Ne lipids were also reported for the first time in green algae [68]. Ragonese et al. [107] used HILIC- ESI/IT-TOF-MS to analyze lipid extracts from different macroalgae and provide a reliableMar. Drugs 2016, 14,17 ofidentification of lipid classes. Pterocladiella capillacea showed the most complex profile, containing several lipids classes (PG, PC, PI, LPI, PS, LPE, DGDG, SQDG, SQMG), while MS023 solubility Asparagopsis taxiformis contained a single sulquinovosyl monoacylglycerol (SGMG). The lipidome of the red macroalgae Dictyota dicotoma was mainly represented by PLs (PC, LPE, SQMG, SQDG). This study enhances the use of the MS-base profiling of polar lipids towards the classification of marine organisms and, in general, the classification of complex lipid matrices. Although HILIC has become a relevant technique in this field, RPLC is still widely used in lipidomics, namely, in plant lipidomics to separate GLs [80,117?20]. Kendal et al. [50] used a C18 column to discover which glycolipid species obtained from Ulva armaricana and Solieria get trans-4-Hydroxytamoxifen chordalis displayed anti-proliferative properties against lung tumor. RPLC was also applied to identify eicosanoids in the red macro algae Gracilaria asiatica [121] and other oxylipins [122]. In fact, LC-MS platforms have greatly improved the resolution, sensitivity and mass range, solving problems of complex lipid separation and characterization. Due to the structural variety of polar lipids, resolving lipids in their representative classes and species rely on the combined use of MS and chromatographic approaches; moreover, it also allows for the possibility of separating and concentrating different classes, taking into account their physicochemical properties [116].Mar. Drugs 2016, 14,18 ofTable 2. Molecular ions formed during electrospray ionization (ESI) and MS/MS fragmentation fingerprint data of each polar lipid class (in bold the most formed ion in ESI-MS).Lipid Class Negative Phosphatidylcholine (PC) Phosphatidylethanolamine (PE) Phosphatidylglycerol (PG) Phosphatidylinositol (PI) Phosphatidylserine (PS) Monogalactosyldiacylglycerol (MGDG) Digalactosyldiacylglycerol (DGDG) Sulfoquinovosildiacylglycerol (SQDG) Ceramide (Cer) Galactosylceramide (GalCer) [M + Ac-H]?H]?H]?H]?[M + [M + Detect Ions in MS Positive H]+ , H]+ , [M + [M + Na]+ Na]+ Na]+ Precursor Ion Scan Negative (m/z) 241 225 Na]+ Positive (m/z) 184 243 347 365 264 264 223 241 259 236 162 180 162 180 87 74 59 87 74 59 Neutral Loss Scan Negative (Da) 74 87 Positive (Da) 141 185 179 162 341 -[M ?[M ?[M ?[M + NH]+ ,[M + ]+[M + NH[M ?H]?[M ?H]?[M ?H]?[M ?H]?[M ?H]?[M +[M + H]+ [M + NH4 ]+ , [M + Na]+ [M + NH4 ]+ , [M + Na]+ [M + NH4 ]+ , [M + Na]+ H]+ , [M + NH4 ]+ , [M +[M ?H]?[M ?H]?[M + H]+ , [M + NH4 ]+ , [M + Na]+ [M + H]+ , [M + NH4 ]+ , [M + Na]+Inositolphosphoceramide (IPC)-Diacylglyceryl-N,N,N-trimethylhomoserine (DGTS) Diacylglycerylhydroxymethyl-N,N,N-trimetyl -alanine (DGTA) Diacylglycerylcarboxyhydroxy methylcholine (DGCC)[M + H]+–[M + H]+ [M + H]+Mar. Drugs 2016, 14,19 ofTable 3. Polar lipid classes in marine macrophytes analyzed by MS-based approaches.Species Name Green macroalgae Codium tomentosum Enteromorpha intestinalis Ulva armaricana Ulva fasciata Ulva lactuca Ulva rigida Red macroalgae Asparagopsis taxiformis Chondria armata Chondrus crispus (cultured) Chondrus crispus (cultured) Galaxoura cylindriea Laurencia papillose Palmaria palmata Pterocodiella capillacea Osmundaria obtusiloba Solieria chordalis Brown