title: Validated RP-HPLC method for the simultaneous analysis of gemcitabine and LY-364947 in liposomal formulations.
abstract: Combined use of gemcitabine (Gem) and LY-364947 (LY), a TGF-β1 receptor inhibitor, has shown promise for the treatment of fibrotic pancreatic cancer, by reducing collagen production and improving tumor drug penetration. The preparation and optimization of novel Gem and LY formulations, including co-encapsulation in liposomes, require a validated method for the simultaneous quantification of both drugs, a method that had yet to be developed. Here we demonstrate an RP-HPLC protocol for the simultaneous detection of Gem and LY at 266 and 228 nm with retention times of 3.37 and 11.34 mins, respectively. The method, which uses a C18 column and a KH2PO4 (10 mM)-methanol mobile phase, was validated for linearity, precision, accuracy, limits of detection, and robustness. Co-loaded liposomes with both Gem and LY (Gem/LY liposomes) were developed to investigate the protocol applicability to pharmacokinetic analysis and formulation characterization. The method specificity was evaluated in presence of liposomal components in fetal bovine serum (FBS). Finally, the method was demonstrated by quantifying Gem/LY liposomal encapsulation efficiency and concentration liposomes-spiked FBS.
authors.affiliation: Department of Nanomedicine, The Methodist Hospital Research Institute, 6670 Bertner Ave., Houston, TX 77030, USA.
authors.forename: Shyam S
authors.initials: SS
authors.lastname: Bansal
authors.forename: Christian
authors.initials: C
authors.lastname: Celia
authors.forename: Silvia
authors.initials: S
authors.lastname: Ferrati
authors.forename: Erika
authors.initials: E
authors.lastname: Zabre
authors.forename: Mauro
authors.initials: M
authors.lastname: Ferrari
authors.forename: Ganesh
authors.initials: G
authors.lastname: Palapattu
authors.forename: Alessandro
authors.initials: A
authors.lastname: Grattoni
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics; D000971:Antineoplastic Combined Chemotherapy Protocols; D002417:Cattle; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis / Q000493:pharmacokinetics; D004355:Drug Stability; D008081:Liposomes; D010190:Pancreatic Neoplasms / Q000188:drug therapy; D011720:Pyrazoles / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics; D011758:Pyrroles / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics; D000093542:Gemcitabine; D011804:Quinolines
chemical_list: D000970:Antineoplastic Agents; D003841:Deoxycytidine; D008081:Liposomes; D011720:Pyrazoles; D011758:Pyrroles; D000093542:Gemcitabine; C506615:HTS 466284; D011804:Quinolines