A study has been made of a novel fluorescent and
phosphorescent label 6-coumarinsulphonyl chloride (6-CSCl)
based an the coumarin nucleus with a reactive group
similar to dansyl chloride. The synthesis, analysis,
derivatisation and application of the label to certain
analytical problems have been studied.
The label was obtained in a high state of purity by a
straightforward sulphonation of the counarin nucleus.
Characterisation of the label was carried out using mass
spectrometry, NMR and IR, its purity was verified by
elemental analysis. The label was found to be highly
stable and react at room temperature with phenolic and
The luminescence properties of the label and its
derivatives were investigated both at room temperature and
at 77K. The room temperature fluorescence showed a strong
pH dependence which was not observed at 77K.
Environmental effects on the fluorescence properties of
the derivatives at room temperature were investigated.
Linearity range, quantum yields and the detection limits
of the derivatives were determined.
The optimum derivatisation conditions for various
compounds were determined by HPLC. The analytical
applications of the label were investigated by studying
the behaviour of the derivatives in HPLC and TLC systems.
In the area of HPLC, both pre-c6lumn and post-column
derivatisation were attempted. The ion-pair pre-column
derivatisation technique was found to be more suitable for
TLC analysis was performed on both normal phase and
HPTLC plates. The separated components were detected
using absorbance and phosphorimetry. Synthesis of the
labels with substituents at the 4 and 7 position of the
coumarin nucleus were carried out. The behaviour of the
modified labels and their derivatives were compared with
the original label and its derivatives.
Doctoral Thesis. Submitted in partial fulfilment of the requirements for the award of Doctor of Philosophy of Loughborough University.