAMP), geranyl monophosphate (GMP), 5-triphosphomevalonate (MTP), and isopentenyl triphosphate or dimethylallyl triphosphate (ITP/DMATP), were identified for the first time using a knowledge-driven prediction strategy. We further explored the tissue distribution of these novel metabolites. Overall, this work developed a sensitive quantification method for all MVA intermediates, which will enhance our understanding of the role of this pathway in various health and disease conditions. The novel metabolites we discovered warrant further investigations into their biosynthesis and biological functions.

Keywords:  Chemical derivatization; Drug resistance; LC-MS/MS; Mass spectrum calculation; Mevalonate pathway

DOI:  https://doi.org/10.1016/j.chroma.2024.465163
J AOAC Int. 2024 Jul 17. pii: qsae062. [Epub ahead of print]

Development and Validation of A GC-MS Method Based on Solid Phase Extraction and Derivatization for Analysis of Free and Glucuronide-Conjugated Bisphenol F in Biological Samples.

Xu-Liang Cao, Svetlana Popovic, Santokh Gill.

   BACKGROUND: As one of the speculated bisphenols to replace bisphenol A (BPA), bisphenol F (BPF), naturally present in mustard, is structurally similar to BPA and may have similar estrogenic activity, but information on its toxicity is very limited compared to BPA.
OBJECTIVE: In order to support the toxicology study of BPF at Heath Canada, a gas chromatography (GC) with mass spectrometry (MS) method based on solid phase extraction and derivatization was developed for analysis of BPF in liver samples.
METHODS: Samples were treated with β-glucuronidase to convert BPF glucuronide to free BPF for analysis of total BPF.
RESULTS: The method was validated for free BPF at different spiking levels, and recoveries ranged from 90.0-97.5% with relative standard deviations from 0.11-5.54%. The method was also validated for glucuronide-conjugated BPF at different spiking levels of BPF mono-β-D-glucuronide, recoveries ranged from 72.3-93.3% with relative standard deviations from 1.7-8.94%. The method was used to analyse 60 liver tissue samples from the rats dosed with BPF at different levels in a toxicology study. Free and glucuronide-conjugated BPF were not detected in any of the control samples which were not dosed with BPF (average method detection limit: 0.31 ng/g), but detected in all the other liver tissue samples with levels increasing at higher doses. The percent of glucuronide-conjugated BPF in total BPF varied among the liver samples, from as low as 9.8% to as high as 77.9%, indicating the importance of analysing biological samples for BPF in both free and conjugated forms for total exposure.
CONCLUSION: A GC-MS method based on solid phase extraction and derivatization was developed for analysis of both free and glucuronide-conjugated BPF in liver samples. This method was validated not only for free BPF, but also for mono-β-D-glucuronide conjugated BPF for the first time to confirm the efficiency of the deconjugation procedure with enzyme.
HIGHLIGHTS: This method can be adapted and applied for analysis of free and glucuronide-conjugated BPF in other biological samples with appropriate validation in target sample matrices.

Keywords:  BPF glucuronide; BPF mono-β-D-glucuronide; bisphenol F (BPF); liver

DOI:  https://doi.org/10.1093/jaoacint/qsae062