Pharmacy

GIOVANNA POCE

This course provides the student with the theoretical knowledge and technical skills to perform drug
analysis and identification through analytical chemistry and spectroscopic methods.
By completing Drug Analysis I, the student acquires:
1. Theoretical knowledge of the most common methods of separating and purifying chemical
substances, of the most common instrumental chemical analysis techniques to determine

physicochemical measurements and of qualitative chemical structural analyses including
molecular spectroscopy through theoretical lecture-based classes and video viewing.
2. The ability to apply that theoretical knowledge. The student attends individual teaching
laboratory lectures where he applies purification and identification methods to
pharmaceutical substances. Moreover, the student attends additional group work exercises in
which he practices on how to read infrared and NMR spectra to identify chemical
substances.
3. Self-direction and to act independently to a level where the student has a practical
understanding of the learned techniques. At the end of the teaching laboratory lectures, the
student has to identify and characterize unknown chemical substances given in the
Pharmacopeia.
4. Written and oral communications skills, to a level where the student can clearly
communicate the conclusions of a chemical and spectroscopic analysis. This is evaluated
during both the group work exercises and the final test.
5. To develop subject-related practical and cognitive skills. The course provides the student
with the core elements for both further courses and the final test of the Pharmacy program.

On successful completion of the course students will be able to develop the right analytical procedure for purification and characterization of drugs with the skills and knowledge gained during the course.

For a better understanding of the theoretical knowledge and technical skills, the student must have learned:
- General chemistry and organic chemistry, necessary prerequisites for understanding the theoretical knowledge of the separation and purification methods.
- General chemistry and organic chemistry, necessary prerequisites for understanding the theoretical knowledge of the chemical analysis techniques and molecular spectroscopy.
- Physics, important prerequisites for understanding the theoretical knowledge of analysis techniques and molecular spectroscopy.
For attending Drug Analysis I, student must have attended Pharmaceutical Analytical Chemistry.

This course provides the student with the theoretical knowledge and technical skills to perform drug analysis and identification through analytical chemistry and spectroscopic methods.
Curriculum content of lecture-based classes:
1. Aspects of security and safety in a chemistry laboratory. (1 hour)
2. Pharmacopeia contents and monographs. (1 hour)
3. Separation and purification methods (10 hours)
- Solvent extraction and pH dependency; liquid-liquid extraction; continuous extraction; solid-liquid extraction; solid-liquid extraction with Soxhlet.
- Mixture separation through liquid-liquid extraction.
- Chromatography: introduction; classification of chromatographic methods; mostly used solid and liquid stationary phases; separation mechanism (partition, expanded bed absorption, ion exchange, displacement, affinity). Planar chromatography on paper and TLC. Column chromatography.
- Distillation: isobar and isotherm diagrams of ideal mixtures; simple distillation, vacuum distillation and fractional distillation of ideal and real mixtures; homogeneous and heterogeneous azeotropes; azeotropic distillation, steam distillation.
- Crystallization: principles and methods.
- Sublimation: principles and methods.
4. Chemical molecular analysis. (8 hours)
- Melting point: theoretical and technical principles, effect of impurities on melting point, methods.
- Boiling point: methods. Effect of impurities on boiling point.
- Refraction index: methods, Abbe refractometer.
- Absolute density and relative density: methods, pycnometer and Mohr-Westphal scale.
- Polarimetry.
5. Chemical structural analyses (14 hours)
- Preliminary assays: solubility, solvents, pH dependency. Solubility in acidic or basic solutions.
- Organoleptic assay.
- Calcination.
- Lassaigne assay.
- Identification assay for aromaticity and saturation.
- Carboxylic acid identification: physical properties, solubility, acidity, identification assays.
- Esters identification: physical properties, solubility, identification assays. Hydrolysis.
- Lactones identification: physical properties, solubility, identification assays.
- Amides identification: Esters identification: physical properties, solubility, identification assays. Hydrolysis.
- Nitriles identification: physical properties, solubility, identification assays.
- Amines identification: physical properties, solubility, basicity, identification assays. Hinsberg method for the separation of primary, secondary and tertiary amines.
- Amino acids identification: physical properties, solubility, identification assays.
- Ketones and aldehydes identification: physical properties, solubility, carbonyl group reactivity, identification assays.
- Carbohydrates identification: physical properties, solubility, identification assays.
- Alcohols identification: physical properties, solubility, identification assays.
- Ethers and phenol identification: physical properties, solubility, identification assays.
- Halogenated compounds identification: halogenated carboxylic compounds, halogenated alkyls and aryls (physical properties, solubility, identification assays).
- Sulphurated compounds identification: sulphates, sulphonic acids, thiols, thiophenols, thioethers, disulphures (physical properties, solubility, identification assays).
6. Spectroscopic methods. (14 hours)
- UV-Vis spectroscopy: electronic transitions, Lambert-Beer law. Single beam and double beams instruments. Factor influencing the wavelength and intensity of absorbance. Application of UV-Vis spectroscopy to qualitative analyses. Solvent and pH dependency on phenols and aromatic amine determination. Derivative spectroscopy.
- IR spectroscopy: introduction, mechanic and harmonic models. Vibrational modes of linear and nonlinear compounds. Number of vibrational modes. Factors that influence IR resonance. Instruments. FT-IR. Sample preparation. Spectra interpretation. Characteristic bands of functional groups.
- NMR spectroscopy: introduction, instruments, solvents for sample preparation. 1H NMR: chemical shift, TMS as zero, number of signal, intensity and integrals, spin-spin coupling, multiplets analysis. Spectra interpretation. 13CNMR: principles.
12 hours of flipped classroom for analysis of IR and NMR spectra.

