TY - JOUR
T1 - Identification of oxidized phospholipids by electrospray ionization mass spectrometry and LC-MS using a QQLIT instrument
AU - Spickett, Corinne M.
AU - Reis, Ana
AU - Pitt, Andrew R.
PY - 2011/12/15
Y1 - 2011/12/15
N2 - Phospholipids are complex and varied biomolecules that are susceptible to lipid peroxidation after attack by free radicals or electrophilic oxidants and can yield a large number of different oxidation products. There are many available methods for detecting phospholipid oxidation products, but also various limitations and problems. Electrospray ionization mass spectrometry allows the simultaneous but specific analysis of multiple species with good sensitivity and has a further advantage that it can be coupled to liquid chromatography for separation of oxidation products. Here, we explain the principles of oxidized phospholipid analysis by electrospray mass spectrometry and describe fragmentation routines for surveying the structural properties of the analytes, in particular precursor ion and neutral loss scanning. These allow targeted detection of phospholipid headgroups and identification of phospholipids containing hydroperoxides and chlorine, as well as the detection of some individual oxidation products by their specific fragmentation patterns. We describe instrument protocols for carrying out these survey routines on a QTrap5500 mass spectrometer and also for interfacing with reverse-phase liquid chromatography. The article highlights critical aspects of the analysis as well as some limitations of the methodology.
AB - Phospholipids are complex and varied biomolecules that are susceptible to lipid peroxidation after attack by free radicals or electrophilic oxidants and can yield a large number of different oxidation products. There are many available methods for detecting phospholipid oxidation products, but also various limitations and problems. Electrospray ionization mass spectrometry allows the simultaneous but specific analysis of multiple species with good sensitivity and has a further advantage that it can be coupled to liquid chromatography for separation of oxidation products. Here, we explain the principles of oxidized phospholipid analysis by electrospray mass spectrometry and describe fragmentation routines for surveying the structural properties of the analytes, in particular precursor ion and neutral loss scanning. These allow targeted detection of phospholipid headgroups and identification of phospholipids containing hydroperoxides and chlorine, as well as the detection of some individual oxidation products by their specific fragmentation patterns. We describe instrument protocols for carrying out these survey routines on a QTrap5500 mass spectrometer and also for interfacing with reverse-phase liquid chromatography. The article highlights critical aspects of the analysis as well as some limitations of the methodology.
KW - Chlorinated phospholipids
KW - Free radicals
KW - Lipidomics
KW - Liquid chromatography
KW - Neutral loss scanning
KW - Phospholipid oxidation
KW - Precursor ion scanning
UR - http://www.scopus.com/inward/record.url?scp=81855176005&partnerID=8YFLogxK
U2 - 10.1016/j.freeradbiomed.2011.09.003
DO - 10.1016/j.freeradbiomed.2011.09.003
M3 - Short survey
AN - SCOPUS:81855176005
SN - 0891-5849
VL - 51
SP - 2133
EP - 2149
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
IS - 12
ER -