Hai folks......... today I am starting a "knowledge sharing program".
As a part of this, today I am sharing some interesting topic about............
" Storing of HPLC columns when not in use"
'Storing and using HPLC columns in a proper manner' can extend the lifetime of the columns.
I can used to say a quation that " Sparing a little extra time can save more investments".
The same point can applicable to this article.
Thinks to avoid (Simply can say Don'ts)
(X) Storage in Buffers
As a part of this, today I am sharing some interesting topic about............
" Storing of HPLC columns when not in use"
'Storing and using HPLC columns in a proper manner' can extend the lifetime of the columns.
I can used to say a quation that " Sparing a little extra time can save more investments".
The same point can applicable to this article.
Thinks to avoid (Simply can say Don'ts)
(X) Storage in Buffers
HPLC columns should not store in buffer solutions. During storage, buffers can precipitate, can effect worsely on the packing material as well as plugging fittings and
causing excessive back-pressure. This will not only affect the frits and
packing material, but can affect analytes and even generate peaks from organic
materials.
(X) Storage in THF
Solvents which degrade
easily, such as THF (Tetrahydrofuran), TEA (Triethylamine) and TFA
(Trifluoracetic Acid), can degrade the column and may even change the
chromatography.
(ü) Procedure for Short Term Column Storage
For overnight storage
when the column is to be used the next day, the columns should be flushed with
the operational mobile phase only without any buffer (if one has been used).
For example, if the mobile phase is 60% Methanol and 40% Buffer, then simply
replace this with 60% Methanol and 40% Water. This eliminates the risk of
precipitation of salts and also reduces the time taken to re-equilibrate the
column the next time it is used.
Ideally, mobile phase
should be continuously pumped through the column at a low flow rate
(0.1-0.2ml/min is enough). This will minimise equilibration time and not use a
lot of solvent.
(ü) Procedure for Longer Term Storage
If the column is to be
disconnected from the HPLC system and stored then it can either be stored in
the mobile phase it was shipped in, or, for reverse phase columns, stored in
100% Acetonitrile. If a mobile phase containing buffer was used, then first
replace the buffer with water, wash the column, and then flush with the
shipping solvent. A flush with 100% ACN before storage can also remove a lot of
contamination from the column.
(ü) Physical Storage
Before storage, ensure
that an end-fitting plug is inserted into each end of the column to prevent the
packing from drying out.
Do not store columns
loose in a drawer or on the bench. If a column is knocked or worse, dropped on
the floor, the physical trauma can damage the bed of the packing material,
leading to voids and a degradation in performance.
Columns should be
stored either in their original boxes, or ideally, using a column storage
system This enables a large number of columns to be stored
efficiently, in a compact space and in the best conditions.
I think this will be useful to the new joinees or beginners to analytical chromatography techniques...
I will be back with new and fresh content soon........
Thank you for visiting......
V.Suresh
I am requesting the viewers to post their question in my blog....
ReplyDeletethanx, nice topic.
ReplyDeleteHow we will do proper column regeneration ?
suchi