The temporal rise of the spectral bleaching of crystal violet (CV)
in alcohol solutions was studied using
femtosecond pump-probe spectroscopy with tunable pump pulses. From
the pump-wavelength dependence of transient
differential absorption (ΔOD) spectra, we concluded that CV has two
ground states. The isomerization rate, not
dependent on solvent viscosity, was ∼500 fs at 295 K. This shows
that the structural difference between the isomers
is not as large as a phenyl ring rotation, which has been regarded as
the origin of the ground-state isomers. We
proposed a ground-state model of CV, in which the molecular structure
of CV has D
3 or C
3
symmetry, based on
molecular orbital calculations. Our model explains not only our
findings but also almost all of the previous findings
that seemed to conflict with each other over the last 50
years.
We introduce a novel approach in highly selective and sensitive fluorescence derivatization of polyamines. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase high-performance liquid chromatography (HPLC). Polyamines, having two to four amino moieties in a molecule, were converted to the corresponding dipyrene- to tetrapyrene-labeled derivatives by reaction (100 degrees C, 20 min) with PSE. The derivatives afforded intramolecular excimer fluorescence (450-520 nm), which can clearly be discriminated from the monomer (normal) fluorescence (360-420 nm) emitted from PSE, its hydrolysate and monopyrene-labeled derivatives of monoamines. The structures of the derivatives were confirmed by HPLC with mass spectrometry, and the emission of excimer fluorescence could be proved by spectrofluorometry and time-resolved fluorometry. The PSE derivatives of four polyamines [putrescine (Put), cadaverine (Cad), spermidine (Spd), and spermine (Spm)] could be separated by reversed-phase HPLC on a C8 column with linear gradient elution. The detection limits (signal-to-noise ratio of 3) for the polyamines were 1 (Put), 1 (Cad), 5 (Spd), and 8 (Spm) fmol on the column. Furthermore, the present method was so selective that biogenic monoamines gave no peak in the chromatogram.
We previously confirmed the existence of two ground-state isomers of crystal violet (CV) in alcohol (Maruyama,
Y.; Ishikawa, M.; Satozono, H. J. Am. Chem. Soc.
1996, 118, 6257). These isomers are called “solvation
isomers” because they are differentiated by the solvation of alcohol. Here, to investigate if solvation isomers
are possible in triphenylmethane (TPM) dyes other than CV, we measured transient differential absorption
(ΔOD) spectra of two classes of TPM dyes: symmetric (ethyl violet and parafuchsin) and asymmetric
(malachite green and brilliant green) using a femtosecond spectral hole-burning technique. The symmetric
TPM dyes that we studied (including CV) showed two spectral holes in the early part of photobleaching,
whereas the asymmetric dyes showed three spectral holes. The ΔOD spectra of all of the TPM dyes depended
on the pump wavelength. This dependence is the key experimental observation that confirms the existence of
ground-state isomers, and thus the asymmetric TPM dyes have three ground-state isomers. Solvation isomers
are possible in TPM dyes in alcohol. To obtain insight into the deactivation mechanism of TPM dyes, we
examined a plateau region that appears in the early part of photobleaching recovery. The plateau is evidence
for the existence of an intermediate state between the lowest singlet excited state (S1) and the ground state.
The absence of the plateau only in parafuchsin, which is due to the difference in a positive charge distribution
on a nitrogen atom, suggests that the intermediate state is a distorted excited state in which a positive charge
is partially shifted to a nitrogen atom in an amino group.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.