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A simple sensing methodology for rapid detection of multiple biomolecules in human health diagnostics remains a significant challenge. Herein, we developed a novel dispersant-free supra single-walled carbon nanotube (supra SWCNT)-coated glassy carbon electrode (GCE) for simultaneous sensing of biomolecules, such as ascorbic acid (AA), dopamine (DA), and uric acid (UA), in ex vivo mouse tissues. Supra SWCNTs with stable hydrogen bonds were synthesized via a simple chemical modification, after which they were characterized and confirmed using different physicochemical and electrochemical techniques. The supra SWCNT-coated GCE exhibited simultaneous sensing of AA, DA, and UA at linear dynamic ranges with lower detection limits of 12, 0.02, and 0.36 μM, respectively. Furthermore, they demonstrated high selectivity (with co-existing interference) and excellent reproducibility for effective sensing of the target biomolecules in different real samples. The rapid functioning, low cost, scalable preparation, and enhanced applicability for simultaneous sensing of target biomolecules in Parkinson's disease-induced mouse liver, kidney, and brain tissues suggest the suitability of the developed sensor for real-world applications. [Display omitted] • Dispersant-free supra SWCNT was prepared to facilitate step chemical modifications. • Supra SWCNTs freely dispersed in water without the addition of dispersing agents. • Supra SWCNTs/GCE were tested for highly sensitive simultaneous biomolecule sensing. • Real-time biomolecule sensing in PD mouse models achieved satisfying results. [ABSTRACT FROM AUTHOR]