An Improved HPLC Method Of Analysis Of Rofecoxib
An improved, simple, specific and more sensitive method for quantitative determination of rofecoxib from the human plasma is described. The method consists of pre-column photoactivation of rofecoxib in acetonitrile by exposure to UV light at 366 nm. The drug probably undergoes a stilbene-phenanthrene like photocyclization reaction with resulting formation of a highly fluorescent species. Subsequent injection on C18e column in HPLC with fluorescent detector at 260 nm excitation and emission at 381 nm with a mobile phase consisting of water:acetonitrile (55:45 v/v) could determine rofecoxib quantitatively in the concentration range of 10 to 350 ng/ml using 20 Î¼l of samples. The assay has been validated and has been successfully utilized in clinical studies.