CCCCD Spectrophotometry Spectrophotometer Parts Wavelength and Color Lab Report

Question Description

SPECTROPHOTOMETRY REPORT SHEET

Download the report sheet, complete it, and then submit it via Blackboard email as an attachment.Do not change the format.

All blue shaded areas require answers.

I.Parts of the spectrophotometer

Experiment (dry lab)

Label the parts of the spectrophotometer (digital display, wavelength readout, wavelength knob, mode button, %Transmission/Absorbance readout, 100% T knob, 0%T knob, sample holder)

a.
b.
c.
d.
e.
f.
g.
h.

Relationship between wavelength and color
Using the spectrophotometer to determine %transmittance (%T) and Absorbance (A)
Development of a standard curve

Experiment (dry lab)

Record the wavelength of where you find the deepest color.

Color	red	orange	yellow	green	blue	violet
Wavelength

Experiment (dry lab)

Outline the steps to calibrating the spectrophotometer. Fill in the blanks.

Adjust the ____ knob Set machine to 0% T with the ____ knob Place blank tube in the ____. Set machine to 100%T with ____knob

Question:

What is the purpose of the Blank tube and why is it set at 100%T?

Measuring %T and A

At 540 nm sample tube measures
% T
absorbance

Experiment (dry lab)

List Absorbance readings (from the lab unit slides) and calculate mg/ml. Use three significant figures.

Tubes	A	Protein concentration mg/ml
1		10.0
2
3
4
5
blank
unknown

Take data from your table and plot protein concentration in mg/ml vs. absorbance. Draw the best fit line through your points. Plot the unknown with a different color and show extrapolation to determine protein concentration (see last slide lab unit). Complete the graph using Microsoft insert shapes (or manually graph, scan your work and embed image into report).

End of Lab J

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