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Normal‐phase versus reversed‐phase thin‐layer chromatography (TLC) to monitor oxidized phosphatidylcholines by TLC/mass spectrometry
Author(s) -
Engel Kathrin M.,
Griesinger Hans,
Schulz Michael,
Schiller Jürgen
Publication year - 2019
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8237
Subject(s) - chemistry , chromatography , popc , thin layer chromatography , phospholipid , mass spectrometry , phosphatidylcholine , electrospray ionization , silica gel , membrane , biochemistry
Rationale Normal‐phase thin‐layer chromatography (NP‐TLC) is an established method for the separation of all major phospholipid classes according to the different polarities of the head groups. In contrast, reversed‐phase (RP)‐TLC is much less frequently used for this purpose. This study aimed to compare the NP and the RP approach regarding their separation potential of phospholipid oxidation products. Methods Commercially available 1‐palmitoyl‐2‐oleoyl‐ sn ‐phosphatidylcholine (POPC) (PC 16:0/18:1) and 1‐palmitoyl‐2‐linoleoyl‐ sn ‐phosphatidylcholine (PLPC) (PC 16:0/18:2) were oxidized by NaMnO 4 . Oxidation products were subsequently separated by NP‐ and RP‐TLC and analyzed by electrospray ionization mass spectrometry. Results In comparison with NP‐TLC, RP‐TLC was clearly superior regarding the separation of oxidation products of phospholipids. RP‐TLC enabled the separation not only of primary oxidation products of POPC such as alcohols and ketones but also of secondary oxidation products. Furthermore some oxidation products, such as aldehydes, were only detectable by ESI after RP‐TLC but not after NP‐TLC. Conclusions RP‐TLC is the method of choice to characterize oxidized PL such as oxidized phosphatidylcholines.