method development and validation of newer analytical methods as per ich guidelines

330 Pages·2013·25.24 MB·English

METHOD DEVELOPMENT AND VALIDATION OF NEWER
ANALYTICAL METHODS AS PER ICH GUIDELINES

Thesis Submitted to
THE TAMILNADU DR. M.G.R. MEDICAL UNIVERSITY
Chennai-600 032
As a partial fulfillment of the requirement for the award of Degree of

DOCTOR OF PHILOSOPHY
(Faculty of Pharmacy)

Submitted by
Mrs.B. UMADEVI, M.Pharm.

Under the Guidance of
Prof.Dr.T. VETRICHELVAN, M.Pharm., Ph.D.
PRINCIPAL

ADHIPARASAKTHI COLLEGE OF PHARMACY
(Accredited by “NAAC” with CGPA of 2.74 on a four point scale at “B” Grade)
MELMARUVATHUR-603 319, TAMILNADU, INDIA.

SEPTEMBER-2013
DECLARATION

I hereby declare that the thesis entitled “METHOD DEVELOPMENT AND
VALIDATION OF NEWER ANALYTICAL METHODS AS PER ICH
GUIDELINES” submitted by me for the degree of DOCTOR OF PHILOSOPHY
(Faculty of Pharmacy), is a record of research work carried out by me during the
period from 2008-2013, under the guidance of Prof.Dr.T. VETRICHELVAN,
M.Pharm., Ph.D., Head, Department of Pharmaceutical Analysis and Principal,
Adhiparasakthi College of Pharmacy, Melmaruvathur- 603 319, TamilNadu, India
and has not formed the basis for the award of any degree, diploma, associateship,
fellowship, title in this or any other university or similar institute of higher learning.

Place: Melmaruvathur (B. UMADEVI)
Date: 05.09.2013
CERTIFICATE

This is to certify that the thesis entitled “METHOD DEVELOPMENT AND
VALIDATION OF NEWER ANALYTICAL METHODS AS PER ICH
GUIDELINES” submitted to The TamilNadu Dr. M.G.R. Medical University,
Guindy, Chennai- 600 032, Tamil Nadu, India as a partial requirements for the award
of Degree of DOCTOR OF PHILOSOPHY (Faculty of Pharmacy) is a record of
research work done by Mrs.B. UMADEVI, M.Pharm., during 2008-2013 under my
guidance and supervision at Adhiparasakthi College of Pharmacy, Melmaruvathur-
603 319, Tamil Nadu, India and that the thesis has not formed the basis for the award
of any other degree, diploma, associateship, fellowship, or any other similar title to
the candidate and the thesis represents independent work of the candidate.

Prof.Dr. T.VETRICHELVAN, M.Pharm., Ph.D.,
Place: Melmaruvathur
Head, Department of Pharmaceutical Analysis
Date: 05.09.2013
and Principal,

