Course

UNIT-I  Chromatography                                                                                      30 Hours

Principles and tecniques of Chromatography, Planar and Columnar chromatography, theories of chromatography. Choice of chromatographic condition. Concept and need for derivatization in pharmaceutical analysis (5 hrs)

Introduction to chromatographic techniques

• Paper Chromatography (1 hr)
• Thin Layer Chromatography (1 hr)
• High Performance Thin Layer Chromatography (2 hr)
• Column Chromatography: (1 hr)
• Gas Chromatography (2 hrs)
• Ion-exchange chromatography (1 hr)
• Gel filtration and affinity chromatography (1 hr)
• Chiral Chromatography(1 hr)

• High Performance Liquid Chromatography (10 hrs)

Introduction, theory, instrumentation, selection of mobile phase, stationary phase, detectors, analytical and bioanalytical applications. Sample preparation of non-biological and biological samples. HPLC- method development, Data interpretation, HPLC in Bioavailability and Equivalence Studies. Hyphenation techniques, Ultra and nano-HPLC. AI technologies used in hyphenated instruments for data interpretation, method optimization, and decision support.

• Electrophoresis (5 hrs)

Principles of separation, gel electrophoresis, SDS PAGE, Clinical and Pharmaceutical Applications. Quality control of protein-based drugs. Identifying and characterizing biomarker proteins. Studying protein-protein or DNA-protein interactions. Genomic and proteomic studies for novel drug targets.

UNIT-II  Spectroscopy                                                                                       21 Hours

• Absorption Spectroscopy: (7 hrs) Introduction to Spectroscopy, Theory of electronic, atomic and molecular spectra, Fundamental laws of photometry, Beer-Lambert’s Law, Principle and Application, Deviations and Limitations, Chromophores and Auxochromes in Biomolecules, Spectral Shifts and Solvent Effects, Instrumentation, Types of Spectrophotometers: Single-beam and double-beam spectrophotometers, Radiation Sources

and Monochromators, Detectors, Advanced Techniques like Derivative Spectroscopy, Derivatisation and Difference Spectroscopy, Applications in Pharmacy: Quality Control in Pharmaceuticals Single and Multi-Component Analysis: Spectroscopy in Biochemical and Pharmacokinetic Studies Equilibrium and Kinetics,Spectroscopic methods in Diagnosis and Monitoring glucose, cholesterol, and other critical parameters in clinical labs,

• Fluoresence Spectroscopy (7 hrs)

Introduction to Fluorescence Spectroscopy, Theory of Luminescence, Factors Affecting Fluorescence and Quenching, Instrumentation, Applications of Fluorescence Spectroscopy in Pharmacy and Clinical Settings, study of pharmaceutically important compounds estimated by fluorimetry. Fluorescence techniques employed for DNA/RNA quantification and enzyme activity assays. Fluorescence imaging and microscopic techniques: imaging tissues and monitoring drug absorption and distribution, deep tissue imaging with minimal damage using NIR. Application of NIR fluorophores in robotic surgery.

Inroduction to structural elucidation techniques and its clinical relevance

• Infrared Spectroscopy (3 hrs)
• NMR & ESR (2 hrs)
• X-RAY Diffraction (2 hrs)

UNIT-III SpectrometricTechniques                                       12 Hours

• Mass Spectrometry: (8 hrs)

Basic principles of Mass Spectrometry Instrumentation: Ionization techniques: Electron ionization, Chemical ionization, Atmospheric pressure ionization (Electrospray ionization, APCI, and APPI), other sources: MALDI, ICP, etc. Mass Analyzers: Quadrupole, Time of flight, Orbitrap, High Resolution Mass Spectrometry, Hyphenated Mass Spectrometry: GC/MS, HPLC/UPLC-MS and Tandem Mass Spectrometry, Mass spectrum and applications. Over view of LCMS as an analytical technicque. A brief outline of omics study using hyphenated tandem MS including the scope of biomarkers study. AI-driven advancements in mass spectrometry, including machine learning models for enhancing spectral clarity, optimizing analytical methods, improving peak identification accuracy, and providing intelligent decision support in data interpretation.

