ANALYSIS OF TOXICOLOGICAL CHEMISTRY II

LEARNING OUTCOMES

The aim of the course is to provide the student with:

-the general rules for good laboratory practices and safety in compliance with Italian law requirements;

-the ability to perform qualitative inorganic analysis;

-the knowledge of the theory of chromatographic separations;

-the ability to develop instrumental high-performance liquid chromatography methods for identification and determination of toxic compounds in complex matrices. The course consists of lectures, seminars and laboratory exercises.

The laboratory exercises are individual when applied to the analysis of species of toxicological interest, and in a group, when making the instrumental analysis.

Attendance at lectures and workshops is mandatory.

The teacher uses three different teaching methods:

1. exhibition method: the lesson;

2. active method: the use of procedures that actively involve the student in the learning process;

3. operative method: laboratory activity with single and group experiments, manual and instrumental.

COURSE SYLLABUS

Safety in laboratory

General rules for good laboratory practices and safety. Italian legislative decrees in safety on health and in workplaces. Types of risks. Material safety data sheets. R-, S-, H- and P-phrases. Safety pictograms. Personal protective equipment. Handling of chemicals. Disposal of chemicals. Acute and chronic intoxication. Hazardous chemicals. Devices aid and behavior in case of accidents.

Analysis inorganic species of toxicological interest

Environmental Chemical Analysis. Sampling . Analysis of water samples . atmospheric monitoring : oxidants, hydrocarbons, particulates, oxides of carbon, sulfur and nitrogen.

Qualitative systematic analysis of cations in mixture. Analytical group of cations: silver (I), copper-arsenic (II), aluminum-iron (III-IV), alkaline earth (V) and alkali metals group (VI). Qualitative anion analysis. Dry tests.

Chemical-analytical profile of inorganic species of toxicological interest: mercury, lead, silver, arsenic, chlorides, bromides, iodides, sulfates, phosphates, nitrates, oxalates, acetates, borates, carbonates, sulfites.

Sample preparation

Solvent extraction: liquid/solid and liquid/liquid extraction. Partition coefficient. Extraction efficiency: pH, temperature and salts addition. Application to pharmaceutical formulations and biological material.

Introduction to chromatography

General concepts and principal classification of chromatographic methods: partition and adsorption chromatography, ion exchange and size exclusion chromatography, affinity chromatography. Chromatographic techniques: thin layer chromatography (TLC), gas chromatography (GC), high pressure liquid chromatography (HPLC). Instrumentation. Detectors. Theory of chromatographic separations: retention time, distribution constant, capacity factor, selectivity. Efficiency: plate theory; van Deemter equation; extra-column factors. Peak asymmetry. Resolution. Direct and reversed phase HPLC methods. Fast HPLC, UPLC (ultra pressure liquid chromatography). Optimization of chromatographic separations: selection of the separation techniques based on the chemical-physical properties of the drug; approaches for the selection of the stationary phase; solvent selectivity groups; Isocratic and gradient elution, Quali-quantitative analysis by chromatographic techniques: external standard and internal standard methods, calibration curves. Peaks normalization. Method validation in drug analysis: linearity, precision, accuracy, reproducibility, detection and quantification limits (LOD, LOQ).

READINGS/BIBLIOGRAPHY

D. A. Skoog, F.J. Holler, S. R. Crouch, Chimica analitica strumentale, Edises;

R.L Snyder, J.J. Kirkland, J.L. Giajch, Practical HPLC method development, Wiley;

C. F. Poole, The essence of chromatography, Elsevier.

T. R. Hogness, W. C. Johnson, A. R. Armstrong, Analisi qualitativa ed equilibrio chimico, Piccin;

A. Araneo Chimica analitica qualitativa, Ambrosiana

Selected articles and reviews on specific arguments.

Lecture slides, analytical laboratory guideline and methods.