Ocean Optics R-2000 Raman Spectrometer Setup and Operating Instructions Arlen Viste and Deanna Donohoue April 2000 Update 2003, DEW

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1 Ocean Optics R-2000 Raman Spectrometer Setup and Operating Instructions Arlen Viste and Deanna Donohoue April 2000 Update 2003, DEW References Raman Systems R-2000 Operating Manual, Version 1.6, Ocean Optics, Inc. Ocean Optics web site, URL Kazuo Nakamoto, Infrared and Raman Spectra of Inorganic and Coordination Compounds, Part A and Part B. 5th ed. Wiley- Interscience, New York, NY, Joseph B. Lambert, Herbert F. Shurvell, David A. Lightner, and R. Graham Cooks. Organic Structural Spectroscopy. Prentice- Hall, Upper Saddle River, NJ, 1998, Chapter 7 and 8. Laser Safety! Wear laser safety goggles while working with the Raman spectrometer.! Post the Caution sign on the door to warn others of the laser hazard and remove it when you are finished with the Raman Start up the R-2000 software by clicking the R2000 icon on the desktop. Intro to the Display Main Screen Toolbar (left side of main screen) 1

2 Laser Operation! Obtain the Remote Interlock and Power Switch Key (Inline Filter Module also required for analysis of solids) from Dr. Weisshaar.! Insert the Remote Interlock into its socket, aligning the red dots on the socket and Interlock.! Turn on the System Power Switch on the front panel of the Raman.! Insert the Power Switch Key and turn it to the on position.! On the main program screen check the Laser On box. In about 20 seconds the laser will fire and the red light on the front panel of the Raman will come on. Unchecking the Laser On box turns the laser off, but again there is a 20 second delay. The red light on the panel accurately indicates when the laser is on and off.! For solids and other efficient scatterers it is necessary to reduce the intensity of the input signal to reduce background in the spectrum. To install the Inline Filter Module unscrew the Spectrometer Input cable. Screw the Inline Filter Module onto the cable, then screw the other end of the Module into the Spectrometer Input on the front panel. Set the Laser Wavelength! Make sure everyone in the room is wearing laser safety goggles.! Make sure the laser is turned off.! Remove the plastic cover from the laser probe tip.! Place the laser probe tip into 2-propanol in a brown bottle. Adjust the septum so it covers the bottle mouth and holds the laser probe tip just below the liquid surface. Orienting the probe perpendicular to the bottle bottom gives the best results (minimizes reflections from the bottle sides).! Put the bottle into the black can and slide the black, slotted cardboard cover around the cable to cover the can. (This minimizes stray light from interfering.)! Set Integration Time = 1000 ms, Spectra to Average = 1, Boxcar Smoothing Width = 7, and Acquisition Mode Selection = Wavelength (nm). The x-axis on the graph should read Wavelength.! Click Laser On. Click the Peak Labels button on the toolbar. You should see a display similar to the figure below.! Measure the laser peak position (approximately 785 nm), and enter that value as the Laser Wavelength. ( nm in the sample screen shown.) 2

3 ! Turn off the laser (uncheck Laser On).! Set Acquisition Mode = Raman Shift (cm-1). The x-axis on the graph should now be labeled Raman Shift. Collect and Store the Dark Current Spectrum A dark spectrum is the signal at the detector when no light is being received and should be subtracted from all spectra obtained. The magnitude of the dark spectrum varies with integration time, so a new dark spectrum must be stored each time the integration time is changed.! Make sure the laser is turned off. It is best to turn off the laser key and uncheck Laser On. Place the plastic cap over the probe to block out all incident light.! Set Integration Time to the desired level. After a time which depends on the integration time chosen, a smoothed dark current spectrum will appear on screen. It should be similar to the figure below.! Click the Store Dark icon on the toolbar to store it.! Click on the Subtract Dark icon on the toolbar. The button will stay depressed.! Each time you change the Integration Time, store a new dark spectrum. Sample Prep! Liquids may be neat or as solutions; concentrations of around 10% are required to give a reasonable signal.! Dissolved gases may show up in the Raman spectrum, so they should be removed. " Pour your sample into a filter flask and stopper the top. Use an aspirator to pull a vacuum on the sample for several minutes. Agitation in an ultrasonic bath while pulling the vacuum will speed the process. Taking a Spectrum section below illustrates the effect of degassing on the spectrum. Aqueous samples can be quite difficult to completely degas. " The effect of dissolved gases can be seen in the sample spectrum shown below. 3

