ANALYTICAL CHEMISTRY- CLINICAL-BIOCHEMICAL ANALYSIS
Learning outcomes of the course unit
One of the main objectives of the Analytical Chemistry module (MODULE 1) is to make the student aware of the main analytical methods, and the potential of modern techniques in the various fields of application, in particular the pharmaceutical and clinical fields.
It will be fundamental to understand the theoretical bases on which the main analysis systems and their operating principles are founded.
In addition, the student will develop the ability to correctly process data according to the laws of statistics, in order to provide a significant analytical result
Regarding the skills that the student will acquire,
they include the application of the theoretical foundations concerning the development of analytical methodologies and the choice of the technique suitable for solving an analytical problem, considering possible advantages and disadvantages.
The student must develop the necessary critical skills to set up an analysis protocol, starting from sampling, taking into account the possible phases of sample preparation.
Furthermore, the cognitive tools will be possessed for the interpretation of analytical data with the help of statistical principles in order to be able to evaluate the reliability of the data obtained from a series of analyzes.
The course of Clinical Biochemical Analysis (MODULE 2) aims to provide information and scientific support for a correct methodological approach that develops knowledge and the necessary skills in understanding the topics of Clinical Biochemistry, specific for the student of the CdLS in Pharmacy.
This knowledge and skills will be put to the test to be applied during work practice by a professional figure who places himself at the interface of the National Health System, (between patient and health facilities, where appropriate communication skills are needed); furthermore, given the specificity and complexity of diagnostic products, self-analysis devices and materials for collection, collection and treatment of biological samples, commonly available in pharmacies, the course is structured so as to provide the means to integrate in autonomous way the knowledge and manage the complex situations that will arise in the carrying out of the profession.
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 contents summary
For the first Module (5 CFU) of Analytical Chemistry (MODULE 1) the following contents are foreseen: A first part of the course concerns the presentation of the role of analytical chemistry, underlining the importance that its application finds in various sectors and in particular in the pharmaceutical sector. This section will deal with topics such as sampling, treatment techniques of a sample and the theory of errors related to obtaining a reliable analytical data.
In a second part will be shown the application of some basic concepts of general chemistry that the student already knows, by reviewing fundamental references to topics such as chemical balance, pH, buffer solutions, salt hydrolysis, titrations, solubility and electrochemistry. This in order to provide the basis for the understanding of analytical methods based on these principles, such as volumetric and gravimetric analysis.
The third part is dedicated to analytical instrumental chemistry for qualitative and quantitative determination. The main topics that will be explored are the spectrophotometric methods and the chromatography.
The main instruments and potentials of modern analytical techniques will be described, focusing attention on applications in the pharmaceutical field.
For the second module (MODULE 2):
The first part of the course concerns the study of biological samples to know their characteristics and possible alterations in relation to the preparation of the subject to the sampling as well as to the collection, treatment and preservation of the sample (pre-analytical variability).
Subsequently, the basic principles of the methods and related instruments used in the dosage techniques most frequently used in clinical biochemistry will be recalled: analytical and post-analytical variables that can influence the data, principles and procedures of quality control with particular reference to outside the centralized laboratory (POCT: Point Of Care Testing). Evolution of POCT technology - Preparations for diagnostic tests.
In the third part of the course we will consider the different situations in which it is possible to apply the POCT technology, with particular reference to the skills that are required to professionals working in the pharmacy (Legislative Decree 153/2009): hyperglycemia and hypoglycemia, diabetic disease and complications ; (auto) glucose monitoring; dyslipidemias, diagnostic products in the monitoring of triglycerides, cholesterol and lipoproteins in the blood; self-monitoring and cardiovascular risk. Urine analysis.
Furthermore, the bases will be provided to understand and treat the most common function and organ lesion markers: cytolysis indexes; clinical enzymology; cardiac function parameters; parameters of hepatic and renal function; tumor markers; therapeutic monitoring of drugs and drugs of abuse.
The Analytical Chemistry module (MODULE 1) has the following program:
The role of Analytical Chemistry. Concept of analytical procedure and qualitative and quantitative analysis.
Stages of a typical quantitative analysis. Sampling. Sample treatment. Extraction.
Uncertainty associated with a measure: types of error; sensitivity, accuracy and precision. Expression of the analytical data.
Gaussian distribution. Standard deviation. Significance test. Quality of the analytical data.
Basic general concepts
Unit of measurement of solution concentration. Preparation of weighing and dilution solutions. Calculation exercises.
Ionic product of water. Chemical equilibrium.
