The feasibility of the use of colloidal silica in combination with a number of divalent or trivalent cations for the removal of plasma phospholipids was evaluated by sequentially adding the two reagents (i.e., colloidal silica and a cation) directly to blank plasma samples or plasma samples spiked with analytes. Three representative plasma phospholipids were monitored to determine the efficiency of the phospholipids removal under different reagent combinations. The recovery of each spiked analyte was also monitored under each condition in order to determine if any of the analyte was removed along with the phospholipids. By optimizing the amounts of the reagents used and the sequence of the addition of the reagents, quantitative and reproducible removal of the phospholipids was achieved. Using the finally selected lanthanum cation, the removal of phospholipids was achieved with minimal concomitant loss of the ten investigated analytes which were carefully selected to incorporate functional groups that could potentially interact with the added reagents and hence could be removed along with the phospholipids. Copyright © 2008 John Wiley & Sons, Ltd.