ANALISI STRUMENTALE DEI FARMACI A - L

At the end of the course the student: - acquires theoretical knowledge of chromatographic and capillary electrophoresis separations; - will be able to apply the acquired knowledge for the development of chromatographic methods for the quali-quantitative identification of drugs in quality control, in medicinal chemistry and in pre-clinical and clinical research fields; - acquires basic notions on the enantioseparation of chiral drugs; - acquires knowledge for the validation of analytical methods for registration and marketing of new drugs; - acquires basic notions on the use of HPAC analytical methodologies for the study of drug/protein interaction in the development of new drugs.

Lectures (35 hrs) and laboratory practical classes (12 hrs) on high performance liquid chromatographs (HPLC, HPLC-IEC).

LECTURES

Introduction to instrumental methods of analysis: classification and selection of analytical methods. Chromatographic techniques. General principles of the chromatographic process and chromatographic parameters. Distribution constant. Retention time. Capacity factor. Selectivity. Efficiency: plate theory; van Deemter's equation; extra-column factors. Asymmetry of chromatographic peaks. Resolution. Liquid chromatography: stationary phase, mobile phase. Liquid-solid chromatography, liquid-liquid chromatography, ion exchange chromatography, ion pair chromatography, molecular exclusion chromatography, affinity chromatography. Column chromatography, thin layer chromatography (TLC). High performance liquid chromatography (HPLC): instrumentation. Gas chromatography (GC): instrumental aspects, types of stationary phase, detection in GC. Enatioselective chromatography separation: direct method with use of enantioselective stationary phases; indirect method with derivatization with defined stereochemical reagents. 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). Qualitative and quantitative analysis in HPLC (external and internal standard methods). Sample preparation and extraction methods. Traditional extraction methods (liquid/liquid and liquid/solid). Extraction with supercritical fluids (SFE). Solid phase extraction (SPE). Preparation of samples from complex biological matrices. Validation of the analytical method: linearity, precision, accuracy, reproducibility, limit of detection or detention (LOD) and the limit of quantification (LOQ). Development of analytical methods for qualitative and quantitative identification of drugs in quality control and in pre-clinical and clinical research. Determination of drug/protein interactions, using high performance affinity chromatography (HPAC), useful for the development of new drugs.

LABORATORY PRACTICAL CLASSES

High performance liquid chromatography: development of an HPLC method for the separation of drugs in complex matrices. Study of efficiency, selectivity and resolution. Calculation of chromatographic parameters (capacity factor, selectivity factor, resolution, number of theoretical plates, peak asymmetry). Qualitative/quantitative determination of drugs in pharmaceutical forms present on the market.

• V. Cavrini. Principi di Analisi Farmaceutica, terza edizione. Società Editrice Esculapio, Bologna.
• Veronika R. Meyer. Practical High-Performance Liquid Chromatography, 5th Edition. Wiley, New York (testo di consultazione)
• Teaching material (slides and pubblications) provided by the teacher and available on the teacher's personal website (www.angelospadaro.it).