Assessment of the Quality of Doping Substances Identification in Urine by GC/MS/MS

Direct
control of doping in sports is based on the analysis of
active substances and/or their metabolites in urine samples of the
athletes by GC/MSn or LC/MSn. The World Anti-Doping
Agency, WADA, defined criteria for the agreement between retention
times, RT, or relative retention times, RRT, and abundance ratios,
AR, of characteristic ions of the mass spectrum of the analyte in
a calibrator (positive control) and the sample. Strict criteria for
confirming analyte presence were defined to reduce false positive
results rates, FP. However, these criteria can lead
to high rates of false negative results, FN. This
work presents a methodology to define statistically sound criteria
for the agreement between RRT and AR that allow keeping the FN under control. This work also determined the FP of identifications. The statistical criteria were set
from Monte Carlo simulations of correlated RT and ion abundances.
The simulation of AR and signal noise was also used to estimate the FN and FP of identifications based on the
criteria defined by WADA. The developed tools were successfully applied
to the control of nine doping substances in urine samples by GC/MS/MS.
The estimated FN were tested from independent experimental
tests proving estimates are accurate. The criteria defined by WADA
are associated with extremely low FP but, in some
cases, associated with FN much larger than 50%. The
statistically sound identification criteria allow a more convenient
balance between FN and FP. The user-friendly
spreadsheet used in this work is made available as Supporting Information.