ANALISI DEI FARMACI E DEI PRODOTTI PER LA SALUTE 3 M - Z

1. Expected learning outcomes

Knowledge and understanding

On completion of the course, students will know the theoretical principles of the main instrumental analytical techniques (spectroscopic, chromatographic, spectrometric and electrophoretic) applied to the identification and quantification of organic compounds of pharmaceutical interest listed in the monographs of the official pharmacopoeias. They will also know the criteria for selecting an analytical method, sample preparation strategies for complex biological matrices, and the validation parameters of an analytical method according to ICH guidelines.

Ability to apply knowledge and understanding

On completion of the course, students will be able to:

• select the appropriate analytical technique for a defined analytical objective;
  
• develop and optimise a protocol for the analysis of an active ingredient in a pharmaceutical dosage form or a biological matrix;
  
• interpret UV-Vis, IR, MS and NMR spectra for structural identification purposes;
  
• apply validation criteria to a chromatographic method.
  

Making judgements

Students will be able to critically interpret experimental data, assess the fitness for purpose of an analytical method, and recognise anomalies and artefacts.

Activities contributing to this outcome: laboratory sessions, analysis of real analytical case studies (problem-based learning), and critical discussion of the data obtained within the group project.

Communication skills

Students will be able to present analytical methods and results using appropriate scientific terminology, both in writing and orally.

Activities contributing to this outcome: writing laboratory reports and presenting the group project in class.

Learning skills

Students will have acquired the theoretical and practical skills needed to independently update their knowledge and address new analytical problems.

Activities contributing to this outcome: guided consultation of pharmacopoeia monographs and primary literature within the group project.

7. Delivery mode

Delivery mode

Traditional classroom teaching, with laboratory activities. English-friendly course. (Field pre-filled by the teaching office according to the Academic Regulations of the degree programme.)

Descriptive text for delivery mode

The course comprises a total of 58 hours, divided into 28 hours of lecture-based teaching and 30 hours of interactive teaching.

Lecture-based teaching — 28 hours. Classroom lectures supported by the projection of slides, spectra and demonstration videos, devoted to presenting the theoretical foundations of analytical techniques. It contributes primarily to the achievement of knowledge and understanding outcomes.

Interactive teaching — 30 hours, distributed as follows:

• individual-bench laboratory sessions (22 hours): qualitative and quantitative analysis of active ingredients, chemical identification tests, determination of physico-chemical constants and chromatographic separations, with individual written reports;
  
• problem-based learning sessions on real analytical cases (6 hours) drawn from pharmacopoeia monographs and primary literature, in which students, working in small groups, define the analytical objective and propose a solution strategy, subsequently discussed in plenary;
  
• presentation and discussion in class of the group project (2 hours) for the development and validation proposal of an analytical method;
  
• formative self-assessment quizzes at the end of the main thematic blocks, with immediate feedback, carried out within the hours indicated above.

Students may use Artificial Intelligence (AI) tools as a means of supporting their learning and study activities, provided that such use complies with all applicable laws and regulations, including those governing copyright and the use of intellectual works. Students are expected to use AI tools responsibly and in accordance with the principles and provisions set out in Section 5.1 of the University guidelines (https://www.unict.it/it/ateneo/cidia-centro-l%E2%80%99informatica-la-digitalizzazione-e-l%E2%80%99intelligenza-artificiale).

Consistency between teaching methods and expected learning outcomes. Interactive teaching activities contribute to the achievement of ability to apply knowledge and understanding (laboratory sessions, PBL), making judgements (critical interpretation of data, reasoned method selection), communication skills (laboratory reports, project presentation) and learning skills (independent consultation of monographs and primary sources).

If the course is delivered in blended or remote mode, appropriate adjustments may be made to the above, in order to ensure consistency with the syllabus.

