bims-metlip 2024-12-08 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2024–12–08
fourteen papers selected by
Sofia Costa, Matterworks

Development and Validation of a Sensitive LC-MS/MS Method for Determination of Lenvatinib and Its Major Metabolites in Human Plasma and Its Application in Hepatocellular Carcinoma Patients.
Rapid quantification of murine bile acids using liquid chromatography-tandem mass spectrometry.
Simultaneous analysis of reduced and oxidized forms of multiple biological aminothiols in cells and tissues by double derivatization combined with liquid chromatography‒mass spectrometry.
[Development of the Analysis of Lysophospholipids and Photo-sensitive Pharmaceuticals Based on LC-MS/MS for Elucidation of Their Behaviors].
Development and Validation of a UHPLC-MS/MS Method for the Simultaneous Quantification of Candesartan and Bisoprolol Together with Other 16 Antihypertensive Drugs in Plasma Samples.
Effective data visualization strategies in untargeted metabolomics.
Development of a UPLC-MS/MS Method for Bioanalysis of Ethoxysanguinarine and Its Application in Pharmacokinetic Study of Ethoxysanguinarine Nanoemulsion.
A highly sensitive UHPLC-MS/MS method for determining 15 designer LSD analogs in biological samples with application to stability studies.
From Hydrocarbons to Highly Functionalized Molecules in a Single Measurement: Comprehensive Analysis of Complex Gas Mixtures by Multi-Pressure Chemical Ionization Mass Spectrometry.
Quantitative Endogenous Polyamine Analysis via Capillary Electrophoresis/Mass Spectrometry: Characterization and Practical Considerations.
Integration of feature-based molecular networking and high-definition data-dependent acquisition for the comprehensive multicomponent characterization of Honghua Xiaoyao Tablet.
Development and validation of a sensitive and fast bioanalytical LC-MS/MS assay for the quantitation of venetoclax and azacitidine in Rat Plasma: Application to pharmacokinetic study.
Mass Spectrometry Imaging for Spatial Toxicology Research.
High-resolution targeted mass spectrometry for comprehensive quantification of sphingolipids: clinical applications and characterization of extracellular vesicles.

J Sep Sci. 2024 Dec;47(23): e70042

Development and Validation of a Sensitive LC-MS/MS Method for Determination of Lenvatinib and Its Major Metabolites in Human Plasma and Its Application in Hepatocellular Carcinoma Patients.

Shaoxing Guan, Guosheng Yuan, Guofang Bi, Qingqing Yu, Jian-Hong Fang, Jinzhang Chen, Huichang Bi.

  Lenvatinib has been demonstrated effective in advanced hepatocellular carcinoma (HCC), but the pharmacokinetic-pharmacodynamics behavior of lenvatinib and its metabolites remains unclear. To investigate the pharmacokinetic-pharmacodynamics behavior of lenvatinib and its active metabolites in advanced HCC patients, it is important to develop a simple and rapid method to analyze the exposures of lenvatinib and its metabolites in human samples. Here, we established and validated a simple and rapid method for determining lenvatinib and its three major metabolites, descyclopropyl lenvatinib (M1), O-demethyl lenvatinib hydrochloride (M2), and lenvatinib N-Oxide (M3) by liquid chromatography-tandem mass spectrometry method. Lenvatinib and its main metabolites were separated on an X-Terra RP18 column (50 × 2.1 mm, 3.5 µm) at 35°C within 3 min, and the analytes were isocratically eluted with the mobile phase of methanol-water (10:90, v/v) containing 0.1% of formic acid at a flow rate of 0.15 mL/min. The calibration range was 1-1000 ng/mL for lenvatinib, while 0.1-100 ng/mL for M1-M3 under positive electrospray ionization mode. The inter- and intra-batch precisions and accuracy were acceptable for lenvatinib and its metabolites. This method was successfully applied to measure lenvatinib and its metabolites in plasma samples from HCC patients, which provides a robust tool for pharmacokinetic-pharmacodynamics studies of lenvatinib.

Keywords:  HCC patients; lenvatinib; liquid chromatography‐tandem mass spectrometry; metabolites; method validation

DOI:  https://doi.org/10.1002/jssc.70042
Anal Bioanal Chem. 2024 Dec 02.

