Rapid method for analysis of sphingomyelin by microwave derivatisation for gas chromatography–mass spectrometry

Abstract

Esterification and transesterification of lipids using boron trifluoride in methanol is widely used in lipid chemistry to study fatty acid (FA) profiles of various lipids by GC-MS. Transesterification of the FAs in sphingolipids is a difficult and time-consuming task. A rapid and convenient method for preparation of FAMEs from sphingomyelin (SM) using microwaves (MWs) was developed for GC-MS analysis. Experiments with three levels of power were performed and the yields were compared at regular time intervals. The maximum yields for all power levels were the same, but the time to reach these yields varied with the power. It was found that at a power of 70 W the maximum yield was reached after 27 min having a stable time window of 8 min for this yield. Practical applications:The technique presented here enables a rapid and easy method for preparing FAMEs from SM by the use of a MW reactor. The reaction time has been reduced from several hours down to only 27 min, with excellent yields.

Plasmalogens: Free Radical Reactivity and Identification of Trans Isomers Relevant to Biological Membranes

“…The results of the FAME composition of the same RBC membrane pellet treated by the two transesterification reaction conditions evidenced clear and statistically relevant differences, as reported in Table 1. This can be expected from the well-known chemical reactivity of lipid structures which contains amide-, ether, and ester bonds to link the fatty acid moiety [58][59][60][61]. In the acidic condition, it was seen the enrichment of SFA residues, explained by the transformation of SFA residues belonging to amide-containing lipids, such as sphingolipids, which need a such strong protocol to be converted [59][60][61][62].…”

“…This can be expected from the well-known chemical reactivity of lipid structures which contains amide-, ether, and ester bonds to link the fatty acid moiety [58][59][60][61]. In the acidic condition, it was seen the enrichment of SFA residues, explained by the transformation of SFA residues belonging to amide-containing lipids, such as sphingolipids, which need a such strong protocol to be converted [59][60][61][62]. Acidic conversion is the most frequently used method for automated protocols of blood lipidomic analysis for epidemiological studies [62,63]; however, it must be clear to the analyst that in this case sphingomyelins are transformed (rich in SFA), whereas plasmalogens, as well as other acid-sensitive PUFA-containing lipids, are not stable.…”

Sphingolipids: promising lipid-class molecules with potential applications for industry. A review

“…Hydrolysis of fatty acids from sphingolipids can be achieved by cleavage of the amide bond, with the use of methanolic solution of HCl (Hidaka et al, 2012), BF 3 (Devle et al, 2011) or NaOH (Groener et al, 2007), and further with the support of microwaves (Groener et al, 2007;Devle et al, 2011). Another method to release fatty acids from sphingolipids, is the enzymatic hydrolysis via sphingolipid ceramide N-deacylase (SCDase) action that breaks the N-acyl linkage (Lee et al, 2012b).…”