Synopsis

Abstract

Comprehensive structural characterisation of complex lipids, such as Glycerophospholipids (GPEs) by tandem mass spectrometry (MS/MS) continues to present a considerable challenge. Although there have been a number of studies which have used high-resolution mass spectrometry with various fragmentation methods, detailed structural and spatial distribution of these complex lipids still remains relatively unknown. Isomeric and isobaric species have also created immense difficulty in the elucidation and identification of these lipid components, and conventional dissociation methods do not generate fragmentation that permit the isomeric discrimination relative to the position of the double bonds on the acyl chains and the branching points on the glycerol backbone. In this investigation, direct infusion by nano-electrospray ionisation tandem-in-time MS/MS with collisionally-activated/induced dissociation (CAD/CID), electron induced dissociation (EID) and infrared multiphoton dissociation (IRMPD) were used for the detailed structural characterisation of L-α-Phosphatidylethanolamine, dioleoyl (GPEtn). This method enabled site-specific localisation of the double bonds on the acyl chains to be determined and provided sn-positional information of the di-acyl branching point on the glycerol backbone.

Abstract

Solution-state proton nuclear magnetic resonance spectroscopy (NMR) is a highly sensitive method used for the elucidation of various organic complexes, such as D-(+)-Glucose which undergoes anomerisation in solution. However, spectral overlapping from multiple proton (¹H) resonances in 1D NMR continues to present a considerable challenge when analyzing such complex mixtures, making it difficult to accurately assign the chemical shifts (δ/ppm) and J-couplings (²J and ³J_H-H) values to the associated spins. In this investigation a 1D-selective total correlation spectroscopy (sel-TOCSY) with a single Gaussian (π) pulse and decoupling in the presence of scalar interactions (DIPSI-2) mixing sequence was used in conjunction with 1D ¹H-NMR, 2D-hetero nuclear single quantum coherence spectroscopy (HSQC) (¹³C-¹H) and 2D-correlation spectroscopy (COSY) (¹H-¹H) to determine the molecular connectivities and enable a complete assignment of the overlapped proton resonances within α- and β-D-(+)-Glucose. Implementation of DIPSI-2 ‘spin-lock’ further provided an indication of the experimental cross-peak (I_2z) intensities and magnetization transfer within the D-(+)-Glucose spin-network relative to the TOCSY mixing time (τ_mix) between 10-100 ms. The experimental pulse sequence, internal Hamiltonian and spin system parameters were then compiled using a scripting interface (tcl), where the cross-peak intensities were simulated using a simulation package for solid-state NMR spectroscopy (SIMPSON), that works on the exploitation of the matrix-matrix multiplications and the numerical integration of the Liouville-von- Neumann equation, providing a comparable understanding of the magnetization transfer efficiency between J-coupled spins within α- and β-D-(+)-Glucose.

Code

SIMPSON 1D-TOCSY Code Available at Link opens in a new window

MAS-CDT MSc Mini-Project Groups

• Project 1: FT-ICR MS Group (Department of Chemistry).
• Project 2: Solid-State NMR Group (Department of Physics).

Previous BSc and MSc Projects

• MSc Dissertation.

Title: Interactions of Organic Molecules and Ceramic Clays: Method Development for the Utilisation of Direct Insertion Probe-Mass Spectrometry and Dispersive Confocal Raman Spectroscopy.

Supervised by Dr Ben Stern and Dr Richard Telford

• BSc Dissertation.

Title: Determination of Polycyclic Aromatic Hydrocarbons (PAHs) in Top Soil by means of Gas Chromatography-Mass Spectrometry.

Supervised by Dr Ben Stern and Professor Carl Heron (British Museum)

Professional Experience

• HPLC and Mass Spectrometry Laboratory Technician in Environmental Fate and Metabolism at Smithers Viscient (ESG), Harrogate, North Yorkshire, United Kingdom, 2018.
• Chemist and Mass Spectrometry Technician - Light Isotope Laboratory, University of Bradford, Department of Archeological and Environmental Science, United Kingdom, 2015.
• Biomedical Laboratory Technician - University of Leeds, School of Biomedical Science, United Kingdom, 2007.

Conferences

• Scientific Advisory Board (SAB) Meeting. University of Warwick, Coventry, United Kingdom. 12^th February, 2024 - Oral Presentation.

Title: Fingerprinting Structural Sequences in Glycerophospholipids via Kendrick Mass Defect (KMD) and Tandem Mass Spectrometry

• Structural Biology at Warwick Symposium. University of Warwick, Coventry, United Kingdom. 15^th November 2023 - Oral Presentation.

