Integrated Laboratories II

Code:

EBE0110

Acronym:

LI2

Keywords
Classification	Keyword
OFICIAL	Basic Sciences

Instance: 2009/2010 - 2S

Active?	Yes
Course/CS Responsible:	Master in Bioengineering

Cycles of Study/Courses

Acronym	No. of Students	Study Plan	Curricular Years	Credits UCN	Credits ECTS	Contact hours	Total Time
MIB	72	Syllabus	1	-	4	120	108

Teaching language

Portuguese

Objectives

In this course unit students will have a chance to carry out laboratory assignments, which will make them apply the knowledge acquired in the various course units of the 2nd semester of the 1st year of the Masters in Bioengineering.

In some classes it will also be discussed some specific topics related to the assignments, namely ultraviolet-visible spectroscopy and the operation of a pH meter.

Program

1. Calorimetric determination of enthalpy of standard formation of solid NH4Cl; Enthalpy formation of ammonium chloride (NH4Cl) can be calculated by adding the enthalpies of formation of HCl(aq) and NH3(aq) with the enthalpy of neutralization of NH3(aq)/HCl(aq) and symmetric of enthalpy of neutralization of solubilisation of NH4Cl(s). Previously, it is determined the heat capacity of a calorimeter using neutralization reaction of HCl(aq)/NaOH(aq), whose molar enthalpy is known.
2. Spectrophotometric and thermodynamic characterization of DNA; This assignment is divided into two parts: in the first part it is determined the hyperchromic effect at 260 nm in aqueous solutions of natural DNA, which is the result of heat denaturing and its dependence of NaCl concentration. In the second part is used the hyperchromic effect to follow the heat denaturing growth of a small DNA oligomer in solution
3. Potentiometric titration of amino acids; grading sensitive glass electrode to H+(aq) using three standard pH solutions; potentiometric titration of a solution of glycine amino acid with NaOH and HCl. It is determined not only the concentration, but also pKa values of glycine.
4. Extraction, identification and purification of caffeine from black tea leaves. This assignment aims to familiarise students with some basic laboratory techniques in organic chemistry, by using a well know composite, which will be extracted from black tea leaves.
5. Extraction and identification of total lipids from a biological sample. In this laboratory assignment are used various biological matrices, which contain different levels and different types of lipids (peanut, milk, egg yolk, etc). Students will have to deal with processes of samples manipulation and extraction, which are different from the previous assignment, namely grinding and ultracentrifugation.
6. Kinetic controls vs. thermodynamic control of a reaction. In this practical assignment students will make a chemical reaction using two carbonyl compounds (a ketone: cyclohexanone and an aldehyde: 2-furaldehyde) and semicarbazide in order to obtain the respective semicarbazones. This practical assignment consists in carrying out reactions at different temperatures and subsequently the recovery, purification and drying the crystals obtained, which will be analysed by ultraviolet-visible spectroscopy.
7. Study of proteins: quantification, separation, analysis and structural characterization
7.1 Separation of a mixture of proteins (haemoglobin and albumin) by chromatography of ionic change and analysis by native electrophoresis. Quantification of proteins by spectrophotometry – adsorption of UV-radiation (280 nm) and Bradford method; Lambert-Beer law and molar extinction coefficient
7.2 Protein analysis by electrophoresis in SDS-PAGE, Immunoblotting and ELISA to detect and separate proteins based on their molecular mass – electrophoresis in SDS-PAGE; number of protein subunits and disulphide connections; application of immunological techniques to identify and quantify proteins; immunoenzymatic tests (ELISA) and electrophoretic transference and immunodetection (immunoblotting); isoelectric titration of a protein – study of protein charge depending on the pH.
7.3 Simulation of protein isolation using the program «ProteinLab»; Using a program of simulation of different strategies to protein isolation (ProteinLab), in order to familiarise students with techniques previously taught and analysis of results.
8. Preparation of films and collagen sponges
8.1 Measurement of mechanical properties of collagen films
8.2 Enzymatic degradation of collagen films
8.3 Porosity measurement of collagen sponges

Evaluation Type

Distributed evaluation with final exam

Assessment Components

Description	Type	Time (hours)	Weight (%)	End date
Attendance (estimated)	Participação presencial	96,00
	Total:	-	0,00