OPERATION OF UV VISIBLE SPECTROPHOTOMETER

• OPERATING PROCEDURE OF UV VISIBLE SPECTROPHOTOMETER :

• Make: SHIMADZU UV-180

• Start-up procedure:
• Turn on instrument.
• Allow instrument to warm up for fifteen minutes.
• Turn on the computer and login.
• Configure a network printer and set it as default.
• Make sure you are connected to the network.
• Open UV Probe 2.34 (Start>Programs>Shimadzu>UV Probe or Select Desktop Icon)
• Control of instrument to computer:On the instrumentAfter initialization of instrument, the mode screen will be displayed.
• Press F4 key (PC Control) on the instrument keypad to control through the computer.
• On the computer Connect to instrument (Instrument >Commends >Connect)
• Create /Edit Method:
• Load method (Edit>Method)
• To create new method select spectrum module icon in menu bar.
• In Edit Method parameters,
• Select Measurement Tab.
• Set the Wavelength range –starting and ending values
• Set scan speed.
• Set auto sampling interval.
• Under Scan Mode click auto on the file name icon and give product Analytical Reference Number as file name.
• Instrument parameters Tab.
• Select absorbance as measurement mode.
• Click Ok
• Change Y-axis range.
• Go to the overlay (select tab at top of graph or Graph>overlay) and click on the minimum and maximum absorbance values.
• Enter a new value
• When finished, click anywhere on the screen to get out of this mode
• Save method (File > Save as)
• Enter file name.
• In the Save as Type list, click Method file (*.smd)
• Click Save
• Collect spectrum
• Perform baseline correction
• Make sure sample and reference compartments are clear.
• Place the two UV cells with blank solution.
• Collect baseline (Instrument>Commands>Baseline).  
• When the Baseline parameter box appears, enter the starting and ending values.
• Click OK.
• Insert sample (front cell holder) and reference (back cell holder).
• Start (Instrument >Commands> Start).
• When scan is complete, save (File >save All).
• Remove sample and reference from compartment.
• Analyze spectrum:
• Perform peak pick (Operation >peak pick)
• The operation screen will open to display results.
•  Right click on the peak pick table and make sure mark peaks. and show peaks are selected.
• Adjust peak threshold (if necessary )
• Right click on peak pick table and select properties.
• Enter new threshold value.
• Close Operation screen (view operation).
• Print peak pick results (File>print>Current View).

• Print Spectrum:
• Select report icon from menu bar and select spectrum report format.Print spectrum (File >print).
• For Photometric measurement
• Load method (Edit >Method)
• To create new method, select photometric module icon in menu bar.
• In Edit method parameter parameters,
• Select measurement Tab.
• Set wavelength type and wavelength (nm)
• Click on add Button for wave length and click on next button.
• Select type formula, units, Wavelength and Standard concentration and Click on next button for properties.
• Click on file name side icon and give product AR No as a file name. title is product name comments are Batch Number
• Click on finish button
• Save method (file >save as).
• Enter file name.
• In the save type list, click method File (*.smd).
• Click on the save icon for save file.

• Collect photometric data:
• Perform base line correction.
• Make sure sample and reference compartments are clear.
• Place two UV cells with a blank solution.
• Click AUTO ZETO Replace the blank solution in sample cell with sample solution Give sample ID in sample table (active) Click on the READ UNK button.
• Wait for 20 seconds till the show a constant value After completion of measurement click on the save button the data will be saved Select the icon form menu bar and select the photometric report format
• Take printout.
• Enter the details in logbook.
• Shutdown procedure:
• Press disconnect on computer to disconnect from instrument (Instrument >commands >disconnect).
• Turn off instrument.

• Log off computer.

• ANNEXURES: 

Annexure-I