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Agilent 1260 Infinity II

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Agilent 1260 Infinity II#

Note

The below procedure was taken from the old HPLC. It will need to be adapted to the new instrument.

Procedure#

  1. The HPLC is a POS, so it should remain on all the time. Make sure that it’s on by noting all of the LEDs on each module (bottom = pump, middle = autosampler and column, top = detector).

  2. Open the ChromQuest software and then double-click HPLC. (Not HPLC/man!)

  3. Go to File ‣ Method ‣ Open and find 321Caffeine.met

  4. On the toolbar, click the icon that says “Single Run”. You may have to hover the cursor over each icon to find out what each one is.

  5. Name your sample something fun and interesting. Click the folder icon next to Method and find 321Caffeine.met if the correct method is not loaded. Change the data path to C:/Data/Practice. Change the Data file to the Sample ID - Method - Date and Time by clicking on the arrow to the right of the field. This allows data files to be automatically named based on what you place in here.

  6. Open the autosampler door and inspect the samples available. If you took 321 last semester, some of the vials may seem familiar! Pick your favorite and note its position - you will need a letter and a number. The letter is printed on the front of the instrument. The number is on the sample holder. You may need to pull the sample holder out to get a better view. Close the door.

  7. Under “Autosampler” on the computer, type in your favorite vial in the Vial field using the format “Letter;#”. So, if I wanted to inject the vial in C3, I would type C;3.

  8. Click start.

  9. As the run starts, you may want to play around with the windows. Click the Window menu at the top of the screen to see what options are available to you.

  10. When the run finishes, you may get some error messages about not being able to integrate. Ignore them.

Questions#

  1. What type of detector is this instrument using? Unsure? Take a look at the wavelengths being used and consider what region of the electromagnetic spectrum that falls under.

  2. You should have noticed that your data was very noisy in the beginning, but after your first peak, it became a lot smoother. What happened to all of the noise in the beginning?

  3. We’re using the mobile phase in position “D”. What is this mobile phase?

  4. Were your peaks clearly resolved? If not, what might you be able to do to increase the resolution between caffeine and benzoic acid?

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