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Antiepileptic drugs determination in serum by hplc

Posted: Thu Jan 31, 2013 10:09 am
by parya_m
Dear colleagues,
Can anybody kindly tell me the HPLC method for simultaneous determination of Antiepileptic drugs (lamotrigin, ethusoximide, primidon, phenobarbital, phenytoin, carbamazepin) in serum using c18 150x4.6mm column and UV detection that can separate them completely ?
what is the mobile phase used and other HPLC parameters?
Sincerely,

Re: Antiepileptic drugs determination in serum by hplc

Posted: Thu Jan 31, 2013 10:17 am
by Gerhard Kratz
Please check out the websites from Waters, Phenomenex, Agilent etc. where you will find tons of applications. Just a little google will help you.
If you face problems with reproducibility of a method you found we will be happy to give you as much as possible support.

Re: Antiepileptic drugs determination in serum by hplc

Posted: Thu Jan 31, 2013 10:19 am
by prepcolumn
there are some paper abstacts in google for simultaneous determination of these molecules

Re: Antiepileptic drugs determination in serum by hplc

Posted: Fri Feb 01, 2013 6:01 pm
by parya_m
Please check out the websites from Waters, Phenomenex, Agilent etc. where you will find tons of applications. Just a little google will help you.
If you face problems with reproducibility of a method you found we will be happy to give you as much as possible support.
thank you so much for your kind reply, I check out above websites, there is an application method in phenomenex website, but they don't explain anything about sample preparation.

I found this method and be setting it up on my column :

Column: c18 150x4.6mm
Detection: 205nm
Mobile Phase: Buffer/ ACN (80:20)
Buffer: H3PO4 20 mmol
pH= 2.1 (adjusted by KOH 1M)
Temp.=50
sample preparation: 1 to 2 of serum and Acetonitile/H3PO4 0.1 M (1000:1) then vortexed for 1 min and centrifuged
at 9700 rpm for 5 min. The organic (upper) layer was diluted to half by water.

I have been struggling to separate carbamazepin and phenytoin appropriately, peaks are very close and broad
what I really can do to obtain sharp and specific peaks and decrease their retention time?