Lima Paulo Marcelo Andrade, Prado Nathália Duque, Silva Maria-Luiza Martins, Diniz Rosiane Dias Lopes, Queiroz Kelly Marques, César Isabela da Costa, Pianetti Gérson Antônio, Santos Daniel Assis
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.
J AOAC Int. 2009 Jul-Aug;92(4):1076-81.
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.
为分析胶囊制剂中的酮康唑，开发并验证了一种高效液相色谱 - 紫外检测法（HPLC - UV）和一种微生物测定法。微生物测定法采用特定的琼脂扩散技术，以白色念珠菌ATCC 18804菌株作为测试生物来开发。在色谱法中分析了流动相pH值在2.5 - 7.5范围内对酮康唑峰的保留因子和拖尾因子的影响，认为pH值4.5是合适的。两种方法的前瞻性验证均显示出足够的线性（两种方法的r2均>0.99）、精密度（微生物测定法日内精密度RSD = 2.42%，日间精密度RSD = 2.69%；HPLC - UV法日内精密度RSD = 0.74%，日间精密度RSD = 0.66%）和准确度（微生物测定法平均回收率为103±1.0%，HPLC - UV法平均回收率为99±1.0%）。学生t检验显示两种方法所得结果之间无显著差异（P < 0.05）。三个胶囊样品的含量结果显示出很强的相关性，皮尔逊系数值（r = 0.9998）证明了这一点，这也证明了所研究方法之间的一致性。
