Negative ion electrospray high-performance liquid chromatography-mass spectrometry method development for determination of a highly polar phosphonic acid/sulfonic acid compound in plasma. Optimization of ammonium acetate concentration and in-source collision-induced dissociation.

A method, based on negative ion electrospray ionization (ESI) single-stage mass spectrometry coupled with HPLC, was developed for the determination of a squalene synthase inhibitor, BMS-187745, in human plasma. BMS-187745, a highly polar compound with both phosphonic acid and sulfonic acid groups, presented difficulties in developing plasma extraction and HPLC procedures. Precipitation of the plasma protein with methanol was finally chosen as the basis for sample preparation since extraction with water-immiscible solvents or with solid-phase extraction columns failed. It was essential to add ammonium acetate to the HPLC mobile phase, not only to enhance the retention of BMS-187745 but also to ensure a well-shaped chromatographic peak. While the use of ammonium acetate had the desired chromatographic effects, it had the undesirable consequence of suppressing the negative ion ESI signal. With the plasma extracts, the [M-H2O-H]- ion (m/z 367) showed significantly lower chemical noise than the [M-H]- ion (m/z 385), and was thus chosen as the analytical ion for the selected ion monitoring. The signal of the m/z 367 ion was significantly enhanced by the optimization of the in-source collision-induced dissociation (CID) of m/z 385 to m/z 367.