Development of novel formulations for mucosal delivery of protein ...

3.2.1 Materials .................................................................................................................. 79
3.2.2 Structural elucidation of chitosan and TG-chitosan- 1 H-NMR ................................ 79
3.2.3 Quantification of immobilised thiol group .............................................................. 80
3.2.4 Molecular weight monitoring of chitosan and TG-chitosan by gel permeation
chromatography (GPC).................................................................................................... 80
3.2.5 ATR-FT-IR analysis ................................................................................................. 81
3.2.6 X-ray powder diffraction (XRPD) .......................................................................... 81
3.2.7 Differential Scanning Calorimetry (DSC) ............................................................... 82
3.2.8 Physical stability of model drugs by far-UV CD spectroscopy ............................... 82
3.2.9 Thermogravimetric analysis for moisture content .................................................. 82
3.2.10 Calibration curves for BSA and INS ..................................................................... 83
3.2.11 Drug loading capacity ........................................................................................... 83
3.2.12 Scanning electron microscopy (SEM) ................................................................... 83
3.2.13 Stability evaluation ................................................................................................ 84
3.2.13.1 Preparation and storage of BSA/INS xerogels for stability studies................ 84
γ.β.1γ.β Assay of BSA and INS content by Bradford’s assay ..................................... 85
3.2.13.3 HPLC assay of BSA and INS content. ........................................................... 85
3.3 RESULTS AND DISCUSSION .............................................................................................. 86
3.3.1 Structural elucidation of chitosan and TG-chitosan- 1 H-NMR ................................ 86
3.3.2 Quantification of immobilised thiol group .............................................................. 88
3.3.3 Molecular weight monitoring of chitosan and TG-chitosan - GPC ........................ 88
3.3.4 ATR-FT-IR analysis ................................................................................................. 89
3.3.5 X-ray powder diffraction (XRPD) ........................................................................... 93
3.3.6 Differential scanning calorimetry (DSC) ................................................................ 96
3.3.7 Physical stability of BSA and INS by far-UV CD spectroscopy .............................. 97
3.3.8 Thermogravimetric analysis - Moisture content ................................................... 100
3.3.9 Calibration curves for BSA and INS in 1% acetic acid, 0.01 PBS and 0.01 M HCl.
........................................................................................................................................ 101
3.3.10 Drug loading capacity ......................................................................................... 103
3.3.11 Scanning electron microscopy (SEM) ................................................................. 103
3.3.12 Stability evaluation .............................................................................................. 108
γ.γ.1β.1 BSA and INS content by Bradford’s assay .................................................. 108
3.3.12.2 HPLC determination of BSA and INS content. ............................................ 109
3.4 CONCLUSIONS ................................................................................................................ 113