BACKGROUND: In bioanalysis, phospholipids may affect the precision and accuracy of LC-MS/MS methods and compromise the quality of the results, especially when samples in complex biomatrices are extracted by protein precipitation techniques. RESULTS: It was found that the retentive behavior of both common pharmaceuticals and physiologically relevant phospholipids under bare silica hydrophilic-interaction LC (HILIC) is more predictable than under reversed-phase conditions. In particular, the retention time of phospholipids was not significantly affected by varying the salt and acid modifiers in the mobile phases, but common pharmaceuticals can be shifted away from these phospholipid interferences through mobile phase modifiers. Several mass spectrometric techniques were applied to confirm this finding. CONCLUSION: HILIC chromatography is a valued tool in the development of robust bioanalytical assays with minimal and predictable phospholipid interferences. Furthermore, addition of a small amount of ion-pairing additives can reliably move pharmaceutical compounds away from these suppressive regions.
BACKGROUND: In bioanalysis, phospholipids may affect the precision and accuracy of LC-MS/MS methods and compromise the quality of the results, especially when samples in complex biomatrices are extracted by protein precipitation techniques. RESULTS: It was found that the retentive behavior of both common pharmaceuticals and physiologically relevant phospholipids under bare silica hydrophilic-interaction LC (HILIC) is more predictable than under reversed-phase conditions. In particular, the retention time of phospholipids was not significantly affected by varying the salt and acid modifiers in the mobile phases, but common pharmaceuticals can be shifted away from these phospholipid interferences through mobile phase modifiers. Several mass spectrometric techniques were applied to confirm this finding. CONCLUSION: HILIC chromatography is a valued tool in the development of robust bioanalytical assays with minimal and predictable phospholipid interferences. Furthermore, addition of a small amount of ion-pairing additives can reliably move pharmaceutical compounds away from these suppressive regions.