Cleaning for SUGAR SP0810 HPLC Column

Record: 895

Category: Column Cleaning
Product: SP0810
Sample:

Question
OK Melissa thank you for the explanation. The analysis that I am trying to optimize is Xylitol Assay via the USP36/NF31 Monograph; I don’t believe we are using any acids when we perform this but if I am missing something please let me know. The mobile phase is 20:80 acetonitrile to water, that is also the solvent for the standard and sample prep. The preps are simply to dissolve so much Xylitol in the mobile phase.

We have seen system suitability failures and abnormal peak shapes with columns that have been previously used; replacing the column with a fresh one always results in better chromatography. Cleaning-wise I am not sure what would be left on the column but I don’t want to improperly clean it, store it, etc. I want to start with a brand new column and closely monitor this analysis and the handling of the system and column so I can better understand how to optimize it. I noticed on your shodex.com website you have an example chromatogram for the Xylitol analysis; is there any technical info you could provide that might help us?

Answer
Some questions: Are you dissolving solid xylitol in the mobile phase? How do the xylitol samples differ? Could you provide a chromatogram of a xylitol peak at the beginning and one with abnormal peak shapes? After about how many injections do the peak shapes become abnormal?

For the SP0810 column, the abnormal peaks may be caused by the removal of the lead ions; however this happens in the presence of acid. Could you check the pH of your samples?

I did some looking around on the internet, and xylitol binds Ca2+ ions and other polyvalent ions effectively. Perhaps the xylitol in the samples are leaching the Pb ions from the packing material over time and hence the peak shape becomes abnormal over time. Please try the regeneration procedure to restore the column performance.

We have several applications for xylitol analysis with the SP0810:
http://shodex.net/index.php?seitenid=1&applic=1010
http://shodex.net/index.php?seitenid=1&applic=1009

Here is a list of applications specifically for the SP0810 column:
http://shodex.net/index.php?seitenid=1&reiter=r_prod&prodid=742

Best of luck. Please send me any additional data, so we can make some progress on resolving your issues.

 
 
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