A direct LC/MS/MS method for the determination of ciclopirox penetration across human nail plate in in vitro penetration studies.
J Pharm Biomed Anal
; 51(1): 230-5, 2010 Jan 05.
Article
em En
| MEDLINE
| ID: mdl-19744810
ABSTRACT
Due to severe chelating effect caused by N-hydroxylpyridone group of ciclopirox, there is no published direct HPLC or LC/MS/MS method for the determination of ciclopirox in any in vitro or in vivo matrix. Instead, the time-consuming pre-column derivatization methods have been adapted for indirect analysis of ciclopirox. After overcoming the chelating problem by using K(2)EDTA coated tubes, a direct, sensitive and high-throughput LC/MS/MS method was successfully developed and validated to determine the amount of ciclopirox that penetrated across the nail plate during in vitro nail penetration studies. The method involved adding a chemical analog, chloridazon as internal standard (IS) in K(2)EDTA coated tubes, mixing IS with ciclopirox in a 96-well plate and then proceeding to LC/MS/MS analysis. The MS/MS was selected to monitor m/z 208.0-->135.8 and 221.8-->77.0 for ciclopirox and IS, respectively, using positive electrospray ionization. The method was validated over a concentration range of 8-256 ng/mL, yielding calibration curves with correlation coefficients greater than 0.9991 with a lower limit of quantitation (LLOQ) of 8 ng/mL. The assay precision and accuracy were evaluated using quality control (QC) samples at three concentration levels. Analyzed concentrations ranged from 101% to 113% of their respective nominal concentration levels with coefficients of variation (CV) below 10.6%. The average recovery of ciclopirox from nail matrix was 101%. The validated method was successfully used to analyze the ciclopirox formulation and in vitro nail penetration samples.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Piridonas
/
Cromatografia Líquida
/
Espectrometria de Massas em Tandem
/
Antifúngicos
Limite:
Humans
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article