Rapid quantification of murine bile acids using liquid chromatography-tandem mass spectrometry.

Sven Hermeling, Johannes Plagge, Sabrina Krautbauer, Josef Ecker, Ralph Burkhardt, Gerhard Liebisch.

  Interest in bile acids (BAs) is growing due to their emerging role as signaling molecules and their association with various diseases such as colon cancer and metabolic syndrome. Analyzing BAs requires chromatographic separation of isomers, often with long run times, which hinders BA analysis in large studies. Here, we present a high-throughput method based on liquid chromatography-tandem mass spectrometry to quantify BAs in mouse samples. After acidic protein precipitation in the presence of a comprehensive mixture of stable isotope-labeled internal standards (SIL-ISs), BAs are separated on a biphenyl column by gradient elution at basic pH. Quantification is performed using a six-point calibration curve. Except for the separation of β- and ω-muricholic acid (MCA) species, a rapid separation of 27 BA species was achieved in a run time of 6.5 min. Plasma quality controls (QCs) were used to evaluate intra- and inter-day precision. The CV was less than 10% for most BA species and exceeded 20% only for glycohyodeoxycholic (GHDCA) and taurohyodeoxycholic acid (THDCA) due to the lack of a corresponding SIL-IS. The limit of quantification (LoQ) was tested using diluted QCs and was found to be compromised for some BA species as a result of insufficient isotopic purity of the SIL-IS, leading to significant interference with the respective analyte. Finally, we tested the mouse sample material requirements for plasma, bile, and liver samples and determined BA concentrations in C57/BL6N wild-type mice. In conclusion, the LC-MS/MS method presented here permits a rapid and reproducible quantification of the major murine BAs.

Keywords:  Bile; Bile acid; LC–MS/MS; Lipidomics; Liver; Plasma

DOI:  https://doi.org/10.1007/s00216-024-05668-0
J Chromatogr A. 2024 Nov 30. pii: S0021-9673(24)00930-0. [Epub ahead of print]1740 465557

Simultaneous analysis of reduced and oxidized forms of multiple biological aminothiols in cells and tissues by double derivatization combined with liquid chromatography‒mass spectrometry.

Mindan Wang, Li Liu, Xialian Cui, Xiuping Chen, Yixin Wei, Wenwen Zhang, Hang Yin, Chenguo Feng, Fang Zhang.

  Biological aminothiols (BATs) typically exist in both reduced and oxidized forms, each exhibiting diverse biological activities. Monitoring the levels and ratios of the two forms is crucial for clinical diagnosis and understanding their roles in biological systems. In this study, we developed a method for simultaneous analysis of both reduced and oxidized BATs using a double derivatization approach combined with liquid chromatography-mass spectrometry (LC-MS). The method employed a sequential derivatization strategy: initially, 2‑bromo-N,N-dimethylacetamide (Br-DMA) reacted with the thiol groups of reduced BATs, followed by derivatization of both reduced and oxidized BATs with stable isotope labeling reagents, [d0]-/[d3]-6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate ([d0]-/[d3]-DMMIC). The methodology validation showed excellent linearity (R2 > 0.99), accuracy (85.07-119.94 %), precision (intraday: 5.26-18.78 %; interday: 6.52-19.01 %), recovery (70.09-119.27 %), and matrix effect (92.69-126.79 %). Finally, the method was successfully applied to nontargeted BAT screening in lung A549 cells, assessing changes in BAT levels in A549 cells upon treatment with the anticancer compounds triptolide and bufalin, and comparing differences in BAT levels between lung adenocarcinoma and paracarcinoma tissues. The results indicated that the developed method could be a comprehensive practical protocol and serve as a platform for profiling reduced and oxidized BATs in biological samples while meeting various analysis demands.

Keywords:  A549 cells; Biological aminothiols; Double derivatization; LC-MS; Lung adenocarcinoma

DOI:  https://doi.org/10.1016/j.chroma.2024.465557
Yakugaku Zasshi. 2024 ;144(12): 1063-1074

[Development of the Analysis of Lysophospholipids and Photo-sensitive Pharmaceuticals Based on LC-MS/MS for Elucidation of Their Behaviors].

Kohei Kawabata.

