By Luca on Tuesday, May 13, 2003 - 03:51 am:
hello!
I was wondering if anyone of you have ever tried to run an HPLC assay of Kanamycin Sulphate or similar antibiotics, by means of derivatization (possibly, pre-column rather than post-column). Also, I read that this substance should absorb at 249 and 306 nm in acidic acqueous solution, but I have not been able to find proof of that so far. What's more, on attempting to separate Kanamycin by HPLC at pH 3.5 with IPRs, I cannot detect any UV absorption in the 200 - 400 range. That's why I am thinking about derivatization and asking for help...
Thanks, Luca
By Jan on Tuesday, May 13, 2003 - 06:19 am:
Hi Luca,
Do you have an electrochemical detector ? I know there is a nice method out there that separates kanamycine from its related substances without derivatisation. Detection is by PED.
Jan
By Anonymous on Tuesday, May 13, 2003 - 08:46 am:
Try this as a starting place.
http://www2.waters.com/watprod.nsf/wwwWebList/?SearchView&Query=Kanamycin
By chempara on Sunday, May 18, 2003 - 11:21 am:
HI
I HAVE WORKED WITH SIMILAR ANTIBIOTICS, SUCH AS AMIKACIN, TOBRAMYCIN ETC
NONE OF THOSE ARE UV ABSORBING.
THE ONLY WAY TO RESOLVE THIS PROBLEM IS TO DERIVATIZE THEM WITH REAGENTS THAT REACT WITH THE AMINO GROUP.
THE CHARACTERISTIC OF SUCH ANTIBIOTICS IS THAT THEY BELONG TO AMINOSACHARIDES AND EXHIBIT A FREE AMINO GROUP.
I RECOMEND OF USING FDNB (2,4-DINITRO-FLUORO BENZENE) TO DERIVATIZE THEM IN SLIGHTLY ALKALINE ENVIROMENT.
THE DERIVATE IS COLORED WITH MAXIMUM ABSORBANCE AROUND 350 - 400 nm.