An important feature of molecular lipid species is the position o

An important feature of molecular lipid species is the position of fatty acid double bonds. Gas phase reaction of ozone with

double bonds results in primary and secondary ozonides, which fragment further to aldehydes, carboxylates and hydroperoxides indicative of the position of the double bond in the fatty acyl chain [60]. Recently, the group of Blanksby introduced custom modified instrumentation for ozone induced dissociation (OzID), at which either a linear ion trap [61] or a quadrupole Inhibitors,research,lifescience,medical collision cell [62] are able to be filled with ozone. Either sequential multistage dissociation with an inert collision induced dissociation (CID) gas and ozone, or single stage dissociation by a mixture of ozone and CID gas, results in a double bond position specific fragmentation pattern. The main limitation of this method is the specialized non-commercial equipment needed. Additionally, no high throughput standardized data analysis software is available for such an approach. Inhibitors,research,lifescience,medical An interesting alternative for obtaining

more structural details by MS/MS is the pseudo MS3 approach on a 4000QTrap proposed Inhibitors,research,lifescience,medical by the Merrill group [63]. Negative ESI tandem MS/MS spectra on sphingomyelin usually result in just one predominant head group specific fragment. But if the selected sphingomyelin precursor is transmitted through Q2 at low collision energies (5–10 eV) and then fragmented by the linear ion trap function in Q3, a much richer abundance of fragment ions indicative for the sphingosine backbone is to be found. 4. MALDI-TOF Although not as widely used as ESI instruments, MALDI-TOF is a good complementary choice for lipids in the mass range above m/z 500. The soft ionization properties of MALDI result in intact molecular adduct ions. Paired with the speed of MALDI-TOF analysis Inhibitors,research,lifescience,medical this fact renders the technology very suitable for fast screening of lipids. MALDI-TOF Inhibitors,research,lifescience,medical instruments equipped with a reflectron nowadays regularly achieve 10,000 resolution and 30 ppm mass accuracy, which is sufficient for assigning intact molecular ions of lipid species.

Choice of the right MALDI matrix is an important step for good sensitivity. 2,5-Dihydroxy benzoic acid, α-cyano-4-hydroxy-cinnamic Resminostat acid, 9-amino-acridine and 2-mercaptobenzothiazole are often used matrix compounds. On the downside of this technology, the mass range below m/z 500 is usually not amenable due to matrix interferences. MALDI-TOF has been used for analysis of various lipid classes [64], but, similar to ESI, MALDI also has certain quantitative limits for crude mixtures due to ion suppression effects [65]. This effect can become quite severe, particularly as MALDI does not allow any chromatographic separation to be coupled directly to the instrument. Recently, TLC/MALDI was proposed by several groups as an interesting alternative [66,67]. RAD001 price Instead of a MALDI target, a developed TLC plate with separated lipid spots is used as target.

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