bims-metlip 2021-02-07 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2021‒02‒07
nineteen papers selected by
Sofia Costa
Cold Spring Harbor Laboratory

Ion mobility mass spectrometry in the omics era: Challenges and opportunities for metabolomics and lipidomics.
Public LC-Orbitrap Tandem Mass Spectral Library for Metabolite Identification.
Evaluation of Fumaric Acid and Maleic Acid as Internal Standards for NMR Analysis of Protein Precipitated Plasma, Serum, and Whole Blood.
Update on LIPID MAPS classification, nomenclature, and shorthand notation for MS-derived lipid structures.
A simple and fast method for metabolomic analysis by gas liquid chromatography-mass spectrometry.
Reevaluating the Role of Polarizability in Ion Mobility Spectrometry.
Challenges and Strategies of Chemical Analysis of Drugs of Abuse and Explosives by Mass Spectrometry.
[Simultaneous determination of 13 antibiotics in disinfection products by ultra-high performance liquid chromatography-tandem mass spectrometry].
A sensitive HPLC-MS/MS method for the detection, resolution and quantitation of cathinone enantiomers in horse blood plasma and urine.
Mass spectrometry techniques for imaging and detection of metallodrugs.
Dual derivatization strategy for the comprehensive quantification and double bond location characterization of fatty acids by ultra-high performance liquid chromatography-tandem mass spectrometry.
Label-free spectral imaging to study drug distribution and metabolism in single living cells.
tidyMicro: a pipeline for microbiome data analysis and visualization using the tidyverse in R.
Application of Predicted Collisional Cross Section to Metabolome Databases to Probabilistically Describe the Current and Future Ion Mobility Mass Spectrometry.
Simultaneous enantioselective separation method for thyroid hormones using liquid chromatography-tandem mass spectrometry and its applications.
LICAR: An Application for Isotopic Correction of Targeted Lipidomic Data Acquired with Class-Based Chromatographic Separations Using Multiple Reaction Monitoring.
Identification of the metabolites of tofacitinib in liver microsomes by liquid chromatography combined with high resolution mass spectrometry.
Determination of bisphenols, parabens, and benzophenones in placenta by dispersive liquid-liquid microextraction and gas chromatography-tandem mass spectrometry.
Determination of 3-iodothyronamine (3-T1AM) in mouse liver using liquid chromatography-tandem mass spectrometry.

Mass Spectrom Rev. 2021 Feb 01.

Ion mobility mass spectrometry in the omics era: Challenges and opportunities for metabolomics and lipidomics.

Giuseppe Paglia, Andrew J Smith, Giuseppe Astarita.

  Researchers worldwide are taking advantage of novel, commercially available, technologies, such as ion mobility mass spectrometry (IM-MS), for metabolomics and lipidomics applications in a variety of fields including life, biomedical, and food sciences. IM-MS provides three main technical advantages over traditional LC-MS workflows. Firstly, in addition to mass, IM-MS allows collision cross-section values to be measured for metabolites and lipids, a physicochemical identifier related to the chemical shape of an analyte that increases the confidence of identification. Second, IM-MS increases peak capacity and the signal-to-noise, improving fingerprinting as well as quantification, and better defining the spatial localization of metabolites and lipids in biological and food samples. Third, IM-MS can be coupled with various fragmentation modes, adding new tools to improve structural characterization and molecular annotation. Here, we review the state-of-the-art in IM-MS technologies and approaches utilized to support metabolomics and lipidomics applications and we assess the challenges and opportunities in this growing field.

Keywords:  CCS; DTIM-MS; FAIM-MS; IM-MS; MS imaging; MSI; TIM-MS; TWIM-MS; cIM-MS; collision cross-section; lipids; metabolites; spatial lipidomics; spatial metabolomics

DOI:  https://doi.org/10.1002/mas.21686
J Proteome Res. 2021 Feb 02.

Public LC-Orbitrap Tandem Mass Spectral Library for Metabolite Identification.

Prasad Phapale, Andrew Palmer, Rose Muthoni Gathungu, Dipali Kale, Britta Brügger, Theodore Alexandrov.

  Liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics studies require high-quality spectral libraries for reliable metabolite identification. We have constructed EMBL-MCF (European Molecular Biology Laboratory-Metabolomics Core Facility), an open LC-MS/MS spectral library that currently contains over 1600 fragmentation spectra from 435 authentic standards of endogenous metabolites and lipids. The unique features of the library include the presence of chromatographic profiles acquired with different LC-MS methods and coverage of different adduct ions. The library covers many biologically important metabolites with some unique metabolites and lipids as compared with other public libraries. The EMBL-MCF spectral library is created and shared using an in-house-developed web application at https://curatr.mcf.embl.de/. The library is freely available online and also integrated with other mass spectral repositories.

