Advanced analysis of active principles in herbal drugs

A.Y. 2019/2020
8
Max ECTS
80
Overall hours
SSD
CHIM/08
Language
Italian
Learning objectives
The main objective of the course is to illustrate and discuss the most important analytical approaches used for plant drug analysis. In particular, the first objective of the course is the transfer of knowledge of the main methods for the analysis of plant drugs and derivatives present in the current Pharmacopeia. The second objective is to provide students with a general understanding of the most modern analytical instrumental approaches and their applications to identify, characterize and dose the analytes in plant matrices. The course aims therefore not only to transfer to the student the knowledge necessary to understand the analytical methods and their applications for the analysis of plant drugs and extracts according to the current pharmacopoeia but also to provide those methodological tools useful to develop analytical methods for qualitative and quantitative analysis, on the basis of the structure of the analytes, the matrix and of the sensitivity required. In order to make the notions provided by the theoretical course understandable, part of the methodologies discussed will be experimentally performed in teaching lab, which will allow a concrete knowledge of the analytical techniques and the functioning of the analysis tools.
Another goal of the practical lessons will be the learning of the correct use of the main laboratory instruments.
Expected learning outcomes
At the end of the learning process the student must know the most important analytical applications used in the processes of research, development and production of the drug. The student must also understand the main analytical problems and design possible solutions applying not only the knowledge acquired during the course but also that obtained in the modules of advanced methodologies in pharmaceutical chemistry. The student will therefore have to demonstrate a full capacity to approach and solve a series of analytical problems that will be presented during the exam, also demonstrating communication skills using correct and appropriate terminology.
Course syllabus and organization

Single session

Responsible
Lesson period
First semester
Course syllabus
Introduction to the course
- the main instrumental techniques for the qualitative and quantitative analysis of the active ingredients of plant drugs

UV-Visible Spectroscopy
- theoretical principles (electromagnetic radiation and Planck equation, molecular orbitals and types of transitions, Jablonski diagram, absorption characteristics of organic compounds, selection rules, absorption law, quantitative aspects (std. external, standard addition method, mixture analysis), absorption prediction (Fieser and Woodward, Fieser Kuhn's rules), instrumentation (source, monochromator, prisms and gratings, detectors, photodiodes, single and double beam spectrophotometers, diode array spectrophotometers).
- Application of UV/VIS spectroscopy to the analysis of the active principles of plant drugs: qualitative (identification and characterization) and quantitative aspects.
- Examples of spectroscopic analysis according to European Pharmacopoeia (9th edition): cranberry anthocyanosides, lemon essential oil, hypericin in the hypericum, dry extract of aloe vera.

Chromatographic separation techniques
- Introduction to chromatography analysis (historical notes, chromatographic classification and techniques, chromatogram, retention factor, chromatographic resolution, plate theory, efficiency and selectivity, Van Deemter's equation, general problem of elution, symmetry of peaks).

Gas chromatography
- General principles: classification of gas chromatographic techniques, instrumentation and stationary phases, derivatization, quantitative analysis,
- applications of GC for the qualitative and quantitative analysis of the active principles of plant drugs; examples of analysis in GC according to European Pharmacopoeia (9th edition): essential oils, residual solvents and pesticides.

High performance liquid chromatography
- General principles: classification of HPLC techniques, stationary and mobile phases, UHPLC and nano-HPLC, instrumentation and detectors, quantitative analysis in HPLC;
- applications of chromatography in quantitative analysis of the active principles of plant drugs (examples from the current European Pharmacopoeia).
Analysis of catechins in green tea, ginsenosides in dry extract of ginseng, juniper acids and terpene lactones in ginkgo biloba.

Mass spectrometry
- General principles: Principles of ionisation and dissociation of ions, isotopic composition and accurate mass, instrumentation (sources and analysers), nitrogen rule.
- Application of mass spectrometry in identification, quantization characterization of active ingredients of plant drugs. Determination of molecular formula from mass spectrum and isotopic pattern, identification of analytes by database, fragmentation pattern studies,.

Infrared spectroscopy
- Vibration of the diatomic molecules, the model of the harmonic oscillator. Vibrations of polyatomic molecules: normal modes, vibrational transitions and related absorption bands. Instrumentation and sample preparation. General characteristics of an infrared spectrum and absorption bands. Interpretation of infrared spectra. IR absorption bands characteristic of certain classes of organic compounds.

Development and validation of an analytical method
- criteria for the choice of the optimal analytical method, development and optimization of the instrumental method, validation of the analytical method.

Sample preparation
- sampling, sample preparation and storage, sample preparation techniques, liquid-liquid extraction, liquid-solid extraction, solid phase extraction, experimental and application aspects.