  Liquid chromatography tandem mass spectrometry (LC-MS/MS) is an essential tool for drug discovery that enables simple and rapid identification and quantification of chemical substances. A combination of a mobile phase and column makes it possible to analyze a wide range of target substances from low-molecular weight high-polar substances, such as amino acids and peptides, to low-polar substances such as lipids, and even macromolecular substances such as proteins. In this paper, we describe the results of applying LC-MS/MS to the analysis of phospholipids and related substances in biological samples and the analysis of photoproducts of pharmaceuticals. First, MS conditions were optimization using several standards, and a system that enables measurement of a vast number of molecular species with different carbon chain lengths and degrees of unsaturation. Its application to the lipid profiling of influenza A virus-infected cells suggested that viral infection triggered the increase of intracellular levels of diacylglycerols and ceramides at the later stages of infection concomitant with viral replication. In addition, the analysis of lysophospholipids in several cell lines revealed partial functions of several types of glycerophosphodiester phosphodiesterase, which metabolize lysophospholipids. Next, the chemical structures of several photoproducts of pharmaceuticals were elucidated. Novel photoproducts of photo-exposed pharmaceuticals were identified, and the photodegradation pathways were suggested. Based on the photodegradation mechanism, the photodegradability of naproxen was regulated by the addition of several additives such as polyphenols.

Keywords:  liquid chromatography tandem mass spectrometry; lysophospholipid; photodegradation

DOI:  https://doi.org/10.1248/yakushi.24-00119
J Med Chem. 2024 Dec 04.

Development and Validation of a UHPLC-MS/MS Method for the Simultaneous Quantification of Candesartan and Bisoprolol Together with Other 16 Antihypertensive Drugs in Plasma Samples.

Alice Palermiti, Amedeo De Nicolò, Alessandra Manca, Miriam Antonucci, Jacopo Mula, Martina Billi, Jessica Cusato, Alessandro Carta, Marco Pappaccogli, Lara Ponsa, Chiara Fasano, Francesco Giuseppe De Rosa, Franco Rabbia, Franco Veglio, Antonio D'Avolio.

Antihypertensive pharmacological therapy is often characterized by a coadministration of different classes of drugs. Therefore, analytical methods allowing the simultaneous quantification of many drugs are needed for therapeutic drug monitoring (TDM) purposes in this context. In particular, TDM represents a useful tool to discriminate poor adherence from real cases of resistant hypertension. For this reason, the aim of this study is to validate, following the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) guidelines, an ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of 18 antihypertensive drugs in human plasma. A LX-50 coupled with a QSight 220 UHPLC-MS/MS system with electrospray ionization and multiple reaction monitoring mode was used, after a binary gradient separation (13 min) on a reverse-phase Acquity UPLC HSS T3 [1.8 μm, 2.1 mm × 150 mm] column. Method validation showed a stable and acceptable matrix effect, recovery, high accuracy, and precision, assessing the eligibility of this method for routine use in the clinical context.

DOI: https://doi.org/10.1021/acs.jmedchem.4c02045
Nat Prod Rep. 2024 Dec 02.

Effective data visualization strategies in untargeted metabolomics.

Kevin Mildau, Henry Ehlers, Mara Meisenburg, Elena Del Pup, Robert A Koetsier, Laura Rosina Torres Ortega, Niek F de Jonge, Kumar Saurabh Singh, Dora Ferreira, Kgalaletso Othibeng, Fidele Tugizimana, Florian Huber, Justin J J van der Hooft.

Covering: 2014 to 2023 for metabolomics, 2002 to 2023 for information visualizationLC-MS/MS-based untargeted metabolomics is a rapidly developing research field spawning increasing numbers of computational metabolomics tools assisting researchers with their complex data processing, analysis, and interpretation tasks. In this article, we review the entire untargeted metabolomics workflow from the perspective of information visualization, visual analytics and visual data integration. Data visualization is a crucial step at every stage of the metabolomics workflow, where it provides core components of data inspection, evaluation, and sharing capabilities. However, due to the large number of available data analysis tools and corresponding visualization components, it is hard for both users and developers to get an overview of what is already available and which tools are suitable for their analysis. In addition, there is little cross-pollination between the fields of data visualization and metabolomics, leaving visual tools to be designed in a secondary and mostly ad hoc fashion. With this review, we aim to bridge the gap between the fields of untargeted metabolomics and data visualization. First, we introduce data visualization to the untargeted metabolomics field as a topic worthy of its own dedicated research, and provide a primer on cutting-edge visualization research into data visualization for both researchers as well as developers active in metabolomics. We extend this primer with a discussion of best practices for data visualization as they have emerged from data visualization studies. Second, we provide a practical roadmap to the visual tool landscape and its use within the untargeted metabolomics field. Here, for several computational analysis stages within the untargeted metabolomics workflow, we provide an overview of commonly used visual strategies with practical examples. In this context, we will also outline promising areas for further research and development. We end the review with a set of recommendations for developers and users on how to make the best use of visualizations for more effective and transparent communication of results.

