Bioanalytical RP-HPLC Method Development and Validation for Simultaneous Estimation of Temozolomide and Resveratrol: A Pharmacokinetic Analysis in Rat Plasma and Brain.

Temozolomide, a chemotherapy drug used to treat glioblastoma, has high-dose and dose-related side effects, limiting its use. Resveratrol, a natural polyphenol, showed potential in the treatment of glioblastoma. Many studies showed the synergistic activity of resveratrol and temozolomide against glioblastoma, but no analytical method for simultaneous estimation in a biological matrix is available till date. This study aimed to develop and validate a simple, rapid, and sensitive bioanalytical method using HPLC technique for simultaneous estimation of temozolomide and resveratrol in rat plasma and brain for pharmacokinetic study. Isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method using C18 column was used. Theophylline and caffeine were used as internal standards for Temozolomide and Resveratrol, respectively. The mobile phase methanol: 0.1% glacial acetic acid (30:70) with flow rate 1.0 mL/min and 310-nm wavelength was used. The LOD for temozolomide and resveratrol in the plasma was 0.96 and 1.29 μg/mL, respectively, whereas LOQ was 2.91 and 3.89 μg/mL, respectively. The LOD for temozolomide and resveratrol in brain homogenate was 1.01 and 1.24 μg/mL, respectively, whereas LOQ was 3.07 and 3.76 μg/mL, respectively. This analytical method can be used for simultaneous estimation of temozolomide and resveratrol in biological samples, with higher sensitivity, resulting in more meaningful and appropriate pharmacokinetic analysis.