Profiling and quantification of aminophospholipids based on chemical derivatization coupled with HPLC-MS
Autor: | Bangfu Wu, Hong Chen, Chen Yang, Zongyuan Wu, Fang Wei, Xin Lv, Ya Xie, Hui-fang Ma |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Male Calibration curve High selectivity QD415-436 030204 cardiovascular system & hematology High-performance liquid chromatography Biochemistry Mass Spectrometry Acetone Rats Sprague-Dawley 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Endocrinology Limit of Detection Animals Derivatization polyphenols Chromatography High Pressure Liquid Phospholipids Research Articles Phosphatidylethanolamine Chromatography high-performance liquid chromatography-mass spectrometry Ms analysis Analytic Sample Preparation Methods Cell Biology Rats Chromatographic separation 030104 developmental biology chemistry Liver |
Zdroj: | Journal of Lipid Research, Vol 60, Iss 1, Pp 121-134 (2019) |
ISSN: | 1539-7262 |
Popis: | In this study, a novel strategy based on acetone stable-isotope derivatization coupled with HPLC-MS for profiling and accurate quantification of aminophospholipids (phosphatidylethanolamine and phosphatidylserine) in biological samples was developed. Acetone derivatization leads to alkylation of the primary amino groups of aminophospholipids with an isopropyl moiety; the use of deuterium-labeled acetone (d6-acetone) introduced a 6 Da mass shift that was ideally suited for profiling and quantification analysis with high selectivity and accuracy. After derivatization, significantly increased column efficiency for chromatographic separation and detection sensitivity for MS analysis of aminophospholipids was observed. Furthermore, an accuracy quantification method was developed. Aminophospholipids in biological samples were derivatized with d0-acetone; while more than two aminophospholipid standards were selected for each class of aminophospholipid and derivatized with d6-acetone, which were then used as the internal standards to typically construct a calibration curve for each class to normalize the nonuniformity response caused by the differential fragmentation kinetics resulting from the distinct chemical constitution of individual aminophospholipid species in the biological samples. The excellent applicability of the developed method was validated by profiling and quantification of aminophospholipids presented in liver samples from rats fed with different diets. |
Databáze: | OpenAIRE |
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