• Flame Photometry (2 hrs)

Theory, nebulisation, flame and flame temperature, interferences, flame spectrometric techniques instrumentation and pharmaceutical applications.

• Atomic Absorption Spectrometry: (2 hrs)

Introduction, Theory, types of electrodes, instrumentation and applications.

UNIT-IV Quality Assurance                                                                               12 hours

• Concept of Quality Control and Quality assurance (2 hr)
• Total quality management, quality review and (2 hr)
• GLP, NABL, ISO (2 hrs)

• ICH- international conference for harmonisation (4hrs)
• Qualification of equipment, validation of analytical instruments and (2 hrs)

SCOPE:

This course focuses on comprehensively exploring the principles, techniques, and practical applications of diverse analytical techniques used in the pharmaceutical sector. The students will better understand various analytical methods employed in modern pharmaceutical laboratories for qualitative and quantitative analysis of dosage forms and biological samples. The curriculum is designed to provide students with essential knowledge concerning the principle, instrumentation, and application of spectroscopic and chromatographic techniques. Through theoretical explanations, participants will gain insight into the underlying phenomena of these analytical methods while delving into the hands-on practical aspects of operating different instruments. Moreover, this course goes beyond traditional techniques and acquaints students with emerging analytical methods, including hyphenated techniques and their real-world applications (SDG 9). Such exposure to cutting-edge technologies will undoubtedly foster a deeper understanding of analytical practices within the pharmaceutical sector.

To ensure a well-rounded learning experience, the course incorporates hands-on laboratory sessions and practical training, allowing students to gain proficiency in instrument operation and data analysis. The students will be equipped with the necessary expertise in measuring drug concentrations in dosage forms and biological fluids, enabling healthcare professionals to monitor drug levels and adjust dosages accordingly. This practical competence contributes directly to ensuring drug quality, safety, and efficacy, which supports public health initiatives and patient-centered healthcare (SDG 3).

Furthermore, the course nurtures analytical thinking and scientific inquiry, equipping students with essential competencies to contribute meaningfully to pharmaceutical research, regulatory affairs, and clinical diagnostics. By promoting a strong foundation in pharmaceutical analysis, this program supports equitable and inclusive education and lifelong learning opportunities (SDG 4), preparing graduates to address global healthcare challenges responsibly and innovatively.Upon successful completion, students will be empowered with scientifically grounded knowledge and practical expertise to ensure the quality and safety of pharmaceuticals, thereby playing a vital role in advancing healthcare standards and sustainable pharmaceutical practices.

COURSE LEARNING OUTCOMES:

Upon completion of the course, the student shall be able to

KNOWLEDGE

K1. Define common terminologies used in instrumental analysis for a comprehensive understanding of their significance in clinical contexts. (Understanding)

K2. Discuss the interaction of matter with electromagnetic radiation and its wide-ranging applications in drug analysis (Understanding)

K3. Illustrate the fundamental principles and diverse applications of various spectroscopic and chromatographic techniques relevant to clinical research and drug development. (Applying) K4. Apply essential practical skills using instrumental techniques, emphasizing the importance of proper sample preparations and solvent selection. (Applying)

K5. Employ appropriate instrumental techniques proficiently for both qualitative and quantitative drug analysis to make informed analytical decisions crucial for clinical diagnosis and treatment. (Analysing)

K6. Evaluate the reliability and accuracy of analytical results, identifying potential limitations and sources of error in the analysis process, particularly concerning clinical relevance and patient care. (Analysing)

Skill

S1. Compare operational techniques of UV, HPLC, fluorimeter, flame photometer, etc., focusing on their clinical applications and significance in healthcare settings. (Understanding) S2. Develop basic practical skills using instrumental techniques relevant to clinical drug analysis and diagnostic applications. (Applying)

