Ctrl+K

Agilent 1260 Infinity II HPLC

Contents

Agilent 1260 Infinity II HPLC#

Procedure#

  1. If the HPLC is not on (as indicated by the illuminated power lights on the instrument), turn on all three modules. The power buttons are at the bottom left of each module.

  2. Open the Control Panel software. Click HPLC 1 in the pane on the left followed by the Launch button in the window that opens to open the program:Acquisition software.

  1. In the top ribbon of Acquisition, click Single Sample.

  2. Name your sample something fun and interesting — this is just practice. Click the … icon next to Acq. method and find C:\CDSProjects\Agilent Project\Methods\CHEM341-Beverages-iso.amx if the correct method is not loaded. Change the Result path to C:\CDSProjects\Agilent Project\Results\Practice. Change the Data file to <D> - <S> - <V> 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.

  3. Open the autosampler door and inspect the samples available. If you took CHEM341 last semester, some of the vials may seem familiar! Pick your favorite and note its position — you will need the following information:

    Tray number

    Either “1” or “2” printed on the white plastic of the autosampler.

    Vial number

    Note the column (A, B, C, D, E, or F) and row (1–11) of your vial as printed on the black sample tray.

    You may need to pull the sample holder out to get a better view. The trays easily slide out of the autosampler. If you encounter any resistance, stop and ask for help. Close the door.

  4. Under “Autosampler” on the computer, in the “Vial” field, type in your favorite vial using the format p<Tray Number>-<Vial Column><Vial Row>. In other words, if I wanted to inject the vial in position Tray 1, C3, I would type p1-c3.

  5. Click Run.

  6. Take note of the signal graph at the bottom. You should begin to see data collected. The entire run will take 20 min.

  7. If you are the last group of the day, click on Status in the Acquisition ribbon. Find the Shutdown Method field and click the … button. Find and Open the Water to Methanol Rinse.amx method. Click Submit Shutdown.

    Note

    Note that submitting a shutdown method does not end your current run. Instead, it adds on a new run immediately after the last run in the queue. This allows you to generate a long run queue of experiments followed by a method to safely shutdown the instrument. This is useful for when you want to run something overnight but don’t want to stick around until it ends!

  8. When your run is finished, click on Status in the Acquisition ribbon. Under the Run Queue, click the History tab. Find the sample you submitted, right click the row, and select Review Selected Run In Data Analysis.

  9. In the Data Analysis program, select the GC/LC area Percent method configuration and then click Link and Process. Please be patient while the computer integrates all of your peaks.

  10. Under the Home tab of the ribbon, click the Results button under Layouts. This will provide you with a summary of the peaks detected and their relative areas.

    Note

    This information could be written down and manually worked up. During the semester, we will attempt to prepare a processing method to automate the analysis.

  11. If you are not the last group of the day, you are all set — close all windows and move on to another station! If you are the last group of the day, did you submit the shutdown method?

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. Our mobile phase is a combination of the solvents in positions “C” and “B”. What is this mobile phase? Inspect the method to retrieve the actual ratio of solvents used.

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

Contents