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Transcript of Chapter No. Title Page No.shodhganga.inflibnet.ac.in/bitstream/10603/9096/2... · 9. 4C.1...
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Table of Contents
Chapter No. Title Page No.
1 INTRODUCTION 1-10
1.1 Introduction to topic
1.2 Biopharmaceutics Classification system
1.3 Importance of bioavailability
1.4 Basics about bioavailability and absorption
1.5 Problems due to low bioavailability
1.6 Methods for enhancing bioavailability
1.7 Aims and objectives
2 LITERATURE REVIEW 11-43
2.1 Reported methods for enhancing solubility and dissolution of the drug
2.2 Selection of drug
2.3 Drug profile
2.4 Drug characterization
2.5 Formulation patents related to drug
2.6 Selection of techniques for solubility enhancement
3 RESEARCH ENVISAGED 44
4 EXPERIMENTAL 45-108
4.1 List of equipment’s used during project
work
4.2 List of materials used during project work
4A Preformulation of drug
4B Development of analytical method
4B.1 Ultraviolet spectroscopic (UV) method development for drug release
4B.2 Development of high performance liquid chromatographic (HPLC) method for assay
4C Solubility enhancement with cyclodextrins
4D Solubility enhancement using “liquisolid compacts” technology
4E Solubility enhancement using self microemulsifying drug delivery systems
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Chapter No. Title Page No.
4F Solubility enhancement by preparation of nanoparticles
4F.1 Preparation of nanoparticles with supercritical Antisolvent precipitation
4F.2 Preparation of nanoparticles using ion gelation technique
4F.3 Preparation of nanoparticles using nanoencapsulation technique
4G Optimization of formulation
4H In –vivo studies
4H.1 Development and validation of analytical method for analysis of candesartan cilexitil in plasma (LC-MS/MS)
4H.2 In Vivo bioavailability of selected formulations in rats
4I Accelerated stability testing
5 RESULTS AND DISCUSSION 109-245
6 SUMMARY AND CONCLUSION 246-256
7 APPENDIX 257-258
8 REFERENCES 259-280
9 LIST OF RESEARCH OUTPUT 281
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Sr. No Table
No. Name of table
Page
No.
1. 1.1 GI Physiology and drug Absorption 8
2. 2.1 List of antihypertensive agents with their solubility and
bioavailability
22
3. 2.2 Characterization of solid forms of Candesartan cilexetil 36
4. 2.3 Crystal properties of Form I and Form II of
candesartan cilexetil
37
5. 2.4 Formulation patents related to drug 39-43
6. 4.1 List of equipments 45
7. 4.2 List of materials 46
8. 4B.1 Optimized chromatographic conditions for analysis of
candesartan cilexetil by HPLC method
50
9. 4C.1 Applications of cyclodextrins in marketed
pharmaceutical products
56
10. 4C.2 Effect of cyclodextrin complexation on various
classes of drugs in BCS classification
59
11. 4C.3 Formulations prepared with cyclodextrins 66
12. 4D.1 Excipients used for formulation of liquisolid compacts 69
13. 4D.2 Formulations with liquisolid technology 71
14. 4E.1 Example of surfactants, co-surfactant, and co-solvent
used in commercial formulations.
