Abstract
Plasmalogens (alkenylacyl glycerophospholipids) are important lipid constituents of many tissues and cells (e.g., selected spermatozoa). Since the molecular weights of plasmalogens overlap with that of diacyl- or alkyl acyl lipids, sophisticated mass spectrometry (MS; including MS/MS) analysis is normally used for the unequivocal identification of plasmalogens. We will show here that a simple matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (without MS/MS capability) in combination with acidic hydrolysis and subsequent derivatization with 2,4-dinitrophenylhydrazine (DNPH) and/or digestion with phospholipase A2 (PLA2) is sufficient to determine the contributions of ether lipids in spermatozoa extracts. As neither diacyl nor alkylacyl lipids are sensitive to acids and do not react with DNPH, the comparison of the mass spectra before and after treatment with acids and/or DNPH addition readily provides unequivocal information about the plasmalogen content. Additionally, the released aldehydes are readily converted into the 2,4-dinitrophenylhydrazones and can be easily identified in the corresponding negative ion mass spectra. Finally, PLA2 digestion is very useful in confirming the presence of plasmalogens. The suggested method was validated by analyzing roe deer, bovine, boar, and domestic cat spermatozoa extracts and comparing the results with isolated phospholipids.
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Abbreviations
- DNPH:
-
2,4-Dinitrophenylhydrazine
- DHB:
-
2,5-Dihydroxybenzoic acid
- GPC:
-
Glycerophosphorylcholine
- MS:
-
Mass spectrometry
- MALDI-TOF:
-
Matrix-assisted laser desorption and ionization time-of-flight
- PL:
-
Phospholipid
- PLA2 :
-
Phospholipase A2
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Acknowledgment
This work was supported by the German Research Council (DFG Schi 476/12-1, SFB 1052/B6, Mu 1520/4-1, Fu 771/1-2, and Ni 1396/3-1.
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Ariane Nimptsch and Beate Fuchs contributed equally to this work.
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Nimptsch, A., Fuchs, B., Süß, R. et al. A simple method to identify ether lipids in spermatozoa samples by MALDI-TOF mass spectrometry. Anal Bioanal Chem 405, 6675–6682 (2013). https://doi.org/10.1007/s00216-013-7147-z
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DOI: https://doi.org/10.1007/s00216-013-7147-z