DOI: https://doi.org/10.1039/d4np00039k
Biomed Chromatogr. 2025 Jan;39(1): e6055

Development of a UPLC-MS/MS Method for Bioanalysis of Ethoxysanguinarine and Its Application in Pharmacokinetic Study of Ethoxysanguinarine Nanoemulsion.

Jianhui Luo, Songshen Chen, WenYang Song, Yongtong Huang, Song Gao, Jiu Wang.

  Ethoxysanguinarine (ETSG), a benzophenanthridine alkaloid, exhibits diverse biological activities, including antibacterial, antifungal, anti-inflammatory, antioxidant, and anti-tumor effects. Despite these properties, limited research exists on ETSG in vivo pharmacokinetics due to its poor solubility and low bioavailability. In this study, we developed a rapid and specific UPLC-MS/MS method for ETSG bioanalysis. Sample preparation involved one-step protein precipitation using methanol and phellodendrine as an internal standard (IS). The Waters HSS T3 column (2.1 * 50 mm, 1.8 μM) employed a gradient elution with mobile phases A (2 mmol/L ammonium formate aqueous solution-formic acid [99.8:0.2, v/v]) and B (methanol-formic acid [99.8:0.2, v/v]). Mass analysis via Waters Q-mass spectrometer utilized positive scan mode and multiple reaction monitoring. ETSG and IS were detected at m/z 332.0 → 274.0 and 342.0 → 177.0, respectively, within 7.0 min. The method demonstrated excellent precision, accuracy, recovery, and stability, with a linear calibration curve (1.1-560 ng/mL) and strong correlation coefficient (0.9984). Successful pharmacokinetic evaluation in Sprague-Dawley rats included intravenous ETSG administration and intragastric ETSG nanoemulsion/suspension. This method enables steroidal saponin analysis from ETSG in biological samples.

Keywords:  UPLC‐MS/MS; ethoxysanguinarine; nanoemulsion; pharmacokinetic

DOI:  https://doi.org/10.1002/bmc.6055
Analyst. 2024 Dec 05.

A highly sensitive UHPLC-MS/MS method for determining 15 designer LSD analogs in biological samples with application to stability studies.

Olga Wachełko, Karolina Nowak, Kaja Tusiewicz, Marcin Zawadzki, Paweł Szpot.

In recent years, the rise in the synthesis and distribution of LSD analogs in illicit drug markets, commonly referred to as "designer psychedelics", has contributed to increased recreational use. This trend has resulted in a rising number of global reports, with law enforcement increasingly detecting these compounds in blotter papers and biological samples. In the presented paper, an UHPLC-QqQ-MS/MS method was developed for trace determination (fg mL-1) of LSD, its designer analogs (ALD-52, AL-LAD, LAMPA, LSM-775, LSZ, MiPLA, 1B-LSD, 1cP-LSD, 1cP-MiPLA, 1P-LSD, 1P-MiPLA, 1V-LSD and 2-Bromo-LSD) and its metabolite (2-oxo-3-OH-LSD) with simultaneous separation of structural isomers. Biological samples were prepared using liquid-liquid extraction (LLE) at pH 9 (with ethyl acetate); quantification was performed in multiple reaction monitoring (MRM) mode. LSD-d3 was used as an internal standard. The limit of quantification (LOQ) for all substances was 0.5 pg mL-1. Precision and accuracy did not exceed 15.8% and ±14.4%, respectively. Recovery and matrix effect values were 80.6-118.6% and ±19.4%. A stability study was conducted over 30 days under different storage conditions (25 °C, 4 °C and -20 °C) for blood, urine, plasma, and serum, collected in various test tube configurations and with different preservative agents. It was found that the collection of samples in NaF can effectively stabilize LSD analogs and minimize the conversion of N1-substituted compounds to LSD or MiPLA. The presented method is the most sensitive to date for analyzing designer LSD analogs in biological samples, with potential for routine clinical and forensic use, enhancing detection of emerging illicit compounds. By examining the mass spectra (QTOF-MS/MS) obtained in this study and reviewing the literature on analytical characterization of LSD analogs, we proposed fragmentation patterns to aid in future identification of new designer LSD analogs (NPS).

