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Effective Date:

24February2014 Review Date: 24February2014


SOP #003
Modified by: A. Marvelle
Version # 0001 Page 1 of 2

Background
The spectrophotometer is used to measure light being absorbed by a sample. Each color that we see
absorbs light at different wavelengths. The spectrophotometer shoots light through a sample and sees
which wavelength is absorbed.


Verifying Results with Spectroscopy

1.0 Purpose
Test each batch of GrapeX with the spectrophotometer to verify the liquid chromatography results.
2.0 Scope
This SOP describes the methods used to run a spectroscopy sample.
3.0 Responsibility
All scientists in the QC-Microbiology group will be responsible for following the correct protocol for
testing drug GrapeX for purity.
4.0 References
Graphs at end of this SOP
5.0 Safety
Safety glasses/goggles, lab coats, gloves must be worn at all times and all materials must be handled
with care.
6.0 Materials and equipment
6.1 GrapeX Testing Sample
6.2 Cuvette
6.3 Transfer pipette
6.4 Spectrophotometer

7.0 Procedure
7.1 Pipetting the test sample into cuvette
7.1.1 Using the transfer pipette, squeeze the air out of the bulb and insert the end
into the test sample.
7.1.2 Release the bulb and move the test sample into the transfer pipette
7.1.3 Place the pipette end in the cuvette and squeeze the bulb to transfer the
sample into the pipette.
7.1.4 Check to make sure the cuvette is almost full. If it is not repeat steps 7.1.1 to
7.13.
7.2 Testing the sample in the spectrophotometer
7.2.1 Place the cuvette in the spectrophotometer
7.2.2 Run the GrapeX program
7.2.3 View the results of the absorbance graph and record below


Effective Date:
24February2014 Review Date: 24February2014
SOP #003
Modified by: A. Marvelle
Version # 0001 Page 2 of 2


7.3 Compare you graph to the GrapeX standard graph below and answer the questions

Questions
1. Do you notice a difference between the standard GrapeX and your batch?


2. Does the spectrophotometer result match up with your chromatography results? Why?


0
0.5
1
1.5
2
2.5
3
400 500 600 700
GrapeX Standard
Wavelength (nm)
Wavelength (nm)
A
b
s
o
r
b
a
n
c
e

A
b
s
o
r
b
a
n
c
e



Effective Date:
24February2014 Review Date: 24February2014
SOP #003
Modified by: A. Marvelle
Version # 0001 Page 3 of 2

3. Look at the graphs below of each color molecule. Label the colors on the peaks of your graph on
the previous page.





Based on the chromatography and spectrophotometer results, do you
recommend using this batch of GrapeX to treat patients with MS?
___________ YES __________ NO


0
0.5
1
1.5
2
2.5
400 500 600 700
Red
0
0.5
1
1.5
2
2.5
400 500 600 700
Blue
0
1
2
3
4
5
400 500 600 700
Yellow
Wavelength (nm) Wavelength (nm)
Wavelength (nm)
A
b
s
o
r
b
a
n
c
e

A
b
s
o
r
b
a
n
c
e

A
b
s
o
r
b
a
n
c
e

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