77
15. 4E.2 Excipients used for formulation of SMEDDS 84
16. 4E.3 Combinations for formulation of SMEDDS 85
17. 4F.1 Critical conditions for some solvents 89
18. 4F.2 Excipients used for formulation of nanoparticles by ion
gelation method
98
19. 4F.3 Formulation design for preparation of nanoparticles by
ion gelation method
98
20. 4F.4 Excipients used for formulation of nanoparticles by
nanoencapsulation method
101
21. 4F.5 Formulation design for preparation of nanoparticles by
nanoencapsulation method
102
22. 5A.1 Physicochemical characterization of drug 109
23. 5A.2 pH solubility profile of drug 112
24. 5A.3 Percentage cumulative release of drug in multimedia
dissolution
112
25. 5A.4 Percentage cumulative release of marketed formulation
in multimedia dissolution
113
26. 5B.1 Precision of UV method 118
27. 5B.2 Accuracy of UV method 119
28. 5B.3 System precision of HPLC method 120
LIST OF TABLES
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29. 5B.4 Linearity of HPLC method 124
30. 5B.5 Limit of detection and Limit of quantification of CC by
HPLC method
125
31. 5B.6 Accuracy of HPLC method 126
32. 5B.7 Precision of HPLC method 126
33. 5B.8 Robustness of HPLC method 127
34. 5C.1 Saturation solubility testing of drug-cyclodextrin
complexes
130
35. 5C.2 Drug content of drug-cyclodextrin complexes 131
36. 5C.3 Multimedia dissolution of F1 132
37. 5C.4 Multimedia dissolution of F2 132
38. 5C.5 Multimedia dissolution of F3 132
39. 5C.6 Multimedia dissolution of F4 133
40. 5C.7 Multimedia dissolution of F5 133
41. 5C.8 Multimedia dissolution of F6 133
42. 5C.9 Multimedia dissolution of F7 134
43. 5C.10 Multimedia dissolution of F8 134
44. 5C.11 Multimedia dissolution of F9 134
45. 5C.12 Multimedia dissolution of F10 135
46. 5C.13 Multimedia dissolution of F11 135
47. 5C.14 Multimedia dissolution of F12 135
48. 5C.15 Multimedia dissolution of F13 136
49. 5C.16 Multimedia dissolution of F14 136
50. 5C.17 Multimedia dissolution of F15 136
51. 5C.18 Multimedia dissolution of F16 137
52. 5C.19 Multimedia dissolution of F17 137
53. 5C.20 Multimedia dissolution of F18 137
54. 5C.21 Multimedia dissolution of F19 138
55. 5C.22 Multimedia dissolution of F20 138
56. 5C.23 Multimedia dissolution of F21 138
57. 5C.24 Multimedia dissolution of F22 139
58. 5C.25 Multimedia dissolution of F23 139
59. 5C.26 Multimedia dissolution of F24 139
60. 5C.27 Percentage cumulative release of drug-cyclodextrin
complexes in 0.1 N HCl
140
61. 5C.28 Percentage cumulative release of drug-cyclodextrin
complexes in phosphate buffer pH 6.8
141
62. 5C.29 Percentage cumulative release of drug-cyclodextrin
complexes in water
142
63. 5C.30 Percentage cumulative release of drug-cyclodextrin
complexes in OGD media
143
64. 5D.1 Incompatability testing of liquisolid excipients(Physical
observation)
166
65. 5D.2 Incompatability testing of liquisolid excipients (Assay) 166
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66. 5D.3 Incompatability testing of liquisolid excipients(FTIR) 167
67. 5D.4 Key formulation characteristics of liquisolid compacts of
CC with tween 20
167
68. 5D.5 Key formulation characteristics of liquisolid compacts of
CC with transcutol
168
69. 5D.6 Saturation solubility testing of liquisolid compacts 168
70. 5D.7 Drug content of liquisolid compact 169
71. 5D.8 Multimedia dissolution of Lst1 170
72. 5D.9 Multimedia dissolution of Lst2 170
73. 5D .10 Multimedia dissolution of Lst3 170
74. 5D .11 Multimedia dissolution of Lst4 171
75. 5D .12 Multimedia dissolution of Lst5 171
76. 5D .13 Multimedia dissolution of Lst6 171
77. 5D .14 Multimedia dissolution of Lst7 172
78. 5D .15 Multimedia dissolution of Lst8 172
79. 5D .16 Multimedia dissolution of Lst9 172
80. 5D .17 Multimedia dissolution of Lstc1 173
81. 5D .18 Multimedia dissolution of Lstc2 173
82. 5D .19 Multimedia dissolution of Lstc3 173
83. 5D .20 Multimedia dissolution of Lstc4 174
84. 5D .21 Multimedia dissolution of Lstc5 174
85. 5D .22 Multimedia dissolution of Lstc6 174
86. 5D .23 Multimedia dissolution of Lstc7 175
87. 5D .24 Multimedia dissolution of Lstc8 175
88. 5D.25 Multimedia dissolution of Lstc 9 175
