555 results found
Brookhart A, Arora M, McCullagh M, et al., 2023, Understanding mobile phase buffer composition and chemical structure effects on electrospray ionization mass spectrometry response., J Chromatogr A, Vol: 1696
Mobile phase selection is of critical importance in liquid chromatography - mass spectrometry (LC-MS) based studies, since it affects retention, chromatographic selectivity, ionization, limits of detection and quantification, and linear dynamic range. Generalized LC-MS mobile phase selection criteria, suitable for a broad class of chemical compounds, do not exist thus far. Here we have performed a large-scale qualitative assessment of the effect of solvent composition used for reversed-phase LC separations on electrospray ionization (ESI) response for 240 small molecular weight drugs, representing various chemical compound classes. Of these 240 analytes 224 were detectable using ESI. The main chemical structural features affecting ESI response were found to all be surface area or surface charge-related. Mobile phase composition was found to be less differentiating, although for some compounds a pH effect was noted. Unsurprisingly, chemical structure was found to be the dominant factor for ESI response for the majority of the investigated analytes, representing about 85% of the replicating detectable complement of the sample data set. A weak correlation between ESI response and structure complexity was observed. Solvents based on isopropanol, and those containing phosphoric or di- and trifluoracetic acids, performed relatively poorly in terms of chromatographic or ESI response, whilst the best performing 'generic' LC solvents were based on methanol, acetonitrile using formic acid and ammonium acetate as buffer components, consistent with current practice in many laboratories.
Metabolomics is an interdisciplinary field with applications in many areas. Analytes include metabolites involved in pathways such as glycolysis, amino acid metabolism, the Krebs cycle, etc. However, the metabolites involved in these biosynthetic pathways are typically highly polar molecules, which represent a major challenge for chromatographic analysis. Whilst there have been significant efforts to address the difficulties involved in the determination of polar metabolites the comprehensive profiling of the polar metabolome remains problematic. The current review summarizes current approaches, advances and trends in the use of liquid chromatography/mass spectrometry as it attempts to address the major instrumental and intellectual challenges involved in mapping the polar metabolome.
Tanna N, Plumb RS, Molloy BJ, et al., 2023, Enhanced chromatographic efficiency obtained with vacuum jacketed columns facilitates the rapid UHPLC/MS/MS-based analysis of fasiglifam in rat plasma, TALANTA, Vol: 254, ISSN: 0039-9140
Trovato FM, Zia R, Artru F, et al., 2023, Lysophosphatidylcholines modulate immunoregulatory checkpoints in peripheral monocytes and are associated with mortality in people with acute liver failure., Journal of Hepatology, Vol: 78, Pages: 558-573, ISSN: 0168-8278
BACKGROUND AND AIMS: Acute liver failure (ALF) is a life-threatening disease characterised by high-grade inflammation and immunoparesis with a high incidence of death from sepsis. Here, we aimed to describe the metabolic dysregulation in ALF and determine whether systemic immune responses are modulated via the lysophosphatidylcholine(LPC)-autotaxin(ATX)-lysophosphatidylcholinic acid (LPA) pathway. METHODS: 96 ALF patients, 71 healthy controls (HC), 104 patients with cirrhosis and 31 septic patients were recruited. The pathways of interest were identified based on multivariate statistical analysis of proton nuclear magnetic resonance (1HNMR) spectroscopy, untargeted ultraperformance liquid chromatography-mass spectrometry (UPLC-MS)-based lipidomics and validated with a targeted metabolomics panel. Peripheral blood mononuclear cells were cultured with LPA 16:0, 18:0, 18:1, and their immune checkpoint surface expression was assessed by flow cytometry. LPA receptor (LPAR) transcript-level expression of monocytes was investigated and the effect of LPAR antagonism was also examined in vitro. RESULTS: LPC 16:0 was found highly discriminant between ALF and HC. There was an increase in ATX and LPA in ALF compared to HC and sepsis. LPCs 16:0, 18:0 and 18:1 were reduced in ALF patients with poor prognosis. Treatment of monocytes with LPA 16:0 increased their PD-L1 expression and reduced CD155, CD163, MerTK levels, without effect on T and NK/CD56+T cells immune checkpoints. LPAR1 and 3 antagonism in culture reversed the LPA effect on monocyte expression of MerTK and CD163. MerTK and CD163, but not LPARs genes, were differently expressed and upregulated in monocytes from ALF patients compared to controls. CONCLUSION: Reduced amounts of LPCs are biomarkers of poor prognosis in patients with ALF. The LPC-ATX-LPA axis appears to modulate innate immune response in ALF via LPAR1 and LPAR3. Further investigations are required to identify novel therapeutic agents targeting these recept