4 ! Liquids must be filtered to removed suspended particles which increase backscatter and therefore cause problems. " A filter holder like the one in the figure on the right and glass fiber filters should be stored on the shelf with the sample bottles by the Raman. Unscrew it and make sure gasket ring is in the upper part of the filter (right side of figure). Insert a filter disk and screw pieces together. Using an appropriate sized syringe, push your sample through the filter into a brown sample bottle.! Solids should be finely divided and free-flowing enough to provide easy insertion and good interaction with the laser probe tip. Grind in a mortar if necessary. Put the solid in a small vial with a mouth wide enough to accept the laser probe tip. Check the drawers on the back bench in the stockroom. Taking a Raman Spectrum! Make sure the laser is off and everyone in the room has laser goggles on.! Rinse off the laser probe tip and gently dry it with tissue.! Place your liquid sample in a brown bottle. Insert the laser probe tip and adjust the septum so it covers the bottle mouth and holds the laser probe tip just below the liquid surface. Orienting the probe perpendicular to the bottle bottom gives the best results (minimizes reflections from the bottle sides).! Put the bottle into the black can and slide the black, slotted cardboard cover around the cable to cover the can. (This minimizes stray light from interfering.)! Solid samples are handled similarly using a small vial (5-10 dram vials work well), but require insertion of a Inline Filter Module in the laser fiber optic line (See Laser Operation section above for details of installation).! Turn the Power Switch Key on and click Laser On. After 20 seconds or so the Raman spectrum will appear on screen.! Click on the Manual Scale button on the toolbar. Set the minimum x value to something other than 0 (say 200 nm) so the laser line itself is not displayed with the spectrum. Set the maximum y value just greater than the height of the largest peak. Sample spectra and the effect of degassing are illustrated below. Broad background envelope and sloping baseline mean not well degassed. Spectra on horizontal baseline means well degassed.! Alternately, hold the shift key and drag a box around the section of spectrum that you want to display. Click on the Autoscale icon on the toolbar to resize this spectrum segment to fit the screen.! Adjust Acquisition Time, Number of Spectra to Average, and Boxcar Smoothing Width as necessary to 4

5 obtain a good quality spectrum.! Peak selection can be done automatically with menu choice Peaks Mark All Peaks, which is sensitive to the settings in Peaks Peak Settings dialog box. Experiment with settings to provide the labeling you desire.! Alternately, turn the cursor on (blue vertical line on the spectrum) by clicking the Cursor View icon on the toolbar. Move the cursor to a peak you want to label and then menu select Peaks Mark Peak. Shutdown and Cleanup! Turn off the laser (uncheck Laser On and turn Power Switch Key to the off position.! Turn off the System Power Switch on the Raman.! Remove your sample bottle from the black can and remove the laser probe tip.! Rinse off the laser probe tip with a solvent that dissolves your sample and wipe gently with a tissue.! Replace the plastic cover on the laser probe tip.! Clean up your sample bottles and return them to the collection on the shelf by the Raman or the stockroom, as appropriate. Recycle or dispose of your sample appropriately; simply pouring down the drain is often not acceptable.! Remove the Caution sign from the door.! Return the Laser Switch Key, Remote Interlock and Inline Filter Module to Dr. Weisshaar. Troubleshooting Check Hardware Configuration! Menu select Setup Configure Hardware and verify or restore settings to those shown on the left below. Then click on OK. Check Spectrometer Configuration! Menu Select Setup Configure Spectrometer and verify or restore settings to those shown on the right above. Then click on OK.! See Dr. Weisshaar. 5

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