LABORATORY OF DRUG ANALYSIS III A - L
Academic Year 2022/2023 - Teacher: Angelo SPADAROExpected Learning Outcomes
Applying knowledge and understanding. The student will have to use the knowledge acquired to perform analytical methods that involve analysis in solution and by instrumental chromatographic techniques.
Making judgments. The studentwill have to interpret the experimental results obtained relating to the analysis of unknown mixtures and active principles present in complex matrices.
Communication skills. At the end of the course the student will have to explain what has learned using appropriate terminology for a correct scientific presentation regarding the analytical techniques and the analysis of compounds of pharmaceutical interest.
Lifelong learning skills. The student will have to developed useful theoretical/practical skills in order to update and expand his knowledge with the aim of face new analytical problems.
Course Structure
Frontal lessons (28 hrs) and laboratory training (30 hrs).
Should teaching be carried out in mixed mode or remotely, it may be necessary to introduce changes with respect to previous statements, in line with the programme planned and outlined in the syllabus.
Information for students with disabilities and / or SLD: To guarantee equal opportunities and in compliance with the laws in force, interested students can ask for a personal interview in order to plan any compensatory and / or dispensatory measures, based on the teaching objectives and specifications needs. It is also possible to contact the CInAP contact person (Center for Active and Participatory Integration - Services for Disabilities and / or SLD) of the DSFS, Prof. Teresa Musumeci.
Required Prerequisites
The basic knowledge necessary for understanding the course relates to General and Inorganic Chemistry, Organic Chemistry, Physical Chemistry, Physics, and Pharmaceutical Analysis Laboratory I and II.
Attendance of Lessons
Attendance in the classroom is mandatory. The maximum limit of absences allowed is 30% as reported in the teaching regulations of the CdS which can be consulted on the website of the Department of Pharmaceutical and Health Sciences (http://www.dsf.unict.it/corsi/lm-13/orario-delle-lezioni ). Once this limit has been exceeded, it will not be possible to access the practical laboratory exercises.
Detailed Course Content
Identification of compounds registered in the FUI and in the Pharm. Eur. Separation of mixtures (solvent extraction, crystallization, distillation, evaporation, filtration, centrifugation). Preliminary tests: organoleptic characteristics, combustion behavior of substances, qualitative (Lassaigne's test) and quantitative (CHNS, minimum formula determination) elementary analysis. Solubility in water and in reactive solvents; classification of substances based on their solubility.
Functional groups and assays for recognition: carboxylic acids, amides, esters, lactones, anhydrides, hydroxy acids, phenols, alcohols, aldehydes and ketones, ethers, halogen derivatives, amines, nitro compounds, aromatic hydrocarbons. Analysis of carbohydrates and amino acids.
Determination of physical constants: melting point, boiling point, density, refractive index, optical rotation.
Extractive methods for sample preparation. Classic extraction methods (liquid / liquid and liquid / solid). Extraction with supercritical fluids (SFE). Solid phase extraction (SPE). Preparation of samples from complex biological and pharmaceutical matrices.
Chromatographic techniques. General principles of the chromatographic process and chromatographic parameters. Constant of distribution. Retention time. Capacity factor. Selectivity. Efficiency: plate theory; van Deemter equation. Resolution. Chromatographic peak asymmetry.
Column chromatography, thin layer chromatography (TLC). High performance liquid chromatography (HPLC, UPLC). Ion exchange chromatography (IEC). Molecular exclusion chromatography (GPC, GFC). Affinity chromatography. Determination of drug / target protein interactions, by high performance affinity chromatography (HPAC), for the development of new drugs.
Enatioselective chromatographic separations. Gas chromatography (GC): instrumental aspects, types of stationary phase, detection in GC. Capillary electrophoresis (EC): instrumentation, theoretical principles, analytical parameters. Combined techniques: liquid chromatography and photodiode UV-VIS detector (HPLC-DAD), liquid chromatography and mass spectrometry (HPLC-MS), gas chromatography and mass spectrometry (GC-MS).
Qualitative and quantitative analysis in chromatography (methods of the external and internal standard). Validation of the analytical method: linearity, precision, accuracy, reproducibility, limit of detection (LOD), limit of quantification (LOQ).
Development of chromatographic methods for drug dosage in quality control and pre-clinical and clinical research.
Textbook Information
Manuale di analisi qualitativa a cura di Giuseppe Caliendo – Edises I/2020 edizione (1 gennaio 2019) D. G. Watson - Analisi farmaceutica – Edra 2014.- V. CAVRINI, V. ANDRISANO. Principi di Analisi Farmaceutica. Società Editrice Esculapio, Bologna. IV edizione 2018.
