Pharmaceutical

Analysis
A Textbook for Pharmacy Students
and Pharmaceutical Chemists
EDITION

D a v i d G. W a t s o n

PGCE

Senior Lecturer in Pharmaceutical Sciences, Strathclyde Institute of Pharmacy

and Biomedical Sciences, University of Strathclyde, Glasgow, UK

by

RuAngelie

BSc

Lecturer, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of

Strathclyde, Glasgow, UK

CHURCHILL
LIVINCSTONE

ELSEVIER
EDINBURGH

LONDON

NEW YORK OXFORD

PHILADELPHIA

ST LOUIS SYDNEY TORONTO

2012

Contents

1. Control of the quality of analytical

1
Introduction /
Control of errors in analysis 2
and
5
Validation of analytical procedures 7
Standard operating
(SOP) for the assay of Paracetamol tablets 10
Compound
errors
Reporting of results
Other terms used in the control of analytical procedures 14
Basic calculations in pharmaceutical analysis 19
Additional problems 24

2.

and

properties of drug molecules 26

Introduction 26
of pH
of aqueous
acids and bases 27
and
strength and p/Ca 29
Henderson-Hasselbalch
29
Ionisation of drug molecules
Buffers 33
Salt
36
Activity,
strength and
Partition
Drug stability 41
of drugs 43
of optical rotation 49
Profiles of physico-chemical properties of
Additional problems 57

of

and

37

drug molecules 50

3. Titrimetric and chemical analysis methods 60

Keypoints 60
Introduction 6/
Instrumentation and reagents 61
acid/base titrations in the aqueous phase 62
Titrations of the
of weak bases in
aqueous/non-aqueous media
Indirect titrations in the aqueous phase 66
titrations
titrations 70
titrations 70
Redox titrations 71

65

analysis
titrations 73
Ion pair titrations 75
Diazotisation titrations 76
Potentiometrie titrations 77
Karl Fischer titration
end-point
Automation of wet chemical methods 82
Applications of
in pharmaceutical analysis 84
Additional problems 87

81

4. Ultraviolet and visible spectroscopy 90

Keypoints 90
Introduction 91
Factors governing absorption of radiation in the
region 92
Law 94
Instrumentation 95
Diode array
96
Instrument calibration 96
UV spectra of
representative drug molecules 98
Use of UV/visible
to
values 102
Applications of UV/visible spectroscopy to pharmaceutical quantitative
analysis 103
Difference spectrophotometry 707
Derivative spectra 109
Applications of UV/visible spectroscopy in
and
Additional problems
5.

spectrophotometry 115
Keypoints
Introduction
Factors
intensity and energy
of absorption in
spectra 777
Instrumentation
Sample preparation 720
Application of IR spectrophotometry in structure elucidation 723
Examples of IR spectra of drug molecules 724
IR spectrophotometry as a fingerprint technique 727
Infrared spectrophotometry as a
for identifying
130
Near-infrared analysis
Keypoints
Introduction
Examples of NIRA applications
Additional problems

6.

spectrophotometry

130

737

138

spectrophotometry (AES)
Keypoints
Introduction 138
Instrumentation

Contents
Examples of quantitation by AES 140
in AES analysis 142
Assays based on the
of Standard additions 143
Atomic absorption spectrophotometry (AAS)
Keypoints
Introduction
Instrumentation
Examples of assays using AAS 146
examples of
AAS
Inductively coupled plasma emission spectroscopy
7. Molecular emission spectroscopy 152

Fluorescence spectrophotometry
Keypoints 752
Introduction 753
Instrumentation 154
Molecules which exhibit fluorescence 754
Factors interfering with fluorescence intensity 755
Applications of fluorescence spectrophotometry in
analysis 756

spectroscopy
Keypoints /59
Introduction
Applications
8. Nuclear magnetic resonance spectroscopy
Keypoints 165
166
Instrumentation 167

765

Application of NMR to
in
Carbon NMR 192
NMR spectra
Application of NMR to quantitative analysis
Other specialised
of NMR 200
9. Mass
204
Keypoints 204
Introduction 205
Ion generation 205
Other ionisation methods
Ion Separation
A
detailed consideration of
spectra 219
Molecular fragmentation patterns 220
Gas
spectrometry (GC-MS)
Applications of GC-MS with
232

drug molecules 188

Pharmaceutical analysis
Tandem

spectrometry 236
mass spectrometry 244
Mass spectrometry of proteins 246
Mass spectrometry in drug discovery 248

10.

theory 252
Introduction 252
Void volume and
factor 252
Calculation of column
253
Origins of band broadening in HPLC 254
Parameters used in evaluating column Performance
Data acquisition 262
Report generation 263

258

Gas
265
Keypoints 265
Introduction 266
Instrumentation 266
Selectivity of liquid stationary phases 272
Use of
in GC 280
Summary of parameters governing
GC Performance 283
GC
284
Applications of GC in quantitative analysis 284
Determination of manufacturing and degradation residues by GC 297
Determination of residual solvents 294
microextraction (SPME) 297
Applications of GC in bioanalysis 298
Additional problems 299
12. High-performance liquid chromatography 307
Keypoints 302
Introduction 302
Instrumentation 302
Stationary and mobile phases 303
Structural factors which govern rate of elution of Compounds from HPLC
306
More advanced consideration of solvent selectivity in reverse-phase
chromatography 373
Effect of temperature on HPLC
Summary of stationary phases used in HPLC 37 7
A more advanced consideration of reverse-phase stationary phases 320
Summary of detectors used in HPLC 322
Performance of a diode array detector 323
Applications of HPLC to the quantitative analysis of drugs in
327
Assays involving more specialised HPLC
340
Additional problems 355

Contents
13.

chromatography 358
Keypoints 358
Introduction 359
Instrumentation 359
TLC chromatogram 360
Stationary phases 36/
Elutropic series and mobile phases 36/
of TLC adsorbant 365
of Compounds on TLC plates following
Applications of TLC analysis 367
TLC (HPTLC) 372

366

14. High-performance capillary electrophoresis 376

376
Introduction 377
Instrumentation 380
Control of Separation 382
Applications of CE in pharmaceutical analysis 384
Use of additives in the running buffer 388
Additional problems 396
15. Extraction methods in pharmaceutical analysis 398
Keypoints 398
Introduction 399
used excipicnts in
399
Tablets and
399
Solvent extraction methods 400
Microdialysis extraction 404
extraction (SPE) 404
Keypoints 404
Introduction 405
Methodology 405
Types of adsorbants used in SPE 407
Typical extraction
using lipophilic
408
Adaptation of SPE for automated
extraction prior to HPLC analysis
Recent
in solid-phase and on-line extraction 414

Index