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.

本研究旨在建立并验证一种紫外分光光度法，用于测定包合物及脂质体包封的LPSF/AC04含量。本方法的验证参数参照国际人用药品注册技术协调会（ICH）与巴西国家卫生监督局（ANVISA）的指导原则制定。采用该紫外分光光度法，在甲醇溶剂中于250 nm波长处对LPSF/AC04进行测定，其在0.3~2 µg·mL⁻¹范围内呈良好线性关系，回归方程为吸光度=0.18068×[LPSF/AC04 µg·mL⁻¹]+0.00348，决定系数r²=0.9995。检测限与定量限分别为0.047 µg·mL⁻¹和0.143 µg·mL⁻¹。该方法具备准确、精密、重现性好且耐用的特性，所有待测样品的变异系数均小于5%，理论浓度与实测浓度间未检测到统计学显著性差异。综上，本研究建立并验证了一种快速、简便、低成本且灵敏的分光光度法，可用于测定含LPSF/AC04的包合物与脂质体的含量。