89. 5D.26 Percentage cumulative release of liquisolid systems in
0.1N HCl
176
90. 5D.27 Percentage cumulative release of liquisolid systems in
phosphate buffer pH 6.8
177
91. 5D.28 Percentage cumulative release of liquisolid systems in
water
178
92. 5D.29 Percentage cumulative release of liquisolid systems in
OGD media
179
93. 5E.1 Solubility studies of CC in modified oils , surfactants
and co surfactants
184
94. 5E.2 Stability and visual observation of SMEDDS 185-
191
95. 5E.3 Selection of SMEDDS on the basis of particle size 192
96. 5E.4 Thermodynamic stability testing of SMEDDS 192
97. 5E.5 Final selected SMEDDS 192
98. 5E.6 Drug content of SMEDDS 195
99. 5E.7 Saturation solubility testing of SMEDDS 195
100. 5E.8 Multimedia dissolution of SMEDDS 1 196
101. 5E.9 Multimedia dissolution of SMEDDS 2 197
102. 5E.10 Multimedia dissolution of SMEDDS 3 198
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103. 5E.11 Multimedia dissolution of SMEDDS 4 199
104. 5E.12 Particle size distribution and polydispersity index of
SMEDDS
200
105. 5E.13 Effect of drug loading on particle size of SMEDDS. 201
106. 5F.1 Key formulation characteristics of nanoparticles with
antisolvent precipitation
204
107. 5F.2 Saturation solubility of nanoparticles with antisolvent
precipitation
204
108. 5F.3 Multimedia dissolution of nanoparticles with antisolvent
precipitation
205
109. 5F.4 Particle size distribution of nanoparticles with antisolvent
precipitation
207
110. 5F.5 Selection of nanoparticles using ion gelation technique 209
111. 5F.6 Evaluation of selected nanoparticle systems prepared by
ion-gelation technique
209
112. 5F.7 Particle size distribution of selected nanoparticle systems
prepared by ion-gelation technique
210
113. 5F.8 Saturation solubility of nanoparticles prepared by ion-
gelation technique
212
114. 5F.9 Multimedia dissolution of nanoparticles prepared by ion-
gelation technique
213
115. 5F.10 Selection of nanoparticles using nanoencpsulation
technique
215
116. 5F.11 Evaluation of selected nanoparticle systems prepared by
nanoencapsulation technique
215
117. 5F.12 Particle size distribution of selected nanoparticle systems
prepared by nanoencapsulation technique
216
118. 5F.13 Saturation solubility of nanoparticles prepared by
nanoencapsulation technique
218
119. 5F.14 Multimedia dissolution of nanoparticles prepared by
nanoencapsulation technique
219
120. 5G.1 Optimized formula of F16 221
121. 5G.2 Drug content and weight variation of optimized F16
formulation
221
122. 5G.3 Multimedia dissolution of optimized F16 formulation 221
123. 5G.4 Optimized formula of SMEDDS 222
124. 5G.5 Drug content and weight variation of optimized
SMEDDS formulation
222
125. 5G.6 Multimedia dissolution of optimized SMEDDS
formulation
223
126. 5H.1 Specificity for Candesartan by LCMS 226
127. 5H.2 Calibrant samples for candesartan 227
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128. 5H.3 Curve parameter summary for Candesartan 227
129. 5H.4 Between run precision and accuracy for candesartan 228
130. 5H.5 Within run precision and accuracy for candesartan 229
131. 5H.6 Percentage extraction yield 230
132. 5H.7 Long term stock solution stability for candesartan 231
133. 5H.8 Freeze thaw stability for candesartan 232
134. 5H.9 Long term stability in matrix of candesartan 233
135. 5H.10 Variation due to change in column 234
136. 5H.11 Variation due to change in analyst 234
137. 5H.12 Variation in days for analysis (Interday variation) 235
138. 5H.13 Descriptive statistics of pharmacokinetic parameters of
candesartan(marketed formulation)
236
139. 5H.14 Descriptive statistics of pharmacokinetic parameters of
candesartan(F16 formulation)
236
140. 5H.15 Descriptive statistics of pharmacokinetic parameters of
candesartan(SMEDDS formulation)
237
141. 5I.1 Drug content of stability batches of F16 240
142. 5I.2 Dissolution of stability batches of F16 241
143. 5I.3 Drug content of stability batches of SMEDDS 243
144. 5I.4 Dissolution of stability batches of SMEDDS 244
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Sr.
No
Figure
No.
Name of figure Page
No.
