Corrigendum to "Utility of Experimental Design in Pre-Column Derivatization for the Analysis of Tobramycin by HPLC-Fluorescence Detection: Application to Ophthalmic Solution and Human Plasma".

Abstract

A novel, selective, and sensitive reversed phase high-performance liquid chromatography (HPLC) method coupled with ­fluorescence­detection­has­been­developed­for­ the­determination­of­ tobramycin­ (TOB)­ in­pure­ form,­ in­ophthalmic­solution­and­ in­ spiked­human­plasma.­Since­TOB­lacks­UV­absorbing­chromophores­and­native­fluorescence,­pre-column­derivatization­of­TOB­was­ carried­out­using­fluorescamine­reagent­(0.01%,­1.5­mL)­and­borate­buffer­(pH­8.5,­2­mL).­Experimental­design­was­applied­for­optimization­of­the­derivatization­step.­The­resulting­highly­fluorescent­stable derivative was chromatographed on C18 column and eluted using­methanol:water­(60:40,­v/v)­at­a­flow­rate­of­1­mL­min.­A­fluorescence­detector­(λex­390­and­λem­480­nm)­was­used.­The­method­ was­linear­over­the­concentration­range­20–200­ng­mL−1.­The­structure­of­the­fluorescent­product­was­proposed,­the­method­was­then­ validated­and­applied­for­the­determination­of­TOB­in­human­plasma.­The­results­were­statistically­compared­with­the­reference­method,­ revealing­no­significant­difference.