Pharmaceutical Analysis and Spectroscopy

Code

PHAR2055

School

Pharmacy

Level

2

Credits

20

Semesters

Autumn Malaysia

Summary

The module is designed to provide second year students with a sound fundamental knowledge of aspects of pharmaceutical analysis and spectroscopy. The theory and applications of spectroscopic and analytical techniques that may be encountered during the course either in practical classes or lectures are described. This includes infrared and ultra-violet spectroscopy, mass spectrometry and nuclear magnetic resonance spectroscopy. Advanced separation techniques are covered in this module and their linkage to mass spectrometry detection systems is described.  The important concepts of analytical validation and quality assurance are introduced and developed in relation to pharmaceutical analysis.  
This module also included a practical component, with the experiments chosen to complement many of the topics covered in the lectures of year-1 and year-2 modules and serve to illustrate some key concepts in pharmaceutical and biological chemistry. The experiments broadly fit into three main categories: (a) Synthetic medicinal chemistry (experiment 2) build upon the practical skills and theoretical aspects acquired during PHAR1026. These experiments further encompass and reinforce topics covered in this module and PHAR2014. (b) The application of analytical techniques to pharmaceuticals (experiments 4) reinforces and builds on the knowledge acquired from this module. (c) The isolation and/or analysis of natural products from natural sources (experiments 2 and 3) provides practical experience and reinforces concepts covered in PHAR2014.
INTRODUCTORY LECTURE (1 lecture)
•    General introduction to spectroscopy. The electromagnetic spectrum.  Electromagnetism. Electrons and orbitals. 

INFRARED SPECTROSCOPY (3 lectures)
•    Origin of IR spectroscopy. Modes of molecular vibration and associated absorption bands.
•    IR bands associated with functional groups: carbonyl, alcohol, ether, amine, and nitrile groups. Factors that cause bands to shift.
•    Sample presentation.  Nujol mulls, KBr discs and solutions position of IR absorption band. Instrument design and operation: the dual-beam optical-null IR spectrometer, Fourier transform infrared (FTIR), and attenuated total reflection-Fourier transform infrared (ATR-FTIR).

ULTRAVIOLET AND FLUORESCENCE SPECTROSCOPY (3 lectures)
•    Beer-Lamberts law. The construction of an UV/visible spectrophotometer. The concept of chromophores and chromophore length, red and blue shifts in spectra.  The extinction coefficient.  Solvent effects.
•    Chemical introduction of chromophores and auxochromes. Pharmaceutical analysis of compounds, assays directed towards the identification of contaminants and assays of concentration. Pharmacopoeial standards and sources of deviation.
•    Fluorescence as a spectroscopic technique. How it arises, its properties and uses in the analysis of pharmaceutical compounds.

NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY (8 lectures)
•    Nuclear magnetic resonance. The theory of nuclear spin and generation of magnetic spin states.  Effect of excitation of spin and the creation of NMR spectra.
•    Effect of different environments on proton resonance. Proton equivalence. The NMR spectrum and standard NMR calibration materials. Shielded and deshielded protons.
•    Characteristic proton chemical shifts. The integration of NMR resonances and spectral interpretation.
•    The origin of spin-spin coupling. Spin-spin coupling in methyl, methylene and methine protons.
•    Examples and predictions of NMR spectra of simple organic and pharmaceutical compounds. Exchangeable protons. Spin-coupling calculations - the use of the Karplus equations.
•    Long range NMR spin-spin coupling. Aromatic rings and the effect of para disubstitution.
•    The effect of electron donating and withdrawing groups on aromatic proton resonance positions. Mono-, di- and tri-substituted aromatic rings. 
•    13C NMR spectroscopy. Natural abundance carbon spectra and the role of proton decoupling.

FLAME SPECTROSCOPY (2 lectures) LKH
•    Techniques of flame spectroscopy.  Metals and their analysis.  Types of flame spectroscopy.  Atomic absorbance spectroscopy, technique and instrumentation.
•    Ionization of metals in flames.  Method of standard additions.  Atomic emission spectroscopy.  Analysis of metals in pharmaceuticals.

MASS SPECTROMETRY (2 lectures)
•    Mass spectrometry: how it works, what students should undertake, what students should learn.

PHARMACEUTICAL ANALYSIS AND CHROMATOGRAPHY (8 lectures)
•    Introduction to assay development and methodology. Quality assurance and validation in pharmaceutical analysis.
•    Adsorption and partition column chromatography theory. Paper and thin-layer chromatography.
•    High performance liquid chromatography 1: instrumentation and basic mechanisms.
•    High performance liquid chromatography 2: modes of analyte separation.
•    Electrophoresis, capillary electrophoresis-instrumentation, separation mechanisms and modes.
•    Gas chromatography instrumentation and separation mechanisms. 
•    Chromatography linked to mass spectrometry: LC-MS, GC-MS, CE-MS.

LABORATORY PRACTICALS (4 x 6 hours)
There are four practical classes associated with this module.
•    Extraction and Analysis of Piperine from Pepper (techniques: extraction, staining, crystallisation, TLC, IR)
•    Synthesis of -Lactam Antibiotics (techniques: synthesis, extraction, crystallisation, TLC, IR, NMR)
•    GC Evaluation of Volatile Oils (techniques: extraction, GC, quantitative analysis)
•    High Performance Liquid Chromatography (HPLC) Assay of Panadol® Extra Caplets (Group Practical) (techniques: sample preparation, HPLC, quantitative analysis)

WORKSHOPS (4 x 2 hours)
There are four problem workshops associated with the module content. They involve undertaking preliminary work prior to the class, which is initially reviewed by the lecturer. This is followed by problem solving exercises. Staff are available to offer help and advice. 
Workshop 1: IR, UV-vis and fluorescence spectroscopy
Workshop 2: NMR spectroscopy
Workshop 3: Exposure to NMR data processing software
Workshop 4: Mass spectrometry and pharmaceutical analysis & chromatography

REVISION LECTURE (1 lecture)

Target Students

BSc Pharmaceutical and Health Sciences students

Classes

• One 2-hour workshop each week for 4 weeks
• One 1-hour lecture
• Two 1-hour lectures each week for 5 weeks
• One 2-hour lecture each week for 6 weeks
• One 6-hour laboratory each week for 4 weeks

Assessment

• 20% Practical write up: In module (Pro-forma / report)
• 20% ExamSys: In module (45 minutes)
• 60% Written Exam (2-hour): 2 hours - end of semester

Assessed by end of autumn semester

Educational Aims

Learning Outcomes

After successful completion of this module, you should be able to:
A. Knowledge and understanding
• A4 Molecular basis of drug structure, design and activity, including the application of fundamental concepts in chemistry, chemical biology and molecular biology

B. Intellectual skills
• B2 Undertake independent, reflective, self-directed learning
• B3 Understand principles of experimental and research methodology
• B5 Analyse and critically evaluate scientific data

C. Professional/practical skills
• C2 Undertake practical experimental work using appropriate materials, operating procedures and instruments
• C3 Undertake numerical tasks and carry out quantitative analysis

D. Transferable/key skills
• D1 Effective communication in a variety of forms, including written, verbal and visual
• D2 Critically use information resources, including library and electronic sources
• D3 Work effectively as an individual or as part of a team

Conveners

• Prof Kuan Hon Lim