Adhiparasakthi College of Pharmacy,

Melmaruvathur- 603 319,
Tamil Nadu.
ACKNOWLEDGEMENT
I honestly acknowledge HIS HOLINESS “ARULTHIRU AMMA” for his ever-
growing blessings in each step of the study. I am grateful to Thirumathi Lakshmi
Bangaru Adigalar, Vice President, ACMEC Trust, Melmaruvathur, for having given
me an opportunity and encouragement all the way in completing the study.
I extend my thanks to Dr.T.Ramesh, M.D., Managing Director, MAPIMS,
Melmaruvathur, for providing all the necessary facilities to carry out this work.
I feel immensely delighted expressing my copious sincere thanks from the core of my
heart and deep sense of my indelible gratitude to my guide Prof.Dr.T.Vetrichelvan,
M.Pharm., Ph.D., Principal and Head, Department of Pharmaceutical Analysis,
Adhiparasakthi College of Pharmacy, Melmaruvathur, for the active guidance,
innovative ideas, creative works, indulgement and enthusiastic guidance, valuable
suggestions, a source of inspiration where the real treasure of my work.
I owe my sincere thanks with heartful pleasure to my Doctoral Committee members
Dr.P.Perumal, Professor, JKK Nataraja College of Pharmacy, Kumarapalayam and
Dr.G.Krishnamoorthy, Periyar College of Pharmaceutical Sciences for Girls,
Trichy. Without their encouragement and moral support it would have been absolutely
impossible to bring out the work in this manner.
My heartful thanks to Mrs.G.Shankari, M.Pharm., Central Instrumentantion in-
charge and Miss.R.Radha, M.Pharm., Central Instrumentation in-charge for their
persuasive support to complete this work.
My sincere thanks to Mrs.S.Karpagavalli, Lab in-charge and Mr.M.Gomathi
Shankar, Lab in-charge for their kind help throughout this work.
I am indeed thanks to the Librarian Mr.M.Suresh, M.L.I.S., for providing all
reference books and to make this project a great success.
I wish to thank Shine Pharmaceuticals Pvt. Ltd., Chennai, Centaur
Pharmaceuticals Pvt. Ltd., Mumbai, CaplinPoint Laboratories, Pvt. Ltd.,
Pondicherry and Apex Pharmaceuticals, Pvt. Ltd., Chennai for providing raw
materials for carrying out my project work.
Finally I take this opportunity to express my gratitude to all the people involved
directly or indirectly in the successful completion of this dissertation work.

