B2904 - Instrumental Methods in Drug Analysis

Learning outcomes

At the end of the course, which includes frontal lessons and single-place laboratory exercises, the student acquires the knowledge on instrumental methods (UV-vis, fluorimetry, HPLC, UPLC, GC and combined techniques such as HPLC-MS) for the quantitative determination of analytes of pharmaceutical interest, including chiral drugs, in medicinal products; - owns the knowledge necessary for method development and validation; - knows new-generation stationary phases and recently developed analytical approaches for rapid and highly efficient analysis of medicines, including biological/biotechnological ones; -is able to apply the acquired knowledge to analytical problems related to the quantification of APIs in formulations and to the quality control of medicines of industrial origin.

Course contents

Instrumental quantitative analysis. Method validation: linearity, accuracy, precision, selectivity, LOD and LOQ.

Sample preparation. Solid phase extraction. Approches (polar, apolar and ion exchange materials). Workflow. Applications.

Use of UV-Visible Spectroscopy in the quantitative analysis of drugs. Selection of the optimal wavelength, selectivity, sensitivity, construction of the calibration curve. Derivative spectrophotometry, differential spectrophotometry and multicomponent spectrophotometry.

Use of fluorimetry in the quantitative analysis of compounds of pharmaceutical interest. Quali- and quantitative analysis of fluorescent drugs. Chemical derivatization aimed at introducing fluorophoric residues on drugs.

Quantitative analysis of drugs and validation with chromatographic techniques. References to chromatographic parameters. External and internal standard methods, standard additions method. Van Deemter equation for standard stationary phases. How to improve chromatographic efficiency and to decrease the time of the analysis: (i) The role of the dimension of the packing material; (ii) High temperature chromatography; (III) Monolithic column technology; (iv) UHPLC (Ultra High Performance Liquid Chromatography); (v) Fused core particle technology. Polar drugs analysis: ionic suppression HPLC and ion pairing chromatography, HILIC (Hydrophilic Interaction Chromatography).

Gas chromatography (GC): instrumentation. Chromatographic parameters optimization. GC applications to the analysis of drugs.

Hyphenated techniques: HPLC-MS, GC-MS. Ion sources for LC-MS: ESI, APCI and APPI. Analyzers: ion trap, quadrupole, triple quadrupole and time of flight analyzers. MALDI mass spectrometry.

Introduction to quality control of biological/biotechnological drugs by HPLC. Dedicated stationary phases.

Enantioselective analysis of chiral drugs using chromatographic techniques. Indirect method: pre-column derivatization using reagents with defined stereochemistry. Direct method: use of enantioselective stationary phases. Pirkle-type, ligand exchange, polysaccharides, cyclodextrins and protein-based chiral stationary phases.

PRACTICAL CLASSES: Quantitative analysis of drugs using instrumental techniques

Readings/Bibliography

V. Cavrini, V. Andrisano. Principi Analisi farmaceutica. Società editrice Esculapio ed. 2013.

Lecture slides

Teaching methods

The lessons and practical classes will delivered in presence.

In consideration of the types of activities and teaching methods adopted, the attendance of this training activity requires the performance of all students of Modules 1 and 2 in e-learning mode [https://www.unibo.it/it/servizi-e-opportunita/salute-e-assistenza/salute-e-sicurezza/sicurezza-e-salute-nei-luoghi-di-studio-e-tirocinio] and the participation in Module 3 of specific training on safety and health in the places of study. Information on dates and methods of attendance of Module 3 can be consulted in the specific section of the degree program website

Assessment methods

The final exam consists in an oral examination, which aims at evaluating the acquisition of sufficient knowledge on the topics of the course.

In particular, the student should demonstrate

- he has acquired sufficient knowledge on the instrumental methods used for the quantitative determination of compounds of pharmaceutical interest;

- to be able to apply the gained knowledge to real problems and new issues related to the analysis of pharmaceutical formulations.

The daily positive qualification during the laboratory practical classes, the preparation of the laboratory reports and the discussion on the reports themselves are key parts of the exam.

Teaching tools

Video projector, PC. Educational material available on virtuale – Insegnamenti online

Office hours

See the website of Manuela Bartolini