Enhancing Coverage of Phosphatidylinositol Species in Canola Through Specialised Liquid Chromatography-Mass Spectrometry Buffer Conditions

Gertner, DS; Bishop, DP; Oglobline, A; Padula, MP

Enhancing Coverage of Phosphatidylinositol Species in Canola Through Specialised Liquid Chromatography-Mass Spectrometry Buffer Conditions

Gertner, DS Bishop, DP Oglobline, A Padula, MP

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Publisher:: Elsevier
Publication Type:: Journal Article
Citation:: Journal of Chromatography A, 2021, 1637, pp. 461860-461860
Issue Date:: 2021

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Field	Value	Language
dc.contributor.author	Gertner, DS
dc.contributor.author	Bishop, DP
dc.contributor.author	Oglobline, A
dc.contributor.author	Padula, MP
dc.date.accessioned	2021-03-01T03:09:18Z
dc.date.available	2020-12-26
dc.date.available	2021-03-01T03:09:18Z
dc.date.issued	2021
dc.identifier.citation	Journal of Chromatography A, 2021, 1637, pp. 461860-461860
dc.identifier.issn	0021-9673
dc.identifier.issn	1873-3778
dc.identifier.uri	http://hdl.handle.net/10453/146552
dc.description.abstract	Phosphatidylinositols (PIs) constitute a minor class of phospholipid with wide-spread influence throughout various cellular functions. Monitoring the distribution of these lipids can therefore provide insight as to the state of cellular processes or reveal the development of various pathologies. The speciation of these compounds is often performed either as part of a comprehensive characterisation of lipids, or specifically targeted using the same methods, however, such methods were intended to maximise coverage of lipid classes rather than provide an in-depth analysis of any single class. In the particular case of PIs, the majority of reported molecular diversity is limited to a small proportion of the already minor class, as such the cursory glance enabled by such methods is insufficient. Therefore, this work compared the suitability of both established and novel LC-MS buffers with the aim of maximising the ionisation efficiency of PIs, in an attempt to enhance coverage of the class. Through experimentation, it was determined that a 0.25 mM ammonium fluoride buffer provided up to a 6-fold increase in signal intensity, and on average a 38-fold increase in the signal-to-noise ratio. Using these new conditions, 14 PI species, and 12 PI candidates were identified within a dilute lipid extract sourced from canola seed, compared to 0 species identified using the generalised method. As a result, it is suggested that this procedure has yielded the highest number of PI species identifications for a sample of this concentration. Methods which therefore intend to characterise PI species in dilute quantities, such as those extracted from mammalian cells, are henceforth provided with the means to conduct more comprehensive characterisations.
dc.format	Print-Electronic
dc.language	en
dc.publisher	Elsevier
dc.relation	http://purl.org/au-research/grants/arc/DE180100194
dc.relation.ispartof	Journal of Chromatography A
dc.relation.isbasedon	10.1016/j.chroma.2020.461860
dc.rights	info:eu-repo/semantics/embargoedAccess
dc.subject	03 Chemical Sciences, 09 Engineering, 10 Technology
dc.subject.classification	Analytical Chemistry
dc.subject.mesh	Animals
dc.subject.mesh	Phosphatidylinositols
dc.subject.mesh	Buffers
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Mass Spectrometry
dc.subject.mesh	Signal-To-Noise Ratio
dc.subject.mesh	Animals
dc.subject.mesh	Buffers
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Mass Spectrometry
dc.subject.mesh	Phosphatidylinositols
dc.subject.mesh	Signal-To-Noise Ratio
dc.title	Enhancing Coverage of Phosphatidylinositol Species in Canola Through Specialised Liquid Chromatography-Mass Spectrometry Buffer Conditions
dc.type	Journal Article
utslib.citation.volume	1637
utslib.location.activity	Netherlands
utslib.for	03 Chemical Sciences
utslib.for	09 Engineering
utslib.for	10 Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
utslib.copyright.status	open_access	*
pubs.consider-herdc	false
utslib.copyright.embargo	2023-01-01T00:00:00+1000Z
dc.date.updated	2021-03-01T03:09:12Z
pubs.publication-status	Published
pubs.volume	1637

Abstract:

Phosphatidylinositols (PIs) constitute a minor class of phospholipid with wide-spread influence throughout various cellular functions. Monitoring the distribution of these lipids can therefore provide insight as to the state of cellular processes or reveal the development of various pathologies. The speciation of these compounds is often performed either as part of a comprehensive characterisation of lipids, or specifically targeted using the same methods, however, such methods were intended to maximise coverage of lipid classes rather than provide an in-depth analysis of any single class. In the particular case of PIs, the majority of reported molecular diversity is limited to a small proportion of the already minor class, as such the cursory glance enabled by such methods is insufficient. Therefore, this work compared the suitability of both established and novel LC-MS buffers with the aim of maximising the ionisation efficiency of PIs, in an attempt to enhance coverage of the class. Through experimentation, it was determined that a 0.25 mM ammonium fluoride buffer provided up to a 6-fold increase in signal intensity, and on average a 38-fold increase in the signal-to-noise ratio. Using these new conditions, 14 PI species, and 12 PI candidates were identified within a dilute lipid extract sourced from canola seed, compared to 0 species identified using the generalised method. As a result, it is suggested that this procedure has yielded the highest number of PI species identifications for a sample of this concentration. Methods which therefore intend to characterise PI species in dilute quantities, such as those extracted from mammalian cells, are henceforth provided with the means to conduct more comprehensive characterisations.

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http://hdl.handle.net/10453/146552