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