HPLC SOP

What is the correct order of purging a pump?

Open drain valve -> Press Purge -> Close the Drain Valve

Press Purge -> Open drain valve -> Close Drain valve

Close Drain valve -> Press Purge -> Open Drain valve

Why is the maximum pressure limit important?

It changes the behaviour of the analytes.

Too high of a backpressure can be safety issue

What function does the mobile phase play in a HPLC?

It removes dissolved gases in the solvent which can interfere with operation of the HPLC

Supplies the force necessary to provide a constant flow rate regardless of the resistance met in the system.

Interacts with the sample to retain the compound in the column.

Moves the sample through the column.

When I start a new data run, a prompt shows up on the LabSolutions Software telling me to press "Start". Do I follow the instruction?

Yes

What do I have to look out for when equilibrating the column?

Constant pressure

10 column volumes have passed

Desired temperature has been reached

Check for leaks

When I am doing Peak Integration of my data run. The LabSolutions software can detect more than 50 peaks! I am only expecting 7 compounds. How do I fix this?

Rerun the sample

Calculate by Height and change the minimum height to 10000 AND change slope to 10000

Change slope to 10000 only.

Change min/area height to 10000.

I want to save a chromatogram and be able to modify it. How do I do that? Tick the options that apply

This is called a metafile

Right click on chromatogram and click "Copy as Metafile"

In a Powerpoint slide, click the arrow under Paste under the Home toolbar.

Click Paste Special then a window should appear named ‘Paste Special’. Then select Picture (Enhanced Metafile).

Ungroup the image twice.

Where do I change %B and the Oven temperature respectively?

LC Time Prog, Detector

Data Acquisition, Detector

Pump, Column Oven

Detector, Column Oven

What do I expect when I increase the oven temperature?

Increase in backpressure

Increased retention time and increase in backpressure

Decrease in backpressure and decreased retention times

Decrease in retention times.

Which of these responses are not calculated?

Resolution

Retention Time

Retention Factor

Selectivity

True or False: Manual HPLCs have a UV VIS detector and UPLCs have a PDA detector.

True

False

HPLC SOP

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