Title: Tandem Mass Spectrometry and Multimodal Fragmentation of Phosphatidylthreonine (PT) and Phosphatidylserine (PS) Species in A549 Human Adenocarcinoma Epithelial Cells

• 43^rd British Mass Spectrometry Society (BMSS) Annual Meeting (43^rd Conference). Royal Northern College of Music, Manchester, United Kingdom. 12th-14th September, 2023 - Oral Presentation.

Title: Tandem Mass Spectrometry and Multimodal Fragmentation of Phosphatidylthreonine (PT) and Phosphatidylserine (PS) Species in A549 Human Adenocarcinoma Epithelial Cells.

(Awarded John Beynon Travel and Conference Fund, BMSS- September 2023).

• Scientific Advisory Board (SAB) Meeting. University of Warwick, Coventry, United Kingdom. December, 2022 - Oral Presentation.

Title: Application of 1-and 2 Dimensional Tandem Mass Spectrometry, and Multimodal Fragmentation of Glycerophospholipids.

• 42^nd British Mass Spectrometry Society (BMSS) Annual Meeting (42^nd Conference), Royal Northern College of Music, Manchester, United Kingdom. 13^th-15^th September, 2022 - Oral Presentation.

Title: 1- and 2-Dimensional Tandem Mass Spectrometry of Lipids Extracted from Human Placental Tissue. 

• 24^th International Mass Spectrometry Conference (IMSC), Maastricht Exhibition and Congress Centre (MECC), Maastricht, The Netherlands. 27^th August-2^nd September, 2022 - Poster Presentation.

Title: Multimodal Fragmentation and 1-, and 2-Dimensional Mass Spectrometry of Lipids Extracted from Human Placental Tissue. 

• 70^th American Society of Mass Spectrometry (ASMS) Conference on Mass Spectrometry and Allied Topics, 2022, Minneapolis Convention Center, Minneapolis, United States of America. 5^th-9^th June, 2022 - Poster Presentation.

Title: Multimodal Fragmentation and 1-, and 2-Dimensional Mass Spectrometry of Lipids Extracted from Human Placental Tissue.

• 41st British Mass Spectrometry Society (BMSS) Annual Meeting (41^st Conference), Sheffield Hallam University, Sheffield, United Kingdom. 8^th-9^th September, 2021- Flash Oral Presentation.

Title: Multimodal Fragmentation and Two-Dimensional Mass Spectrometry Characterisation of Human Placental Lipid Extracts.

• University of Warwick, MAS-CDT Annual Conference (Online), United Kingdom. June 2020 - Flash Oral Presentation.

Title: FT-ICR MS Analysis of Glycerophospholipids.

• 68^th American Society of Mass Spectrometry (ASMS) Conference on Mass Spectrometry and Allied Topics, 2020 (Reboot, Online), United States of America. 1^st-12^th June, 2020.

• 4th Short Course of the European Network of Fourier-Transform Ion-Cyclotron-Resonance Mass Spectrometry (EU FT-ICR MS), University of Warwick, Coventry, United Kingdom. 21^st-23^rd August 2019.

• University of Warwick, MAS-CDT Annual Conference, Somerset, United Kingdom. June 2019 - Flash Oral and Poster Presentation.

Title: Analysis of GPEtn using CAD, EID, and IRMPD FT-ICR MS.

Academic Societies

• Member of the British Mass Spectrometry Society (BMSS).
• Member of the American Society for Mass Spectrometry (ASMS).
• Member of the Royal Society of Chemistry (RSC).
• Member of the Chromatographic Society (ChromSoc)

Grants and Awards

• Awarded John Beynon Travel and Conference Fund, BMSS - September, 2023.
• Engineering and Physical Sciences Research Council (EPSRC) Scholarship for Integrated MSc and PhD in Molecular Analytical Science, University of Warwick - September, 2018.

Project Collaborations

• NHS Wales
• Swansea University
• Cardiff University
• Syngenta
• Verdel Instruments

Publications/Manuscripts

Basim R. Hussain
PhD Researcher

MAS-CDT and FT-ICR MS Group

Ion Cyclotron Resonance Laboratory
Department of Chemistry.

Contact

Email

Basim.Hussain@warwick.ac.uk

ResearchGate/BasimHussain

Link opens in a new windowLink opens in a new window

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Address

MOAC DTC, Senate House
University of Warwick,
Coventry, United Kingdom
CV4 7AL.