DOI: https://doi.org/10.1039/d4an01361a
Anal Chem. 2024 Dec 03.

From Hydrocarbons to Highly Functionalized Molecules in a Single Measurement: Comprehensive Analysis of Complex Gas Mixtures by Multi-Pressure Chemical Ionization Mass Spectrometry.

Aleksei Shcherbinin, Henning Finkenzeller, Jyri Mikkilä, Jussi Kontro, Netta Vinkvist, Juha Kangasluoma, Matti Rissanen.

Chemical Ionization Mass Spectrometry (CIMS) is a well-established analytical method in atmospheric research, process monitoring, forensics, breathomics, and food science. Despite significant advancements in procedural techniques, several instrument configurations, especially operating at different ionization pressures, are typically needed to analyze the full range of compounds from nonfunctionalized parent compounds to their functionalized reaction products. For polar, functionalized compounds, very sensitive detection schemes are provided by high-pressure adduct-forming chemical ionization techniques, whereas for nonfunctionalized, nonpolar compounds, low-pressure chemical ionization techniques have consistently demonstrated superior performance. Here, using a MION2 chemical ionization inlet and an Orbitrap Exploris 120 mass spectrometer, we present multi-pressure chemical ionization mass spectrometry (MPCIMS), the combination of high- and low-pressure ionization schemes in a single instrument enabling quantification of the full distribution of precursor molecules and their oxidation reaction products from the same stream of gas without alterations. We demonstrate the performance of the new methodology in a laboratory experiment employing a-pinene, a monoterpene relevant to atmospheric particle formation, where MPCIMS allows us to measure the spectrum of compounds ranging from the volatile precursor hydrocarbon to highly functionalized condensable reaction products. MPCIMS carries the potential as an all-in-one method for the analysis of complex gas mixtures, reducing technical complexities and the need for multiple instruments without compromise of sensitivity.

DOI: https://doi.org/10.1021/acs.analchem.4c03859
Electrophoresis. 2024 Dec 06.

Quantitative Endogenous Polyamine Analysis via Capillary Electrophoresis/Mass Spectrometry: Characterization and Practical Considerations.

Cameron J Kaminsky, Jericha Mill, Thomas Raife, Lingjun Li.

  Commonly used analytical techniques for polyamine analysis, including derivatization and mixed-mode liquid chromatography (LC), have inherent disadvantages. Capillary electrophoresis (CE) is uniquely suited to analyze small, highly charged molecules because analytes are separated on the basis of their electrophoretic mobility, not polarity or association with a stationary phase. Microfluidic CE-mass spectrometry (mCE-MS) is a relatively recent addition to commercially available CE offerings that streamlines traditional CE-MS interfacing and has the potential to improve upon classic CE challenges to robustness and reproducibility. MS instrument choice and scanning parameters are strongly influenced by a need for high acquisition rate to adequately sample CE peaks. Alternatively, isotachophoresis on loading can be intentionally avoided to produce sufficiently wide peaks. The mCE platform utilized here performed very well in many metrics; a limit of detection (LOD) as low as 0.25 ng/mL was achieved for spermidine, and endogenous spermidine was easily detected in blood with this method. Both of these are challenging tasks for any separation technique and demonstrate a strong use case for the platform. During experimentation, various idiosyncrasies in the commercial CE-MS interface resulted in extensive chip-to-chip variability in both peak shape and LOD, complicating the application to robust absolute quantitation. Practical guidance for similar analyses is provided.

Keywords:  microfluidic; polyamine; quantitation; spermidine

DOI:  https://doi.org/10.1002/elps.202400165
Talanta. 2024 Nov 28. pii: S0039-9140(24)01680-1. [Epub ahead of print]285 127298

Integration of feature-based molecular networking and high-definition data-dependent acquisition for the comprehensive multicomponent characterization of Honghua Xiaoyao Tablet.

