Lipidomic approaches now gaining acceptance are based on the rapid, comprehensive mass spectrometry (MS) analysis of carbon length and degree of unsaturation in fatty acyl groups and their positions (regiospecificity) at the sn 1,2, or 3 positions of glycerolipids. The most important analytical issue facing lipidomic analysis is that no general method has emerged that delivers double bond position in the fatty acyl groups simultaneously with regiospecific information, We propose to exploit a novel mass spectrometric method developed in our laboratory that should enable complete, automated characterization of all acylated lipids in crude lipid extracts, Ion-molecule reactions that result in adducts during chemical ionization (Cl) mass spectrometry (MS) are effectively a gas phase derivatization process. In a series of 8 papers since 1999, we reported co-discovery of gas derivatization of unsaturated fatty acid methyl esters (FAME) by acetonitrile CI/MS/MS. Fragmentation of the resulting adducts provides the first rapid, convenient structural analysis of FAME. Acetonitrile CI produces an ion at m/z 54 which adds covalently across double bonds; the resulting [M+54]+ ion undergoes charge-driven fragmentation upon collisional activation to yield diagnostic ions that unequivocally establish the position of double bonds in polyunsaturates. Recently, this reaction has been observed for atmospheric pressure CI in a liquid stream. We propose a two pronged approach where we 1) develop APCI based on acetonitrile for analysis of double bond position for fatty acyl groups within lipid classes derived from crude lipid extracts, and 2) complete development of acetonitrile GClCIIMSlMS to include analysis of all polyunsaturated non-methylene-interrupted fatty acids. These goals will eventually be coupled, and together with conventional MS/MS modes (precursor, product, and neutral loss scans), analysis of gas phase derivatized acyl-containing compounds should provide complete structural and quantitative information for most glycerolipids and can be automated for lipidomic analysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM071534-04
Application #
7268633
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Edmonds, Charles G
Project Start
2004-08-01
Project End
2009-07-31
Budget Start
2007-08-01
Budget End
2009-07-31
Support Year
4
Fiscal Year
2007
Total Cost
$237,662
Indirect Cost
Name
Cornell University
Department
Nutrition
Type
Other Domestic Higher Education
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Brenna, J Thomas; Kothapalli, Kumar S D; Park, Woo Jung (2010) Alternative transcripts of fatty acid desaturase (FADS) genes. Prostaglandins Leukot Essent Fatty Acids 82:281-5
Park, Woo Jung; Reardon, Holly T; Tyburczy, Cynthia et al. (2010) Alternative splicing generates a novel FADS2 alternative transcript in baboons. Mol Biol Rep 37:2403-6
Park, Woo Jung; Kothapalli, Kumar S D; Lawrence, Peter et al. (2009) An alternate pathway to long-chain polyunsaturates: the FADS2 gene product Delta8-desaturates 20:2n-6 and 20:3n-3. J Lipid Res 50:1195-202
Stroud, Chad K; Nara, Takayuki Y; Roqueta-Rivera, Manuel et al. (2009) Disruption of FADS2 gene in mice impairs male reproduction and causes dermal and intestinal ulceration. J Lipid Res 50:1870-80
Gómez-Cortés, Pilar; Tyburczy, Cynthia; Brenna, J Thomas et al. (2009) Characterization of cis-9 trans-11 trans-15 C18:3 in milk fat by GC and covalent adduct chemical ionization tandem MS. J Lipid Res 50:2412-20
Park, Woo Jung; Kothapalli, Kumar S D; Reardon, Holly T et al. (2009) Novel fatty acid desaturase 3 (FADS3) transcripts generated by alternative splicing. Gene 446:28-34
Ran-Ressler, Rinat R; Devapatla, Srisatish; Lawrence, Peter et al. (2008) Branched chain fatty acids are constituents of the normal healthy newborn gastrointestinal tract. Pediatr Res 64:605-9
Liang, Xiaorong; Liu, Jian; LeBlanc, Yves et al. (2007) Electron transfer dissociation of doubly sodiated glycerophosphocholine lipids. J Am Soc Mass Spectrom 18:1783-8
Xu, Yichuan; Brenna, J Thomas (2007) Atmospheric pressure covalent adduct chemical ionization tandem mass spectrometry for double bond localization in monoene- and diene-containing triacylglycerols. Anal Chem 79:2525-36
Brenna, J Thomas; Diau, Guan-Yeu (2007) The influence of dietary docosahexaenoic acid and arachidonic acid on central nervous system polyunsaturated fatty acid composition. Prostaglandins Leukot Essent Fatty Acids 77:247-50

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