In the present investigation, Rheum emodi roots extract mediated magnesium hydroxide nanoparticles [Mg(OH)2 NPs] through the bio-inspired experimental technique were synthesised. Mg(OH)2 NPs were characterised by using various characterisation techniques such as field emission scanning electron microscopy, transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), and ultraviolet-visible spectroscopy. The formation of Mg(OH)2 NPs was confirmed by X-ray diffraction. The structural analysis confirmed the hexagonal crystal symmetry of Mg(OH)2 NPs with space group P-3m1 and space group no. 164 using the Rietveld refinement technique. TEM micrographs illustrated the nano-size formation of Mg(OH)2 NPs of spherical shape and size ∼14.86 nm. With the aid of FTIR data, plant metabolites such as anthraquinones have been identified as a stabilising and reducing agent for the synthesis of biogenic Mg(OH)2 NPs. The synthesised Mg(OH)2 NPs showed antimicrobial and cytotoxic potential against Gram-negative and Gram-positive bacteria such as Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) and MDA-MB-231 human breast cancer cell lines.