(a) pH
dependence of the molar absorbance at 20 °C of hydrodabcyl
isopropylamide (4) in an aqueous mixture of buffers CAPS,
CHES, HEPES, MES, and ammonium acetate, each present at 15 mM and
(b) protonation states. The gray arrow represents the experimental
acidification of the solution. The red dashed line indicates the putative
pH range of the protonation of the phenolate, and the red asterisk
indicates the corresponding spectral effect, that is, the isosbestic
point. The blue dashed line indicates the putative pH range of the
protonation of the diazo group, and the corresponding spectral effects
are indicated by the blue arrow showing the appearance of the local
maximum at 350 nm and the blue asterisk showing the shoulder at around
500 nm visible below pH 2.6.