6.2-5
Section 6
ApplicAtions
Pinnacle Operators Manual
Pickering Laboratories Inc.
USING SODIUM HYPOCHLORITE, FOLLOWED BY OPA REAGENT:
Reagent 1: 250 µl of 5% Sodium Hypochlorite in GA116 Diluent
Reagent 2: o-Phthalaldehyde and Thiofluor
™
in OD104 Diluent, plus 30% Brij-35
®
Pump 1 Flow Rate: 0.30 ml/min
Pump 2 Flow Rate: 0.30 ml/min
Reactor 1 Volume: 500 µl
Reactor 2 Volume: 100 µl
Reactor 1 Temp: 55°C
Reactor 2 Temp: Ambient
Procedure
Pickering Laboratories recommends different gradient conditions depending on the column and type of sample.
Use the program recommended on the column data sheet for the initial testing. Do not change this
program until you are sure that the other aspects of the system are functioning properly.
The column oven temperature programming gives additional flexibility when optimizing methods. Using
temperature gradient allows to improve separation, shorten analysis time and fine-tune the method for detecting
compounds of interest. Please refer to page 4.8 for details on how to set up timetable for the column oven.
Set the maximum pressure limit on the HPLC to 220 bar to protect the column. Allow the column to equilibrate
for about 30 minutes under initial conditions. Inject 10µl of the Amino Acid Standard, and collect three
chromatograms. The first chromatogram will not be representative of the systems performance, so use the
second two to evaluate the performance.
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