Waters AccellPlus QMA and CM Bulk Media - Manual (Page 3)

[ Care and Use ManUal ]
Waters AccellPlus QMA and CM Bulk Media 3
The AccellPlus column should now be tested (see Packed Column
Testing). Use methanol as the mobile phase to run the efficiency and
asymmetry test.
d. Packed Column Testing
Waters recommends that columns be tested prior to use. There are
many different ways of doing this, but one of the simplest is to inject
a small volume of a 500:1 buffer:acetone solution. This will absorb
effectively at 280 nm. The peak should be symmetrical. A typical
calculated plate count an AccellPlus IEX slurry packed, 1 x 10 cm
column should be greater than 500 plates. Lower observed values
would be normal for columns that are packed using the less efficient,
dry packing process.
N = 16
Rt 2
W
Rt = retention time in minutes
W = peak width at baseline in minutes
III. bulk medIa use
Determine the amount of bulk ion exchange media required for
application needs (see Section II, Table 2 or Table 3).
Slurry the media with 5 times the media’s volume of dilute buffer
for 2-3 minutes. The media should be allowed to settle and then the
excess buffer drained off. A similar volume of concentrated buffer
(0.1 M) should be added, slurried up 2-3 minutes, and then drained
off. The same procedure should be repeated with dilute buffer until pH
and salt concentration remain at the desired values.
IV. addItIonal InformatIon
a. Chemical Compatibility
AccellPlus is fully stable in aqueous buffers between pH 2-9. The
following reagents are compatible with AccellPlus media for use in
cleaning and as anti-microbials.
• 10% acetic acid (pH 2.5)
• 2.5 M sodium acetate (pH 8) 20% aqueous propanol,
0.1% trifluoroacetic acid 5 M urea
• 0.5% Hibitane™ (chlorohexidine)
• 5 M urea
• 1% Triton™ X-100
• 0.1% sodium azide
• 100% methanol
• 100% acetone
• 100% ethanol
• 0.1% sodium azide
• 70% ethanol
• 2 M sodium chloride
• 0.25 M hydrochloric acid (limited)
• 0.1 M sodium hydroxide (limited)
b. Sanitization
70% ethanol, 0.10% sodium azide or 0.5% Hibitane are effective
sanitizing agents.
c. Regeneration
Clean protein solutions, carefully filtering of buffers, and occasional
column regeneration will yield 100 or more runs. Occasionally, a
reduction in capacity is observed. When this happens, the column
should be regenerated by pumping 5 column volumes of 2 M sodium
chloride at pH 7.5 for the AccellPlus QMA media and pH 4 for the
AccellPlus CM media. More aggressive cleaning solutions may be
found in Table 4.
Table 4. Suggested Cleaning Solutions
Agent Concentration Exposure
Sodium Hypochlorite 1000 ppm 2 hours
Hydrochloric Acid 0.25 M less than 30 minutes
Sodium Hydroxide 0.1 M
3 column volumes pump through
in less than 20 minutes
d. Short-Term Storage (less than 72 hours)
AccellPlus media does not require special storage or treatment for
periods less than 72 hours. We recommend, however, that the
columns be stored at 4 °C.
e. Long-Term Storage (more than 72 hours)
Although AccellPlus media is not susceptible to microbial attack, an
anti-microbial such as Hibitane, sodium azide or 70% ethanol can be
added (see Chemical Compatibility). If the medium is to be stored dry,
pump the medium out of the column, wash with 10 column volumes
of 1 M sodium chloride, 10 column volumes of HPLC-grade water, and
10 column volumes of methanol. Dry the media under vacuum at 80 °C.