title: Assay of amikacin in the skin by high-performance liquid chromatography.
abstract: Amikacin is used in the systemic treatment of serious infections, but also locally for the treatment of skin infections. The aim of this work was to develop and validate a simple procedure for amikacin determination inside the epidermal tissue: this implies a simple method for an efficient drug extraction from the skin and a clean and easy HPLC analysis. Amikacin was extracted from epidermis samples with 500 microl of a mixture methanol-water-0.05 M NaOH (5:5:2 v/v/v) at 60 degrees C for 1 h. After filtration, the obtained solution was derivatized (1-fluoro-2,4-dinitrobenzene at 90 degrees C for 10 min) and analyzed by HPLC, on a C18 microBondapack 300 mmx4.6 mm column thermostatted at 45 degrees C. The mobile phase was a mixture of acetonitrile-water-acetic acid (47:53:0.1 v/v/v) at a flow rate of 1.5 ml/min and the UV detector was set at 365 nm. The derivatization and HPLC analysis were validated in the concentration interval 1.64-49.21 microg/ml. The linearity resulted very good (R=0.9995); the R.S.D.% varied between 0.20% and 3.89% depending on the concentration and the ER% was included between 5.4 and 0.9. The extraction method used demonstrated to be specific and the recovery resulted about 93%. The extraction, derivatization and HPLC assay has good reproducibility, sensitivity and specificity resulting in a reliable method for biopharmaceutical studies of AK distribution in the epidermis.
authors.affiliation: Dipartimento Farmaceutico, Università degli Studi di Parma, Parco Area delle Scienze 27/A, 43100 Parma, Italy. sara.nicoli@unipr.it
authors.forename: Sara
authors.initials: S
authors.lastname: Nicoli
authors.forename: Patrizia
authors.initials: P
authors.lastname: Santi
mesh_terms: D000583:Amikacin / Q000032:analysis*; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015203:Reproducibility of Results; D012867:Skin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000900:Anti-Bacterial Agents; D000583:Amikacin