Pharmacopoeia analysis:
- determination of content of essential oils, determination of water in drugs containing volatiles, alcohol content of tinctures, essential oils

[Program for not attending students with reference to descriptor 1 and 2]:
Introduction to the course
- the main instrumental techniques for the qualitative and quantitative analysis of the active ingredients of plant drugs

UV-Visible Spectroscopy
- theoretical principles (electromagnetic radiation and Planck equation, molecular orbitals and types of transitions, Jablonski diagram, absorption characteristics of organic compounds, selection rules, absorption law, quantitative aspects (std. external, standard addition method, mixture analysis), absorption prediction (Fieser and Woodward, Fieser Kuhn's rules), instrumentation (source, monochromator, prisms and gratings, detectors, photodiodes, single and double beam spectrophotometers, diode array spectrophotometers).
- Application of UV/VIS spectroscopy to the analysis of the active principles of plant drugs: qualitative (identification and characterization) and quantitative aspects.
- Examples of spectroscopic analysis according to European Pharmacopoeia (9th edition): cranberry anthocyanosides, lemon essential oil, hypericin in the hypericum, dry extract of aloe vera.

Chromatographic separation techniques
- Introduction to chromatography analysis (historical notes, chromatographic classification and techniques, chromatogram, retention factor, chromatographic resolution, plate theory, efficiency and selectivity, Van Deemter's equation, general problem of elution, symmetry of peaks).

Gas chromatography
- General principles: classification of gas chromatographic techniques, instrumentation and stationary phases, derivatization, quantitative analysis,
- applications of GC for the qualitative and quantitative analysis of the active principles of plant drugs; examples of analysis in GC according to European Pharmacopoeia (9th edition): essential oils, residual solvents and pesticides.

High performance liquid chromatography
- General principles: classification of HPLC techniques, stationary and mobile phases, UHPLC and nano-HPLC, instrumentation and detectors, quantitative analysis in HPLC;
- applications of chromatography in quantitative analysis of the active principles of plant drugs (examples from the current European Pharmacopoeia).
Analysis of catechins in green tea, ginsenosides in dry extract of ginseng, juniper acids and terpene lactones in ginkgo biloba.

Mass spectrometry
- General principles: Principles of ionisation and dissociation of ions, isotopic composition and accurate mass, instrumentation (sources and analysers), nitrogen rule.
- Application of mass spectrometry in identification, quantization characterization of active ingredients of plant drugs. Determination of molecular formula from mass spectrum and isotopic pattern, identification of analytes by database, fragmentation pattern studies,.

Infrared spectroscopy
- Vibration of the diatomic molecules, the model of the harmonic oscillator. Vibrations of polyatomic molecules: normal modes, vibrational transitions and related absorption bands. Instrumentation and sample preparation. General characteristics of an infrared spectrum and absorption bands. Interpretation of infrared spectra. IR absorption bands characteristic of certain classes of organic compounds.

Development and validation of an analytical method
- criteria for the choice of the optimal analytical method, development and optimization of the instrumental method, validation of the analytical method.

Sample preparation
- sampling, sample preparation and storage, sample preparation techniques, liquid-liquid extraction, liquid-solid extraction, solid phase extraction, experimental and application aspects.

Pharmacopoeia analysis:
- determination of content of essential oils, determination of water in drugs containing volatiles, alcohol content of tinctures, essential oils

Application examples of analytical techniques for the identification of analytes in matrices, chemotaxonomy study, bioactivity explanation, quality control, adulteration, counterfeiting, and pollutant analysis
Prerequisites for admission
Knowledge of general and inorganic chemistry, organic chemistry, analytical toxicological-chemistry and chemistry and phytopharmaceutcial analysis
Teaching methods
Frontal lectures
Teaching Resources
All the teaching material presented during the lessons (constituting the topics of the exam) is made available in pdf format on the Ariel portal of Unimi.
Recommended texts: CAVRINI ANDRISANO - Principi di Analisi Farmaceutica, casa editrice Esculapio
Assessment methods and Criteria
The exam consists of a written test which is related to the topics covered in class, and is articulated in several points with exercises and open-ended questions with a final grade out of thirty. The written test lasts 120 minutes.
In order to be able to take the examination, it is essential to register for the exam session through the online SIFA service of the University. The outcome of the written exam will be published on the Ariel portal of Unimi.
CHIM/08 - PHARMACEUTICAL CHEMISTRY - University credits: 8
Single bench laboratory practical: 32 hours
Lessons: 48 hours
Shifts:
Professor: Aldini Giancarlo
Turno 1
Professor: Sgrignani Jacopo