1. Turn on the UV-1800 UV-VIS spectrophotometer, using the power switch located on the right side of the instrument.
2. Open the UVProbe 2.43 software on the computer desktop
3. Click the Connect button on the bottom of the window, the software will then find the instrument and automatically switch to display the experimental window.
4. Click on the Edit method tab in the toolbar (the green M in a yellow circle).
5. Under the "Measurement" tab, input the Wavelength Range (nm) you wish to scan between (Highest value goes in Start and Lower values go in end, the maximum range is between 190 and 1100 nm) as well as the Scan Speed for the experiment.
6. Under the "Instrument Parameters" tab, the "Measuring Mode" should be set to Absorbance (the slit width for this instrument is fixed at 1nm). Once parameters have been set, click "OK" at the bottom of the window.
7. Zero the instrument by clicking the Auto Zero button at the bottom of the window.
8. Create a reference solution containing everything in your sample besides the analyte (or as best that you can). This will be used to baseline the instrument. Baseline must be done with the same thing in both the sample and reference slots!
NOTE: The reference solution is placed in the rear-most slot. All solutions should be placed in cuvettes (cuvettes have two clear and two opaque sides), with the clear sides of the cuvettes facing to the left and right. With the reference in the sample holder, close the door and click the option for "Baseline" towards the bottom of the window.
9. Once the baseline scan is done, the bottom left of the window (above the control buttons) will display the starting wavelength again. Open the lid and place in your first sample to be analyzed, then again close the lid. To begin analysis, click the "Start" button two over from the Baseline button at the bottom of the window.
10. After the spectrum has been collected, you will see a new window pop up: This is where you will give the sample a file name. To the right of the File blank is three dots, click this to direct where to store the data, making sure to store it in the proper folder. It should be stored in the UVProbe-Data, then the Data folder directory, but be sure to create an individual or lab group folder to keep the data in. Once satisfied with the file location and the name, select "Open" to confirm the input. After all the information is set, click "OK".
Note: This does not save your data, it just sets up the raw data to be manipulated and later saved as the file when you so choose.
11. Once the spectrum has been collected, go to "File" along the top toolbar and select the option for "Save". This step will actually save the data to where it was specified previously.
NOTE: Be sure to save the individual spectra immediately after the run has completed, this will ensure data is saved properly
12. Now your raw data will be displayed on the graph on the right hand side of the window. As long as you do not change any parameters or use a different solvent for your sample you can use the background you have already generated and run more samples by putting in your new sample and hitting the Start button. This will place all of your raw data onto one graph. Remember to save each spectrum as with its own file name and comment.
13. Once you have taken some spectra your window will look something like this:
Notice that many of the solutions were too concentrated, if you have an absorbance that is greater than 2 you are much too concentrated. An absorbance of 2 is telling you that 99% of the light was attinuated, or another way of putting it, only 1% of the light made it through. Notice that all spectra are displayed. If you want to look at only one you can either uncheck all of the spectra but the one you want or better way is to click on the graph menu at the top and click the active button which will only display the spectrum that is active. While in the active graph setup, to observe another spectrum simply double click on its name in the list to the right of the active spectrum.
14. An additional format may wish to be saved, called Data Print. This format provides the wavelength and corresponding absorbance in an easily transferable format. While in the active spectrum mode and having only one spectrum displayed, click on file and save as, make sure you are in the folder that you want your data saved in then save the file (already named) as
"Save as type: Data Print Table (*.txt).
Do both of theses saves to all of your data.
15. Be sure to also save or move your data to another device that is accessible to you, such as a flash drive or upload it to a cloud storage system such as google drive.
16. When done taking data click on the Disconnect button at the bottom of the window.
17. After disconnecting you can exit the program then turn off the instrument.
