37187 - Instrumental Methods in Drug Analysis (GR.A)

Learning outcomes

The student as a result of the frequency and after passing the final exam of the course of instrumental analysis of drugs, should:

- have acquired the knowledge on instrumental methods (HPLC, GC, HPLC-MS, fluorometry, HPLC-DAD, UV-VIS, etc.), which is required for the quantitative determination of compounds of pharmaceutical interest, as reported in monographs of the official Italian Pharmacopoeia (F.U.I.) and European (Ph. Eur.).

- be able to apply the gained knowledge to real problems as well as to new issues related to the pharmaceutical analysis.

Course contents

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). Examples from European Pharmacopeia. Method validation: linearity, accuracy, precision, selectivity, LOD and LOQ.

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 and time of flight analyzers. MALDI mass spectrometry.

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.


Laboratory PRACTICAL CLASSES

Quantitative analysis of drugs using instrumental techniques

1. Colorimetric Analysis of acetylcysteine in a generic drug
2. First order UV-derivative spectrophotometry: assessment of the presence of salicylic acid as an impurity in aspirin tablets.
3. First order UV-derivative spectrophotometry: stability of aspirin.
4. HPLC-DAD analysis of aspirin.
5. Fluorometric analysis of quinine in tonic water.
6. HPLC-DAD analysis of trimethoprim and sulfamethoxazole in tablets.
7. HPLC-DAD analysis of betamethasone in Ecoval cream.
8. GC-FID analysis of camphor and menthol in inhalation ointment.
9. Solid phase extraction (SPE) of racemic ketoprofen from a commercial drug using SAX cartridges. Parte1. Construction of the calibration curve. Part 2. Quantitative analysis after SPE extraction.
10. HPLC analysis on chiral stationary phase of racemic ketoprofen. Determination of the enantiomeric excess and resolution value.
11. HPLC analysis on chiral stationary phase of (S) ketoprofen in Ketesse tablets: quantitative analysis and control of the enantiomeric excess.

Readings/Bibliography

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

Lecture slides.

Teaching methods

Lectures and laboratory practical classes

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 IOL – Insegnamenti online

Office hours

See the website of Manuela Bartolini