1. 1.1 Rate limiting steps in oral absorption 1
2. 1.2 Biopharmaceutical classification system of drugs 3
3. 1.3 ADME pattern of drugs 4
4. 1.4 Factors affecting bioavailability 5
5. 1.5 Barriers for the modification of lipophilic drugs 9
6. 2.1 Chemical structure of candesartan cilexetil 24
7. 2.2 Schematic diagram of solid forms of candesartan cilexetil 36
8. 2.3 Reported DSC spectra of different forms of candesartan cilexetil
36
9. 2.4 Reported XRDspectra of different forms of candesartan cilexetil.
37
10. 2.5 Reported 13C CP/MAS spectra of Form I and Form II of candesartan cilexetil
37
11. 2.6 Reported 13C CP/MAS spectra of amorphous form of candesartan cilexetil
38
12. 2.7 Reported IR spectra of different forms of candesartan cilexetil
38
13. 4C.1 Structures of α,β and γ Cyclodextrins 54
14. 4C.2 Phase solubility profiles and classification of complexes according to Higuchi and Connors.
59
15. 4F.1 Diagram of supercritical region 89
16. 4F.2 RESS technique 90
17. 4F.3 Precipitation with compressed fluid antisolvent 91
18. 4F.4 Schematic representation of Gas antisolvent or SAS laboratory scale apparatus
91
19. 5A.1 FTIR spectra of pure drug 109
20. 5A.2 XRD spectra of pure drug 110
21. 5A.3 DSC thermogram of pure drug 110
22. 5A.4 Microscopy of pure drug 111
23. 5A.5 PH solubility profile of pure drug 111
24. 5A.6 Saturation solubility of drug 112
25. 5A.7 Multimedia dissolution of drug 113
26. 5A.8 Multimedia dissolution of marketed formulation 114
27. 5B.1 UV spectra of Pure drug in acetonitrile 115
28. 5B.2 Linearity by UV of CC in acetonitrile 115
29. 5B. 3 Linearity by UV of CC in methanol 116
30. 5B. 4 Linearity by UV of CC in water 116
31. 5B. 5 Linearity by UV of CC in 0.1 N HCl 117
32. 5B.6 Linearity by UV of CC in phosphate buffer pH 6.8 117
List of Figures
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33. 5B.7 Linearity by UV of CC in OGD medium 118
34. 5B.8 Chromatogram of optimized conditions of HPLC 119
35. 5B.9 Chromatogram of acid degradation(1 hour) of CC 120
36. 5B.10 Chromatogram of acid degradation (15 minutes) of CC 121
37. 5B.11 Chromatogram of degradation in base (1 hour) of CC 121
38. 5B.12 Chromatogram of degradation in base (15 minutes) of CC 122
39. 5B.13 Chromatogram of degradation by hydrolysis of CC 122
40. 5B.14 Chromatogram of placebo interference (Cyclodextrins) of CC
123
41. 5B.15 Chromatogram of placebo interference (SMEDDS excipients) of CC
123
42. 5B.16 Chromatogram of degradation by oxidation of CC 124
43. 5B.17 Linearity by HPLC method 125
44. 5C.1 Phase solubility diagram with βCD 128
45. 5C.2 Phase solubility diagram with HPβCD 128
46. 5C.3 Phase solubility diagram with γCD 129
47. 5C.4 Saturation solubility of drug-cyclodextrin complexes 131
48. 5C.5 Percentage cumulative release of drug-cyclodextrin complexes in 0.1 N HCl
144
49. 5C.6 Percentage cumulative release of drug-cyclodextrin complexes in phosphate buffer pH 6.8
144
50. 5C.7 Percentage cumulative release of drug-cyclodextrin complexes in water
145
51. 5C.8 Percentage cumulative release of drug-cyclodextrin complexes in OGD media
145
52. 5C.9 FTIR spectra of βCD 146
53. 5C.10 FTIR spectra of HPβCD 146
54. 5C.11 FTIR spectra of γCD 146
55. 5C.12 FTIR spectra of F1 147
56. 5C.13 FTIR spectra of F2 147
57. 5C.14 FTIR spectra of F3 147
58. 5C.15 FTIR spectra of F4 148
59. 5C.16 FTIR spectra of F5 148
60. 5C.17 FTIR spectra of F6 148
61. 5C.18 FTIR spectra of F7 149
62. 5C.19 FTIR spectra of F8 149