Keywords:  LC-MS/MS; Orbitrap; fragmentation spectra; high-resolution mass spectrometry; liquid chromatography; mass spectral library

DOI:  https://doi.org/10.1021/acs.jproteome.0c00930
Anal Chem. 2021 Feb 04.

Evaluation of Fumaric Acid and Maleic Acid as Internal Standards for NMR Analysis of Protein Precipitated Plasma, Serum, and Whole Blood.

G A Nagana Gowda, Natalie N Hong, Daniel Raftery.

Significant advances have been made in unknown metabolite identification and expansion of the number of quantifiable metabolites in human plasma, serum, and whole blood using NMR spectroscopy. However, reliable quantitation of metabolites is still a challenge. A major bottleneck is the lack of a suitable internal standard that does not interact with the complex blood sample matrix and also does not overlap with metabolite peaks apart from exhibiting other favorable characteristics. With the goal of addressing this challenge, a comprehensive investigation of fumaric and maleic acids as potential internal standards was made along with a comparison with the conventional standards, TSP (trimethylsilylpropionic acid) and DSS (trimethylsilylpropanesulfonic acid). Both fumaric acid and maleic acid exhibited a surprisingly high performance with a quantitation error <1%, while the TSP and DSS caused an average error of up to 35% in plasma, serum, and whole blood. Further, the results indicate that while fumaric acid is a robust standard for all three biospecimens, maleic acid is suitable for only plasma and serum. Maleic acid is not suited for the analysis of whole blood due to its overlap with coenzyme peaks. These findings provide new opportunities for improved and accurate quantitation of metabolites in human plasma, serum, and whole blood using NMR spectroscopy. Moreover, the use of protein precipitation prior to NMR analysis mirrors the sample preparation commonly used for mass spectrometry based metabolomics, such that these findings further strengthen efforts to combine and compare NMR and MS based metabolite data of human plasma, serum, and whole blood for metabolomics based research.

DOI: https://doi.org/10.1021/acs.analchem.0c04766
J Lipid Res. 2020 Dec;pii: S0022-2275(20)60017-7. [Epub ahead of print]61(12): 1539-1555

Update on LIPID MAPS classification, nomenclature, and shorthand notation for MS-derived lipid structures.

Gerhard Liebisch, Eoin Fahy, Junken Aoki, Edward A Dennis, Thierry Durand, Christer S Ejsing, Maria Fedorova, Ivo Feussner, William J Griffiths, Harald Köfeler, Alfred H Merrill, Robert C Murphy, Valerie B O'Donnell, Olga Oskolkova, Shankar Subramaniam, Michael J O Wakelam, Friedrich Spener.

  A comprehensive and standardized system to report lipid structures analyzed by MS is essential for the communication and storage of lipidomics data. Herein, an update on both the LIPID MAPS classification system and shorthand notation of lipid structures is presented for lipid categories Fatty Acyls (FA), Glycerolipids (GL), Glycerophospholipids (GP), Sphingolipids (SP), and Sterols (ST). With its major changes, i.e., annotation of ring double bond equivalents and number of oxygens, the updated shorthand notation facilitates reporting of newly delineated oxygenated lipid species as well. For standardized reporting in lipidomics, the hierarchical architecture of shorthand notation reflects the diverse structural resolution powers provided by mass spectrometric assays. Moreover, shorthand notation is expanded beyond mammalian phyla to lipids from plant and yeast phyla. Finally, annotation of atoms is included for the use of stable isotope-labeled compounds in metabolic labeling experiments or as internal standards. This update on lipid classification, nomenclature, and shorthand annotation for lipid mass spectra is considered a standard for lipid data presentation.

Keywords:  fatty acyls; glycerolipids; glycerophospholipids; lipidomics; mass spectrometry; sphingolipids; sterols

DOI:  https://doi.org/10.1194/jlr.S120001025
Metabolomics. 2021 Feb 06. 17(2): 22

A simple and fast method for metabolomic analysis by gas liquid chromatography-mass spectrometry.

Diana Cosovanu, Montserrat Llovera, Gemma Villorbina, Ramon Canela-Garayoa, Jordi Eras.

