Validation of a UV-spectrophotometric analytical method for determination of LPSF/AC04 from inclusion complex and liposomes

The aim of this study was to develop and validate a UV spectrophotometric method for determination of LPSF/AC04 from inclusion complex and encapsulated into liposomes. The validation parameters were determined according to the International Conference on Harmonisation (ICH) and National Health Surveillance Agency (ANVISA) guidelines. LPSF/AC04 was determined at 250 nm in methanol by a UV spectrophotometric method, exhibiting linearity in the range from 0.3 to 2 µg.mL−1 (Absorbance=0.18068 x [LPSF/AC04 µg.mL-1] + 0.00348), (r2=0.9995). The limits of detection and quantification were 0.047µg.mL−1 and 0.143µg.mL−1, respectively. The method was accurate, precise, reproducible and robust since all the samples analyzed had coefficient of variation of less than 5% and no statistically significant difference between theoretical and practical concentrations was detected. Thus, a rapid, simple, low cost and sensitive spectrophotometric method was developed and validated for determining the content of inclusion complex and liposomes containing LPSF/AC04.

本研究旨在开发并验证一种紫外分光光度法（UV spectrophotometric method），用于测定包合复合物及脂质体包封的LPSF/AC04的含量。验证参数依据人用药品注册技术要求国际协调会（International Conference on Harmonisation，ICH）与巴西国家卫生监督局（National Health Surveillance Agency，ANVISA）的指导原则进行测定。采用该方法，在250 nm波长下的甲醇溶剂中对LPSF/AC04进行定量测定，其在0.3~2 µg·mL⁻¹浓度范围内呈良好线性关系，标准曲线为吸光度=0.18068×[LPSF/AC04 µg·mL⁻¹]+0.00348，决定系数r²=0.9995。检出限与定量限分别为0.047 µg·mL⁻¹和0.143 µg·mL⁻¹。该方法具备良好的准确度、精密度、重现性与耐用性，所有待测样品的变异系数均小于5%，且理论浓度与实测浓度间无统计学显著差异。综上，本研究开发并验证了一种快速、简便、低成本且灵敏度优异的分光光度法，可用于测定含LPSF/AC04的包合复合物及脂质体的含量。