Learning objectives
The objective of this course is to provide students, through theoretical lectures and exercises in the classroom, knowledge relating to principles of chemical equilibrium, calculation of pH of acid, base and saline solutions and principles and applications of the main techniques of volumetric analysis and instrumental analysis (chromatography and spectrophotometric techniques).
The course of Analisi Biochimiche Cliniche aims to provide the informations and the scientific underground to approach correctly the main topics in Clinical Biochemistry by students of pharmaceutical sciences.
This knowledge is needed especially by a professional like the chemist which is at the connection between patients and medical structures also to orientate between many specific diagnostic devices and tools for specimen’s collection, treatment and managing commonly available in the chemist shop.
Prerequisites
The candidate must have knowledge of General Chemistry exam.
There are no obligatory prerequisites, but Clinical Biochemistry is an interdisciplinary science and needs the basic knowledge in Biology, Anatomy, Physiology, Chemistry and Biochemistry.
Course unit content
Introduction to analytical chemistry: qualitative analysis and quantitative analysis. Traditional methods and instrumental methods of analysis.
Principle of chemical equilibrium. Various types of equilibrium:
auto-ionisation of water, solubility product, dissociation of acids and bases, compound equilibrium formation, oxidation-reduction equilibriums.
Definition of homogeneous and heterogeneous equilibriums.
Equilibrium constant. Dependence of equilibrium constant on temperature.
Principle of mobile equilibrium (Le Chatelier-Braun).
Definition of acids and bases. Auto-ionisation and ionic product of water. Calculation of pH and concentration of the species present in the equilibrium of solutions of strong bases and acids.
Definition of weak bases and acids. Dissociation constant. Calculation of pH and concentration of the species present in the equilibrium of solutions of weak bases and acids.
Polyprotic acids. Calculation of pH and concentration of the species present in the equilibrium of solutions of polybasic acids. Distribution curve of equilibrium species on the basis of pH.
Salt hydrolysis equilibriums formed by weak acid and strong base, strong acid and weak base, weak acid and base. Acid salt hydrolysis equilibriums. Calculation of pH of saline solutions.
Buffer solutions. Calculation of pH of buffer solutions. Buffering
capacity.
Principles of volumetric analysis. Definition of primary standard.
Direct and return titration. Final point and point of equivalence.
Methods for determining point of equivalence.
Titration of strong acids with strong bases and vice versa. Titration curve. Titration of weak acids with strong bases and vice versa. Titration curve. Acid-base titration indicators.
Oxidation-reduction titrations. Oxidation-reduction indicators.
Titration curve.
Complexometric titrations. Titrations with EDTA. Conditional formation constant. Metallochromic indicators. Titration curve.
Precipitation titrations. Argentometric titrations: Mohr method, Volhard method, Fajans method. Titration curve. Spectrophotometry UV, IR. Liquid chromatography. Gaschromatography.
For the second Module:
Pre-analytical, analytical and post-analytical variability. Types of biological samples and optimal procedures to collect them. Variables affecting laboratory data
The concept of normal values in biology and the risk functions (normal values stated by statistical calculations or by biological observations; desirable values and risk function; decisional values for diagnostic or therapeutic purpose).
Interpretation of the complete urinalysis test.
Interpretation of enzymatic and iso-enzymatic tests like auxiliary diagnostic tool in some specific organ disease.
Laboratory test modifications by diseases affecting purine metabolism.
Laboratory test modifications by diseases affecting the hydro-electrolyte balance.
Interpreting the variations of acid-base parameters and possible pathological conditions.
Interpretation of the modifications of plasma protein profile.
Full programme
Introduction to analytical chemistry: qualitative analysis and quantitative analysis. Traditional methods and instrumental methods of analysis. Errors in chemical analysis. Units of measure.
Ways of expressing the concentration of solutions: molarity, percentage by weight and by volume, parts per million and related. Preparation of standard solutions by weight and by dilution. Conversion between the various methods of expressing concentration levels. Exercises.
Principle of chemical equilibrium. Various types of equilibrium:
auto-ionisation of water, solubility product, dissociation of acids and bases, oxidation-reduction equilibriums.
Definition of homogeneous and heterogeneous equilibriums.
Equilibrium constant. Dependence of equilibrium constant on temperature.
Principle of mobile equilibrium (Le Chatelier-Braun).