  INTRODUCTION: The metabolomic profile is an essential tool for understanding the physiological processes of biological samples and their changes. In addition, it makes it possible to find new substances with industrial applications or use as drugs. As GC-MS is a very common tool for obtaining the metabolomic profile, a simple and fast method for sample preparation is required.OBJECTIVES: The aim of this research was to develop a direct derivatization method for GC-MS to simplify the sample preparation process and apply it to a wide range of samples for non-targeted metabolomic analysis purposes.
METHODS: One pot combined esterification of carboxylic acids with methanol and silylation of the hydroxyl groups was achieved using a molar excess of chlorotrimethylsilane with respect to methanol in the presence of pyridine.
RESULTS: The metabolome profile obtained from different samples, such as bilberry and cherry cuticles, olive leaves, P. aeruginosa and E. coli bacteria, A. niger fungi and human sebum from the ceruminous gland, shows that the procedure allows the identification of a wide variety of metabolites. Aliphatic fatty acids, hydroxyfatty acids, phenolic and other aromatic compounds, fatty alcohols, fatty aldehydes dimethylacetals, hydrocarbons, terpenoids, sterols and carbohydrates were identified at different MSI levels using their mass spectra.
CONCLUSION: The metabolomic profile of different biological samples can be easily obtained by GC-MS using an efficient simultaneous esterification-silylation reaction. The derivatization method can be carried out in a short time in the same injection vial with a small amount of reagents.

Keywords:  Derivatization; GC/MS; Hydroxy fatty acids; Metabolomics; Non-targeted analysis

DOI:  https://doi.org/10.1007/s11306-021-01771-w
J Am Soc Mass Spectrom. 2021 Feb 03.

Reevaluating the Role of Polarizability in Ion Mobility Spectrometry.

Cameron N Naylor, Brian H Clowers.

  With the expanding commercial availability of gas-phase separation systems that incorporate gas-phase mobility, there is a concurrent rise in efforts to cast the gas-phase mobility coefficient in terms of an ion-neutral collision cross-section (CCS). The motivating factors for this trend are varied, but many aim to complement experimental results with computationally generated CCS values from in silico structural approximations. Unfortunately, the current paradigm for relating experimental mobility results to computationally derived structures relies upon empirical approaches, including a myriad of variables that do not realistically bound the comparison. In this Critical Insight, we advocate for the development of a self-consistent experimental and computational framework that uses laboratory results to constrain the scope of the modeling effort. This paper aims to prompt discussion, challenge assumptions, and promote the development of more efficient, accurate computational techniques within the gas-phase ion measurement community. Specifically, we postulate whether experimental deviations from Langevin's polarization limit (Kpol) are suitable to estimate the relative contributions of hard-sphere collisions and long-range interactions within CCS values. Not surprisingly, different molecule classes exhibit different trends in the K/Kpol ratio when normalized for reduced mass, and the most common IMS calibrants (e.g., tune mix, polyalanine, tetraalkylammonium salts) follow different polarizability trends than many of the analytes probed in the literature. Succinctly, if gas-phase ion structure is largely invariant based upon the colliding neutral and newly developed experimental efforts can quantitatively capture ion polarizability, then modeling efforts describing a target analyte must be self-consistent as the collision neutral is changed in silico.

Keywords:  Blanc’s law; drift gases; ion mobility spectrometry; open-source; polarizability

DOI:  https://doi.org/10.1021/jasms.0c00338
Front Chem. 2020 ;8 598487

Challenges and Strategies of Chemical Analysis of Drugs of Abuse and Explosives by Mass Spectrometry.

Ahsan Habib, Lei Bi, Huanhuan Hong, Luhong Wen.

  In analytical science, mass spectrometry (MS) is known as a "gold analytical tool" because of its unique character of providing the direct molecular structural information of the relevant analyte molecules. Therefore, MS technique has widely been used in all branches of chemistry along with in proteomics, metabolomics, genomics, lipidomics, environmental monitoring etc. Mass spectrometry-based methods are very much needed for fast and reliable detection and quantification of drugs of abuse and explosives in order to provide fingerprint information for criminal investigation as well as for public security and safety at public places, respectively. Most of the compounds exist as their neutral form in nature except proteins, peptides, nucleic acids that are in ionic forms intrinsically. In MS, ion source is the heart of the MS that is used for ionizing the electrically neutral molecules. Performance of MS in terms of sensitivity and selectivity depends mainly on the efficiency of the ionization source. Accordingly, much attention has been paid to develop efficient ion sources for a wide range of compounds. Unfortunately, none of the commercial ion sources can be used for ionization of different types of compounds. Moreover, in MS, analyte molecules must be released into the gaseous phase and then ionize by using a suitable ion source for detection/quantification. Under these circumstances, fabrication of new ambient ion source and ultrasonic cutter blade-based non-thermal and thermal desorption methods have been taken into account. In this paper, challenges and strategies of mass spectrometry analysis of the drugs of abuse and explosives through fabrication of ambient ionization sources and new desorption methods for non-volatile compounds have been described. We will focus the literature progress mostly in the last decade and present our views for the future study.

