DoE Engineered Development and Validation of an RP-HPLC Method for Simultaneous Estimation of Temozolomide and Resveratrol in Nanostructured Lipid Carrier

Abstract

Abstract Background 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. Objective 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. Method 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. Results 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. Conclusions 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. Highlights 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.