Plumb RS, Gethings LA, Rainville PD, et al., 2023, Advances in high throughput LC/MS based metabolomics: A review, TrAC Trends in Analytical Chemistry, Vol: 160, Pages: 1-9, ISSN: 0165-9936
Properly implemented, metabolic and lipidomic profiling can provide a deeper understanding of mammalian, plant and bacterial biology. These omics-tools have developed and matured over the last 40-years and are now being deployed to provide valuable information in epidemiological studies, drug toxicology and pharmacology, disease biology and progression and patient stratification. LC/MS has become the technology of choice for both metabolic and lipid profiling, due to its speed, sensitivity and structural elucidation capabilities. In the preceding two decades there have been many technological and methodological advances in LC/MS that have facilitated the evolution of the technology into a rugged, reliable, and easily deployed tool. These advances include, but are not limited to, improvements in chromatography (phases, columns, and delivery system), instruments for mass spectrometry, optimization of sample preparation, the introduction of ion mobility, data analysis tools, metabolite databases, harmonized protocols, and the more widespread use of quality control methods and reference standards/matrices. Here, recent developments and advances in high throughput liquid chromatography/high resolution mass spectrometry for metabolic phenotyping are described. These advances which may provide improved feature detection, increased laboratory efficiency and data quality, as well as “biomarker” identification, are discussed in relation to their potential application to the analysis of large clinical studies, or biobank collections.
Theodoridis G, Gika H, Raftery D, et al., 2023, Ensuring fact-based metabolite identification in liquid chromatography-mass spectrometry-based metabolomics., Analytical Chemistry, Vol: 95, Pages: 3909-3916, ISSN: 0003-2700
Metabolite identification represents a major bottleneck in contemporary metabolomics research and a step where critical errors may occur and pass unnoticed. This is especially the case for studies employing liquid chromatography-mass spectrometry technology, where there is increased concern on the validity of the proposed identities. In the present perspective article, we describe the issue and categorize the errors into two types: identities that show poor biological plausibility and identities that do not comply with chromatographic data and thus to physicochemical properties (usually hydrophobicity/hydrophilicity) of the proposed molecule. We discuss the problem, present characteristic examples, and propose measures to improve the situation.
Sarmad S, Viant MR, Dunn WB, et al., 2023, A proposed framework to evaluate the quality and reliability of targeted metabolomics assays from the UK Consortium on Metabolic Phenotyping (MAP/UK), NATURE PROTOCOLS, ISSN: 1754-2189
Molloy BJ, King A, Gethings LA, et al., 2023, Investigation of the pharmacokinetics and metabolic fate of Fasiglifam (TAK-875) in male and female rats following oral and intravenous administration, Xenobiotica: the fate of foreign compounds in biological systems, Vol: 53, Pages: 93-105, ISSN: 0049-8254
The metabolism and pharmacokinetics of fasiglifam (TAK-875, 2-[(3S)-6-[[3-[2,6-dimethyl-4-(3-methylsulfonylpropoxy)phenyl]phenyl]methoxy]-2,3-dihydro-1-benzofuran-3-yl]acetic acid), a selective free fatty acid receptor 1 (FFAR1)/GPR40 agonist, were studied following intravenous (5 mg/kg) and oral administration (10 and 50 mg/kg) to male and female Sprague Dawley rats.Following intravenous dosing at 5 mg/kg, peak observed plasma concentrations of 8.8/9.2 µg/ml were seen in male and female rats respectively.Following oral dosing, peak plasma concentrations at 1 h of ca. 12.4/12.9 µg/ml for 10 mg/kg and 76.2/83.7 µg/ml for 50 mg/kg doses were obtained for male and female rats respectively. Drug concentrations then declined in the plasma of both sexes with t1/2’s of 12.4 (male) and 11.2 h (female). Oral bioavailability was estimated to be 85-120% in males and females at both dose levels.Urinary excretion was low, but in a significant sex-related difference, female rats eliminated ca. 10-fold more drug-related material by this route.Fasiglifam was the principal drug-related compound in plasma, with 15 metabolites, including the acyl glucuronide, also detected. In addition to previously identified metabolites, a novel biotransformation, that produced a side-chain shortened metabolite via elimination of CH2 from the acetyl side chain was noted with implications for drug toxicity.