Keywords:  ambient ionization source; drugs of abuse; explosives; headspace method; hollow cathode discharge ionization; mechanism of ionization and desorption; non-thermal desorption; tribological effect

DOI:  https://doi.org/10.3389/fchem.2020.598487
Wei Sheng Yan Jiu. 2021 Jan;50(1): 121-128

[Simultaneous determination of 13 antibiotics in disinfection products by ultra-high performance liquid chromatography-tandem mass spectrometry].

Feng Zhu, Xiaosong Wu, Fang Li, Wei Wang, Wenliang Ji, Hualiang Liu, Yan Xu.

  OBEJECTIVE: To develop a method for the determination of 13 antibiotics in 8 classes for desinfection products by ulta-high perfomance chromatography-tandem mass spectrometry(UPLC-MS/MS).METHODS: Samples were extracted by methanol or acetonitrile. The target compouds were separated on a Waters HSS T3 column(100 mm×2. 1 mm, 1. 8 μm), and detected by triple quadrupole tandem mass spectrometer.
RESULTS: The 13 selected antibiotics showed good linear relationships in the range of 4-100 μg/L and the correlation coefficients(r~2) were all above 0. 991. The limits of detection ranged from 2 to 25 μg/kg. The recovery rates at three spiked levels(low, medium and high) in three dosage forms of disinfection products were in the range of 71. 2%-130. 4%, and the relative standard deviations(RSD) were all less than 11. 3%, which could meet the detection requirements of illegal addition of antibiotics in disinfection products. Ofloxacin at a concentration of 21. 1 mg/kg was found in a cream disinfection product by the developed method, and no related drugs were detected in other samples.
CONCLUSION: This method is simple, reliable, reproducible, which covers a wide range of antibiotics, and provides technical support for monitoring the illegal addition of antibiotics in disinfection products.

Keywords:  antibiotics; disinfection products; ulta high perfomance chromatography-tandem mass spectrometry

DOI:  https://doi.org/10.19813/j.cnki.weishengyanjiu.2021.01.021
Anal Bioanal Chem. 2021 Jan 31.

A sensitive HPLC-MS/MS method for the detection, resolution and quantitation of cathinone enantiomers in horse blood plasma and urine.

Devan Loganathan, Rong Yi, Bhavesh Patel, Julia Zhang, Noel Kong.

  Resolution of cathinone enantiomers in equine anti-doping analysis is becoming more important to distinguish the inadvertent ingestion of plant-based products from those of deliberate administration of designer synthetic analogs. With this in mind, a rapid and sensitive method was developed and validated for the detection, resolution and quantitative determination of cathinone enantiomers in horse blood plasma and urine. The analytes were recovered from the blood plasma and urine matrices by using a liquid-liquid extraction after adjusting the pH to 9. The recovered analytes were derivatized with Nα-(2,4-dinitro-5-fluorophenyl)-L-valinamide, a chiral derivatizing agent analogous to Marfey's reagent. The resulting diastereoisomers were baseline resolved under a reversed-phase liquid chromatographic condition. Derivatization of the analytes not only allowed the separation of the enantiomers using cost-effective traditional liquid chromatography conditions and reversed-phase columns but also increased the sensitivity, at least to an order of magnitude, when tandem mass spectrometry is used for the detection. A limit of detection of 0.05 ng/mL was achieved for cathinone enantiomers for both matrices. Acceptable intraday and interday precision and accuracy along with satisfactory dilution accuracy and precision were observed during the method validation. The method suitability was tested using the post administration urine samples collected after single doses of cathinone and ephedrine as single-enantiomeric form and methcathinone as racemic form. Finally, a proof of concept of the isomeric ratio in urine samples to distinguish the presence of cathinone as a result of accidental ingestion of plant-based product from that of an illicit use of a designer product is demonstrated. To the best of our knowledge, this is the first such work where cathinone enantiomers were resolved and quantified in horse blood plasma and urine at sub nanogram levels.

