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Development and validation of HPLC method for imiquimod determination in skin penetration studies
Author(s) -
Paula Daniel De,
Martins Carolina Azenha,
Bentley Maria Vitória Lopes Badra
Publication year - 2008
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1075
Subject(s) - imiquimod , chemistry , chromatography , penetration (warfare) , stratum corneum , high performance liquid chromatography , diethylamine , detection limit , elution , dermatology , medicine , pathology , operations research , engineering , organic chemistry
A simple, rapid and sensitive analytical procedure for the measurement of imiquimod in skin samples after in vitro penetration studies has been developed and validated. In vitro penetration studies were carried out in Franz diffusion cells with porcine skin. Tape stripping technique was used to separate the stratum corneum (SC) from the viable epidermis and dermis. Imiquimod was extracted from skin samples using a 7:3 (v/v) methanol:acetate buffer (100 m m , pH 4.0) solution and ultrasonication. Imiquimod was analyzed by HPLC using C 8 column and UV detection at 242 nm. The mobile phase used was acetonitrile:acetate buffer (pH 4.0, 100 m m ):diethylamine (30:69.85:0.15, v/v) with flow rate 1 mL/min. Imiquimod eluted at 4.1 min and the running time was limited to 6.0 min. The procedure was linear across the following concentration ranges: 100–2500 ng/mL for both SC and tape‐stripped skin and 20–800 ng/mL for receptor solution. Intra‐day and inter‐day accuracy and precision values were lower than 20% at the limit of quantitation. The recovery values ranged from 80 to 100%. The method is adequate to assay imiquimod from skin samples, enabling the determination of the cutaneous penetration profile of imiquimod by in vitro studies. Copyright © 2008 John Wiley & Sons, Ltd.