Department of Pharmaceutics, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi 110062, India.
J AOAC Int. 2022 Sep 6;105(5):1258-1267. doi: 10.1093/jaoacint/qsac045.
Temozolomide is drug of choice for the treatment of glioblastoma, but dose-related side effects limit its use. Resveratrol suppresses tumor growth and promotes apoptosis. Many studies showed synergistic activity of resveratrol and temozolomide against glioblastoma. There are methods reported for the assessment of temozolomide and resveratrol individually, but no analytical method has been reported for assessment of temozolomide and resveratrol simultaneously.
Therefore, the present study aimed to develop and optimize an HPLC analytical method for the simultaneous assessment of temozolomide and resveratrol in a developed nanostructured lipid carrier.
A Central composite rotable design was used to optimize the method. The method was developed using a C18 column. The composition of the mobile phase was 30% methanol and 70% glacial acetic acid (0.1% v/v in HPLC grade water); detecting wavelength was 310 nm. Forced degradation test was also performed to demonstrate the proposed HPLC method's ability to indicate stability.
The LOD for temozolomide and resveratrol was found to be 1.10 and 0.83 µg/mL, respectively, while LOQ was 3.33 and 2.52 µg/mL, respectively. The drug loading and entrapment efficacy of the formulation, as determined using the aforementioned method, was found to be 6.73 and 96.28% for temozolomide and 3.45 and 89.39% for resveratrol, respectively.
The developed HPLC method was simple, rapid, economical, precise, accurate, and reproducible, and it had high selectivity with good detection limits. Standard guidelines of ICH Q2 (R1) including linearity, specificity, system suitability, robustness, precision, accuracy, the LOQ, and LOD gave satisfactory results. Forced degradation studies showed a good stability-indicating capacity of the developed HPLC method.
Analytical Quality by Design is a powerful tool that could be used for the development of the analytical method. Central composite rotable design was used for optimizing the method. The percent of methanol and concentration of glacial acetic acid were selected as two independent variables for optimization.
替莫唑胺是治疗胶质母细胞瘤的首选药物，但剂量相关的副作用限制了其使用。白藜芦醇能抑制肿瘤生长并促进细胞凋亡。许多研究表明，白藜芦醇和替莫唑胺联合使用对胶质母细胞瘤具有协同作用。虽然有报道称可以分别评估替莫唑胺和白藜芦醇，但尚未有报道称可以同时评估替莫唑胺和白藜芦醇。
因此，本研究旨在开发和优化一种同时评估替莫唑胺和白藜芦醇在纳米结构脂质载体中的 HPLC 分析方法。
采用中心复合旋转设计优化方法。该方法采用 C18 柱，流动相组成为 30%甲醇和 70%冰醋酸（HPLC 级水，0.1%v/v）；检测波长为 310nm。还进行了强制降解试验，以证明所提出的 HPLC 方法能够指示稳定性。
替莫唑胺和白藜芦醇的检测限分别为 1.10 和 0.83μg/mL，定量限分别为 3.33 和 2.52μg/mL。采用上述方法测定，该制剂的载药量和包封率分别为替莫唑胺 6.73%和 96.28%，白藜芦醇 3.45%和 89.39%。
所开发的 HPLC 方法简单、快速、经济、精确、准确、重现性好，具有高选择性和良好的检测限。ICH Q2（R1）的标准指南，包括线性、特异性、系统适用性、稳健性、精密度、准确度、定量限和检测限，均获得了满意的结果。强制降解研究表明，所开发的 HPLC 方法具有良好的稳定性指示能力。
分析质量源于设计是一种强大的工具，可用于开发分析方法。采用中心复合旋转设计优化方法。甲醇的百分比和冰醋酸的浓度被选为两个独立变量进行优化。