Keywords:  Anti-doping analysis; Cathinone; Chiral analysis; Equine; HPLC-MS/MS

DOI:  https://doi.org/10.1007/s00216-021-03182-1
Curr Opin Chem Biol. 2021 Jan 31. pii: S1367-5931(20)30164-2. [Epub ahead of print]61 123-134

Mass spectrometry techniques for imaging and detection of metallodrugs.

Sarah Theiner, Anna Schoeberl, Andreas Schweikert, Bernhard K Keppler, Gunda Koellensperger.

  Undoubtedly, metallomic approaches based on mass spectrometry have evolved into essential tools supporting the drug development of novel metal-based anticancer drugs. This article will comment on the state-of-the-art instrumentation and highlight some of the recent analytical advances beyond routine, especially focusing on the latest developments in inductively coupled plasma-mass spectrometry (ICP-MS). Mass spectrometry-based bioimaging and single-cell methods will be presented, paving the way to exciting investigations of metal-based anticancer drugs in heterogeneous and structurally, as well as functionally complex solid tumor tissues.

Keywords:  Imaging mass spectrometry; Laser ablation-ICP-MS; Metallodrug development; Metallomics; Single-cell analysis

DOI:  https://doi.org/10.1016/j.cbpa.2020.12.005
J Chromatogr A. 2021 Jan 23. pii: S0021-9673(21)00063-7. [Epub ahead of print]1639 461939

Dual derivatization strategy for the comprehensive quantification and double bond location characterization of fatty acids by ultra-high performance liquid chromatography-tandem mass spectrometry.

Ting Hu, Yuan Sun, Han Li, Ping Du, Lihong Liu, Zhuoling An.

  Comprehensive analysis of fatty acids (FAs) has long been challenging due to their poor ionization efficiency, lack of characteristic fragment ions and difficulty of identifying C=C bond locations. In this study, a high coverage ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the quantification and C=C bond location characterization of FAs using two structural analogues, 2-hydrazinyl-4,6-dimethylpyrimidine (DMP) and 2-hydrazinylpyrimidine (DP), as dual derivatization reagents. DP-labeled FA standards were used as internal standards to reduced matrix effects, which guaranteed the accurate quantification of FAs. The derivatization yields of FAs were larger than 99% and the sensitivities were increased by 400-fold compared with non-derivatized FAs. Pretreatment and instrumental analysis of FAs can be completed in 20 minutes. Only 5 μL rat plasma can satisfy the quantification of 36 FAs with good linearity (r>0.99). Both intra-day and inter-day accuracies were in the range of 85-105%, and the precisions were less than 15%. The extraction recoveries were investigated to be in the range of 88-112%. No obvious matrix effects were observed for the derivatized FAs. In addition, the locations of C=C bonds in DMP-derivatized FAs could be identified by diagnostic fragment ions generated from 1,4-hydrogen elimination and allylic cleavage under low energy collision induced dissociation (CID). The new method was finally employed for FA profiling in plasma from rats with moxifloxacin-induced liver injury. Significant downregulation of butyric acid was observed in moxifloxacin treated model rats, which was believed to be related to the liver injury.

Keywords:  Double bond location; Dual derivatization; Fatty acid; Moxifloxacin; UHPLC-MS/MS

DOI:  https://doi.org/10.1016/j.chroma.2021.461939
Sci Rep. 2021 Feb 01. 11(1): 2703

Label-free spectral imaging to study drug distribution and metabolism in single living cells.

Qamar A Alshammari, Rajasekharreddy Pala, Nir Katzir, Surya M Nauli.

During drug development, evaluation of drug and its metabolite is an essential process to understand drug activity, stability, toxicity and distribution. Liquid chromatography (LC) coupled with mass spectrometry (MS) has become the standard analytical tool for screening and identifying drug metabolites. Unlike LC/MS approach requiring liquifying the biological samples, we showed that spectral imaging (or spectral microscopy) could provide high-resolution images of doxorubicin (dox) and its metabolite doxorubicinol (dox'ol) in single living cells. Using this new method, we performed measurements without destroying the biological samples. We calculated the rate constant of dox translocating from extracellular moiety into the cell and the metabolism rate of dox to dox'ol in living cells. The translocation rate of dox into a single cell for spectral microscopy and LC/MS approaches was similar (~ 1.5 pM min-1 cell-1). When compared to spectral microscopy, the metabolism rate of dox was underestimated for about every 500 cells using LC/MS. The microscopy approach further showed that dox and dox'ol translocated to the nucleus at different rates of 0.8 and 0.3 pM min-1, respectively. LC/MS is not a practical approach to determine drug translocation from cytosol to nucleus. Using various methods, we confirmed that when combined with a high-resolution imaging, spectral characteristics of a molecule could be used as a powerful approach to analyze drug metabolism. We propose that spectral microscopy is a new method to study drug localization, translocation, transformation and identification with a resolution at a single cell level, while LC/MS is more appropriate for drug screening at an organ or tissue level.

