Change in the standard chromatogram
Posted: Wed Sep 29, 2010 2:31 am
I got a question with the HPLC method I am working now and need immediate solution.....
Please help if anyone has an idea for this....
I am running several standards with C30 column in a RP-HPLC. I have 5 standard compounds and when I develop the method I got a good baseline separation for all the compounds. And I used the same method for quantification of samples too...
And yesterday when I tried to inject the standard mixture again, I did not get a baseline separation for two of them. they were half overlapped...and today i tried again and I only got frour peaks!!!! those two compounds are completely overlapped and appeared as one compound!!
I have no idea what is happening inside the column....Has anyone experienced this before????? anybody knows a solution.....Its urgent, please help.....
Please help if anyone has an idea for this....

I am running several standards with C30 column in a RP-HPLC. I have 5 standard compounds and when I develop the method I got a good baseline separation for all the compounds. And I used the same method for quantification of samples too...
And yesterday when I tried to inject the standard mixture again, I did not get a baseline separation for two of them. they were half overlapped...and today i tried again and I only got frour peaks!!!! those two compounds are completely overlapped and appeared as one compound!!

I have no idea what is happening inside the column....Has anyone experienced this before????? anybody knows a solution.....Its urgent, please help.....