Weidolf L, Wilson I, 2023, Minimizing the DILI potential of carboxylic acid-containing drugs: a perspective, Medicinal Chemistry Research, ISSN: 1054-2523
Drugs containing carboxylic acid moieties are associated with a range of toxicities, some related directly to their pharmacology and others to the metabolites that they produce on undergoing biotransformations that reduce biological activity and enhance excretion. The reactions involved in metabolism include a wide range of conjugations to the carboxylic acid moiety plus modifications to the spacers used to attach them to the main body of the drug. Here we provide a metabolic perspective on the biotransformations that have been found to occur with drugs that contain a carboxylic acid and discuss the potential of these to cause toxicity. Based on our current understanding of the metabolism of carboxylic acid-containing drugs we then consider approaches that may mitigate toxicity.
Wilson ID, Poole CF, 2023, Planar chromatography - Current practice and future prospects, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol: 1214, Pages: 1-15, ISSN: 1570-0232
Planar chromatography, in the form of thin-layer or high-performance thin-layer chromatography (TLC, HPTLC), continues to provide a robust and widely used separation technique. It is unrivaled as a simple and rapid qualitative method for mixture analysis, or for finding bioactive components in mixtures. The format of TLC/HPTLC also provides a unique method for preserving the separation, enabling further investigation of components of interest (including quantification/structure determination) separated in both time and space from the original analysis. The current practice of planar chromatography and areas of development of the technology are reviewed and promising future directions in the use of TLC/HPTLC are outlined.
Gika H, Theodoridis G, Plumb RS, et al., 2023, Metabolic phenotyping (metabonomics/metabolomics) by liquid chromatography-mass spectrometry, Liquid Chromatography: Applications, Pages: 403-429, ISBN: 9780323983006
LC–MS is now the primary investigative technology, employing both targeted and untargeted approaches, used to obtain characteristic metabolic phenotypes (metabotypes) in metabonomic/metabolomic studies. Current applications use various modes of LC but reversed-phase (RP) separations remain the first choice. Hydrophilic interaction liquid chromatography (HILIC), and less frequently ion-pair (IP) and ion exchange (IE) LC, provide methods that are suitable for polar compounds poorly retained by RP phases. Ultra (high) performance liquid chromatography [U(H)PLC] is now becoming the predominant methodology in metabonomic/metabolomic applications, replacing the more traditional HPLC format. Allied with this has been a rise in the use of rapid profiling methods in both targeted and untargeted metabolomic profiles as sample numbers rise in application areas such as epidemiology, biobanking, and clinical analysis. However, the requirement for speed has not reduced the need for sensitive and high-resolution separations in order to provide high metabolome coverage. As a result, there is increased interest in the addition of ion mobility (IM) to the LC/MS toolbox. The current status of U(H)PLC/MS and UHPLC/IM/MS for untargeted metabotyping is considered and potential future directions are suggested.
Munjoma N, Isaac G, Muazzam A, et al., 2022, High Throughput LC-MS Platform for Large Scale Screening of Bioactive Polar Lipids in Human Plasma and Serum, JOURNAL OF PROTEOME RESEARCH, Vol: 21, Pages: 2596-2608, ISSN: 1535-3893
Letertre M, Bhatt A, Harvey M, et al., 2022, Characterizing the metabolic effects of the selective inhibition of gut microbial β-glucuronidases in mice, Scientific Reports, Vol: 12, ISSN: 2045-2322
The hydrolysis of xenobiotic glucuronides by gut bacterial glucuronidases reactivates previously detoxified compounds resulting in severe gut toxicity for the host. Selective bacterial β-glucuronidase inhibitors can mitigate this toxicity but their impact on wider host metabolic processes has not been studied. To investigate this the inhibitor 4-(8-(piperazin-1-yl)-1,2,3,4-tetrahydro-[1,2,3]triazino[4′,5′:4,5]thieno[2,3-c]isoquinolin-5-yl)morpholine (UNC10201652, Inh 9) was administered to mice to selectively inhibit a narrow range of bacterial β-glucuronidases in the gut. The metabolomic profiles of the intestinal contents, biofluids, and several tissues involved in the enterohepatic circulation were measured and compared to control animals. No biochemical perturbations were observed in the plasma, liver or gall bladder. In contrast, the metabolite profiles of urine, colon contents, feces and gut wall were altered compared to the controls. Changes were largely restricted to compounds derived from gut microbial metabolism. This work establishes that inhibitors targeted towards bacterial β-glucuronidases modulate the functionality of the intestinal microbiota without adversely impacting the host metabolic system.
