LIST OF TABLES

Table No. Title Page No.

5.1 Phase Wise Distribution of Work 17 - 18
6.1 Solvent Used for Fractionation 20
6.2 Analytical Markers from Extracts for Analytical Method Development 21
6.3 Solvent System for TLC Development of Extracts 22
6.4 HPLC Chromatographic Conditions 27
6.5 Translation of Coded Levels in Actual Units for Cream Base 37
6.6 The 32 Full Factorial Design Layout for Topical Cream Base 37
6.7 Composition Formula for Topical Cream 38
Translation of Coded Levels in Actual Units for Topical Transdermal
6.8 41
Patch Base
6.9 The 32 Full Factorial Design Layout for Topical Transdermal Patch Base 41
6.10 Composition Formula for Topical Transdermal Patch 42
Translation of Coded Levels in Actual Units for Topical Film Forming
6.11 44
Solution
6.12 The 32 Full Factorial Design Layout for Topical Film Forming Solution 44
6.13 Composition Formula for Topical Film Forming Solution 45
6.14 Treatment Protocol for Skin Irritancy Study 47
6.15 Draize Scoring Method 47
6.16 Protocol for Anti-Inflammatory Study 49
6.17 Solvent System for Analytical Method Development using HPTLC 51
6.18 Protocol for Accelerated Stability Study 56
7.1 Preliminary Investigations of Extracts 57
7.2 Phytochemical Screening of Extracts 58
7.3 Physicochemical Evaluation of Extracts 58
7.4 Fractionation of Extracts 59
7.5 Analytical Markers from Extracts for Analytical Method Development 59
7.6 Details of TLC Development for Extracts 60
 LIST OF TABLES

Table No. Title Page No.

7.7 Particulars for Isolation of Phytoconstituents 61
7.8 Results for HPLC Method Development and Validation 65
7.9 Quantification of Analytical & Chemical Markers from Extractsby HPTLC 68
7.10 Particulars for PHPTLC 69
7.11 Preliminary Investigations of Isolated compound 70
7.12 UV Visible Spectral Details for Curcumin 71
7.13 IR Spectral Details for Curcumin 71
7.14 NMR Spectral Details for Curcumin 72
7.15 LCMS Spectral Details for Curcumin 72
7.16 UV Visible Spectral Details for Quercetin 73
7.17 IR Spectral Details for Quercetin 73
7.18 NMR Spectral Details for Quercetin 74
7.19 LCMS Spectral Details for Quercetin 74
7.20 UV Visible Spectral Details for Ellagic acid 75
7.21 IR Spectral Details for Ellagic acid 75
7.22 NMR Spectral Details for Ellagic acid 76
7.23 LCMS Spectral Details for Ellagic acid 76
7.24 UV Visible Spectral Details for CW-1 77
7.25 IR Spectral Details for CW-1 77
7.26 NMR Spectral Details for CW-1 78
7.27 LCMS Spectral Details for CW-1 78
7.28 UV Visible Spectral Details for CW-2 79
7.29 IR Spectral Details for CW-2 79
7.30 NMR Spectral Details for CW-2 80
7.31 LCMS Spectral Details for CW-2 80
7.32 UV Visible Spectral Details for Diosmin 81
7.33 IR Spectral Details for Diosmin 81
 LIST OF TABLES

Table No. Title Page No.

7.34 NMR Spectral Details for Diosmin 82
7.35 LCMS Spectral Details for Diosmin 82
7.36 UV Visible Spectral Details for Rutin 83
7.37 IR Spectral Details for Rutin 83
7.38 NMR Spectral Details for Rutin 84
7.39 LCMS Spectral Details for Rutin 84
7.40 UV Visible Spectral Details for AB-1 85
7.41 IR Spectral Details for AB-1 85
7.42 NMR Spectral Details for AB-1 86
7.43 LCMS Spectral Details for AB-1 86
7.44 UV Visible Spectral Details for AB-2 87
7.45 IR Spectral Details for AB-2 87
7.46 NMR Spectral Details for AB-2 88
7.47 LCMS Spectral Details for AB-2 88
7.48 Structural Details for Analytical and Chemical Markers 89 - 90
7.49 Molecular Docking Details for Analytical and Chemical Markers 91
7.50 Design for Optimization Topical Cream Base 95
7.51 Analysis of Variance for Topical Cream Base 95
7.52 Solutions for 9 Combinations of Categoric Factor Levels for Cream Base 96
7.53 Designs for Optimization Topical Transdermal Patch 97
7.54 Analysis of Variance for Topical Transdermal Patch 97
Solutions for 9 Combinations of Categoric Factor Levels for Topical
7.55 98
Transdermal Patch
7.56 Designs for Optimization Topical Film Forming Solutions 99
7.57 Analysis of Variance for Topical Film Forming Solutions 99
Solutions for 9 Combinations of Categoric Factor Levels for Topical Film
7.58 100
Forming solutions
 LIST OF TABLES

Table No. Title Page No.

