31571 - Phytoderivative Analysis II

Course Unit Page

SDGs

This teaching activity contributes to the achievement of the Sustainable Development Goals of the UN 2030 Agenda.

Good health and well-being Responsible consumption and production Life on land

Academic Year 2021/2022

Learning outcomes

At the end of the course the student possesses an advanced theoretical and practical knowledge to carry out an updated phytoderivative analysis. In particular, the student learns the analytical characteristics of active compounds in different types of phytoderivatives and related products.
Moreover, the students gains the skills to perform a reliable quali-quantitative analysis, by using the most proper techniques and methods applied to herbal origin samples.
The course is composed of both lectures and single seat laboratory activities.

Course contents

PROGRAM OF THE LECTURES
Module 1 - Advanced principles and methods of the phytoderivative analysis (Prof. Laura Mercolini) 

 - Principles, aims and problems of the advanced phytoderivative analysis. Peculiar characteristics of the instrumental analysis applied to herbal matrices and related products.

- Method validation referred to the application to phytoderivatives: validation parameters according to the international guidelines.

- Instrumental separative techniques: liquid chromatography (HPLC, UHPLC, and nano-LC), gas chromatography (GC) and capillary electrophoresis (CE). Specific principles of each analytical technique and related instrumentations.

- Means of detection, principles for their operation, coupling to separation techniques and effects on the analytical performance.

- UV-Vis absorption spectroscopy: instrumentation, quantitative analysis, spectrophotometry.

- Molecular fluorescence spectroscopy: instrumentation, quantitative analysis, analysis by quenching. Coupling to separative techniques: native fluorescence and after derivatization.

- Mass spectrometry: ionization techniques (ESI, APCI, MALDI), types of sources, mass analyzers (QqQ, TOF, Iontrap, Orbitrap) and acquisition Mode (SRM, MRM). High resolution mass spectrometry (HRMS).

- Sampling method and pretreatment, particularly focusing on innovative and miniaturized ones, for the  matrices obtained from phytoderivatives: S/L extraction, L/L extraction, solid phase extraction (SPE), MicroExtraction by Packed Sorbent (MEPS).

 

PROGRAM OF THE LECTURES
Module 2 - Application of instrumental analysis to herbal matrices (Prof. Federica Belluti)

- Analytical profile of some classes of natural molecules: cinnamic acids and esters, anthocyanins, terpenes, flavonoids, alkaloids, curcuminoids.

- UV-Visible absorption spectroscopy: experimental and applicative aspects of UV-Vis spectroscopy for the analysis of the main chemical classes of phytoderivatives (cinnamic acids and esters, anthocyanins, terpenes, flavonoids, alkaloids, curcuminoids). Qualitative (identification and characterization) and quantitative aspects. Examples of spectroscopic analysis according to European Pharmacopoeia and Italian Pharmacopoeia.

- Chromatographic separative techniques: experimental and applicative aspects of chromatographic methods (TLC, HPLC and GC) for the analysis of the main chemical classes of phytoderivatives (cinnamic acids and esters, anthocyanins, terpenes, flavonoids, alkaloids, curcuminoids). Qualitative and quantitative aspects. Examples of chromatographic analysis according to European Pharmacopoeia and Italian Pharmacopoeia.

- Development and validation of an analytical method: criteria for the selection of the most suitable analytical method. Development and optimization of an analytical instrumental method.

- Sample preparation: experimental and applicative aspects of sample preparation through the use of techniques such as: liquid-liquid extraction, liquid-solid extraction, solid phase extraction.

- Examples of application of the analytical techniques for the identification and quantitation of phytoderivatives.



PROGRAM OF THE LABORATORY ACTIVITIES
Module 1 - Advanced principles and methods of the phytoderivative analysis (Prof. Laura Mercolini)

1) Analysis of glycyrrhizin and glycyrrhetinic acid in roots of Glycyrrhiza glabra (licorice) and derived products: identification by TLC

2) Analysis of glycyrrhizin and glycyrrhetinic acid in roots of Glycyrrhiza glabra (licorice) and derived products: quantitation by SPE-HPLC-DAD

3) Analysis of glycyrrhizin and glycyrrhetinic acid in roots of Glycyrrhiza glabra (licorice) and derived products: quantitation by MEPS-HPLC-DAD

 

PROGRAM OF THE LABORATORY ACTIVITIES
Module 2 - Application of instrumental analysis to herbal matrices (Prof. Federica Belluti)

Practical examples of application of the techniques to the analysis and purification of phytoderivatives.
Quantitative (UV-Vis and HPLC-DAD spectroscopy) and qualitative (TLC) analysis of the active ingredients of the following herbal drugs:
Craategus monogyna
Curcuma longa
Cynara scolymus
Paullinia cupana var. sorbilis.