2. Prerequisites

The following knowledge is essential for effective attendance of the course (cultural prerequisites):

1. principles of general and inorganic chemistry: acid-base equilibria, partition equilibria, solutions and their concentration;
   
2. organic chemistry: structure, nomenclature and reactivity of the main functional groups, stereochemistry and chirality;
   
3. principles of classical qualitative and quantitative analysis (gravimetry, volumetry) and basic statistical treatment of analytical data;
   
4. elements of physics: electromagnetic waves, the electromagnetic spectrum, electrostatics.
   

Basic computer skills for processing experimental data are useful.

Formal prerequisites: as established by the Academic Regulations of the degree programme, admission to the examination is subject to passing the examinations listed therein as prerequisites.

8. Attendance requirements

Attendance is compulsory, as established by the Academic Regulations of the degree programme: admission to the examination requires absences not exceeding 30% of the total teaching hours. Attendance of laboratory sessions is a necessary condition for admission to the examination.

Active participation in lectures and interactive teaching activities is decisive for achieving the expected learning outcomes: problem-based learning sessions, the group project and formative quizzes with immediate feedback are designed to develop applied competences and critical skills that are difficult to acquire through individual study alone.

3. Course contents

1. Introduction to instrumental analytical methods. Classification of instrumental analytical methods. Criteria for method selection. Figures of merit.
   
2. Spectroscopic methods. UV-Vis absorption and emission spectroscopy (fluorescence and phosphorescence). IR absorption spectroscopy. Nuclear magnetic resonance spectroscopy (¹H- and ¹³C-NMR). Integrated spectral interpretation.
   
3. Identification of functional groups and determination of physico-chemical constants. Chemical identification tests on compounds of pharmaceutical interest according to pharmacopoeia monographs.
   
4. General principles of the chromatographic process. Distribution constant, retention time, capacity factor. Resolution. Selectivity. Efficiency: plate theory; van Deemter equation. Chromatographic peak asymmetry.
   
5. Liquid chromatography. Stationary and mobile phases. Liquid-solid, liquid-liquid, ion-exchange, ion-pair, size-exclusion and affinity chromatography. Column chromatography and thin-layer chromatography (TLC). HPLC: instrumentation. Enantioselective separation: direct method using enantioselective stationary phases.
   
6. Gas chromatography and capillary electrophoresis. GC: instrumentation, types of stationary phase, detection. CE: instrumentation, theoretical principles, analytical parameters.
   
7. Mass spectrometry and hyphenated techniques. Classes of instrumentation, ionisation techniques, fragmentation analysis, analysers and detectors. HPLC-DAD, HPLC-MS, GC-MS.
   
8. Qualitative and quantitative analysis. External and internal standard methods. Sample preparation and extraction methods: liquid/liquid and liquid/solid extraction, supercritical fluid extraction (SFE), solid-phase extraction (SPE). Preparation of samples from complex biological matrices.
   
9. Analytical method validation. Linearity, precision, accuracy, reproducibility, LOD, LOQ. Development of chromatographic methods for quality control and for pre-clinical and clinical research. 
   
10. Determination of drug/target protein interactions by high-performance affinity chromatography (HPAC).
    

6. Textbook information

Reference textbooks

1. R.M. Silverstein, F.X. Webster, Identificazione spettroscopica di composti organici, Ambrosiana Editrice, Milan, 2016. (English edition available: Spectrometric Identification of Organic Compounds, Wiley.)

2. V. Cavrini, Principi di Analisi Farmaceutica, latest edition, Società Editrice Esculapio, Bologna.

3. D.A. Skoog, D.M. West, F.J. Holler, S.R. Crouch, Fondamenti di chimica analitica, EdiSES, 3rd ed., 2015. (English edition available: Fundamentals of Analytical Chemistry, Cengage.)

Recommended for further study

4. V.R. Meyer, Practical High-Performance Liquid Chromatography, 5th ed., Wiley, New York.

5. Italian Official Pharmacopoeia / European Pharmacopoeia, current edition (for consultation).

6. ICH Q2(R2) Validation of Analytical Procedures.

7. Teaching material and scientific articles provided by the lecturer on the Studium platform.

Note for international students. Slides report technical terminology in English; textbooks 4 and 6 and the primary literature material are in English and may be used as alternative references. English editions of textbooks 1 and 3 are available and may be used in place of the Italian editions.

