31571 - Phytoderivative Analysis II

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

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

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

- Types of detection, principles of their operation, coupling to separation techniques and effects on 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).

- Sampling method and pretreatment, particularly innovative and miniaturized ones, of 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

- Instrumental separative techniques: application of HPLC methods to phytoderivatives and related products.

- Instrumental separative techniques: application of GC methods to phytoderivatives and related products.

- UV-Vis absorption spectroscopy: application to the qualitative-quantitative analysis of phytoderivatives.

- Molecular fluorescence spectroscopy: application to the qualitative-quantitative analysis of phytoderivatives.

PROGRAM OF THE LABORATORY ACTIVITIES
Module 1 - Advanced principles and methods of the phytoderivative analysis

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

1) Analysis of curcumin and analogues in rhizomes of Curcuma longa (curcuma) and products derived by HPLC-DAD (part 1)

2) Analysis of curcumin and analogues in rhizomes of Curcuma longa (curcuma) and products derived by HPLC-DAD (part 2)

3) Qualitative analysis of xanthine alkaloids in seeds of Paullinia cupana (guaranà) and derived products by TLC

4) Quantitative analysis of xanthine alkaloids in seeds of Paullinia cupana (guaranà) and derived products by HPLC-DAD

5) Identification of aloin and congeners in Aloe barbadensis (Aloe vera) and in Aloe ferox by TLC and HPLC-DAD

6) Hydrolysis of aloin to aloe-emodin: confirmation by TLC and HPLC-DAD

 

Readings/Bibliography

STUDY MATERIALS:
- Slides presented and discussed during the lectures
- Papers from the scientific literature 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

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 - 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 - 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 takes 2 oral examinations, 1 for each module, 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;

- 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 of the 2 modules 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.
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 Elisa Uliassi