Sack: HPLC Protocol

HPLC

Chemicals

Precautions

Ensure that you are grounded, touch the door handle, or another surface where you can discharge any static electricity, prior to handling equipment.

Mobile Reservoir, level should be below that of equipment.

Protocol

1. Power on, ensure pumps are connected without creating back-pressure.
2. Fill reservoir with mobile phase, and put suction filter in it.
   • Change the operation if mobile phase is changed out.

Look at the pipe, see if its connected to the multiple phase outlet port

1. Check end of the drain pipe put into the waste liquid bottle.
2. Power on pump
   • Pressure will read 0-5.
   • Max Pressure setting, and the min pressure setting are adequate (limiter settings can be adjusted if needed)

Open drain valve
Press the upper right keys 4 0, and Enter
this sets the flow rate: 40.0.

5.Press Pump key; pump will be turned on, it’ll speed up gradually.

Check liquid running out of the drain pipe ~ 30 secs, see for continual flow

Press Pump key; to turn pump off

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Set desired flow rate

Check outlet of the fraction collector or the detector.

Normalizing Method of Liquid Transfer, purging air from head

• Things to look out
  
  • Irregularities in Liquid Transfer
  • Liquid is not transferred, is not drawn up.
  • Liquid is synced with pump
  • Indicated

Stopping the unit

Ensure that buffer is completely flushed

if drain valve = open, when liquid is high in reservoir, liquid may flow (siphon)
close drain valve when not transferring liquid.

• drain valve left open.
• mobile phase level in the reservoir high
• drain pipe outlet port low

Quanitate amount going in and out...

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