Universidad Pedagógica y

Tecnológica de Colombia
Facultad de Ciencias básicas
Escuela de Ciencias Químicas
Alumno: Gustavo Abraham Valero Pibaral

Profesor: Julie Joseane Murcia Mesa

Asignatura: Química Analítica II
Abstract espectroscopia UV-Vis
The basic tools needed in order to work with a spectrometer are cuvettes, solvent and the sample.
Cuvettes can be made of glass, plastic or quartz according to the type of light supposed to use.
Note that cuvettes possess to sides transparent and two opaque sides. Cuvettes must be handled
in the opaque sides

We must be aware of not touching the transparent parts of the cuvette, light passes through that
parts and could be diffracted if they are dirty. If this happens clean with a Kleenex s

Firstly, fill the cuvette with solvent and put in the blank holder, pick another cuvette with solvent
in the sample holder and run a scan (zeroing).

Remove the solvent of the cuvette in the sample holder and wash it with sample solution twice,
this prevents from diluting the sample.

Put the sample in the sample holder, set up the experiment parameters such as scan range,
measure mode, number of scans, etc. and run the scan.

You will get the maximum absorption peak of the sample.

In order to determinate the amount of an analyte in a sample using a spectrophotometer, we

must create a calibration curve. To do this, we prepare an analytical series made of a standard at
different concentrations, then we run the experiment at the maximum peak of absorption of the
standard with each of the solutions previously prepared, we will get the absorbance at the given
wavelength, after that, graph the absorbance vs concentration and get the curve equation.

Once this is done, run an experiment with the sample, and finally extrapolate the absorbance of
the sample with the equation previously gotten and the concentration of the sample is determined
via spectrophotometry.