Determination of ketoconazole in capsules by high-performance liquid chromatography and microbiological assay.
The development and validation of an HPLC-UV method and a microbiological assay were performed for the analysis of ketoconazole in capsule formulations. The bioassay was developed using a specific agar diffusion technique with the strain of Candida albicans ATCC 18804 as the test organism. The effect of the mobile phase pH in the range of 2.5-7.5 on the retention and tailing factors of the ketoconazole peak was analyzed in the chromatography method and a pH value of 4.5 was considered to be adequate. A prospective validation of both methods showed adequate linearity (r2 > 0.99 for the two methods), precision, (RSD = 2.42% for intraday and 2.69% for interday precision for bioassay; RSD = 0.74% for intraday and 0.66% for interday precision for HPLC-UV), and accuracy (mean recoveries were 103 +/- 1.0% for bioassay and 99 +/- 1.0% for HPLC-UV). Student's t-test revealed no significant difference between the results obtained by the two methods (P < 0.05). The contents found for three capsule samples showed a strong correlation, as attested by Pearson's coefficient value (r = 0.9998), which also evidenced the concordance between the studied methodologies.
Lima
Paulo Marcelo Andrade
PM
Universidade Vale do Rio Doce, Faculdade de Ciências da Saúde, Laboratório de Microbiologia, Rua Israel Pinheiro, 2000, Bairro Universitário, 35020-220, Governador Valadares, Minas Gerais, Brazil.
Prado
Nathália Duque
ND
Silva
Maria-Luiza Martins
ML
Diniz
Rosiane Dias Lopes
RD
Queiroz
Kelly Marques
KM
César
Isabela da Costa
Ida C
Pianetti
Gérson Antônio
GA
Santos
Daniel Assis
DA
D000935:Antifungal Agents / Q000032:analysis* / Q000494:pharmacology*; D001681:Biological Assay; D002176:Candida albicans / Q000187:drug effects; D002214:Capsules; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007654:Ketoconazole / Q000032:analysis* / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000935:Antifungal Agents; D002214:Capsules; D007202:Indicators and Reagents; D007654:Ketoconazole