By Dennis Gere on Monday, August 11, 2003 - 04:45 am:
Does anyone have any experience with the HPLC separation of either of these compounds or any related separations? Dennis Gere - New Bolton Center, Univ of PA School of Vet Medicine, Toxicolgy Lab
Any information will be helpful. Have tried Phenyl phase and C 18 with water Acetonitrile, Ammonium Acetate buffer at pH 4.6. No visible resolution from 25 to 50 deg C. thank you in advance
By Anonymous on Monday, August 11, 2003 - 05:13 am:
Method for rapid separation of liposome-associated doxorubicin from free doxorubicin in plasma
Thies, R.L. et al.
Anal. Biochem,1990,188,65
By Dennis Gere on Monday, August 11, 2003 - 11:23 am:
Thank you for your prompt reply.
Upon careful reading of this article by Theis, et.al., it appears that they did the actual separation of the liposome associated doxorubicin from the "free" doxorubicin uses a method of solid phase extraction ( an ion-exchange spe material). The first fraction is eluted from the spe cartridge with a phosphate buffer adn collected and then injected onto the HPLC column. A second fraction is eluted with hexane and subsequently injected (a separate injection after the first chromatogram is finished). We speculate that the injection of the liposome encapsulated doxorubicin (if it eluted intact from the spe cartridge) comes apart at the head of the HPLC column in the HPLC mobile phase (the micelle sticks and releases the "free" doxorubicin). Alternately they simply did the assays by Fluo. spectroscopy without any separation. A later publication from the same lab compared this to separation on Microcon-30 ultrafilters, again without using the HPLC, but doing separate assays on the individually collected fractions.
Anyone else out there have any luck with HPLC directly?? Thank you again, Dennis Gere
By Anonymous on Tuesday, August 12, 2003 - 04:21 am:
Lots of references, perhaps one will help.
http://www2.waters.com/watprod.nsf/wwwNewWebSearch?SearchView&Query=Doxorubicin