As iobitridol showed a more resolved spectrum in D2O, the aqueous system was further exploited with the purpose of improving signals resolution. In alkaline solution, the proton spectrum of iobitridol FP displayed a distinct signal at about 2.6 ppm accounting for one proton integral which was assigned to the single proton of the propylcarboxamino substituent of the benzene ring (proton 1, Fig. 1). The chemical shift of proton 1 showed dependence on the sodium hydroxide concentration: the more alkaline the solution the more shielded and resolved the proton was (Fig. 2). This trend may be attributable to the partial deprotonation of the adjacent amide group in alkali: the consequent local increase of electron density led to an upfield shift of the proton 1 signal [19]. A 1.4% (w/w) NaOH concentration in D2O represented a good compromise between a satisfactory separation of the selected proton from the neighboring signals of the N-methyl groups and the drawbacks of high alkalinity (potential active standard degradation and lengthening of the NMR pulse). 3,5-Dinitrobenzoic acid was selected as quantitative reference standard because of its deshielded NMR signals which assured no interferences with the iobitridol signals and with the strong water signal.