Abstract
Background:
Anthocyanins from plant sources have attracted considerable attention as natural bioactive compounds with antioxidant and hypocholesterolemic properties. Hibiscus sabdariffa L flowers are rich in anthocyanins, which may serve as potential natural additives in poultry feed to reduce cholesterol levels.

Aim:
This study aimed to determine the optimal solvent type and maceration duration for extracting anthocyanins from Hibiscus sabdariffa L. flowers and characterize their anthocyanin profile as a potential natural hypocholesterolemic agent for poultry feed additives.

Methods:
A factorial experiment with a completely randomized design was conducted. The first factor was the solvent type, and the second factor was the maceration duration. The evaluated parameters included color measurement, anthocyanin extract yield, total anthocyanin content (TAC), thin-layer chromatography (TLC), LC–MS/MS anthocyanin profiling, total phenolic content, and antioxidant activity using the DPPH radical scavenging assay.

Results:
A significant interaction (P < 0.05) between solvent type and maceration duration was observed for color measurement, anthocyanin extract yield, and TAC. TLC analysis revealed five distinct anthocyanin spots. LC–MS/MS analysis identified three major anthocyanin compounds: delphinidin-3-sambubioside, cyanidin-3-sambubioside, and delphinidin. The extract also exhibited considerable total phenolic content, DPPH radical scavenging activity, and specific antioxidant activity. Variations in ΔE color change, anthocyanin extract yield, and TAC were observed among different solvent types and maceration durations.

Conclusion:
Anthocyanins from H. sabdariffa L contain delphinidin-3-sambubioside, cyanidin-3-sambubioside, and phenolics that may inhibit HMG-CoA reductase in the synthesis of cholesterol. DPPH antioxidant activity indicates free radical scavenging ability.

Key words: Anthocyanin; Cholesterol; Feed additive; Hibiscus sabdariffa L; Poultry.