HPTLC Standardization, Phytochemical Profiling and Pharmacological Evaluation of Barleria prionitis Leaf Extract in High-Fat-Diet-Induced Obese Rats

Obesity has emerged as a global health crisis, necessitating the exploration of safe and effective plant-based therapeutic alternatives. Barleria prionitis Linn. (Acanthaceae), a medicinal plant widely used in Ayurvedic medicine, has been traditionally employed for various ailments, though its anti-obesity potential remains underexplored. This study aimed to standardize B. prionitis leaf extract using High-Performance Thin-Layer Chromatography (HPTLC), conduct comprehensive phytochemical profiling, and evaluate its pharmacological efficacy in high-fat-diet (HFD)-induced obese rats. The hydroethanolic leaf extract was subjected to preliminary phytochemical screening, total phenolic and flavonoid content estimation, and HPTLC fingerprinting using a validated method with quercetin, gallic acid, and tannic acid as marker compounds. For pharmacological evaluation, male Wistar rats were divided into six groups: normal control, HFD control, HFD + standard drug (orlistat 30 mg/kg), and HFD + extract at three doses (100, 200, and 400 mg/kg) for 12 weeks. Body weight, food intake, adiposity indices, lipid profile, glycemic parameters, hepatic and renal function markers, oxidative stress parameters, and histopathological changes were assessed. The extract exhibited significant phenolic (86.42 ± 2.14 mg GAE/g) and flavonoid (42.18 ± 1.86 mg QE/g) content. HPTLC analysis confirmed the presence of quercetin (Rf 0.52), gallic acid (Rf 0.64), and tannic acid (Rf 0.36) with good resolution and validation parameters (linearity r² > 0.998, LOD 12.4-28.6 ng/band, recovery 97.8-99.6%). The extract at 400 mg/kg significantly (p < 0.001) reduced body weight gain (42.3%), adiposity index (48.6%), serum total cholesterol (38.2%), triglycerides (41.5%), LDL-C (44.8%), and glucose (36.4%), while increasing HDL-C (52.1%). It also restored hepatic and renal function markers and enhanced antioxidant enzyme activities (SOD, CAT, GSH) with concomitant reduction in MDA levels. Histopathological examination revealed attenuated hepatic steatosis and adipocyte hypertrophy. These findings demonstrate that B. prionitis leaf extract possesses significant anti-obesity, hypolipidemic, hypoglycemic, and antioxidant properties, likely mediated through its rich phytochemical constituents, particularly iridoid glycosides and flavonoids. The validated HPTLC method provides a reliable approach for quality standardization of B. prionitis-based formulations.