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

ABSTRACT

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.