01651 - Analytical Clinical Chemistry

Learning outcomes

The student acquires basic knowledge of quantitative analytical methods and common analytical techniques based on biospecific reagents (enzymatic analysis, immunometric methods). The student is able to: solve in the laboratory a clinical analysis problem by choosing one or more methods; critically evaluate the analytical data.

Course contents

INTRODUCTION TO ANALYTICAL CHEMISTRY

The analytical process

Sampling of biological fluids

STATISTICS

Statistical analyisis of replicates

Accuracy and precision

Statistical paramaters

Parametric significance tests: T test, F test, Q test

Confidence interval

Linear regression

Calibration: external standards, internal standard, standard additions

Validation of an analytical method: accuracy, precision (repeatability, reproducibility), sensitivity, limit of detection, limit of quantification, dynamic range, specificity, selectivity, robustness, solidity.

SPECTROPHOTOMETRY

Lambert-Beer law: definition and use in quantitative analysis

Absorbance and transmittance

Absormance measurements: choice of wavelength and deviations from ideality

Instrumentation for spectrophotometry. Components and instrumental setup

CHROMATOGRAPHY

Basics of chromatography: chromatogram, chromatographic peak, retention time, void time

Optimization of chromatographic separations: capacity factor, selectivity, efficiency, resolution

Instrumentation for chromatography

Gas chromatography: instrumentation and methods

Liquid chromatography (HPLC). Instrumentation

Methods in HPLC

Normal phase and reversed phase

Quantitative analysis in chromatography

MASS SPECTROMETRY

Basics and instrumental setup

Hard and soft ion sources

Mass analyzers

MS of proteins

IMMUNOMETRY

Principles and advantages of immunometric methods

Monoclonal and polyclonal antibodies. Properties and applications

Labels for immunometry. Immunoenzymatic methods

Classification of immunomethods

Homogeneous and heterogeneous, competitive and non-competitive methods

Calibration

Quantitative analysis

ENZYMATIC METHODS

Enzymatic analysis. Basics and applications.

Properties and structure of enzymes

Enzymatic activity

Kintics

Regions of the activity/[substrate] curve

Michaelis-Menten equation

Determination of enzymatic activity

Single-point, double-point and multipoint methods.

Quantitative analysis of antigens.

Endpoint methods

Measurement in enzymatic analysis

 

 

Readings/Bibliography

L. Spandrio "Principi e Tecniche di Chimica Clinica", Piccin

-A.J. Ninfa, D.P. Ballou "Metodologie di Base per la Biochimica e la Biotecnologia", Zanichelli (consultazione)

-J.M. Berg, J.L. Tymoczko, L. Stryer "Biochimica", V Edizione, Zanichelli

Teaching methods

Class lectures with overhead projection. The slides are available on http://iol.unibo.it

Laboratory with supporting tutors. Laboratory protocols are made available on http://iol.unibo.it

The course consists of 6 credits (5 credits room lectures, 1 creditlaboratory). During the lectures the topics of the course will be presented and discussed, with theoretical insights and explanatory examples. The course includes laboratory exercises, which will be illustrated during lectures. The exercises are designed to allow each student to acquire the necessary skills and knowledge of the basic analytical techniques for working in a laboratory according to quality and safety guidelines. Each student will elaborate the results obtained in the laboratory by using the statistical tools learned in the course, and will write down reports on the exercises.

Assessment methods

Final written exam on the whole course program. A detailed list of the topics included in the test will be available at the end of the couse on http://iol.unibo.it

The students have 90 minutes to answer 3 open questions.

The evaluation criterion is based on the demonstration by the student to be able to use and apply the knowledge, information and cultural tools provided by the course, and on the examination of the laboratory reports. The threshold of learning sufficiency is represented in particular by the demonstration of skills and critical understanding of the course topics for discussion.

The teacher is available for further clarification and to verify the level of preparation prior to examination.

Teaching tools

Computer and overhead projector for room lectures.

Analytical chemistry laboratories, tools and glassware, spectrophotometers, plate reader, HPLC-UV.

Material: the material presented in class will be made available to the student in electronic format via the Internet.
To get educational materials: http://iol.unibo.it/  
Access is reserved to the students enrolled on the course.

Office hours

See the website of Andrea Zattoni