Acids and bases. Calculation of the pH and concentration of the species present at the equilibrium of solutions of acids and strong bases. Calculation of the pH and of the concentration of the species present at the equilibrium of weak acid and base solutions. Polyprotic acids. Ionisation according to pH.
Salt hydrolysis. Calculation of the pH of saline solutions.
Buffer solutions. Calculation of the pH of buffer solutions. Buffering capacity.
Precipitation balances. Solubility product. Types of precipitates. Ion to common. Effect of pH on solubility.
Standard potential, eq by Nernst. Stack, pH meter
General principles. Determination of the title of a solution. Definition of primary standard.
Acid-base titrations. Titration curves of strong acids with strong bases and vice versa. Titration curve of weak acids with strong bases and vice versa. Indicators for acid-base titrations.
Complexometric titrations. EDTA.
Precipitation titrations (Mohr method and Volhard method).
Spectroscopic methods of analysis
Electromagnetic waves. Property. Absorption and emission. Origin of spectra, relationship between electronic structure and absorption bands. Spectral fields of analytical interest and types of energy transitions.
UV-visible spectrophotometry. Lambert-Beer Law. Limitations. Instrumentation. Quantitative analysis.
Molecular fluorescence. Characteristics of fluorescent substances. Derivatizing. Instrumentation.
Molecular spectrophotometry in Infrared absorption. Origin of the spectra. Bands characteristics. Instrumentation.
Chromatographic analysis methods
Introduction to chromatographic methods. Principles. General description of the chromatographic techniques. Liquid chromatography. Qualitative and quantitative analysis. Mechanisms of separation. Adsorption and breakdown chromatography. Direct and inverse phase, ion exchange, molecular exclusion.
Chromatographic parameters. Van Deemter's equation.
Instrumentation for HPLC. Injector, columns, pumps. Refractive index detector; UV-DAD; fluorescent. Isocratic and gradient elution.
Gas chromatography. Principles. Instrumentation: injectors, columns, detectors.
Construction of a calibration line. Method to external and internal standards. Linearity. Detection limit, quantification limit.
The Biochemical-Clinical Analysis module (MODULE 2) provides the following program:
Part 1. The clinical chemistry laboratory: general. Analyzes in the clinical chemistry laboratory. The biological sample: subject preparation, collection, treatment and conservation (pre-analytical variability).
Part 2. Analysis of biological samples, separation and measurement techniques. Main biochemical methodologies and evaluation principles of molecules of chemical-clinical interest. Analytical and post-analytical variables that can influence the data. Quality control. Prepared for diagnostic tests.
Part 3. Chemical-clinical analyzes outside the centralized laboratory. (POCT: Point Of Care Testing). Criteria for decentralization.
Harris, Chimica Analitica Quantitativa, Ed. Zanichelli
- Skoog, West, Holler, Crouch, Fondamenti di Chimica Analitica. EdiSES.
- Hage, Carr, Chimica Analitica e Analisi Quantitativa. Ed. Piccin
Powerpoint presentation used at lesson.
- Kellerman G. Valori anormali di laboratorio. Mc Graw Hill
- Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. Elsevier Saunders, St.Louis (Missouri)-USA.
The teaching will be performed in mixed mode or by e-learning via the Teams platform according to the COVID EMERGENCY situation.
For the first part of the program (MODULO 1) 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 (MODULO 2)is mainly based on interactive lessons.
Assessment methods and criteria
The examination of the Analytical Chemistry (module 1) includes an oral test by using the Teams platform. The questions will cover the theoretical principles but also the processing of data by exercises, and the construction of graphs.
For the purposes of evaluation, the teacher's attention will be focused on the training aspect rather than on the descriptive and informative one, in order to verify if the student, in addition to acquiring knowledge, has acquired awareness of the usefulness of them for the solution of common problems that can be encountered in practical laboratory work.
For the overall opinion, reference can be made to the following items: Knowledge and understanding of the subject: 80%
- Ability to apply knowledge and understanding: 15%
- Communication skills: 5%
The written exam will be passed if both the exercises and the theory questions are sufficient. They result sufficient when more than the half of the questions are correct.
For the Clinical Biochemical Analysis module (MODULE 2) a final oral exam is planned, focusing on the topics of the program. The student must demonstrate that he / she has understood and is able to use the fundamental concepts of each topic, answering questions aimed at verifying the technical skills (criteria that allow to obtain a reliable laboratory data), the knowledge on the levels of investigation and management of practical situations. In each of these situations the degree of mastery of the argument will be verified and on this criterion the score will be attributed, useful for the final vote.
The material used for the preparation of teaching activities (video lessons, slides, texts other than the reference ones, different sources accessible on the net, etc.) is explained during the lessons and is available at the request of the student.