GC/MS Determination of N-butyl-N-(3-carboxypropyl) Nitrosamine (BCPN) in Bladder Cancers: The Skewed Molecular Interaction Caused Retention Time Shift

N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) has been widely used in rodents as an invaluable experimental tool for investigation of bladder cancer (BCA). The urinary level of its metabolite, N-butyl-N-(3-carboxypropyl) nitrosamine (BCPN) was reported to be a very reliable predicative parameter of BCA. However, in determination of the urinary BCPN we found the retention time (tR) of BCPN was randomly damping. The tR values of the authentic BCPN at 5, 10, 20, 50, and 100 ppm were 28.48, 27.59, 27.43, 28.00, and 28.32 min comparing with 28.23 min of the urinary BCPN in HPLC analysis, similarly, 17.30 min for the urinary and the 18.00 min for the authentic BCPN in GC/MS analysis. To interpret such a damping, we theoretically proposed that a certain transient skewed molecular interaction could occur during the chromatographic separation, which would cause a certain degree of fluctuation on the tR of target molecules. Conclusively, the retention time of a chemical is not a definite value as often considered in HPLC and GC/MS analyses. In reality it fluctuates depending mainly upon the interaction among a cluster of coexisting molecules, in particular, when operated at higher concentrations.