Storage stability kinetics and preclinical pharmacological evaluation of freeze-dried roselle anthocyanin microcapsules for commercial nutraceutical applications (HIBI®)

This study systematically evaluates standardized freeze-dried roselle anthocyanin microcapsules as a commercial nutraceutical supplement (HIBI®). The study aims to characterize some physical properties, antioxidant contents and activities, assess storage stability of anthocyanin through kinetic modeling, and determine pharmacological suitability for commercialization. The freeze-dried microcapsules exhibited a vivid red color and high solubility (83.6%), supported by a microporous structure that facilitated rapid rehydration, as shown by SEM/TEM analyses. Anthocyanin content reached 0.719 mg/g, alongside an exceptional encapsulation efficiency of 99.03%, resulting in strong antioxidant performance. Storage evaluation for anthocyanin retention using the Weibull kinetic model (R 2 = 0.9885; RMSE = 0.0098) showed a half-life of 176.1 days under ambient conditions, demonstrating commercially acceptable shelf-life for functional food applications. Subsequently, pharmacological evaluation of the commercial nutraceutical supplement HIBI®, containing anthocyanin microcapsules at 5 mg/g, demonstrated favourable preclinical safety and promising biological activity. Acute and 28-day sub-acute toxicity studies in Swiss albino mice revealed no mortality or adverse physiological effects at doses up to 2000 mg/kg, supporting its safety profile. In a cholesterol- and fat-enriched diet -induced metabolic disorder model, HIBI® supplementation (200–300 mg/kg) prevented excess weight gain and hyperlipidaemia, restored hepatic mRNA expression of antioxidant-related genes ( SOD, CAT and GPx-1 ) and suppressed pro-inflammatory cytokine expression ( TNF-α, IL-6 and IL-1β ). The microencapsulated anthocyanin formulation also exhibited strong in vitro radical-scavenging activity as determined by DPPH and ABTS assays. The supplement also normalized elevated serum AST and ALT, reduced fasting glucose, and restored serum creatinine, suggesting comprehensive protection against diet-induced oxidative stress, metaflammation and hepatic injury. Collectively, the findings support HIBI® as a stable formulation with promising potential for further nutraceutical development, pending additional clinical validation.