Kirwan JA, Gika H, Beger RD, et al., 2022, Quality assurance and quality control reporting in untargeted metabolic phenotyping: mQACC recommendations for analytical quality management, Metabolomics, Vol: 18, Pages: 1-16, ISSN: 1573-3882
BackgroundDemonstrating that the data produced in metabolic phenotyping investigations (metabolomics/metabonomics) is of good quality is increasingly seen as a key factor in gaining acceptance for the results of such studies. The use of established quality control (QC) protocols, including appropriate QC samples, is an important and evolving aspect of this process. However, inadequate or incorrect reporting of the QA/QC procedures followed in the study may lead to misinterpretation or overemphasis of the findings and prevent future metanalysis of the body of work.ObjectiveThe aim of this guidance is to provide researchers with a framework that encourages them to describe quality assessment and quality control procedures and outcomes in mass spectrometry and nuclear magnetic resonance spectroscopy-based methods in untargeted metabolomics, with a focus on reporting on QC samples in sufficient detail for them to be understood, trusted and replicated. There is no intent to be proscriptive with regard to analytical best practices; rather, guidance for reporting QA/QC procedures is suggested. A template that can be completed as studies progress to ensure that relevant data is collected, and further documents, are provided as on-line resources.Key reporting practicesMultiple topics should be considered when reporting QA/QC protocols and outcomes for metabolic phenotyping data. Coverage should include the role(s), sources, types, preparation and uses of the QC materials and samples generally employed in the generation of metabolomic data. Details such as sample matrices and sample preparation, the use of test mixtures and system suitability tests, blanks and technique-specific factors are considered and methods for reporting are discussed, including the importance of reporting the acceptance criteria for the QCs. To this end, the reporting of the QC samples and results are considered at two levels of detail: “minimal” and “best reporting practice” le
Kerins A, Koszyczarek M, Smith C, et al., 2022, The in vitro metabolism and in vivo pharmacokinetics of the bacterial beta-glucuronidase inhibitor UNC10201652, XENOBIOTICA, Vol: 52, Pages: 904-915, ISSN: 0049-8254
Wilson ID, Gethings LA, Plumb RS, 2022, Defining The Pharmacometabodynamics of Gefitinib after Intravenous Administration to the Mouse by UHPLC/MS and UPLC-IMMS Study, Publisher: ELSEVIER, Pages: S52-S52
Yang J, Wilson I, Rainville P, 2022, Evaluation of hybrid surface technology for the analysis of the B-group vitamins by LC-ESI-MS/MS, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol: 1204, Pages: 123336-123336, ISSN: 1570-0232
Recently, a novel hybrid surface technology (HST) has been developed to mitigate metal analyte adsorption in liquid chromatography. The HST provides a hybrid organic-inorganic surface on the metal fluidic path, from injection to detector and including the column frits and wall, to mitigate the interaction between analytes and metals. Here the impact of the HST on the analysis of B group vitamins using liquid chromatography coupled with electrospray tandem mass spectrometry (LC-ESI-MS/MS) has been evaluated. Significant improvements in analyte intensity, limit of quantification (LOQ), carry-over, and peak shape were observed using an LC-ESI-MS/MS system and column that incorporated the HST. The key observed improvements include a 3-10 times increase in sensitivity (providing a lower LOQ) for riboflavin, thiamine, nicotinamide, FMN, PLP, and 5MTHF, no carry-over, and a more symmetrical peak for thiamine. When applied to the analysis of B group vitamins in energy drinks and B vitamin dietary supplement samples, the HST system demonstrated excellent accuracy and repeatability.