Siyi Ma, Yuhao Zhang, Renwen Zhu, Shiyu Wu, Shiyu Zhang, Huawu Zeng, Weidong Zhang, Ji Ye.

  Systematically identifying the chemical constituents in complex matrices is a challenge due to the inherent characteristics of compounds. The combination of liquid chromatography-tandem mass spectrometry (LC-MS) and classical molecular networking (CLMN) is a powerful technology for annotating small molecules. However, the low coverage from inappropriate acquisition modes and the inseparability of isomeric compound nodes still hinders the comprehensive metabolite characterization. A novel strategy that integrated high-definition data-dependent acquisition (HDDDA) from traveling-wave ion mobility mass spectrometry (TWIMS) and feature-based molecular networking (FBMN) was developed to improve chemical component characterization and enhance isomeric component discernment. The data-dependent acquisition (DDA) and HDDDA, were effectively and visually evaluated by CLMN and FBMN via the number of nodes, clustered nodes and clusters. Moreover, the efficiency of the three strategies was validated. The results strongly demonstrated that the HDDDA-FBMN strategy improves MS coverage and offers significant advantages for isomer identification. With the assistance of the UNIFI platform, the developed strategy was successfully applied to systematically investigate the chemical profile of Honghua Xiaoyao Tablet (HHXYT), a traditional folk empirical prescription for treating various gynecological diseases. 184 compounds were unambiguously identified or tentatively characterized, including 12 pairs of isomers, and two unreported compounds. In conclusion, this hybrid approach achieves dimensionally enhanced MS data acquisition and visual recognition of isomeric compounds, accelerating the structural characterization in complex systems. We anticipate that HDDDA-FBMN strategies will be a flexible and versatile tool for the chemical components in a complex system of TCMs.

Keywords:  Collision cross section; Feature-based molecular networking; High-definition data-dependent acquisition; Honghua Xiaoyao tablet; Liquid chromatography-tandem mass spectrometry

DOI:  https://doi.org/10.1016/j.talanta.2024.127298
Anal Biochem. 2024 Dec 03. pii: S0003-2697(24)00285-9. [Epub ahead of print] 115741

Development and validation of a sensitive and fast bioanalytical LC-MS/MS assay for the quantitation of venetoclax and azacitidine in Rat Plasma: Application to pharmacokinetic study.

Manal El-Gendy, Mohamed Hefnawy, Sarah Alrubia, Abdulaziz Alnasser, Amsha Alsegiani, Yousef Bin Jardan, Adel El-Azab, Alaa Abdel-Aziz, Mohamed Attwa, Emad Alsarhani.

  The combination of venetoclax plus azacitidine (VTX-AZA) is FDA-approved to treat patients with acute myeloid leukemia (AML) aged ≥ 75 years and has become the standard of care for AML patients. However, the literature has not reported an analytical method for determining VTX-AZA in plasma samples. Therefore, developing an accurate and sensitive bioanalytical assay to quantify VTX-AZA in plasma is important. For the first time, this study describes the development of a new liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for the simultaneous determination of VTX and AZC in plasma samples with its application to pharmacokinetic study in rats. The assay employs repaglinide (RPG) as an internal standard. The chromatographic separations of VTX, AZC, and RPG are achieved within 2.5 min at 25 °C on an Eclipse plus C18 column (100 mm × 2.1 mm, 1.8 μm) and an isocratic mobile phase consisted of water with 0.1% formic acid and acetonitrile (50:50, v/v, pH 3.2) at a flow rate of 0.30 mL/min. VTX and AZC have been extracted from rat plasma using the solid-phase extraction (SPE) procedure without interference from plasma endogenous. The FDA guidelines were followed in the validation of the developed assay, and linearity in rat plasma was observed for AZC and VTX, respectively, ranging from 5-3000 and 5-1000 ng/mL, with r ≥ 0.998. The lower limits of detection (LLOD) were 2 ng/mL for both drugs. In addition, the inter-day and intra-day accuracy were 0.8-6.6% and 2.2-5.7%; the inter-day and intra-day precision were 3-6.6% and 1.5-7.1%, respectively. The validated assay was effectively used in a pharmacokinetic investigation including the simultaneous oral administration of 40 mg/kg of AZA and 100 mg/kg of VTX to rats. The maximum plasma concentration (Cmax) for AZC and VTX was 794 ± 99.6 ng/mL and 641 ± 96.9 ng/mL achieved at 0.5 ± 0.03 hr and 6 ± 0.05 hr, respectively. The AUC0-∞ for AZC and VTX was 1253± 252.6 and 4881 ± 745.4 ng/mL.h; respectively.

