Pickering Laboratories Pinnacle PCX - Post-Column Conditions 6

6.2-4
Section 6
ApplicAtions
Pinnacle Operators Manual
Pickering Laboratories Inc.
Note: TRIONE
®
is air sensitive, and must be kept under Nitrogen. The useful lifetime of T100 is three months*
unopened, and one month in the reservoir. The shelf-life of T200 is one year* unmixed, and one month in the
reservoir.
*From date of manufacture.
OPA PREPARATION
1. Pour 945ml of the OPA Diluent (Cat.No. OD104) into the reagent reservoir. Save approximately 5 ml for
Step 5.
2. Put the cap on the bottle, open the vent valve, and turn on the gas supply. Thoroughly deaerate the contents
by sparging with inert gas. Continue bubbling for at least 10 minutes.
3. Dissolve 300mg of OPA (Cat. No. O120) in 10mL of HPLC-grade methanol in a clean, dry container.
4. Turn off the gas supply and remove the cap from the bottle. Add the OPA solution to the deoxygenated diluent
in the reservoir. Wash any residual mixture into the reservoir with an additional 1–2 ml of methanol.
5. Dissolve 2g of Thiofluor™ (Cat. No. 3700-2000) in the reserved 5 ml of OPA Diluent and add to the
reservoir.
6. Add 3ml of 30% Brij-35
®
(Sigma) solution.
7. Replace the cap and close the vent valve. Gently swirl the reagent to complete the mixing. Turn on the inert gas.
Note: OPA reagent is sensitive to air oxidation and will degrade over time. The Pinnacle PCX system is designed
to minimize this oxidation. When the OPA reagent reservoir is maintained under inert gas pressure, the OPA
reagent can maintain its activity for up to one week without significant loss of activity.
Post Column Conditions
These are the recommended post-column conditions for the most common methods of amino acid analysis.
For the HPLC conditions, refer to the section titled Sample Chromatograms and Gradient Programs.
USING TRIONE NINHYDRIN REAGENT:
Reagent 1: TRIONE
®
(Cat. No. T100 or T200)
Pump 1 Flow Rate: 0.30 ml/min
Reactor 1 Volume: 500 µl
Reactor 1 Temp: 130°C
USING OPA REAGENT:
Reagent 1: o-Phthalaldehyde and Thiofluor
™
in OD104 Diluent, plus 35% Brij-35
®
Pump 1 Flow Rate: 0.30 ml/min
Reactor 1 Volume: 150 µl
Reactor 1 Temp: 45°C