TCNJ Chemistry Instrumentation

Operating Instructions

Ocean Optics USB 2000 UV/VIS Spectrophotometer

1) Open Ocean Optics USB UV/VIS folder.

2) Open OOI Base-32.

3) If the last spectrometer used on the computer is not plugged into the USB port, an error message will appear twice. Click OK both times.

4) Select configure in the spectrometer pull-down menu, or click the corresponding icon in the toolbars on the main screen.

5) Select the A/D Interface tab and open the USB Serial Number pull-down menu.

6) a. If only one spectrometer is plugged into the USB port, verify that the serial number matches the number located on the bottom of the spectrometer. Select it, and then click Apply followed by OK.

b. If more than one spectrometer is plugged in, select the serial number of the spectrometer you wish to use. Then click Apply followed by OK.

7) To switch between spectrometers plugged into the USB ports, repeat steps 3-6b. (NOTE: Do not unplug spectrometer from USB Port while OOI Base-32 is running. This will cause the program to freeze.)

8) Make sure the Lamp On, Vis Lamp On, Shutter Open, and Strobe/Lamp Enable check boxes are checked. Do not change any other instrument parameters at this time.

9) Place OOI Base-32 in Scope Mode by clicking the Scope Mode ("S") icon.

10) Ensure that the entire signal is on scale. The intensity of the reference signal should be about 3500 counts. If necessary, adjust the integration time until the intensity is approximately 3500 counts.

11) If the signal appears noisy, increase the Boxcar value. This will smooth your spectrum.

12) To take a reference spectrum of your solvent, click the Store Reference Spectrum icon (yellow light).

13) Block the light path to the spectrometer by sticking a pen or similar object into the sample holder. To take a dark spectrum, click the Store Dark Spectrum icon (grey light).

14) To take an absorbance measurement of your sample, click the Absorbance Mode ("A") icon.

(NOTE: At this point, if you decide to change any of the sampling variables (integration time, boxcar smoothing, etc.), you must store a new dark and reference spectrum.)

15) To save your spectrum, select File | Save | Processed from the menu bar. Save your spectrum in the appropriate class folder (HONR 321, etc.).

16) To generate a list of absorbance values (and their corresponding wavelengths), select Edit | Copy Spectral Data | All Spectrometer Channels from the menu bar.

17) Go to the WordPad program (Start | Programs | Accessories| WordPad). Click Edit | Paste to paste the data in WordPad. (NOTE: This list of absorbance values is huge! You can highlight and delete the values you do not need.)