title: Quantitative profiling of 4'-geranyloxyferulic acid and its conjugate with l-nitroarginine methyl ester in mononuclear cells by high-performance liquid chromatography with fluorescence detection.
abstract: Oxyprenylated natural products were shown to exert in vitro and in vivo remarkable anti-cancer and anti-inflammatory effects. This paper describes a rapid, selective, and sensitive HPLC method with fluorescence detection for determination of 4'-geranyloxyferulic acid (GOFA) and its conjugate with l-nitroarginine methyl ester (GOFA-L-NAME) in mononuclear cells. Analytes were extracted from cells using methanol and eluted on a GraceSmart RP18analytical column (250×4.6mm i.d., 5μm particle size) kept at 25°C. A mixture of formic acid 1% in water (A) and methanol (B) were used as mobile phase, at a flow-rate of 1.2mL/min in gradient elution. A fluorescence detector (excitation/emission wavelength of 319/398nm for GOFA and GOFA-L-NAME), was used for the two analytes. Calibration curves of GOFA and GOFA-L-NAME were linear over the concentration range of 1.0-50μg/mL, with correlation coefficients (r2)≥0.9995. Intra- and inter-assay precision do not exceed 6.8%. The accuracy was from 94% to 105% for quality control samples (2.0, 25.0 and 40μg/mL). The mean (RSD%) extraction recoveries (n=5) for GOFA and GOFA-L-NAME from spiked cells at 2.0, 25.0 and 40.0μg/mL were 92.4±1.5%, 94.7±0.9% and 93.8±1.1%, for GOFA and 95.3±1.2%, 94.8±1.0% and 93.9±1.3%, for GOFA-L-NAME. The limits of detection and quantification were 0.3μg/mL and 1.0μg/mL for GOFA and GOFA-L-NAME. This method was successfully applied to measure GOFA and GOFA-L-NAME concentrations in a mononuclear cells.
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Vito Alessandro
authors.initials: VA
authors.lastname: Taddeo
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy. Electronic address: s.genovese@unich.it.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Genovese
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Carlucci
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Ferrone
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Antonia
authors.initials: A
authors.lastname: Patruno
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Ferrone
authors.affiliation: Affichem S.A., 9, Rue de Saint Joseph, 31400 Toulouse, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: de Medina
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Fiorito
authors.affiliation: Department of Pharmacy, University "G. d'Annunzio" - Chieti-Pescara, Via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Epifano
mesh_terms: D002138:Calibration; D002478:Cells, Cultured / Q000737:chemistry*; D003373:Coumaric Acids / Q000032:analysis*; D006801:Humans; D057230:Limit of Detection; D019335:Nitroarginine / Q000031:analogs & derivatives* / Q000032:analysis; D015203:Reproducibility of Results; D020298:U937 Cells
chemical_list: C552732:4'-geranyloxyferulic acid; C000590392:4'-geranyloxyferulic acid-N(omega)-nitroarginine methyl ester; D003373:Coumaric Acids; D019335:Nitroarginine
keywords: 4'-Geranyloxyferulic acid; Cancer chemoprevention; HPLC-FLD; Mononuclear cell line U937; Oxyprenylated secondary metabolites