S3. Correlate quantitative and qualitative analysis of drugs using various analytical instruments, emphasizing their implications for patient diagnosis and treatment decisions. (Analysing)

S4. Analyze drug concentrations in the blood and other body fluids to optimize drug dosages for individual patients. (Analysing)

S5. Ensure the best possible outcomes in patient care by interpreting the data generated from analytical instruments and the significance of the results in a clinical context. (Analysing)

S6. Evaluate knowledge of interpretation of data obtained from spectra and of chromatograms (Evaluate)

Attitude

A1. Distinguish the critical role in advancing scientific knowledge and solving real-world clinical problems (Understanding)

A2. Show enthusiasm and curiosity towards exploring newer techniques and their applications. (Applying)

A3. Explore opportunities for continuous learning and staying updated with advancements. (Applying)

A4. Attain confidence in handling and operating analytical instruments effectively and safely. (Applying)

A5. Demonstrate professionalism by adhering to ethical standards and best practices in research and data handling. (Applying)

A6. Illustrate confidence in troubleshooting instrument-related issues and finding appropriate solutions. (Analysing)

TEXT BOOKS:

1. Willard, H.H., Merritt, L.L., Dean, J.A., & Settle, F.A. Instrumental methods of analysis, 7th edition. EWP, East West Press Ltd., Delhi/Madras;1988.
2. Skoog, A., Heller, F.J., Nieman, T.A., Principles of Instrumental Analysis, WBSaunders;1997
3. Becket H. & Stenlake J.B. Practical Pharmaceutical Chemistry Vol. I and II, 4^th edition, the Athlone Press of the University of London;1988.
4. Kenneth A. Connors. A Textbook Of Pharmaceutical Analysis, 3rd Edition. Wiley India Pvt. Limited; 2007.
5. Silverstein, M., Webster, F. X., Kiemle, D. J. Spectrometric Identification of Organic Compounds. 6th edition. Wiley India Pvt. Limited; 2006
6. Sethi, D. Quantitative Analysis of Drugs in Pharmaceutical Formulations. India: CBS Publishers & Distributors; 2015
7. Kemp, Organic Spectroscopy. United States: Palgrave Macmillan; 1991
8. Vogels Textbook Of Quantitative Chemical India: Pearson Education; 2006.

REFERENCE BOOKS:

1. Indian Pharmacopoeia, Indian Pharmacopoeia Commission.The Controller of Publications, New Delhi; 2018.
2. BPC- of Health, U.K. for HMSO.
3. USP – Mack Publishing , Easton, PA.
4. The Extra Pharmacopoeia – The Press, London.
5. Sethi, D. Sethi’s HPLC High Performance Liquid Chromatography: Quantitative Analysis of Pharmaceutical Formulations, Volume 8. India: CBS Publishers & Distributors; 2015
6. Clarke, G. C. Clarke’s Analysis of Drugs and Poisons: In Pharmaceuticals, Body Fluids and Postmortem Material. United Kingdom: Pharmaceutical Press; 2004
7. Remington, P. Remington: The Science and Practice of Pharmacy. United Kingdom: Lippincott Williams & Wilkins; 2006
8. Stahl, Thin-Layer Chromatography: A Laboratory Handbook. Germany: Springer Berlin Heidelberg. Higuchi. T and Hasen. E. B. Text Book of Pharm. Analysis. New York Inter Science Publishers; 2013

JOURNALS:

Biomedical Chromatography. https://analyticalsciencejournals.onlinelibrary.wiley.com/journal/10990801

Journal of Pharmaceutical and Biomedical Analysis. https://www.sciencedirect.com/journal/journal-of-pharmaceutical-and-biomedical- analysis

Clinical Chemistry and Laboratory Medicine (CCLM) https://www.degruyter.com/journal/key/cclm/html#overview