63. 5C.20 FTIR spectra of F9 149
64. 5C.21 FTIR spectra of F10 150
65. 5C.22 FTIR spectra of F11 150
66. 5C.23 FTIR spectra of F12 150
67. 5C.24 FTIR spectra of F13 151
68. 5C.25 FTIR spectra of F14 151
69. 5C.26 FTIR spectra of F15 151
70. 5C.27 FTIR spectra of F16 152
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71. 5C.28 FTIR spectra of F17 152
72. 5C.29 FTIR spectra of F18 152
73. 5C.30 FTIR spectra of F19 153
74. 5C.31 FTIR spectra of F20 153
75. 5C.32 FTIR spectra of F21 153
76. 5C.33 FTIR spectra of F22 154
77. 5C.34 FTIR spectra of F23 154
78. 5C.35 FTIR spectra of F24 154
79. 5C.36 XRD spectra of F2 155
80. 5C.37 XRD spectra of F4 155
81. 5C.38 XRD spectra of F6 155
82. 5C.39 XRD spectra of F8 156
83. 5C.40 XRD spectra of F10 156
84. 5C.41 XRD spectra of F12 156
85. 5C.42 XRD spectra of F14 157
86. 5C.43 XRD spectra of F16 157
87. 5C.44 XRD spectra of F18 157
88. 5C.45 XRD spectra of F20 158
89. 5C.46 XRD spectra of F22 158
90. 5C.47 XRD spectra of F24 158
91. 5C.48 DSC thermogram of BCD 159
92. 5C.49 DSC thermogram of HPBCD 159
93. 5C.50 DSC thermogram of GCD 159
94. 5C.51 DSC thermogram of F2 160
95. 5C.52 DSC thermogram of F4 160
96. 5C.53 DSC thermogram of F6 160
97. 5C.54 DSC thermogram of F8 161
98. 5C.55 DSC thermogram of F10 161
99. 5C.56 DSC thermogram of F12 161
100. 5C.57 DSC thermogram of F14 162
101. 5C.58 DSC thermogram of F16 162
102. 5C.59 DSC thermogram of F18 162
103. 5C.60 DSC thermogram of F20 163
104. 5C.61 DSC thermogram of F22 163
105. 5C.62 DSC thermogram of F24 163
106. 5D.1 Saturation solubility testing of liquisolid compacts 169
107. 5D.2 Percentage cumulative release of liquisolid system in 0.1 N HCl
176
108. 5D.3 Percentage cumulative release of liquisolid system in phosphate buffer pH 6.8
177
109. 5D.4 Percentage cumulative release of liquisolid system in water
178
110. 5D.5 Percentage cumulative release of liquisolid system in OGD media
179
111. 5D.6 Reported XRD spectra of MCC(2θ at 15.5˚ and 22˚) 180
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112. 5D.7 XRD spectra of Lst1 180
113. 5D.8 XRD spectra of LStc1 180
114. 5D.9 Reported FTIR spectra of MCC 181
115. 5D.10 Reported FTIR spectra of silica 181
116. 5D.11 FTIR spectra of Lst1 182
117. 5D.12 FTIR spectra of Lstc1 182
118. 5E.1 Solubility of drug in SMEDDS excipients 184
119. 5E.2 Pseudoternary phase diagram of SMEDDS 1 193
120. 5E.3 Pseudoternary phase diagram of SMEDDS 2 193
121. 5E.4 Pseudoternary phase diagram of SMEDDS 3 194
122. 5E.5 Pseudoternary phase diagram of SMEDDS 4 194
123. 5E.6 Saturation solubility of SMEDDS 195
124. 5E.7 Multimedia dissolution of SMEDDS 1 196
125. 5E.8 Multimedia dissolution of SMEDDS 2 197
126. 5E.9 Multimedia dissolution of SMEDDS 3 198
127. 5E.10 Multimedia dissolution of SMEDDS 4 199
128. 5E.11 Intensity distribution curve of SMEDDS 1 200
129. 5E.12 Intensity distribution curve of SMEDDS 2 200
130. 5E.13 Intensity distribution curve of SMEDDS 3 201
131. 5E.14 Intensity distribution curve of SMEDDS 4 201
132. 5F.1 Saturation solubility of nanoparticles with antisolvent precipitation
204
133. 5F.2 Multimedia dissolution of nanoparticles with antisolvent precipitation
205
134. 5F.3 X-ray diffraction spectra of nanoparticles with antisolvent precipitation
206
135. 5F.4 FTIR spectra of nanoparticles with antisolvent precipitation 206
136. 5F.5 Intensity distribution curve of nanoparticles with antisolvent precipitation
207
137. 5F.6 Intensity distribution curve of S1 210
