title: Development and validation of HPLC method with fluorometric detection for quantification of bisnaphthalimidopropyldiaminooctane in animal tissues following administration in polymeric nanoparticles.
abstract: A simple, sensitive and specific high-performance liquid chromatography method for the quantification of bisnaphthalimidopropyldiaminooctane (BNIPDaoct), a potent anti-Leishmania compound, incorporated into poly(d,l-lactide-co-glycolic acid) (PLGA) nanoparticles was developed and validated toward bioanalysis application. Biological tissue extracts were injected into a reversed-phase monolithic column coupled to a fluorimetric detector (λexc=234nm, λem=394nm), using isocratic elution with aqueous buffer (acetic acid/acetate 0.10M, pH 4.5, 0.010M octanesulfonic acid) and acetonitrile, 60:40 (v/v) at a flow rate of 1.5mLmin(-1). The run time was 6min, with a BNIPDaoct retention time of 3.3min. Calibration curves were linear for BNIPDaoct concentrations ranging from 0.002 to 0.100μM. Matrix effects were observed and calibration curves were performed using the different organ (spleen, liver, kidney, heart and lung) extracts. The method was found to be specific, accurate (97.3-106.8% of nominal values) and precise for intra-day (RSD<1.9%) and inter-day assays (RSD<7.2%) in all matrices. Stability studies showed that BNIPDaoct was stable in all matrices after standing for 24h at room temperature (20°C) or in the autosampler, and after three freeze-thaw cycles. Mean recoveries of BNIPDaoct spiked in mice organs were >88.4%. The LOD and LOQ for biological matrices were ≤0.8 and ≤1.8nM, respectively, corresponding to values ≤4 and ≤9nmolg(-1) in mice organs. The method developed was successfully applied to biodistribution assessment following intravenous administration of BNIPDaoct in solution or incorporated in PLGA nanoparticles.
authors.affiliation: UCIBIO, REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal. Electronic address: msegundo@ff.up.pt.
authors.forename: Marcela A
authors.initials: MA
authors.lastname: Segundo
authors.affiliation: UCIBIO, REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
authors.forename: Vera L R G
authors.initials: VL
authors.lastname: Abreu
authors.affiliation: UCIBIO, REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
authors.forename: Marcelo V
authors.initials: MV
authors.lastname: Osório
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal and IBMC, Rua Campo Alegre, 824, 4150-180 Porto, Portugal.
authors.forename: Sonia
authors.initials: S
authors.lastname: Nogueira
authors.affiliation: School of Pharmacy and Life Sciences, Robert Gordon University, Riverside East, Garthdee Road, Aberdeen AB10 7GJ Scotland, UK.
authors.forename: Paul Kong Thoo
authors.initials: PK
authors.lastname: Lin
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal and IBMC, Rua Campo Alegre, 824, 4150-180 Porto, Portugal; Department of Biological Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
authors.forename: Anabela
authors.initials: A
authors.lastname: Cordeiro-da-Silva
authors.affiliation: UCIBIO, REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal; Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal and IBMC, Rua Campo Alegre, 824, 4150-180 Porto, Portugal.
authors.forename: Sofia A C
authors.initials: SA
authors.lastname: Lima
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D005470:Fluorometry / Q000379:methods* / Q000592:standards; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism*; D053644:Naphthalimides / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism*; D015203:Reproducibility of Results; D014018:Tissue Distribution / Q000187:drug effects / Q000502:physiology
chemical_list: D053644:Naphthalimides; C557289:bisnaphthalimidopropyldiaaminooctane
keywords: Anti-leishmanial compound; Bisnaphthalimido-propyldiamino-octane; Fluorometry; Monolithic column; Poly(d,l-lactide-co-glycolic acid)