Ru(DMSO)₄Cl₂ is a good precursor complex in synthetic inorganic chemistry and also considered to be an efficient anti-cancer drug. Here in, we are reporting ∼80 nm nano-aggregates of Ru(DMSO)₄Cl₂ complex stabilized Nafion membrane modified glassy carbon electrode (GCE/Nf–{RuDMSO–Cl–H₂O}–MME) for simultaneous electro-catalytic oxidations and separation-less detection of purine bases; uric acid (UA), xanthine (X) and hypoxanthine (Hx) in a physiological solution. The MME was characterized using solid state UV–Vis, X-ray photoelectron spectroscopy (XPS) and transmission electron-microscopy (TEM) techniques. It has been found that Ru(DMSO)₄Cl₂ undergoes dissolution reactions within the water rich hydrophilic micro-channels of Nafion membrane and get electro-statically stabilized as [Ru^IICl_x(DMSO)_y(H₂O)_4−(x+y)]ⁿ⁺ through the cationic sulphonic acid structures. The MME film behaves like metallic electronic conductor, investigated with [Math Processing Error] redox couple. This new electrode shows efficient electrochemical sensing of the UA, X and Hx with a calibration windows (detection limit) up to 700 (0.372 μM), 500 (2.351 μM) and 300 μM (2.37 μM) respectively. The electrode shows exceptional stability and applicability to various real samples.