7.59 Evaluation of Prepared Topical Cream Formulations 101
7.60 Evaluation of Prepared Topical Transdermal Patch 102
7.61 Evaluation of Prepared Topical Film Forming Solutions 103
7.62 Skin Irritation Test Rendering to Draize Scoring Method 105
7.63 Anti-Inflammatory Screening for Topical Cream 106
7.64 Anti-inflammatory Screening for Topical Transdermal Patch 107
7.65 Anti-inflammatory Screening for Topical Film Forming Solution 108
Quantification of Extracts, Analytical Markers and Chemical Markers in
7.66 116
Formulation by HPTLC
Quantification of Analytical Markers from Extracts by UV Visible
7.67 118
Spectroscopy
Quantification of Extracts, Analytical Markers and Chemical Markers in
7.68 119
Formulation by UV Visible Spectroscopy
7.69 Drug Release Kinetics. 122
7.70 Evaluation of Prepared Topical Cream Formulations (01 Month) 123
7.71 Evaluation of Prepared Topical Cream Formulations (02 Month) 124
7.72 Evaluation of Prepared Topical Cream Formulations (03 Month) 125
7.73 Evaluation of Prepared Topical Cream Formulations (06 Month) 126
7.74 Evaluation of Prepared Topical Transdermal Patch (01 Month) 127
7.75 Evaluation of Prepared Topical Transdermal Patch (02 Month) 128
7.76 Evaluation of Prepared Topical Transdermal Patch (03 Month) 129
7.77 Evaluation of Prepared Topical Transdermal Patch (06 Month) 130
7.78 Evaluation of Prepared Topical Film Forming Solutions (01 Month) 131
7.79 Evaluation of Prepared Topical Film Forming Solutions (02 Month) 132
7.80 Evaluation of Prepared Topical Film Forming Solutions (03 Month) 133
7.81 Evaluation of Prepared Topical Film Forming Solutions (06 Month) 134
 LIST OF FIGURES

Figure No. Title Page No.

4.1 Formulation of Research Aim and Objectives 15
5.1 Flow Chart of Research Envisaged 16
6.1 Setting image for Molecular Docking 34
7.1 Herbal Extracts 57
7.2 Analytical Markers of Herbal Extracts 59
7.3 HPLC Chromatogram of Standard Curcumin Solution 62
HPLC Chromatogram of Sample Solution {Curcuma longa Extracts
7.4 62
(CME)}
7.5 HPLC Chromatogram of Standard Ellagic acid Solution 63
HPLC Chromatogram of Sample Solution {Epilobium angustifolium
7.6 63
Extract (EAE)}
7.7 HPLC Chromatogram of Standard Rutin Solution 64
7.8 HPLC Chromatogram of Sample Solution {Agathosma betulina (ABE)} 64
HPTLC Image and Scanning Graph for CME, Curcumin and Isolated
7.9 66
Curcumin Fraction
HPTLC Image and Scanning Graph for EAE, Analytical Markers and
7.10 66
Elutes (Chemical Markers)
HPTLC Image and Scanning Graph for ABE, Analytical Markers and
7.11 67
Elutes (Chemical Markers)
HPTLC Image and Scanning Graph for MEX, Extracts, Analytical
7.12 67
Markers and Elutes (Chemical Markers)
7.13 UV Visible Spectra for Curcumin 71
7.14 IR Spectra for Curcumin 71
7.15 NMR Spectra for Curcumin 72
7.16 LCMS Spectra for Curcumin 72
7.17 UV Visible Spectra for Quercetin 73
7.18 IR Spectra for Quercetin 73
 LIST OF FIGURES

Figure No. Title Page No.

7.19 NMR Spectra for Quercetin 74
7.20 LCMS Spectra for Quercetin 74
7.21 UV Visible Spectra for Ellagic acid 75
7.22 IR Spectra for Ellagic acid 75
7.23 NMR Spectra for Ellagic acid 76
7.24 LCMS Spectra for Ellagic acid 76
7.25 UV Visible Spectra for CW-1 77
7.26 IR Spectra for CW-1 77
7.27 NMR Spectra for CW-1 78
7.28 LCMS Spectra for CW-1 78
7.29 UV Visible Spectra for CW-2 79
7.30 IR Spectra for CW-2 79
7.31 NMR Spectra for CW-2 80
7.32 LCMS Spectra for CW-2 80
7.33 UV Visible Spectra for Diosmin 81
7.34 IR Spectra for Diosmin 81
7.35 NMR Spectra for Diosmin 82
7.36 LCMS Spectra for Diosmin 82
7.37 UV Visible Spectra for Rutin 83
7.38 IR Spectra for Rutin 83
7.39 NMR Spectra for Rutin 84
7.40 LCMS Spectra for Rutin 84
7.41 UV Visible Spectra for AB-1 85
7.42 IR Spectra for AB-1 85
7.43 NMR Spectra for AB-1 86
7.44 LCMS Spectra for AB-1 86
7.45 UV Visible Spectra for AB-2 87
 LIST OF FIGURES

Figure No. Title Page No.