138. 5F.7 Intensity distribution curve of S5 210
139. 5F.8 Intensity distribution curve of S6 211
140. 5F.9 TEM images of Nanoparticles prepared by ion gelation technique
211
141. 5F.10 Saturation solubility of nanoparticles prepared by ion gelation technique
212
142. 5F.11 Multimedia dissolution of nanoparticles prepared by ion gelation technique
213
143. 5F.12 Intensity distribution curve of NE 3 216
144. 5F.13 Intensity distribution curve of NE 6 216
145. 5F.14 Intensity distribution curve of NE 9 217
146. 5F.15 TEM images of nanoparticles prepared by nanoencapsulation technique
217
147. 5F.16 Saturation solubility of nanoparticles prepared by 218
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nanoencapsulation technique
148. 5F.17 Multimedia dissolution of nanoparticles prepared by nanoencapsulation technique
219
149. 5G.1 Multimedia dissolution of optimized F16 formulation 222
150. 5G.2 Multimedia dissolution of optimized SMEDDS formulation
223
151. 5H.1 Representative LC-MS/MS spectra of drug (a) and internal standard(b)
225
152. 5H.2 Plasma concentration V/S time curve for marketed formulation
237
153. 5H.3 Plasma concentration V/S time curve for FD 16 formulation
237
154. 5H.4 Plasma concentration V/S time curve for SMEDDS formulation
238
155. 5H.5 Comparative data for Cmax of candesartan cilexetil 238
156. 5H.6 Comparative data for Tmax of candesartan cilexetil 238
157. 5H.7 Comparative data for AUC(0-t) of candesartan cilexetil 239
158. 5I.1 Dissolution of stability batches (real time) of F16 242
159. 5I.2 Dissolution of stability batches (accelerated) of F16 242
160. 5I.3 Dissolution of stability batches (real time) of SMEDDS 244
161. 5I.4 Dissolution of stability batches (accelerated) of SMEDDS 245
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LIST OF ACRONYMS
Sr.No. Term Acronym
1 Celcius C
2 Minutes min
3 Fourier transform Infra red FTIR
4 Differential Scanning Calorimeter DSC
5 Potassium bromide KBr
6 Kilo Voltage kV
7 mili Ampere mA
8 X-ray diffraction XRD
9 Hydrochloric acid HCl
10 Normal N
11 United states pharmacopoeia USP
12 Ultra violet UV
13 Over gastric dissolution OGD
14 Molar M
15 Mili gram mg
16 Mili litre ml
17 Parts per million ppm
18 Revolutions per minute rpm
19 Nano meter nm
20 High performance liquid
chromatography
HPLC
21 Relative standard deviation RSD
22 Mili molar mM
23 Candesartan cilexitil CC
24 Micro gram µg
25 Sodium hydroxide NaOH
26 Self microemulsifying drug delivery
system
SMEDDS
27 International conference on
hormonisation
ICH
28 Beta cyclodextrin βCD
29 Hydroxypropylated beta cyclodextrin HPβCD
30 Gamma cyclodextrin √CD
31 Cyclodextrin CD
32 Supercritical Fluid SCF
33 Supercritical fluid extraction SCFE
34 Supercritical anti solvent SAS
35 Supercritical carbon dioxide SC-CO2
36 Gram gm
37 Hour hr
38 Sodium tripolyphosphate STTP
39 High density polyethylene HDPE
40 Child resistant cap CRC
41 Concentration maximum Cmax
42 Time to reach concentration maximum Tmax
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43 Area under curve AUC
44 Liquid chromatographic mass
spectrophotometry mass
spectrophotometry
LC-MS/MS
45 Percent coefficient of variation %CV
46 Ethylene diamine tetra acetic acid EDTA
47 High quality control HQC
48 Low quality control LQC
49 Middle quality control MQC
50 Lower limit of quantification LLOQ
51 Upper limit of quantification ULOQ
52 Standard deviation SD
52 Certificate of analysis COA
53 Hour h