Wednesday, 17 September 2014

Storing HPLC columns( simple and small)

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


 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.

(üPhysical 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




                             

2 comments:

  1. I am requesting the viewers to post their question in my blog....

    ReplyDelete
  2. thanx, nice topic.
    How we will do proper column regeneration ?
    suchi

    ReplyDelete