7.46 IR Spectra for AB-2 87
7.47 NMR Spectra for AB-2 88
7.48 LCMS Spectra for AB-2 88
7. 49 (A) Molecular Docking Details for Analytical and Chemical Markers 92
7. 49 (B) Molecular Docking Details for Analytical and Chemical Markers 93
7. 49 (C) Molecular Docking Details for Analytical and Chemical Markers 94
7.50 3D Surface Response for Topical Cream Base 96
7.51 3D Surface Response for Topical Transdermal Patch 98
7.52 3D Surface Response for Topical Film Forming Solutions 100
7.53 Prepared Topical Cream Formulations 101
7.54 Prepared Topical Transdermal Patch 102
7.55 Prepared Topical Film Forming Solutions 103
7.56 Skin Irritation Study 104-105
HPTLC image and Scanning Graph for CMEC, CME and Analytical
7.57 109
Markers

7.58 HPTLC image and Scanning Graph for EAEC, EAE, Analytical Markers 110
and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for ABEC, ABE, Analytical Markers
7.59 110
and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for MEXC, CME, EAE, ABE,
7.60 111
Analytical Markers and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for CMEP, CME and Analytical
7.61 112
Markers
HPTLC image and Scanning Graph for EAEP, EAE, Analytical Markers
7.62 112
and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for ABEP, ABE, Analytical Markers
7.63 113
and Elutes (Chemical Markers)
 LIST OF FIGURES

Figure No. Title Page No.

HPTLC image and Scanning Graph for MEXP, CME, EAE, ABE,
7.64 113
Analytical Markers and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for CMEF, CME and Analytical
7.65 114
Markers
HPTLC image and Scanning Graph for EAEF, EAE, Analytical Markers
7.66 114
and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for ABEF, ABE, Analytical Markers
7.67 115
and Elutes (Chemical Markers)
HPTLC image and Scanning Graph for MEXF, CME, EAE, ABE,
7.68 115
Analytical Markers and Elutes (Chemical Markers)
 LIST OF GRAPHS

Graph No. Title Page No.

7.1 In vitro Drug Release Pattern for Topical Creams 120
7.2 In vitro Drug Release Pattern for Topical Transdermal Patches 120
7.3 In vitro Drug Release Pattern for Topical Film Forming Solutions 121
 ABBREVIATIONS

 ABE : Agathosma betulina extract

 ABEC : Agathosma betulina extract Cream
 ABEF : Agathosma betulina extract Topical Film Forming Solution
 ABEP : Agathosma betulina extract Transdermal Patch
 Abs : Absorbance
 CCT : Caprylic/Capric Triglyceride
 CME : Curcuma longa Extract
 CMEC : Curcuma longa Extract Cream
 CMEF : Curcuma longa extract Topical Film Forming Solution
 CMEP : Curcuma longa extract Transdermal Patch
 COA : Certificate of Analysis
 COX-1 : Cyclooxygenase-I
 COX-II : Cyclooxygenase-II
 EAE : Epilobium angustifolium Extract
 EAEC : Epilobium angustifolium Extract Cream
 EAEF : Epilobium angustifolium Extract Topical Film Forming Solution
 EAEP : Epilobium angustifolium Extract Transdermal Patch
 EW : Emulsifying Wax
 FTIR : Fourier Transform Infra-Red
 HPLC : High Performance Liquid Chromatography
 HPMC : Hydroxy Propyl Methyl Cellulose
 HPTLC : High Performance Thin Layer Chromatography
 ICH : International Council for Harmonisation
 LCMS : Liquid Chromatography Mass Spectroscopy
 MEXC : Cream of Mixture of Curcuma longa Extract, Epilobium
angustifolium Extract and Agathosma betulina Extracts
 MEXF : Topical Film Forming Solution of Mixture of Curcuma longa
Extract, Epilobium angustifolium Extract and Agathosma
Betulina Extracts
 ABBREVIATIONS

 MEXP : Transdermal Patch of Mixture of Curcuma longa extract,

Epilobium angustifolium extract and Agathosma betulina Extracts
 NMR : Nuclear Magnetic Resonance
 PEG : Polyethylene Glycol
 PG : Propylene Glycol
 PTLC : Preparative Thin Layer Chromatography
 PVP : Polyvinyl Povidone
 PVP-K30 : Polyvinylpyrrolidone K 30
 QA : Quality Assurance
 QbD : Quality By Design
 QC : Quality Control
 RSD : Relative Standard Deviation
 SEM : Standard Error Mean
 TDDS : Transdermal Drug Delivery System
 TFFS : Topical Film Forming Solution
 TLC : Thin Layer Chromatography
 TNF-A : Tumor Necrosis Factor -Alpha
 UV : Ultra Violet
 v/v : Volume / Volume
 w/w : Weight / Weight
 WHO : World Health Organization