Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS
Phospholipids, including ether phospholipids, are composed of numerous isomeric and isobaric species that have the same backbone and acyl chains. This structural resemblance results in similar fragmentation patterns by collision-induced dissociation of phospholipids regardless of class, yielding com...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-12-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752060030X |
| _version_ | 1818408723788333056 |
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| author | Junhwan Kim Charles L. Hoppel |
| author_facet | Junhwan Kim Charles L. Hoppel |
| author_sort | Junhwan Kim |
| collection | DOAJ |
| description | Phospholipids, including ether phospholipids, are composed of numerous isomeric and isobaric species that have the same backbone and acyl chains. This structural resemblance results in similar fragmentation patterns by collision-induced dissociation of phospholipids regardless of class, yielding complicated MS/MS spectra when isobaric species are analyzed together. Furthermore, the presence of isobaric species can lead to misassignment of species when made solely based on their molecular weights. In this study, we used normal-phase HPLC for ESI-MS/MS analysis of phospholipids from bovine heart mitochondria. Class separation by HPLC eliminates chances for misidentification of isobaric species from different classes of phospholipids. Chromatography yields simple MS/MS spectra without interference from isobaric species, allowing clear identification of peaks corresponding to fragmented ions containing monoacylglycerol backbone derived from losing one acyl chain. Using these fragmented ions, we characterized individual and isomeric species in each class of mitochondrial phospholipids, including unusual species, such as PS, containing an ether linkage and species containing odd-numbered acyl chains in cardiolipin, PS, PI, and PG. We also characterized monolysocardiolipin and dilysocardiolipin, the least abundant but nevertheless important mitochondrial phospholipids. The results clearly show the power of HPLC-MS/MS for identification and characterization of phospholipids, including minor species. |
| first_indexed | 2024-12-14T09:48:16Z |
| format | Article |
| id | doaj.art-8b047f209cfe4677b69f3988f3eb3661 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T09:48:16Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-8b047f209cfe4677b69f3988f3eb36612022-12-21T23:07:35ZengElsevierJournal of Lipid Research0022-22752020-12-01611217071719Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MSJunhwan Kim0Charles L. Hoppel1Center for Mitochondrial Diseases, Case Western Reserve University, Cleveland, OH, USA; Department of Pharmacology, Case Western Reserve University, Cleveland, OH, USACenter for Mitochondrial Diseases, Case Western Reserve University, Cleveland, OH, USA; Department of Pharmacology, Case Western Reserve University, Cleveland, OH, USA; Department of Medicine, Case Western Reserve University, Cleveland, OH, USA; For correspondence: Charles L. Hoppel; For correspondence: Charles L. HoppelPhospholipids, including ether phospholipids, are composed of numerous isomeric and isobaric species that have the same backbone and acyl chains. This structural resemblance results in similar fragmentation patterns by collision-induced dissociation of phospholipids regardless of class, yielding complicated MS/MS spectra when isobaric species are analyzed together. Furthermore, the presence of isobaric species can lead to misassignment of species when made solely based on their molecular weights. In this study, we used normal-phase HPLC for ESI-MS/MS analysis of phospholipids from bovine heart mitochondria. Class separation by HPLC eliminates chances for misidentification of isobaric species from different classes of phospholipids. Chromatography yields simple MS/MS spectra without interference from isobaric species, allowing clear identification of peaks corresponding to fragmented ions containing monoacylglycerol backbone derived from losing one acyl chain. Using these fragmented ions, we characterized individual and isomeric species in each class of mitochondrial phospholipids, including unusual species, such as PS, containing an ether linkage and species containing odd-numbered acyl chains in cardiolipin, PS, PI, and PG. We also characterized monolysocardiolipin and dilysocardiolipin, the least abundant but nevertheless important mitochondrial phospholipids. The results clearly show the power of HPLC-MS/MS for identification and characterization of phospholipids, including minor species.http://www.sciencedirect.com/science/article/pii/S002222752060030Xcollision-induced dissociationacyl chainether phospholipidscardiolipinodd-numbered acyl chainhigh-performance liquid chromatography-tandem mass spectrometry |
| spellingShingle | Junhwan Kim Charles L. Hoppel Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS Journal of Lipid Research collision-induced dissociation acyl chain ether phospholipids cardiolipin odd-numbered acyl chain high-performance liquid chromatography-tandem mass spectrometry |
| title | Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS |
| title_full | Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS |
| title_fullStr | Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS |
| title_full_unstemmed | Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS |
| title_short | Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS |
| title_sort | identification of unusual phospholipids from bovine heart mitochondria by hplc ms ms |
| topic | collision-induced dissociation acyl chain ether phospholipids cardiolipin odd-numbered acyl chain high-performance liquid chromatography-tandem mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S002222752060030X |
| work_keys_str_mv | AT junhwankim identificationofunusualphospholipidsfrombovineheartmitochondriabyhplcmsms AT charleslhoppel identificationofunusualphospholipidsfrombovineheartmitochondriabyhplcmsms |