7.6
Section 7
TroubleshooTing
Pinnacle Operators Manual
Pickering Laboratories Inc.
COMMON COLUMN PROBLEMS
OBSERVED PROBLEM COMMON CAUSE ACTION TO TAKE NOTES
Loss of Resolution Guard column dirty
Bad tubing connection: wrong style
nut, too large tubing, wrong type
union
Column worn out
Poor Peak Shape - General Column worn out
Guard column dirty
Deposits in post-column flow path
Partial obstruction of flowcell
Too strong a solvent
Too large a sample injected
Reagent flow rate too high.
Improper tubing connection.
Protein contamination on amino acid
column
Start by replacing filter, then guard.
Replace column as last resort.
If new tubing connections have been
made, check connections.
Ensure that protein is completely
removed form column
Send a chromatogram to Pickering
Laboratories’ Technical Support
Department
Improper tubing connections are:
wrong style nut, too large tubing,
wrong type union, improper swaging
of ferrule.
Reverse and flush ion-exchange
column at elevated temperature
Reagent backflows into column Improper Shutdown procedures
Improper maintenance Procedures
Leaking fittings between column and
HPLC pump
Defective reagent control valves.
If NaOH is on column, follow
procedure on Page 7.9
If Reagent backflows onto column,
follow the procedure on Page 7.8
High Column pressure Guard column is blocked.
Worn HPLC seal or rotor seal.
Particulate matter in eluant reservoirs
Column is damaged
Organic solvent in ion exchange
column
NaOH on Carbamate column
Excessive eluant flow rate through
column
For reagent back-flow onto column,
see above.
If the column back-pressure is high
(> 2800psi), isolate the source
of the high pressure and replace
appropriate part. The analytical
column can be back-flushed to clear
partial blockage. Disconnect the
outlet of the column during the back
flush operation.
If organic solvent is on the cation-
exchange column, follow the
procedure on Page 7.8
Unfiltered samples
Pressure from filter and guard should
be < 200psi).
Organic contaminants can be washed
off the carbamate column by first
washing with methanol then with
dichloromethane. Wash again with
methanol before use
.
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