DOI: https://doi.org/10.1038/s41598-021-81817-0
BMC Bioinformatics. 2021 Feb 01. 22(1): 41

tidyMicro: a pipeline for microbiome data analysis and visualization using the tidyverse in R.

Charlie M Carpenter, Daniel N Frank, Kayla Williamson, Jaron Arbet, Brandie D Wagner, Katerina Kechris, Miranda E Kroehl.

  BACKGROUND: The drive to understand how microbial communities interact with their environments has inspired innovations across many fields. The data generated from sequence-based analyses of microbial communities typically are of high dimensionality and can involve multiple data tables consisting of taxonomic or functional gene/pathway counts. Merging multiple high dimensional tables with study-related metadata can be challenging. Existing microbiome pipelines available in R have created their own data structures to manage this problem. However, these data structures may be unfamiliar to analysts new to microbiome data or R and do not allow for deviations from internal workflows. Existing analysis tools also focus primarily on community-level analyses and exploratory visualizations, as opposed to analyses of individual taxa.RESULTS: We developed the R package "tidyMicro" to serve as a more complete microbiome analysis pipeline. This open source software provides all of the essential tools available in other popular packages (e.g., management of sequence count tables, standard exploratory visualizations, and diversity inference tools) supplemented with multiple options for regression modelling (e.g., negative binomial, beta binomial, and/or rank based testing) and novel visualizations to improve interpretability (e.g., Rocky Mountain plots, longitudinal ordination plots). This comprehensive pipeline for microbiome analysis also maintains data structures familiar to R users to improve analysts' control over workflow. A complete vignette is provided to aid new users in analysis workflow.
CONCLUSIONS: tidyMicro provides a reliable alternative to popular microbiome analysis packages in R. We provide standard tools as well as novel extensions on standard analyses to improve interpretability results while maintaining object malleability to encourage open source collaboration. The simple examples and full workflow from the package are reproducible and applicable to external data sets.

Keywords:  Microbiome; Open source; Pipeline; R; Tidyverse; Visualization

DOI:  https://doi.org/10.1186/s12859-021-03967-2
J Am Soc Mass Spectrom. 2021 Feb 04.

Application of Predicted Collisional Cross Section to Metabolome Databases to Probabilistically Describe the Current and Future Ion Mobility Mass Spectrometry.

Corey D Broeckling, Linxing Yao, Giorgis Isaac, Marisa Gioioso, Valentin Ianchis, Johannes P C Vissers.

Metabolomics is a powerful phenotyping platform with potential for high-throughput analyses. The primary technology for metabolite profiling is mass spectrometry. In recent years, the coupling of mass spectrometry with ion mobility spectrometry (IMS) has offered the promise of faster analysis time and greater resolving power. Our understanding of the potential impact of IMS on the field of metabolomics is limited by availability of comprehensive experimental data. In this analysis, we use a probabilistic approach to enumerate the strengths and limitations, the present and future, of this technology. This is accomplished through use of "model" metabolomes, predicted physicochemical properties, and probabilistic descriptions of resolving power. This analysis advances our understanding of the importance of orthogonality in resolving (separation) dimensions, describes the impact of the metabolome composition on resolution demands, and offers a system resolution landscape that may serve to guide practitioners in the coming years.

DOI: https://doi.org/10.1021/jasms.0c00375
J Pharm Biomed Anal. 2021 Jan 18. pii: S0731-7085(21)00016-9. [Epub ahead of print]196 113904

Simultaneous enantioselective separation method for thyroid hormones using liquid chromatography-tandem mass spectrometry and its applications.

Yunhee Lee, Eunjung Bang, Wonjae Lee, Yun-Cheol Na.