(UMADEVI. B)
Dedicated to
My
Beloved Parents
and
My Family
CONTENTS
SECTION TITLE PAGE
NO
1. INTRODUCTION 1-11
1.1 Introduction to Analytical Chemistry 1
1.2 Why Validation Analytical Procedure 1
1.3 Process of Analytical Method Validation 2
1.4 ICH Guidelines for Analytical Method Validation 2
1.5 System Suitability test 6
1.6 Ultraviolet Spectroscopy 8
1.7 High Performance Liquid Chromatography 10
2. AIM AND OBJECTIVES OF THE STUDY 12
3. LITERATURE REVIEW 13-82
3.1 Drug Profile 13
3.1.1 Drug Profile for Doxofylline 13
3.1.2 Drug Profile for Ambroxol Hydrochloride 15
3.1.3 Drug Profile for Metolazone 18
3.1.4 Drug Profile for Spironolactone 21
3.1.5 Drug Profile for Metoprolol 24
3.1.6 Drug Profile for Olmesartan Medoxomil 27
3.1.7 Drug Profile for Aspirin 30
3.1.8 Drug Profile for Rosuvastatin Calcium 35
3.2 Reported Methods 39
3.2.1 Reported methods of Doxofylline 39
3.2.2 Reported methods of Ambroxol Hydrochloride 45
3.2.3 Reported methods of Metolazone 56
3.2.4 Reported methods of Spironolactone 59
3.2.5 Reported methods of Metoprolol 64
3.2.6 Reported methods of Olmesartan Medoximil 68
3.2.7 Reported methods of Aspirin 73
3.2.8 Reported methods of Rosuvastatin 76
4. SCOPE AND PLAN OF WORK 83
5. MATERIALS AND METHODS 84-113
5.1 Instrument Specification 84
5.2 Reagents and chemicals used in the study 86
5.3 Materials and methods for Doxofylline and Ambroxol 87
Hydrochloride combination dosage form
5.3.1 UV Spectrophotometric Methods 88
5.3.2 High Performance Liquid Chromatographic Method 92
5.4 Materials and methods for Metolazone and Spironolactone 95
combination dosage form
5.4.1 UV Spectrophotometric Methods 96
5.4.2 High Performance Liquid Chromatographic Method 101
5.5 Materials and methods for Metoprolol and Olmesartan 104
Medoximil combination dosage form
5.6 Materials and methods for Aspirin and Rosuvastatin 109
combination dosage form
6. RESULTS AND DISCUSSION 114-152
6.1 UV- Spectroscopic Methods for Doxofylline and Ambroxol 114
Hydrochloride combination dosage form
6.1.1. Simultaneous Equation Method 115
6.1.2. Absorbance Correction Method 118
6.1.3. Absorbance Ratio Method 120
6.2 Reverse Phase High Performance Liquid Chromatography 121
Method for Doxofylline and Ambroxol Hydrochloride
combination dosage form
6.3 UV Spectroscopic Methods for Metolazone and 125
Spironolactone combination dosage form
6.3.1. Simultaneous Equation Method 126
6.3.2. Absorbance Correction Method 128
6.3.3. Derivative Spectroscopic Method 131
6.4 Reverse Phase High Performance Liquid Chromatography 133
Method for Metolazone and Spironolactone combination dosage
form
6.5 UV Spectroscopic methods for Metoprolol and Olmesartan 136
combination dosage form.
6.5.1 Simultaneous Equation Method 137
6.5.2 Area Under the Curve Method 141
6.5.3 Derivative Spectroscopic Method 144
6.6 UV Spectroscopic methods for Aspirin and Rosuvastatin 148
combination dosage form.
6.6.1 Simultaneous Equation Method 149
6.6.2 Absorbance Ratio Method 151
7. SUMMARY AND CONCLUSION 153-163
7.1. UV Spectroscopic methods for Doxofylline and Ambroxol 153
Hydrochloride
7.2. Reverse Phase High Performance Liquid Chromatography 155
method for Doxofylline and Ambroxol Hydrochloride
7.3. UV Spectroscopic methods for Metolazone and 156
Spironolactone
7.4. Reverse Phase High Performance Liquid Chromatography 157
method for Metolazone and Spironolactone
7.5. UV Spectroscopic methods for Metoprolol and Olmesartan 159
7.6 UV Spectroscopic methods for Aspirin and Rosuvastatin 162
8. IMPACT OF THE STUDY 164
9. APPENDIX
List of figures
List of tables
Copies of research articles
10. BIBLIOGRAPHY
LIST OF FIGURES
FIGURE SUBJECT
NO.
1 UV spectrum of Doxofylline in distilled water
2 UV spectrum of Ambroxol Hydrochloride in distilled water
3 Overlaid spectrum of Doxofylline and Ambroxol Hydrochloride in distilled
water
4 Calibration curve of Doxofylline in distilled water at 244.5 nm
5 Calibration curve of Doxofylline in distilled water at 274 nm
6 Calibration curve of Ambroxol Hydrochloride in distilled water at 244.5 nm
7 Calibration curve of Ambroxol Hydrochloride in distilled water at 274 nm
8 Overlaid spectrum of Doxofylline and Ambroxol Hydrochloride (Absorbance
Correction Method)
9 Calibration curve of Ambroxol Hydrochloride in distilled water at 308 nm
10 Overlaid spectrum of Doxofylline and Ambroxol Hydrochloride
(Absorbance Ratio Method)
11 Calibration curve of Doxofylline in distilled water at 233.5 nm
12 Calibration curve of Ambroxol Hydrochloride in distilled water at 233.5 nm
13 Linearity chromatogram of Doxofylline and Ambroxol Hydrochloride
(7, 0.5 µg ml-1)- First Set [1/3]
14 Linearity chromatogram of Doxofylline and Ambroxol Hydrochloride
(14, 1 µg ml-1)- First Set [1/3]
15 Linearity chromatogram of Doxofylline and Ambroxol Hydrochloride
(21, 1.5 µg ml-1)- First Set [1/3]
16 Linearity chromatogram of Doxofylline and Ambroxol Hydrochloride
(28, 2 µg ml-1)- First Set [1/3]
17 Linearity chromatogram of Doxofylline and Ambroxol Hydrochloride
(35, 2.5 µg ml-1)- First Set [1/3]
18 Calibration curve of Doxofylline by RP-HPLC
19 Calibration curve of Ambroxol Hydrochloride by RP-HPLC
20 Chromatogram for analysis of formulation [SYNASMA-AX] Repeatability -1
21 Chromatogram for analysis of formulation [SYNASMA-AX] Repeatability-2