SOP for Shimadzu Spectrofluorophotometer

OBJECTIVE

This SOP describes the procedure for the operation of the Shimadzu Spectrofluorophotometer RF-5301PC.

SCOPE

Spectrofluorophotometer is used for qualitative and quantitative analysis of fluorescent organic and inorganic compounds.

RESPONSIBILITIES

It is the responsibility of the QC Analyst to ensure adherence to this procedure.

It is the responsibility of the QC Manager to follow the instructions of this procedure.

PRECAUTIONS

Handle the fluorimeter cells with care;

• Always handle on the corners, or at the top and bottom.
• Ensure that the cell is washed thoroughly after each solution and then rinsed out with the new solution before taking its spectrum.
• Ensure that the same cell face is used for each solution.
• Ensure that the cell windows are clean, dry, and not contaminated.

PROCEDURE

Starting the Instrument

• Turn on the computer,
• Turn on the instrument, The on/off switch is located in the lower right corner
• Starting the software, Double click the icon PC 150X

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Software Operation

• To open the menu, Select acquire mode drop-down menu
• Ensure the spectrum is checked
• Select configure drop-down menu
• Select spectrum parameter

• Select spectrum type
• When excitation is selected, the emission grating is in a fixed position
• When emission is selected, the excitation grating is in a fixed position

Enter desired parameters

• Excitation/Emission wavelength: 0, 220-900nm
• Excitation/Emission wavelength range: 220-900nm
• The minimum scanning range is 25
• Sensitivity can be set to high or low
• Recording range – limit of the graph
• The allowable range is -100 to 1000
• Repeat Scan - allows automatic consecutive scanning from 2-100 scans
• Press OK.

Running an Experiment

• Loading a Sample
• Flip door down
• Insert a cuvette
• Close the door
• Click on the start button

• When the scan is completed the file name dialogue box appears
• Type in a file name
• Maximum of 8 characters
• A comment may be typed into the dialogue box

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Select the presentation drop-down menu

• Press graph
• Press radar
• Both axes

Zoom In

• Click a drag a box around the area you wish to enlarge
• To return to full screen
• Right mouse click
• Select limits
• Press reset
• Press ok

Cross Hair Coordinates

• Right mouse click
• Select cross hair
• Select display
• To remove the cross-hair coordinates press display again
• Select the presentation drop-down menu
• Select plot to display the plot layout window
• Under the graph column
• Click on the first box in the row
• If it is already checked, uncheck the box then recheck the box
• The default for plot data is a screen, this will print what appears on the screen
• Select Ok
• Select the box under Params

Printing a Spectrum

• Click boxes 3 and 4 under quadrants
• The boxes are on the right side of the menu
• Select preview to display what will be printed

Select Print

• Shutting down the instrument
• Remove cuvette
• Close PC-1501 program
• Turn off instrument

ABBREVIATION

QC: Quality Control

ANNEXURE

Nil

REVISION HISTORY

Nil

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