macroalgae Colpomenia sinu.Ne lipids were also reported for the first time in green algae [68]. Ragonese et al. [107] used HILIC- ESI/IT-TOF-MS to analyze lipid extracts from different macroalgae and provide a reliableMar. Drugs 2016, 14,17 ofidentification of lipid classes. Pterocladiella capillacea showed the most complex profile, containing several lipids classes (PG, PC, PI, LPI, PS, LPE, DGDG, SQDG, SQMG), while Asparagopsis taxiformis contained a single sulquinovosyl monoacylglycerol (SGMG). The lipidome of the red macroalgae Dictyota dicotoma was mainly represented by PLs (PC, LPE, SQMG, SQDG). This study enhances the use of the MS-base profiling of polar lipids towards the classification of marine organisms and, in general, the classification of complex lipid matrices. Although HILIC has become a relevant technique in this field, RPLC is still widely used in lipidomics, namely, in plant lipidomics to separate GLs [80,117?20]. Kendal et al. [50] used a C18 column to discover which glycolipid species obtained from Ulva armaricana and Solieria chordalis displayed anti-proliferative properties against lung tumor. RPLC was also applied to identify eicosanoids in the red macro algae Gracilaria asiatica [121] and other oxylipins [122]. In fact, LC-MS platforms have greatly improved the resolution, sensitivity and mass range, solving problems of complex lipid separation and characterization. Due to the structural variety of polar lipids, resolving lipids in their representative classes and species rely on the combined use of MS and chromatographic approaches; moreover, it also allows for the possibility of separating and concentrating different classes, taking into account their physicochemical properties [116].Mar. Drugs 2016, 14,18 ofTable 2. Molecular ions formed during electrospray ionization (ESI) and MS/MS fragmentation fingerprint data of each polar lipid class (in bold the most formed ion in ESI-MS).Lipid Class Negative Phosphatidylcholine (PC) Phosphatidylethanolamine (PE) Phosphatidylglycerol (PG) Phosphatidylinositol (PI) Phosphatidylserine (PS) Monogalactosyldiacylglycerol (MGDG) Digalactosyldiacylglycerol (DGDG) Sulfoquinovosildiacylglycerol (SQDG) Ceramide (Cer) Galactosylceramide (GalCer) [M + Ac-H]?H]?H]?H]?[M + [M + Detect Ions in MS Positive H]+ , H]+ , [M + [M + Na]+ Na]+ Na]+ Precursor Ion Scan Negative (m/z) 241 225 Na]+ Positive (m/z) 184 243 347 365 264 264 223 241 259 236 162 180 162 180 87 74 59 87 74 59 Neutral Loss Scan Negative (Da) 74 87 Positive (Da) 141 185 179 162 341 -[M ?[M ?[M ?[M + NH]+ ,[M + ]+[M + NH[M ?H]?[M ?H]?[M ?H]?[M ?H]?[M ?H]?[M +[M + H]+ [M + NH4 ]+ , [M + Na]+ [M + NH4 ]+ , [M + Na]+ [M + NH4 ]+ , [M + Na]+ H]+ , [M + NH4 ]+ , [M +[M ?H]?[M ?H]?[M + H]+ , [M + NH4 ]+ , [M + Na]+ [M + H]+ , [M + NH4 ]+ , [M + Na]+Inositolphosphoceramide (IPC)-Diacylglyceryl-N,N,N-trimethylhomoserine (DGTS) Diacylglycerylhydroxymethyl-N,N,N-trimetyl -alanine (DGTA) Diacylglycerylcarboxyhydroxy methylcholine (DGCC)[M + H]+–[M + H]+ [M + H]+Mar. Drugs 2016, 14,19 ofTable 3. Polar lipid classes in marine macrophytes analyzed by MS-based approaches.Species Name Green macroalgae Codium tomentosum Enteromorpha intestinalis Ulva armaricana Ulva fasciata Ulva lactuca Ulva rigida Red macroalgae Asparagopsis taxiformis Chondria armata Chondrus crispus (cultured) Chondrus crispus (cultured) Galaxoura cylindriea Laurencia papillose Palmaria palmata Pterocodiella capillacea Osmundaria obtusiloba Solieria chordalis Brown macroalgae Colpomenia sinu.