Abstract
Understanding the biological roles and mechanisms of lipids and glycolipids is challenging due to the vast number of possible isomers that may exist. Mass spectrometry (MS) measurements are currently the dominant approach for studying and providing detailed information on lipid and glycolipid presence and changes. However, difficulties in distinguishing the many structural isomers, due to the distinct lipid acyl chain positions, double bond locations or specific glycan types, inhibit the delineation and assignment of their biological roles. Here we utilized ultra-high resolution ion mobility spectrometry (IMS) separations by applying traveling waves in a serpentine multi-pass Structures for Lossless Ion Manipulations (SLIM) platform to enhance the separation of selected lipid and glycolipid isomers. The multi-pass arrangement allowed the investigation of paths ranging from ~16 m (one pass) to ~60 m (four passes) for the distinction of lipids and glycolipids with extremely small structural differences. These ultra-high resolution SLIM IMS-MS analyses provide a foundation for exploring and better understanding isomer-specific biological activities and disease processes.
Highlights
Evaluating the role of lipids in diverse biological systems has important implications in understanding disease initiation and progression [1]
We examined gangliosides which are some of the most complex glycosphingolipids with glycan head groups containing one or more sialic acids
This manuscript provides an initial demonstration of a multi-pass Structures for Lossless Ion Manipulations (SLIM) ion mobility spectrometry-mass spectrometry (IMS-Mass spectrometry (MS)) platform achieving the ultra-high resolution necessary to distinguish lipids and glycolipids with very similar structures
Summary
Evaluating the role of lipids in diverse biological systems has important implications in understanding disease initiation and progression [1]. ((AA)) IIlllluussttrraatteess ppootteennttiiaall ddoouubbllee bboonndd ppoossiittiioonnss iinn aa 1166::11 ffaattttyy aacciidd;; ((BB)) SShhoowwss ppoossssiibbllee ffaattttyy aacciidd lliinnkkaaggee ccoonnnneeccttiivviittyy ssiitteess ((ssnn--33 iiss sshhoowwnn ffoorraapphhoosspphhoolliippiiddwwhheerree XX cchhaannggeess ddeeppeennddiinnggoonntthheeggllyycceerroopphhoosspphhoolliippiiddttyyppee));;((CC))EElluucciiddaatteesstthhee hhyyddrrooxxyyllggrroouupp ddiiffffeerreenncceess bbeettwweeeenn ggaallaaccttoossee aanndd gglluuccoossee wwhhiicchh aarree hhiigghhlliigghhtteedd iinn bblluuee aannddoorraannggee aanndd ((DD)) IIlllluussttrraatteess tthhee tthhrreeee mmaaiinn ccaatteeggoorriieess ooff ggllyyccoolliippiiddss. For the isomers with different double bond placements, each was distinguished in the one- and two-pass analyses with the isomer having the double bonds at the 9th carbon having a larger structure than the isomer with the double bonds closer to the head group at the 6th carbon. For the isomers with different double bond placements, each was distinguished in the one- and two-pass analyses with the isomer having the double bonds at the 9th carbon having a larger structure than the isomer with the double bonds closer to the hIenat.dJ. TThhuuss,, aafftteerr oonneeccyycclele,,oonnlylyppaarrtitaiallsseeppaarraatitoionnwwaass oobbsseerrvveedd;;hhoowweevveerr, ,aannaadddditiitoionnaallccyyccleleaalllolowweeddththeeppeeaakkssfrfroommththeetwtwooccoommppoonneenntststotobbeebbaasseelilninee reresosolvlveedd. .WWhhilieleththeetrtraannssddoouubblelebboonnddssoonnlylyccaauusseessaammininoorrcchhaannggeeininssttrruucctuturerefrfroommththaattoofftthheettoottaalllyly ssaatuturarateteddlilpipidi,dt,htehceiscidsoduobulebbleobnodns dsisgsniigfinciafnictalyntclhyacnhgaentghee gtheeomgeeotrmy,eftorlyd,ifnogldthinegfatthtye afactidtycahcaiidn cbhaackin inbwacakrdinswonaridtsseolfnaintsdellfeaadnidnlgeatodiansgmtoalalesrmtharleleer-dthimreeen-dsiiomneanl ssitoruncatlusrter.uHctouwree.vHeor,wperevveiro,upsrewvoiorkusuwsinorgk aucsoinngveantcioonnavlelnotwionparel slsouwrepIrMesSsudrreiftIMtubSedprliafttftourbme opnlalytfpoarmrtiaolnlylyrepsaorlvtieadllythreesseoplveeadksthfoerseinpdeivaikdsufaolr sitnadndivairddusal[2s]t.anTdhaursd, sth[e2]b. aTsheulisn,etsheepabraasteiloinnefosrepbaortahtitohne fdoorubbolethbothned dpoousbitlieonbsonanddpoorsiietniotnatsioannsd porroiveindteastitohnesbparsoivs ifdoersththeeirbeavsaislufaotriothneiinr ecovmalupaletxiobnioinlocgoimcapl lseaxmbpiolelso.gical samples
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