Isotopic analogs as internal standards for quantitative analyses by GUMS - evaluation of cross-contribution to ions designated for the analyte and the isotopic internal standard

Isotopic analogs of the analytes are currently preferred internal standards (IS) for quantitative analyses of drugs and their metabolites in biological matrices by GC/MS procedures. Contributions of the analyte and the IS to the intensities of ions designated for the IS and the analyte, respectively - an undesirable phenomenon termed "cross-contribution" - greatly weakens the effectiveness of this approach. The cross-contribution phenomenon has been, in the past, evaluated by a "direct measurement" approach, in which intensities of interested ions were measured in two separate experiments using equal quantities of the analyte and the IS. Alternate procedures that may generate improved results are hereby studied. For the "improved direct measurement" approach, ion intensity data derived from the previously reported direct measurement procedure are first normalized before being used to calculate the extent or cross-contribution. An "internal standard" approach is also developed, in which a set amount of a third compound is incorporated into these two separate experiments, thus allowing corrections of ion intensity data that are imbedded with variations inherent to separate experiments. Finally, a "standard addition" approach, involving a series "addition" of "standards", generates multiple data points, thus, providing a mechanism to validate the resulting cross-contribution data. Secobarbital/H-2(5)-secobarbital and secobarbital/C-13(4)-secobarbital pairs are adapted as the exemplar systems for this study. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.