Plackett-Burman Design and Fragmentation Studies to Assist the Comparison of Techniques used to Extract Phospholipids Prior to Regiospecific Characterization by Liquid Chromatography Mass Spectrometry

A Plackett-Burman design was used for screening 12 variables that could affect the mass spectrometry signals of phosphatidylcholines (PtdCho) and proposing optimal fragmentation conditions to study the fragmentation mechanisms of PtdCho standards. Under optimal conditions, three well-established methods for extracting phospholipids from biological samples, specifically liquid-liquid extraction (LLE), normal phase liquid chromatography (NPLC) and high performance thin layer chromatography (HPTLC), were compared in terms of the number of regiospecifically characterized PtdCho by means of liquid chromatography three-stage mass spectrometry (LCMS 3 ). The same number of PtdCho species (35) were characterized by LCMS3 in the LLE and NPLC fractions, whereas only 15 in the HPTLC fraction.