Open AccessA simple solid‐phase extraction method for the analysis of red cell phospholipids by liquid chromatography‐ tandem mass spectrometry
Author(s) -
Nguyen Van Long
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22417
Subject(s) - chromatography , chemistry , phospholipid , liquid chromatography–mass spectrometry , tandem mass spectrometry , extraction (chemistry) , solid phase extraction , mass spectrometry , lysis , coefficient of variation , analytical chemistry (journal) , membrane , biochemistry
Background There has been increasing interest in the analysis of phospholipids in red blood cells as potential long‐term biomarkers of different disease states. Here, we describe a simple method for the analysis of two phospholipids: 1‐Palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphoethanol ( PE 16:0/18:1) and 1‐Palmitoyl‐2‐linoleoyl‐sn‐glycero‐3‐phosphoethanol ( PE 16:/0/18:2) in erythrocytes by liquid chromatography‐tandem mass spectrometry ( LC ‐ MS / MS ). Methods Whole blood samples were removed free of plasma and washed in isotonic saline. Red cells were lysed with ultrapure water. Lysate samples were processed using a hybrid solid‐phase extraction ( SPE ) phospholipid cartridge (1 mL , 30 mg). Both PE 16:0/18:1 and PE 16:0/18:2 and their deuterated internal standards were separated on an ACE C4 (150 mm × 2.1 mm, 2.7 μm particle size) by gradient elution at a flow rate of 0.5 mL per minute using mobile phases consisting of 0.01 mol/L ammonium acetate in: water (A), methanol (B), and isopropanol (C). The phospholipid species were quantified by the following transitions: PE 16:0/18:1: 701.5→281.3 and PE 16:0/18:2: 699.5→279.3. Results Both PE species displayed linearity ranging from 10 to 500 μg/L. The coefficient of variation ( CV %) of PE 16:0/18:1 concerning intraday and interday precision was between 1.9%‐2.6% and 3.0%‐4.3%, respectively. For PE 16:0/18:2, this was between 1.8%‐3.4% and 3.7%‐4.1%, respectively. Both phospholipid species had accuracy ( PE 16:0/18:1: 91%‐98% and PE 16:0/18:2: 94%‐103%) and extraction recovery ( PE 16:0/18:1: 95%‐106% and PE 16:0/18:2: 92%‐102%) exceeding 90% over the analytical range. The limit of detection was 5 μg/L. Conclusion Here we propose a simple SPE LC ‐ MS / MS method for analyzing phospholipids in erythrocytes, which can be easily adopted.