Waters Protein-Pak SEC Columns - Manual (Page 5)

[ CARE AND USE MANUAL ]
Protein-Pak SEC Columns 5
Figure 3: 5 Sigma Method
c. Test Conditions
Columns are thoroughly testing in our quality control laboratories for adherence to our specifications. Since slight variations in your results will
occur depending on the equipment used, test your sample makeup and equipment settings and condition, perform the test sample run given here for
your new column and record the results (retention time and the settings used) before attempting the first analysis. Use these results for comparison
throughout the life of your column.
Detectors should run at 254 nm or 280 nm [UV]; otherwise, use intstrument settings that produce an acceptable test peak [this may mean using a
lower sensitivity on UV detectors than normally practiced for a specific analysis].
Note: Be sure to record results and instrument settings (and configurations) to allow exact reproduction and comparison in the future.
Key
Mobile Phase Test Sample Peak
1 100% Methanol A 1,2 Dichlorobenzene
2 0.05% Sodium Azide B Ethylene glycol in water
Column name Part Number Mobile Phase Optimal Flow Rate
(mL/min)
Test Sample Peak
Protein-Pak 60
7.8 mm x 30 cm
WAT085250 1 0.5 A
Protein-Pak 125
7.8 mm x 30 cm WAT084601 1 0.5 A
Protein-Pak 300 SW
7.8 mm (i.d.) x 30 cm, Steel
WAT080013 2 0.5 B
Protein-Pak 200 SW
8.0 mm (i.d.) x 30 cm, Glass
WAT011786 2 0.5 B
Protein-Pak 300 SW
8.0 mm (i.d.) x 30 cm, Glass
WAT011787 2 0.5 B
INJEC
T
W
Vr
N = Column efficiency
Vr = Elution volume (mL)
or distance from INJET (mm)
W 1/2= Volume at 4.4%
of peak height (mL) or peak
width at 4.4% (mm)
N = 25 x
2