Poole C, Wilson ID, 2022, The state of the art in planar chromatography., Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol: 1203, Pages: 1-2, ISSN: 1570-0232
Sagi-Kiss V, Li Y, Carey MR, et al., 2022, Ion-pairing chromatography and amine derivatization provide complementary approaches for the targeted LC-MS analysis of the polar metabolome., Journal of Proteome Research, Vol: 21, Pages: 1428-1437, ISSN: 1535-3893
Liquid chromatography coupled to mass spectrometry is a key metabolomics/metabonomics technology. Reversed-phase liquid chromatography (RPLC) is very widely used as a separation step, but typically has poor retention of highly polar metabolites. Here, we evaluated the combination of two alternative methods for improving retention of polar metabolites based on 6-aminoquinoloyl-N-hydroxysuccinidimyl carbamate derivatization for amine groups, and ion-pairing chromatography (IPC) using tributylamine as an ion-pairing agent to retain acids. We compared both of these methods to RPLC and also to each other, for targeted analysis using a triple-quadrupole mass spectrometer, applied to a library of ca. 500 polar metabolites. IPC and derivatization were complementary in terms of their coverage: combined, they improved the proportion of metabolites with good retention to 91%, compared to just 39% for RPLC alone. The combined method was assessed by analyzing a set of liver extracts from aged male and female mice that had been treated with the polyphenol compound ampelopsin. Not only were a number of significantly changed metabolites detected, but also it could be shown that there was a clear interaction between ampelopsin treatment and sex, in that the direction of metabolite change was opposite for males and females.
Claude E, Lafont R, Plumb RS, et al., 2022, High performance Reversed-Phase Thin-Layer Chromatography-Desorption electrospray ionisation - time of flight high resolution mass spectrometric detection and imaging (HPTLC/DESI/ToFMS) of phytoecdysteroids, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol: 1200, Pages: 1-7, ISSN: 1570-0232
Reversed-phase high performance thin-layer chromatography (RP-HPTLC) on C18 bonded silica gel was combined with desorption electrospray ionization (DESI) and high resolution time of flight mass spectrometry (HRToFMS) to detect, characterize and image (MSI) phytoecdysteroids (plant-derived insect moulting hormones) in ethanolic extracts of members of the Silene plant family. As seen previously for silica gel, DESI provided a simple and convenient method for recovering polar polyhydoxysteroids from RP-HPTLC plates for the purposes of both the MS and MSI of extracts obtained from three species of the Silene family (Silene otites, S. nutans and S. viridiflora). Using RP-HPTLC/DESI/MSI/HRToFMS a number of ecdysteroids, including 20-hydroxyecdysone, polypodine-B, 2-deoxy-20-hydroxyecdysone and 2-deoxyecdysone were identified in these extracts. Differences were noted in the mass spectra obtained depending upon both the stationary phase on which they were separated, and the temperatures used in the heated transfer line used for introduction into the ion source. Ecdysteroids detected after chromatography on C18 bonded silica showed increased fragmentation due to water loss compared to those imaged from silica. In addition, the benefits of the additional resolution provided by 2-dimensional TLC for increasing spectral quality compared to a 1-dimensional separation are demonstrated.
Hughes CJ, Gethings LA, Wilson ID, et al., 2022, Access to the Phospho-proteome via the Mitigation of Peptide-Metal Interactions, JOURNAL OF CHROMATOGRAPHY A, Vol: 1673, ISSN: 0021-9673
Lioupi A, Virgiliou C, Walter TH, et al., 2022, Application of a hybrid zwitterionic hydrophilic interaction liquid chromatography column in metabolic profiling studies, JOURNAL OF CHROMATOGRAPHY A, Vol: 1672, ISSN: 0021-9673
Lippa KA, Aristizabal-Henao JJ, Beger RD, et al., 2022, Reference materials for MS-based untargeted metabolomics and lipidomics: a review by the metabolomics quality assurance and quality control consortium (mQACC), Metabolomics, Vol: 18, ISSN: 1573-3882
IntroductionThe metabolomics quality assurance and quality control consortium (mQACC) is enabling the identification, development, prioritization, and promotion of suitable reference materials (RMs) to be used in quality assurance (QA) and quality control (QC) for untargeted metabolomics research.ObjectivesThis review aims to highlight current RMs, and methodologies used within untargeted metabolomics and lipidomics communities to ensure standardization of results obtained from data analysis, interpretation and cross-study, and cross-laboratory comparisons. The essence of the aims is also applicable to other ‘omics areas that generate high dimensional data.ResultsThe potential for game-changing biochemical discoveries through mass spectrometry-based (MS) untargeted metabolomics and lipidomics are predicated on the evolution of more confident qualitative (and eventually quantitative) results from research laboratories. RMs are thus critical QC tools to be able to assure standardization, comparability, repeatability and reproducibility for untargeted data analysis, interpretation, to compare data within and across studies and across multiple laboratories. Standard operating procedures (SOPs) that promote, describe and exemplify the use of RMs will also improve QC for the metabolomics and lipidomics communities.ConclusionsThe application of RMs described in this review may significantly improve data quality to support metabolomics and lipidomics research. The continued development and deployment of new RMs, together with interlaboratory studies and educational outreach and training, will further promote sound QA practices in the community.
