Waters Pico-Tag Amino Acid Analysis Column - Manual (Page 3)

[ Care and Use ManUal ]
Pico•Tag Amino Acid Analysis Column 3
a. Sample and Eluent Preparation and Filtration
Use HPLC grade solvents, filtered to remove microparticulate matter above
0.45 µm. (Waters Pico•Tag prefiltered Eluents A [P/N 88]108 and B [P/N
88]112 are recommended.) This reduces the problem of plugged filters
and preserves column life. Vacuum filtration or sonification may be used to
remove dissolved gasses which could affect your solvent delivery system.
Always filter prepared samples to prevent excessive pressure buildup due to
particulate matter
• Adequate sample cleanup (using Sep-Pak ® cartridges designed for this
purpose) prevents alteration of the column chemistry by strongly adsorbing
or precipitating sample components.
• Use of an in-line pre-column filter (P/N 84560), is recommended.
b. Precautions
Normal recommended pressure should not exceed 3500 psi.
• For all silica-based packing materials, stay within a pH range of 2-8
(i.e., avoid using concentrated acids or bases). Maximum column life
(pH): 3.5 - 6.5.
• Filter all aqueous buffers (Waters Pico•Tag eluents are prefiltered).
Avoid using turbid or cloudy buffers. Be sure that any solutions contain-
ing buffers, salts, etc. are compatible with the wetted surfaces of the
column and equipment.
• Protect column from vibration, mechanical shock, and rapid changes in
pressure. Column packings are based on a highly porous and delicate
silica gel alignment. Any thermal, physical or chemical shock (such as
changing solvents rapidly or at high flow rates) can cause the particles
to shift and may result in a loss of efficiency.
• When using water, distill or treat with a Milli-Q ® or equivalent system.
De-ionized water is not ac-ceptable because it contains organic com-
pounds which alter column selectivity.
• Protect the column from rapid changes in solvent composition. DO NOT
change the flow rate faster than 0.5 ml/min increments.
c. Storage Considerations (more than 72 hours without use)
Leaving the column unused for less than 72 hours does not generally require
storage procedures.
• DO NOT store the column in water alone; this practice will promote
microbial contamination. Store the column in Pico•Tag Eluent B or in
10% organic in water mixture.
• DO NOT allow buffers or other potentially harmful materials to remain
in the system when not being used. Flush and replace with Eluent B or
10% organic in water mixture.
• Return the column to its box with the end plugs firmly in place for
storage. Allowing steel columns to dry out can result in poor chromato-
graphic performance.
d. Troubleshooting
Table 2: Typical Column Problems and Solutions
e. Service and Applications Information
Waters staff of trained and experienced Service Specialists provides main-
tenance for Waters in-struments on preventative and/or corrective levels.
Contact Waters at 1-800-252-HPLC or your local Waters Representative for
answers to specific chromatography questions in areas such as methods devel-
opment, applications, quality control, and service related matters.
Problem Cause Solution
Excess pressure
buildup
• Filters plugged
with Particulates
• Sample precipi-
tates on column
(sample not
soluble in mobile
phase)
• Clean in an ultrasonic bath
or replace.
• Always alter mobile phases
and samples.
• Slowly purge with a strong
mobile phase that is both
appropriate to dissolve the
contaminate and compat-
ible with the column.
Loss of resolution,
broad peaks, low
plate counts • Contaminated
column, insuffi-
cient equilibration
• Column col-
lapse and void
formation
• Filters partially
plugged
• Slowly purge with very
strong solvent (refer to
Equilibration section)
• Since this may occur the
1st 2 weeks of use, Waters
will replace any column
with this defect. Refer to
warranty for details.
• Replace or clean inlet and
outlet filters.