104
Running an Isocratic Analysis
What You Will Need
What You Will Need
Instruments Agilent 1100 Series isocratic, binary or quaternary pump,
autosampler and a UV-detector.
Column A 125 mm × 4.0 mm Hypersil ODS, 5 µm (Agilent
Technologies part number 7982618-564).
Solvents For the isocratic pump, a solvent mixture of LC grade
bidistilled water (35 %) and acetonitrile (65 %).
Sample The Agilent Technologies isocratic standard sample (Agilent
Technologies part number 01080-68704). This contains
0.15 wt.% dimethylphthalate, 0.15 wt% diethylphthalate,
0.01 wt.% biphenyl and 0.03 wt.% o-terphenyl dissolved in
methanol.
Preparing the LC System
1 For the isocratic pump, fill the solvent bottle with the mixture of LC-grade bidistilled
water (35 %) and acetonitrile (65 %). For the binary or quaternary pump, fill one
solvent bottle with bidistilled water (channel A) and the other with acetonitrile
(channel B).
2 Turn on the detector lamp and pump using the On/Off button [F7] in the Analysis
screen. (Use Action keys then to select the module)
3 For the quaternary pump, turn on the degasser by pressing the line-power switch.
4 Purge the pump. For more information see Chapter 3 “Using the Pump”.
5 Allow the detector at least 15 minutes to provide a stable baseline.
6 Fill the contents of a Agilent Technologies isocratic standard sample ampoule into a
vial and seal the vial with a cap. Place the vial in position 1 of the autosampler tray.
7 Pump the water/acetonitrile (35/65 %) mobile phase through the column for
10 minutes at a flow rate of 2 ml/min.
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