Definition of acids and bases. Auto-ionisation and ionic product of water. Calculation of pH and concentration of the species present in the equilibrium of solutions of strong bases and acids. Exercises.
Definition of weak bases and acids. Dissociation constant. Calculation of pH and concentration of the species present in the equilibrium of solutions of weak bases and acids. Distribution curve of equilibrium species on the basis of pH.
Salt hydrolysis equilibriums. Calculation of pH of saline solutions.
Buffer solutions. Calculation of pH of buffer solutions. Buffering
capacity.
Principles of volumetric analysis. Definition of primary standard.
Direct and return titration. Titration of strong acids with strong bases and vice versa. Titration curve. Titration of weak acids with strong bases and vice versa. Titration curve. Acid-base titration indicators. Oxidation-reduction titrations. Oxidation-reduction indicators.
Titration curve. Complexometric titrations. Titrations with EDTA. Precipitation titrations. Argentometric titrations: Mohr method, Volhard method. Titration curve. Standardization and calibration. Spectrochemical methods. UV-spectrophotometry. Infrared spectrophotometry. Molecular absorption spectrometry. Molecular fluorescence. Atomic spectroscopy. Analytical separations. Liquid chromatography. High-performance liquid chromatography (HPLC). Gaschromatography. Mass spectrometry.
Part 1. General Clinical Chemistry.
Biological specimens consideration: instruction for optimal collection, treatment and conservation (avoiding pre-analytical variation).
Part 2. Analysis of biological samples; analytical techniques and instrumentations.
The main analytical techniques and instrumentations to evaluate the molecules of interest in Clinical Chemistry. Analytical and post-analytical variability affecting the laboratory data. Clinical laboratory quality assurance. Kits for diagnostic tests.
Part 3. Current waived test.
Point of care testing (POCT): use and application. Criteria for test waiving. Quality control in POCT. Implementation and monitoring of POCT. Specimen and its treatment in POCT. Places of POCT utilization. Auto-monitoring of some biochemical parameters.
Part 4. Application of POCT
Control of glycemia; Diabetes and patient auto-monitoring. Lipid metabolism diseases; POCT for blood triglycerides, cholesterol and lipoprotein analysis; auto-monitoring and cardiovascular risk.
Part 5. Outline of Clinical Biochemistry
Markers of function or damage of some organs and systems. Index of cytolysis, clinical enzymology. Main cardiac, hepatic, renal and tumor markers. Toxicology, therapeutic drug monitoring; sport doping and abuse substances control
Bibliography
Harris, Chimica Analitica Quantitativa, Ed. Zanichelli
- Skoog, West, Holler, Crouch, Fondamenti di Chimica Analitica. EdiSES.
- Hage, Carr, Chimica Analitica e Analisi Quantitativa. Ed. Piccin
Presentazioni powerpoint utilizzate a lezione
- Kellerman G. Valori anormali di laboratorio. Mc Graw Hill
- Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. Elsevier Saunders, St.Louis (Missouri)-USA.
Teaching methods
For the first part of the program the explanations will be based on frontal lessons with the help of the chalkboard for the implementation of examples and explanatory charts.
For the instrumental part during the lessons the teacher will use the projection of schemes and photographs that will help the description of the instrumentation.
Exercises: Simple exercises will be proposed in order to reproduce some common procedures that may occur in a laboratory for the preparation of solutions and so on.
Other tutorials will be devoted to showing the use of common software dedicated to analytical data processing, for example for building calibration lines, charts, or for computing results using some of the statistical notions.
The Course of Analisi Biochimico Cliniche is mainly based on interactive lessons.
Assessment methods and criteria
The exam is for a written test and an oral exam. The questions will concern theoretical principles, but also data processing through exercises, and the construction of charts.
The teacher's attention will be focused on the formative aspect rather than the descriptive and informative aspect in order to verify whether the learner, in addition to acquiring knowledge, has become aware of the usefulness of the solution for the solution Of common problems that can be encountered in practical laboratory work.
For the overall judgment, reference may be made to the following items: Knowledge and understanding of matter: 80%
- Ability to apply knowledge and understanding: 15%
- Communicative Skills: 5%
For the Course of Analisi Biochimico Cliniche the final oral examination is about the topics of the course; the student needs to demonstrate to have understood and to be able to manage the essential meanings of every section of the program.
Other information
The iconographic supports shown during the lessons will be available to the students since the beginning of the course
