Appendix 5: Operation of the Shimadzu spectrophotometers

Precautions:

• Clean up spills in the sample compartment immediately.
• Keep spectrophotometer vents (on sides of instrument) free of any obstructions.
• Use clean, dry hands on the keypads.
• Do not place anything on top of the spectrophotometer.

Spectrophotometer setup

• Remove dust cover from instrument and place in a clean, out of the way area.
• Remove any cuvettes from the sample and reference beams.
• Turn the POWER to ON (left side of instrument).  Allow 15 min. for warm-up.
• After initialization, the MAIN MENU will be displayed.

Fixed wavelength measurements

• Press [1] from MAIN MENU to enter Photometric mode
• Select ABS or %T by pressing the [F1] key
• Press the [GOTO WL] key, enter the desired wavelength, and press [ENTER]
• Insert a sample blank into both the front and back slots and press the [AUTO ZERO] key. Be sure to wait until the machine beeps – it takes a minute.
• Insert an unknown sample into the front slot and press [START] to read the sample absorbance (%transmittance) on the digital display.

Single Scan

• Set up the spectrophotometer as described in 1 above. Then:
• Enter the Spectrum mode from the MAIN MENU.
• Select the MODE (ABS, %T) by pressing [1].
• Select the scan speed:
  • Press [4]
  • Choose a speed (1-5)
  • Press [ENTER]
• Enter wavelength range:
  • Press [2]
  • Enter upper limit
  • Press [ENTER]
  • Enter lower limit
  • Press [ENTER]
• Set the (ABS, %T) range:
  • Press [3]
  • Enter upper limit
  • Press [ENTER]
  • Enter lower limit
  • Press [ENTER]
• Perform a background correction
  • Place the appropriate blank and reference samples in position.
  • Press the [F1] key to perform a baseline correction.  This may take a few minutes depending upon settings.
  • Set the sample and reference cells in place.
  • Press the [START] key to start scan.

Time Drive

• Set up the spectrophotometer as described in 1 above.  Then:
• Enter the Time Scan mode from the MAIN MENU
• Select the MODE (ABS, %T) by pressing [1].
• Enter the desired wavelength by pressing the [GOTO WL] key, entering the wavelength, and pressing [ENTER].
• Set the (ABS, %T) range:
  • Press [3]
  • Enter upper limit
  • Press [ENTER]
  • Enter lower limit
  • Press [ENTER]
• Set the time scale (sec, min) by pressing [4]
• Set the measurement time by pressing [2].  The measurement interval will automatically be set relative to the length of measurement chosen.
• Press the [START] key to start measurement.

Shutting down

• Turn the spectrophotometer POWER to OFF.
• Replace dust cover on spectrophotometer and make sure all electronics and wires are covered.