Materials and MethodsThe experiment wascarried out with a Spectrometer.
The values were obtained were throughspectroscopy, an analytical technique which “studies the interactions betweenelectromagnetic radiations and matter. Atoms, molecules and ions can scatter,absorbs, or emit electromagnetic radiation in ways that depend upon thechemical composition and structure of the substance”. (Wang.Qiuyu,A.
,Nessar,G.and Hedley (2010) Biomedical Science Practice, Oxford University Press).Spectroscopy is used to identify which elements are presentin a sample, determine how much of an analyte is present. It is of greatimportance to state that it is a non invasive technique and samples are notdestroyed during spectroscopic analyses. For this particular, the experiment wantedto find out the absorbance which means quantifying how much light of aparticular wavelength is absorbed by a sample.”The absorbance ofan analyte is proportional to its concentration in a manner described by theBeer Lambert Law. The law describes the absorbance of monochromatic light whichis one of a single wavelength, as it passes through a dilute solution. The lawstates that the absorbance is directly proportional to both the concentrationof the solution and also the length of the path that the light takes throughthat solution.
“(Wang. Qiuyu, A., Nessar, G. And Hedley (2010) BiomedicalScience Practice, Oxford University Press).
In our case weused absorbance for quantitative analyses therefore the absorbance of oursolution was determined by the wavelength at which the material investigated,in this case albumin, absorbs most strongly and that is where the absorbancewas at its greatest (562nm).TheSpectrometerwas used in this experiment to obtain values by spectroscopictechnique, which is the study ofinteraction of electromagnetic radiation with matter. When a beam of lightstrikes a triangular prism, it is separated into its various components thatallows production and viewing and to determine how much elements is present inthe spectrum.
In this experiment we would like to identify how much light wasabsorbed in order to calculate the concentration of the samples.as stated inbeers law. ”Beer’s Law is an equation that relates the attenuation of light toproperties of a material. The law states the concentration of a chemical isdirectly proportional to the absorbance of a solution.” In case, theexperimental used absorbance for quantitative analyses. Therefore, theabsorbance of the solution was determined by the wavelength at which thematerial investigated, in this case ferritin, absorbs most strongly and that iswhere the absorbance was at its greatest (562nm).
First of all, makingseries of dilutions of Fe(II) standard solutions. Secondly, the concentration wereprepared in the range 20 to 80?M. Beginning from the provided mother solutionthat has a concentration of (1 mM) the final volume in final volume each tube should be 2 ml.
Next, the sampleswhere thoroughly mixed and mixed and incubated at room temperature for 2 minutes.After, the spectrophotometer was set at 562nm and it was then blanked withwater in to take the absorbance of the chemicals.