  An analytical method for the simultaneous determination of chiral thyroxine and the related iodinated chiral compounds using LC-MS/MS is introduced in this study. D-Thyroid hormones (THs), which are not commercially available, were produced through the racemization reaction of the L-THs in acetic acid solution containing salicylaldehyde. The solution containing D- and L-THs after the reaction was used for optimizing the chiral separation. The D- and L-THs were well separated enantiomerically under isocratic conditions in 70 % acetonitrile containing 0.1 % formic acid on a CROWNPAK® CR-I (+) column, but some peaks, such as those of diiodo-D-tyrosine (D-DIT)/monoiodo-L-tyrosine, diiodo-D-thyronine/diiodo-L-tyrosine and D-thyroxine/triiodo-L-thyronine, overlapped chromatographically, causing misinterpretation in impurity analysis. This was overcome by using the gradient condition providing the best chiral selectivity (α) and resolution (Rs) ranging from 1.14 to 1.37 and from 2.39 to 4.52, respectively. The linearity was above 0.999 and the detection limits ranged from 8.2 to 57.7 ng/mL by the separation method. This method was applied to identify and quantify chiral impurities in authentic standards and pharmaceuticals. As a result, D-enantiomers corresponding to the L-THs standards as well as L-DIT were commonly observed as impurities. In the stability test of DL-thyroxine under acidic conditions for identifying the distribution of chiral products, it was observed that the formation of DIT by hydrolysis increased over time. Additional products formed through esterification, including thyroxine methyl ester and diiodo-tyrosine methyl ester, were newly separated and identified using a C18 column.

Keywords:  Chiral stationary phase; Enantiomer separation; Racemization; Thyroid hormone

DOI:  https://doi.org/10.1016/j.jpba.2021.113904
Anal Chem. 2021 Feb 03.

LICAR: An Application for Isotopic Correction of Targeted Lipidomic Data Acquired with Class-Based Chromatographic Separations Using Multiple Reaction Monitoring.

Liang Gao, Shanshan Ji, Bo Burla, Markus R Wenk, Federico Torta, Amaury Cazenave-Gassiot.

Lipidomics is developing as an important area in biomedical and clinical research. Reliable quantification of lipid species is required for clinical translation of lipidomic studies. Hydrophilic interaction chromatography (HILIC), normal-phase liquid chromatography (NPLC), and supercritical fluid chromatography (SFC) are commonly used techniques in lipidomics and provide class-based separation of lipids. While co-elution of lipid species and their internal standards is an advantage for accurate quantification, it leads to isotopic overlap between species of the same lipid class. In shotgun lipidomics, isotopic correction is typically done based on elemental formulas of precursor ions. In multiple reaction monitoring (MRM) analyses, however, this approach should not be used, as the overall contribution of heavy isotopes to the MRM transitions' intensities depends on their location in the molecule with respect to the fragmentation pattern. We present an algorithm, provided in the R programming language, for isotopic correction in class-based separation using MRM, extracting relevant structural information from MRM transitions to apply adequate isotopic correction factors. Using standards, we show that our algorithm accurately estimates the isotopic contribution of isotopologues to MRM transitions' measured intensities. Using human plasma as an example, we demonstrate the necessity of adequate isotopic correction for accurate quantitation of lipids measured by MRM with class-based chromatographic separation. We show that over a third of the measured phosphatidylcholine species had their intensity corrected by more than 10%. This isotopic correction algorithm and R-implemented application enable a more accurate quantification of lipids in class-based separation-MRM, a prerequisite for successful translation of lipidomic applications.

DOI: https://doi.org/10.1021/acs.analchem.0c04565
Biomed Chromatogr. 2021 Feb 01. e5081

Identification of the metabolites of tofacitinib in liver microsomes by liquid chromatography combined with high resolution mass spectrometry.

Ping Liu, Shuang Wu, Zhaoqin Dong.

  Tofacitinib is an orally available Janus kinase inhibitor. The purpose of this study was to investigate the metabolism of tofacitinib in mouse, rat, monkey and human liver microsomes fortified with β-nicotinamide adenine dinucleotide phosphate tetrasodium salt (NADPH) and uridine diphosphate glucuronic acid (UDPGA). The biotransformation was executed at a temperature of 37 o C for 60 min and the samples were analyzed by ultra-high performance liquid chromatography combined with high resolution mass spectrometry (UHPLC-HRMS) operated in positive electrospray ionization mode. The structures of the metabolites were elucidated according to their retention times, accurate masses and MS/MS spectra. Under the current conditions, a total of thirteen metabolites, including one glucuronide conjugate, were detected and structurally proposed. Oxygenation of the pyrrolopyrimidine ring, oxygenation of piperidine ring, N-demethylation, oxygenation of piperidine ring side chain and glucuronidation were the primary metabolic pathways of tofacitinib. Among the tested species, tofacitinib showed significant species difference. Compared with other species, rat showed similar metabolic profiles to humans. The present study provides some new information regarding the metabolism of tofacitinib in animals and humans, which would bring us considerable benefits for the subsequent studies focusing on the pharmacological effect and toxicity of this drug.

