S14-0011, 1/2 Shipping Release

Record: 863

Category: Applications
Product: NH2P equilibration
Sample:

Question
Retuning the column never has been our choice if you consider the time we spent trying to make thinks work with your help. I have attached a file where you can see sucrose always eluting very fast. If there is something you know that could improve the retention time, please let us know. Or if you believe that you need to check the column yourself and send it back to us, please do. But, as of now, it is not working. Even if ammonium acetate has caused the problem, it is in the manual. Would you please check again, if the volumes I used to regeneate the column (0.4 ml/min) is enough as the manual suggest 2 ml/min. Why the manual is asking us to do things we should not do?

Answer
Could you please send the flow rate, injection size, and sample preparation for the chromatograms you have attached? Our technical support in Japan would appreciate the additional information.

You had asked: Would you please check again, if the volumes I used to regenerate the column (0.4 ml/min) is enough as the manual suggest 2 ml/min. Why the manual is asking us to do things we should not do?

The washing procedure outlined in the operation manual is for the line filter (different and smaller than the column you have). The procedure outlines the molarity general procedure for the washing procedure. You should use the recommended flow rate for the NH2P-50 2D column, and the amount of eluent used for each step should be about 10 column volumes. In some cases, the use of ammonium acetate is needed to equilibrate the columns; however, I have found, through customer experience, that some compounds are not retained once the ammonium acetate has been used. The column can be returned to the shipping conditions through the washing procedure.

 
 
Additional Comments
Could you please send the flow rate, injection size, and sample preparation for the chromatograms you have attached? Our technical support in Japan would appreciate the additional information.

You had asked: Would you please check again, if the volumes I used to regenerate the column (0.4 ml/min) is enough as the manual suggest 2 ml/min. Why the manual is asking us to do things we should not do?

The washing procedure outlined in the operation manual is for the line filter (different and smaller than the column you have). The procedure outlines the molarity general procedure for the washing procedure. You should use the recommended flow rate for the NH2P-50 2D column, and the amount of eluent used for each step should be about 10 column volumes. In some cases, the use of ammonium acetate is needed to equilibrate the columns; however, I have found, through customer experience, that some compounds are not retained once the ammonium acetate has been used. The column can be returned to the shipping conditions through the washing procedure.

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