Individual teaching laboratory lectures (60 hours):
- Separation of a mixture by liquid/liquid extraction.
- Crystallization.
- Sublimation.
- Calcination.
- Solubility assay.
- Melting point determination.
- Functional groups identification according to the Pharmacopeia.
- TLC analysis of Calcium pantothenate.
- Identification of unknown substances given in the Pharmacopeia.

- Caliendo, G. Manuale di Analisi Qualitativa, Ed. EdiSES
For separation and purification methods, molecular analysis, UV-Vis spectroscopy., and for chemical structural analysis.

- Silverstein, R. M.; Webster, F. X.; Kiemle, D. J.; Bryce, D. L. Identificazione spettrometrica di composti organici, Ed. Ambrosiana
For: IR and NMR spectroscopy.

This course provides the student with the theoretical knowledge and technical skills to perform drug analysis and identification through analytical chemistry and spectroscopic methods.
Teaching/learning methods and strategies:
- 48 hours lectures will be used to deliver the core material supplemented with videos.
- 12 hours of flipped classroom for analysis of IR and NMR spectra.
- The practical sessions will complement the material presented in lectures by 60 hours individual teaching laboratory lectures. It will provide guidance and experience of following written experimental procedures and help students to consolidate their practical skills and the subsequent reporting and analysis of practical results.

Mandatory

The assessment strategy is designed to verify student ability in critically evaluating and representing theoretical and practical knowledge acquired during the course.

Assessment strategy:
- At the end of the teaching laboratory lectures, the student has to identify and characterize 3 unknown chemical substances given in the Pharmacopeia. Self-direction and to act independently in choosing the right analytical process will be assessed. The test takes 8 hours divided by two laboratory lectures.
- End-of-course written exam consists of:
o A first part where the student has to interpret simple spectroscopic data (IR, 1H NMR and 13C NMR spectra) in terms of absorption bands, chemical shifts, integration traces and spin-spin coupling patterns.
o A second part where both theoretical and practical knowledge of the most common methods of separating and purifying chemical substances (extraction methods, chromatography, distillation, sublimation and crystallization), of the most common instrumental chemical analysis techniques to determine physicochemical measurements (melting point, boiling point, density, refractive index and specific optical rotation) and of qualitative chemical structural analyses (identification of functional groups) and UV-visible, infrared (IR), nuclear magnetic resonance (NMR) will be assessed.
It is a requirement that the major categories of assessment are passed separately in order to achieve an overall pass.

Exercise 1:
Determine the structure of the compound with the molecular formula C₁₀H₁₂O₂ by analyzing the following spectra.

Exercise 2:
Describe:
• the physical state of the analyte identified in Exercise 1
• the solubility and acid-base characteristics of the analyte identified in Exercise 1, describing the procedures and results of the experimental determination of its solubility and pH.

Exercise 3:
Which tests would respond to the functional group responsible for the IR absorption band at 1691 cm⁻¹? Describe the tests in detail.

Exercise 4:
For the following mixture, indicate the best separation and purification conditions for the individual components once separated. Describe the methods used and the physicochemical constants that could be determined after purification.

Exercise 5:
If an unknown analyte gives a positive result in Fehling’s test, how could one distinguish between an aldehyde and a sugar?