Giloy (Tinospora cordifolia) is a medicinal plant rich in bioactive compounds known for their diverse health benefits. This study examined the nutritional value and biological activity of giloy stems and leaves. Hence, giloy stem (GSE), and leaf extract (GLE) was extracted using green extraction technology, ultrasound, and screening secondary metabolites and bioactive compounds. Using a central composite rotatable design combined with RSM, GSE, and GLE's antioxidant activity, total phenolic content and yield were optimized for solvent-to-solid ratio, sonication time, and solvent concertation. The optimum extraction conditions were found to be 22.5:1, 40 min, and 75%. Additionally, the extract inhibited the growth of S. aureus and E. coli. Screening of optimized extract through LC/MS reported the presence of significant polyphenols. Bioactive substances such as catechin, malic acid, quercetin, kaempferol, ellagic acid, hesperidin, and berberine were found. These findings indicate that ultrasonication, a green extraction method, promotes GSE and GLE bioactive chemical extraction. Giloy can make tasty, long-lasting food and drinks as a functional ingredient.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10068-024-01810-x.