Isaac G, Wilson ID, Plumb RS, 2022, Application of hybrid surface technology for improving sensitivity and peak shape of phosphorylated lipids such as phosphatidic acid and phosphatidylserine, JOURNAL OF CHROMATOGRAPHY A, Vol: 1669, ISSN: 0021-9673
Kodra D, Pousinis P, Vorkas PA, et al., 2022, Is Current Practice Adhering to Guidelines Proposed for Metabolite Identification in LC-MS Untargeted Metabolomics? A Meta-Analysis of the Literature, JOURNAL OF PROTEOME RESEARCH, Vol: 21, Pages: 590-598, ISSN: 1535-3893
Plumb RS, Isaac G, Rainville PD, et al., 2021, High throughput UHPLC-MS-based lipidomics using vacuum jacketed columns, Journal of Proteome Research, Vol: 21, Pages: 691-701, ISSN: 1535-3893
Reversed-phase UHPLC-MS is extensively employed for both the profiling of biological fluids and tissues to characterize lipid dysregulation in disease and toxicological studies. With conventional LC-MS systems the chromatographic performance and throughput are limited due to dispersion from the fluidic connections as well as radial and longitudinal thermal gradients in the LC column. In this study vacuum jacketed columns (VJC), positioned at the source of the mass spectrometer, were applied to the lipidomic analysis of plasma extracts. Compared to conventional UHPLC, the VJC-based methods offered greater resolution, faster analysis, and improved peak intensity. For a 5 min VJC analysis, the peak capacity increased by 66%, peak tailing reduced by up to 34%, and the number of lipids detected increased by 30% compared to conventional UHPLC. The narrower peaks, and thus increased resolution, compared to the conventional system resulted in a 2-fold increase in peak intensity as well a significant improvement in MS and MS/MS spectral quality resulting in a 22% increase in the number of lipids identified. When applied to mouse plasma samples, reproducibility of the lipid intensities in the pooled QC ranged from 1.8–12%, with no related drift in tR observed.
Trovato FM, Zia R, Napoli S, et al., 2021, Dysregulation of the Lysophosphatidylcholine/Autotaxin/Lysophosphatidic Acid Axis in Acute-on-Chronic Liver Failure Is Associated With Mortality and Systemic Inflammation by Lysophosphatidic Acid Dependent Monocyte Activation, Publisher: WILEY, Pages: 907-925, ISSN: 0270-9139
Plumb RS, McDonald T, Rainville PD, et al., 2021, High-throughput UHPLC/MS/MS-based metabolic profiling using a vacuum jacketed column, Analytical Chemistry, Vol: 93, Pages: 10644-10652, ISSN: 0003-2700
In UHPLC, frictional heating from the eluent flowing through the column at pressures of ca. 10–15 Kpsi causes radial diffusion via temperature differences between the center of the column and its walls. Longitudinal dispersion also occurs due to temperature gradients between the inlet and outlet. These effects cause band broadening but can be mitigated via a combination of vacuum jacketed stainless steel tubing, reduced column end nut mass, and a constant temperature in the column from heating the inlet fitting. Here, vacuum jacketed column (VJC) technology, employing a novel column housing located on the source of the mass spectrometer and minimized tubing from the column outlet to the electrospray probe, was applied to profiling metabolites in urine. For a 75 s reversed-phase gradient separation, the average peak widths for endogenous compounds in urine were 1.2 and 0.6 s for conventional LC/MS and VJC systems, respectively. The peak tailing factor was reduced from 1.25 to 1.13 when using the VJC system compared to conventional UHPLC, and the peak capacity increased from 65 to 120, with a 25% increase in features detected in urine. The increased resolving power of the VJC system reduced co-elution, simplifying MS and MS/MS spectra, providing a more confident metabolite identification. The increased LC performance also gave more intense MS peaks, with a 10–120% increase in response, improving the quality of the MS data and detection limits. Reducing the LC gradient duration to 37 s gave peak widths of ca. 0.4 s and a peak capacity of 84.
Gethings LA, Gray N, Plumb RS, et al., 2021, Proteomic consequences of the deletion of cytochrome P450 (CYP450) reductase in mice, JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, Vol: 1179, ISSN: 1570-0232
This data is extracted from the Web of Science and reproduced under a licence from Thomson Reuters. You may not copy or re-distribute this data in whole or in part without the written consent of the Science business of Thomson Reuters.