Nunes Débora, Tavares Tânia G, Malcata Frenacisco Xavier, Loureiro Joana A, Pereira Maria Carmo
LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
Pharmaceuticals (Basel). 2024 Sep 2;17(9):1162. doi: 10.3390/ph17091162.
Memantine, a non-competitive NMDA receptor antagonist, is used to treat Alzheimer's disease. Therefore, loading memantine in nanoparticles (NPs) could be an essential tool to improve the treatment effectiveness while reducing drug toxicity. Even though some approaches have been described to quantify memantine, none reported optimized methods using high-performance liquid chromatography resorting to ultraviolet detection (UV-HPLC) to determine encapsulation in NPs. The present research developed a HPLC method using pre-column derivatization for quantitatively analyzing memantine hydrochloride in NPs. Memantine was derivatized using 9-fluorenylmethyl chloroformate (FMOC). The developed method was fully validated regarding suitability, specificity, linearity, sensitivity, precision, accuracy, and robustness according to the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The retention time of memantine was 11.393 ± 0.003 min, with a mean recovery of 92.9 ± 3.7%. The new chromatographic method was validated and found to respond linearly over 5-140 μg/mL, with a high coefficient of determination. Intraday precision lay between 3.6% and 4.6%, and interday precision between 4.2% and 9.3%. The stability of memantine was also tested at 4 °C and -20 °C, and no signs of decay were found for up to 6 months. The new method was properly validated and proved simple, sensitive, specific, accurate, and precise for determining memantine encapsulation efficiency in lipid NPs. Greenness was evaluated, presenting a final score of 0.45. In the future, this methodology could also be applied to quantify memantine in different nanoformulations.
美金刚是一种非竞争性N-甲基-D-天冬氨酸（NMDA）受体拮抗剂，用于治疗阿尔茨海默病。因此，将美金刚负载于纳米颗粒（NPs）中可能是提高治疗效果同时降低药物毒性的重要手段。尽管已有一些方法用于定量美金刚，但尚无使用高效液相色谱-紫外检测法（UV-HPLC）来确定其在NPs中的包封率的优化方法报道。本研究开发了一种采用柱前衍生化的HPLC方法，用于定量分析NPs中的盐酸美金刚。美金刚用9-芴基甲基氯甲酸酯（FMOC）进行衍生化。根据人用药品注册技术要求国际协调会议指南，对所开发的方法在适用性、特异性、线性、灵敏度、精密度、准确度和稳健性方面进行了全面验证。美金刚的保留时间为11.393±0.003分钟，平均回收率为92.9±3.7%。新的色谱方法经验证，在5-140μg/mL范围内呈线性响应，测定系数较高。日内精密度在3.6%至4.6%之间，日间精密度在4.2%至9.3%之间。还在4℃和-20℃下测试了美金刚的稳定性，在长达6个月的时间内未发现降解迹象。新方法经过了适当验证，证明对于测定脂质NPs中美金刚的包封效率简单、灵敏、特异、准确且精密。对该方法的绿色度进行了评估，最终得分为0.45。未来，该方法也可应用于定量不同纳米制剂中的美金刚。