Processing and critical evaluation of the data obtained.

 

Readings/Bibliography

STUDY MATERIALS:
- Slides presented and discussed during the lectures
- Papers from the scientific literature presented and discussed during the lectures
- Monographs of the Italian Pharmacopoeia and the European Pharmacopoeia presented and discussed during the lectures
- Videos and similar materials
- Handout of the laboratory experiments.



FURTHER MATERIALS:

- Carrieri A. "Manuale di Analisi Quantitativa dei Medicinali"
EDISES, Napoli, 2019.
- Holler J.F., Crouch S.R. “Fondamenti di Chimica Analitica” di Skoog & West, EdiSES s.r.l., 2015.
- Hage D.S., Carr J.D. "Chimica Analitica e Analisi Quantitativa", Piccin, 2012.
- Skoog D.A., Holler J.F., Crouch S.R., Sabbatini L. “Chimica Analitica Strumentale”, EdiSES s.r.l., 2009.

 

Teaching methods

Considering the activities and teaching methods adopted in this course, attending it requires the students to have already completed modules 1 and 2 (in e-learning mode at this web address, as well as module 3 (in person) of the specific "Health and Safety in places of study" course. An updated schedule and the manner of attending module 3 are available in the corresponding section of the Graduation Programme webpage.

 

The course consists of lectures (32 h, 4CFU) during which the main instrumental techniques and methods required for the analysis phytoderivatives are discussed.
The course deals also with single seat laboratory activities (36 h, 3 CFU), during which the student prepares a laboratory notebook. The laboratory activities offer the student the opportunity of carrying out different types of advanced analysis for identification and quantitation of the compounds of interest in phytoderivatives and related products, by firsthand using the techniques and methodologies required to perform such analyses.

The course consists of exactly 2 modules: Module 1 - ADVANCED PRINCIPLES AND METHODS OF THE PHYTODERIVATE ANALYSIS (Prof. Laura Mercolini)- 4 CFU, of which 3 CFU (24 hours) of lectures and 1 CFU (12 hours) of laboratory activities. Module 2 - APPLICATION OF INSTRUMENTAL ANALYSIS TO HERBAL MATRICES (Prof. Federica Belluti) - 3 CFU, of which 1 CFU (8 hours) of lectures and 2 CFU (24 hours) of laboratory activities.

Lecture attendance is very important for a satisfactory learning process.
Laboratory activity attendance is mandatory for the learning process.
The student can take the oral examination for each module in the same day and anyway within the same session, and at the end the student will achieve a single grade for the Phytoderivatives Analysis II course.

 

Assessment methods

The assessment of the learning aims to assess practical skills and theoretical knowledge acquired by the student while attending the lectures and the laboratory activities. In particular, the following skills are evaluated:

- Knowledge of the advanced instrumental analytical terminology and of the theoretical basis of the discussed topics;

- Ability to perform basic laboratory procedures for the quali-quantitative analysis;

- Specific training on the uses, properties and analytical procedures of the examined products and compounds;

- Theoretical and practical familiarity with the main analytical applications described in the Pharmacopoeia for the analysis of phytoderivatives and natural extracts.

- Critical capacity in the evaluation of the laboratory analytical results.

 

The assessment of the learning is performed on the basis of an oral examination for each module and consisting of some questions (5 to 10 questions) about both the theoretical and the practical parts.
The student could also be asked to comment on a paper from the scientific literature and chosen among those presented and discussed during the lectures, with the possibility for the student to consult the paper itself. Similarly, the comment of the Monographs of the Italian Pharmacopoeia and/or of the European Pharmacopoeia may be requested on those presented and discussed during the lectures.
Moreover, during the laboratory activities of each module, it is planned to carry out practical tests the results of which are reported by the student within his/her own laboratory notebook, evaluated by the teacher, and which is part of the final score. A laboratory notebook must therefore be prepared for each module. The file of the laboratory notebook has to be delivered to the teacher of the corresponding module at least one week before taking the oral exam.

 

Teaching tools

Lectures are carried out by means of the common electronic media, such as pc, tablet, smartphone and similar devices connected to a videoprojector and to the wi-fi.

The practical activities are developed in specially equipped didactic laboratories where each student has at disposal a single place of work, all the instrumentations and materials necessary for the advanced phytoderivative analysis.


Office hours

See the website of Laura Mercolini

See the website of Federica Belluti