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  • Title: Assay of amikacin in the skin by high-performance liquid chromatography.
    Author: Nicoli S, Santi P.
    Journal: J Pharm Biomed Anal; 2006 Jun 07; 41(3):994-7. PubMed ID: 16466885.
    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.
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