Keywords:  Tofacitinib; liver microsomes; metabolite identification; species differences

DOI:  https://doi.org/10.1002/bmc.5081
Chemosphere. 2021 Jan 25. pii: S0045-6535(21)00176-4. [Epub ahead of print]274 129707

Determination of bisphenols, parabens, and benzophenones in placenta by dispersive liquid-liquid microextraction and gas chromatography-tandem mass spectrometry.

M F Fernández, V Mustieles, B Suárez, I Reina-Pérez, A Olivas-Martinez, F Vela-Soria.

  Human exposure to endocrine disrupting chemicals (EDCs) is of particular concern during development. Bisphenols, parabens, and benzophenones are EDCs widely used in the manufacture of numerous goods, personal care products, and cosmetics. The aim of this study was to develop a new and practical method for determining three bisphenols, four parabens, and five benzophenones in placenta samples. It uses dispersive liquid-liquid microextraction (DLLME) in combination with gas chromatography-tandem mass spectrometry (GC-MS/MS). Several chemometric approaches were employed to optimize the experimental parameters. Limits of detection ranged from 0.04 to 0.08 ng g-1 and inter-day variabilities (evaluated as relative standard deviation) from 4.2% to 13.4%. The method was validated using matrix-matched standard calibration followed by a recovery assay with spiked samples. Recovery percentages ranged from 87.1% to 113.2%. Finally, the method was used to measure target compounds in 20 placental tissue samples from voluntary donors. This analytical procedure can provide information on the exposure of the fetus to non-persistent EDCs.

Keywords:  Benzophenones; Bisphenols; DLLME; GC–MS/MS; Parabens; Placenta

DOI:  https://doi.org/10.1016/j.chemosphere.2021.129707
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jan 18. pii: S1570-0232(21)00032-5. [Epub ahead of print]1165 122553

Determination of 3-iodothyronamine (3-T1AM) in mouse liver using liquid chromatography-tandem mass spectrometry.

Zhong-Min Li, Manuel Miller, Sogol Gachkar, Jens Mittag, Sonja C Schriever, Paul T Pfluger, Karl-Werner Schramm, Meri De Angelis.

  3-iodothyronamine (3-T1AM) has been suggested as a novel chemical messenger and potent trace amine-associated receptor 1 ligand in the CNS that occurs naturally as endogenous metabolite of the thyroid hormones. Discrepancies and variations in 3-T1AM plasma and tissue concentrations have nonetheless caused controversy regarding the existence and biological role of 3-T1AM. These discussions are at least partially based on potential analytical artefacts caused by differential decay kinetics of 3-T1AM and the widely used deuterated quantification standard D4-T1AM. Here, we report a novel LC-MS/MS method for the quantification of 3-T1AM in biological specimens using stable isotope dilution with 13C6-T1AM, a new internal standard that showed pharmacodynamic properties comparable to endogenous 3-T1AM. The method detection limit (MDL) and method quantification limit (MQL) of 3-T1AM were 0.04 and 0.09 ng/g, respectively. The spike-recoveries of 3-T1AM were between 85.4% and 94.3%, with a coefficient of variation of 3.7-5.8%. The intra-day and inter-day variations of 3-T1AM were 8.45-11.2% and 3.58-5.73%, respectively. Endogenous 3-T1AM liver values in C57BL/6J mice were 2.20 ± 0.49 pmol/g with a detection frequency of 50%. Higher liver 3-T1AM values were found when C57BL/6J mice were treated with N-acetyl-3-iodothyronamine or O-acetyl-3-iodothyronamine. Overall, our new stable isotope dilution LC-MS/MS method improves both the sensitivity and selectivity compared with existing methods. The concomitant possibility to quantify additional thyroid hormones such as thyroxine, 3,5,3'-triiodo-L-thyronine, 3,3',5'-triiodo-L-thyronine, 3,3'-diiodo-L-thyronine, and 3,5-diiodo-L-thyronine further adds to the value of our novel method in exploring the natural occurrence and fate of 3-T1AM in biological tissues and fluids.

Keywords:  3-iodothyronamine; HPLC-MS/MS; Mouse liver; N-acetyl-3-iodothyronamine; O-acetyl-3-iodothyronamine; Thyroid hormones

DOI:  https://doi.org/10.1016/j.jchromb.2021.122553