Chromatography Forum

Hi to all,


I want devlop a new method for estimation of IMIDAZOLE by Reverse Phase HPLC. Already i have done with YMC ODS (150 x 4.6, 5mm) by using mobile phase 50:50 (acetonitrile : water: 0.1% TEA) . I found the peak is comming before second minute then i change the mobile phase concentration to 20:80(acetonitrile : water) eventhen i did not get any improvement.

Anybody of you can suggest good method for the same. I want retention time about 8.0 minute

Thanks in advance

Hi SRINIVASAN,

I doupt that you can bring Imidazole on your YMC ODS-A on a retention time of 8 min.
As far as I remember Imidazole is a base. So try a buffered mobile phase (80 to 90% of water) at pH around 8 to 10 with a column like Waters XTerra or Zorbax Extend.
Any form of ion pairing chromatography should work too.
(Guess, the guy from Alltech Sep will post his suggestion in the next time.)

For such a polar base I always recommend to use HILIC: use a silica column with a mobile phase of about 80% acetonitrile and 20% water. You may need to adjust the pH with a small amount of acetic or formic acid.

From my experience, HILIC is much suitable for imidazole and its derivates. Try Si-gel column with water/ACN at low pH.