Methodology for the development of a drug library based upon collision-induced fragmentation for the identification of toxicologically relevant drugs in plasma samples

The possibility of creating a robust mass spectral library with use of high-performance liquid chromatography-atmospheric pressure-electrospray ionization (HPLC-AP-ESI) for the identification of drugs misused in cases of clinical toxicology has been examined. Factors reported as influencing the fragmentation induced by "source transport region collision induced dissociation" (CID) have been tested in this study (i.e. solvent, pH, different acids or buffer salts and their concentration, different organic modifiers and the modifier concentration). The tests performed on a few "model drugs" were analysed with use of two different single quadrupole instruments. The large number of mass spectra obtained appears to be affected by the mobile phase conditions to only a minor extent. This also holds for the mass spectra obtained at two different instruments (laboratories). Subsequently breakdown curves have been measured for about 20 randomly chosen drugs by variation of the kinetic energy of their ions in the CID zone through changing the fragmenter voltage. These breakdown curves were used to optimize the fragmenter voltage for each drug. The optimized fragmenter voltages were then applied by use of a variably ramped fragmenter voltage to acquire mass spectra for the library. The chromatographic separations were run on a Zorbax Stable bond column using a 10-mM ammonium formate-acetonitrile gradient method. Spiked blank serum and patient samples with a total of 40 different drugs were extracted with use of a standard basic liquid-liquid extraction (LLE) method. A search of significant peaks in the chromatogram by application of the developed mass spectral library is shown to result in a more than 95% positive identification. (C) 2001 Elsevier Science B.V. All rights reserved.