- R.M. SILVERSTEIN, F.X. WEBSTER: "Identificazione spettroscopica di composti organici" - Ambrosiana Editrice, Milano, 2016 (testo di consultazione)
- Teaching material provided by the teacher is available on Studium
Course Planning
| Subjects | Text References | |
|---|---|---|
| 1 | Identification of compounds registered in the FUI and Pharm. Eur. Separation of mixtures (solvent extraction, crystallisation, distillation, evaporation, filtration, centrifugation). Preliminary assays: organoleptic characters, combustion behaviour of substances, qualitative elemental analysis (Lassaigne assay) and quantitative elemental analysis (CHNS, minimum formula determination). Solubility in water and reactive solvents; classification of substances according to their solubility. | Book 1, 2, 3, 4 and teaching material |
| 2 | Functional groups and assays for recognition: carboxylic acids, amides, esters, lactones, anhydrides, hydroxy acids, phenols, alcohols, aldehydes and ketones, ethers, halogen derivatives, amines, nitro compounds, aromatic hydrocarbons. Recognition of carbohydrates and amino acids. | Book 1, 2, 3, 4 and teaching material |
| 3 | Determination of physical constants: melting point, boiling point, density, refractive index, rotational optical power. | Book 1, 2, 3, 4 and teaching material |
| 4 | Extractive methods for sample preparation. Classical extraction methods (liquid/liquid and liquid/solid). Supercritical fluid extraction (SFE). Solid-phase extraction (SPE). Sample preparation from complex biological and pharmaceutical matrices. | Book 1, 2, 3, 4 and teaching material |
| 5 | Chromatographic techniques. General principles of the chromatographic process and chromatographic parameters. Distribution constant. Retention time. Capacity factor. Selectivity. Efficiency: plate theory; van Deemter equation. Resolution. Chromatographic peak asymmetry. | Book 1, 2, 3, 4 and teaching material |
| 6 | Column chromatography, thin layer chromatography (TLC). High performance liquid chromatography (HPLC, UPLC): instrumentation. Ion exchange chromatography (IEC). Molecular exclusion chromatography (GPC, GFC). Affinity chromatography. Determination of drug/target protein interactions using high-performance affinity chromatography (HPAC) for new drug development. | Book 1, 2, 3, 4 and teaching material |
| 7 | Enatioselective chromatographic separations. Gas chromatography (GC): instrumental aspects, stationary phase types, GC detection. Capillary Electrophoresis (EC): instrumentation, theoretical principles, analytical parameters. Combined techniques: liquid chromatography and UV-VIS photodiode detector (HPLC-DAD), liquid chromatography and mass spectrometry (HPLC-MS), gas chromatography and mass spectrometry (GC-MS). | Book 1, 2, 3, 4 and teaching material |
| 8 | Qualitative and quantitative analysis in chromatography (external and internal standard methods). Analytical method validation: linearity, precision, accuracy, reproducibility, limit of detection (LOD), limit of quantification (LOQ). Development of chromatographic methods for drug assays in quality control and pre-clinical and clinical research. | Book 1, 2, 3, 4 and teaching material |
| 9 | Laboratory Exercises. Identification of compounds of pharmaceutical interest reported in the FUI and Pharm. monographs. Eur. Solubility testing. Acid - basic - amphoteric character. Identification of functional groups. Melting point determination. Identification of unknown samples by TLC. Measurement of rotational optical power and enantiomeric excess. Determination of refractive index. Determination of the density of a solid and a liquid sample. | Book 1, 2, 3, 4 and teaching material |
| 10 | Laboratory Exercises. Development of analytical methods for the separation of drugs by HPLC. Qualitative/quantitative determination of molecules of pharmaceutical interest in medicinal products. Extraction, purification and characterisation of an active ingredient from a complex matrix. | Book 1, 2, 3, 4 and teaching material |
Learning Assessment
Learning Assessment Procedures
Practical, written and/or oral exam.
Learning assessment may also be carried out online, should conditions require it.
Information for students with disabilities and / or SLD
To guarantee equal opportunities and in compliance with the laws in force, interested students can ask for a personal interview in order to plan any compensatory and / or dispensatory measures, based on the teaching objectives and specifications needs. It is also possible to contact the CInAP contact person (Center for Active and Participatory Integration - Services for Disabilities and / or SLD) of the Department of Drung and Health Sciences.
Examples of frequently asked questions and / or exercises
Physico-chemical characteristics and amine recognition reactions. Factors Affecting Resolution in HPLC. Capillary electrophoresis: theoretical principles, instrumentation and applications in the pharmaceutical field.