4. Learning assessment

Learning assessment methods

Final written examination, with in-course tests. (Field pre-filled by the teaching office according to the Academic Regulations of the degree programme.)

Descriptive text for learning assessment methods

Learning is assessed through two in-course tests and a final written examination, complemented by the assessment of laboratory activity and of the group project.

In-course tests (optional, reserved for attending students). First test, approximately mid-course: spectroscopic methods and identification of functional groups; duration 90 minutes; 10 multiple-choice questions, 2 open-ended questions and 1 spectral interpretation exercise. Second test, at the end of the course: separation techniques, mass spectrometry, sample preparation and validation; same structure and duration. Each test is marked out of 30. Admission to the second test requires a mark of no less than 18/30 in the first. Passing both tests (average ≥ 18/30) exempts students from the final written examination; validity is limited to the current academic year.

Final written examination (for students who do not take or do not pass the in-course tests). Duration 120 minutes; 15 multiple-choice questions, 3 open-ended questions and 2 applied exercises (spectral interpretation and calculation of chromatographic or validation parameters). Valid only for the session in which it is taken.

Group project. Each group develops and critically discusses an analytical protocol for an assigned real case (for example, the assay of an active ingredient in a pharmaceutical dosage form or in plasma), including a validation proposal. Assessment considers the scientific soundness of the approach, the consistency between the analytical objective and the proposed technique, and the clarity of presentation.

Composition of the final mark: in-course tests or final written examination 70%; group project and laboratory reports 30%.

Criteria for the award of the final mark

Mark Description

Fail The student does not possess the minimum knowledge of the principles of the analytical techniques covered; is unable to select or justify an analytical approach; technical language is poor or absent.

18–21 Minimum knowledge of the main techniques; able to apply known procedures only in previously seen contexts; limited ability to interpret experimental data; poorly developed command of technical language.

22–25 Fair knowledge, limited to the main topics; sets out an analytical protocol correctly but not always coherently; interprets data with some uncertainty; reasonably clear presentation.

26–28 Good knowledge of theoretical and instrumental principles; independently selects the technique suited to the analytical problem; critically interprets experimental data; clear presentation with appropriate language.

29–30 cum laude Thorough knowledge; integrates different techniques to solve complex analytical problems, including in biological matrices; critically assesses the suitability and limitations of methods; excellent communication skills and command of language.

Learning assessment may also be carried out on-line, should the conditions require it.

To ensure equal opportunities and in compliance with current laws, interested students may request a personal interview in order to plan any compensatory and/or dispensatory measures based on educational objectives and specific needs. Students can also contact the CInAP (Centro per l'Integrazione Attiva e Partecipata — Servizi per le Disabilità e/o i DSA) referring teacher within their department (https://www.cinap.unict.it/content/referenti).

5. Examples of frequently asked questions

1. Describe the theoretical principles of chromatographic separations and discuss approaches to improving resolution in liquid chromatography.
   
2. Illustrate the theoretical principles and applications of capillary electrophoresis, highlighting its advantages and limitations compared with HPLC.
   
3. Describe the block diagram of a mass spectrometer and the main ionisation techniques.
   
4. Describe the chemical identification of amines of pharmaceutical interest.
   
5. Illustrate the validation parameters of an analytical method and the experimental procedures used to determine them.
   
6. Exercise: given a chromatogram with two partially resolved peaks (retention times and widths provided), calculate the resolution, capacity factor and number of theoretical plates, and propose two changes to the chromatographic conditions to improve the separation.
   
7. Exercise: given the IR, ¹H-NMR, ¹³C-NMR and MS spectra of an unknown compound of pharmaceutical interest, propose its structure, justifying the assignment of the diagnostic signals.
   
8. Analytical case: a basic, lipophilic active ingredient must be assayed in human plasma at nanomolar concentrations. Propose a complete strategy (sample preparation, separation technique, detection, standards), justifying each choice.