title: Celecoxib determination in different layers of skin by a newly developed and validated HPLC-UV method.
abstract: A simple, rapid and sensitive analytical procedure for the measurement of celecoxib (CXB) levels in skin samples after in vitro penetration studies was developed and validated. In vitro permeability studies in porcine skin were performed for quantification of CXB at different layers of skin, the stratum corneum (SC) and epidermis plus dermis (EP + D) as well as in the acceptor solution (AS) to assess CXB permeation through skin. CXB was quantified by HPLC using a C18 column and UV detection at 251 nm. The mobile phase was methanol-water 72:28 (v/v) and the flow-rate was 0.8 mL/min. The CXB retention time was 5 min. The assay was linear for CBX in the concentration range of 0.1-3.0 μg/mL in the AS (drug permeated through skin) and 5.0-50.0 μg/mL for drug retained in SC and [EP + D] in vitro. The linear correlation coefficients for the different calibration curves were equal or greater than 0.99. Intra- and inter-assay variabilities were below 8.0%. Extraction of CXB from skin samples showed recoveries higher than 95.0% after 15 min of ultrasonic sound and centrifugation at 2500 rpm for 3 min. The method was considered appropriate for the assay of CXB in skin samples, after in vitro cutaneous penetration studies.
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, Ribeirão Preto, SP, Brazil.
authors.forename: Fabíola Silva Garcia
authors.initials: FS
authors.lastname: Praça
authors.forename: Maria Vitória Lopes Badra
authors.initials: MV
authors.lastname: Bentley
authors.forename: Marilisa Guimarães
authors.initials: MG
authors.lastname: Lara
authors.forename: Maria Bernadete Riemma
authors.initials: MB
authors.lastname: Pierre
mesh_terms: D000818:Animals; D000068579:Celecoxib; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D020405:Dermis / Q000737:chemistry* / Q000378:metabolism; D004817:Epidermis / Q000737:chemistry* / Q000378:metabolism; D016014:Linear Models; D011720:Pyrazoles / Q000032:analysis* / Q000493:pharmacokinetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012869:Skin Absorption; D013056:Spectrophotometry, Ultraviolet; D013449:Sulfonamides / Q000032:analysis* / Q000493:pharmacokinetics*; D013552:Swine
chemical_list: D011720:Pyrazoles; D013449:Sulfonamides; D000068579:Celecoxib