Keywords:  Acute myeloid leukemia; Azacitidine; LC–MS/MS; Pharmacokinetics; Rat plasma; Venetoclax

DOI:  https://doi.org/10.1016/j.ab.2024.115741
J Mass Spectrom. 2024 Dec;59(12): e5104

Mass Spectrometry Imaging for Spatial Toxicology Research.

Tian Autumn Qiu.

  The spatial information of xenobiotics distribution, metabolism, and toxicity mechanisms in situ has drawn increasing attention in both pharmaceutical and environmental toxicology research to aid drug development and environmental risk assessments. Mass spectrometry imaging (MSI) provides a label-free, multiplexed, and high-throughput tool to characterize xenobiotics, their metabolites, and endogenous molecules in situ with spatial resolution, providing knowledge on spatially resolved absorption, distribution, metabolism, excretion, and toxicity on the molecular level. In this perspective, we briefly summarize applications of MSI in toxicology on xenobiotic distribution and metabolism, quantification, toxicity mechanisms, and biomarker discovery. We identified several challenges regarding how we can fully harness the power of MSI in both fundamental toxicology research and regulatory practices. First, how can we increase the coverage, sensitivity, and specificity in detecting xenobiotics and their metabolites in complex biological matrices? Second, how can we link the spatial molecular information of xenobiotics to toxicity consequences to understand toxicity mechanisms, predict exposure outcomes, and aid biomarker discovery? Finally, how can we standardize the MSI experiment and data analysis workflow to provide robust conclusions for regulation and drug development? With these questions in mind, we provide our perspectives on the future directions of MSI as a promising tool in spatial toxicology research.

Keywords:  ADMET; drug distribution; environmental pollutant; mass spectrometry imaging; spatial toxicology; xenobiotics

DOI:  https://doi.org/10.1002/jms.5104
Anal Biochem. 2024 Nov 30. pii: S0003-2697(24)00276-8. [Epub ahead of print]698 115732

High-resolution targeted mass spectrometry for comprehensive quantification of sphingolipids: clinical applications and characterization of extracellular vesicles.

Thiago V D Felippe, Diana M Toro, Jonatan C S de Carvalho, Pedro Nobre-Azevedo, Luiz F M Rodrigues, Bianca T M Oliveira, Pedro V da Silva-Neto, Adriana F L Vilela, Fausto Almeida, Lúcia H Faccioli, Carlos A Sorgi.

  Sphingolipids (SL), a class of membrane lipids, play important roles in numerous biological processes. Their significant structural diversity poses challenges for accurate quantification. To address this, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has emerged as a powerful tool for sphingolipidomics, capable of profiling these lipids comprehensively. In this study, we utilized LC-MS/MS with high-resolution mass spectrometry (MRMHR) to develop a targeted method for the identification and quantification of various SL species. This method, based on validated parameters such as precursor/fragment ions (m/z) and retention time, demonstrated high sensitivity and accuracy, successfully identifying SL species across 12 distinct classes. Its open-panel design also facilitates the analysis of new SL-species targets. Notably, using this approach, we identified 40 SL species in plasma samples from COVID-19 patients, and we determined the influence of matrix metalloproteinase-3 (MMP-3) expression on the positive downstream of SL metabolism. Beyond plasma analysis, this method has potential applications in other biomedical contexts, such as extracellular vesicles (EVs), describing the cargo of sphingosine-1-phosphate (S1P) on macrophage-derived EVs. The establishment of this targeted workflow enabling precise quantification of a wide range of SL species, holds promise for identifying novel biomarkers and therapeutic targets.

Keywords:  COVID-19; Extracellular vesicles; LC-MS/MS; Sphingolipids

DOI:  https://doi.org/10.1016/j.ab.2024.115732