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Assessment of Antioxidant Potential and Standardization of Fermented Biomedicine Balarista by High-Performance Thin-Layer Chromatography and Fourier-Transform Infrared Spectroscopy Fingerprint Analysis

Author(s): C. Das, S. Mukhi, A. Nanda and D. Das*
Department of Pharmacognosy, 1Department of Pharmaceutical Analysis and Quality Assurance, School of Pharmaceutical Sciences, Siksha ‘O’ Anusandhan Deemed to be University, Bhubaneswar, Odisha 751003, 2Department of Pharmacology, Centurion University of Technology and Management, Bhubaneswar, Odisha 761211, India

Correspondence Address:
D. Das, Department of Pharmacognosy, School of Pharmaceutical Sciences, Siksha ‘O’ Anusandhan Deemed to be University, Bhubaneswar, Odisha 751003, India, E-mail:

Balarista, a traditional fermented biomedicine, is prescribed for treating rheumatoid arthritis. The present work was undertaken to standardize the Balarista formulation using Fourier-transform infrared fingerprinting and high-performance thin-layer chromatography analysis and evaluate its antioxidant properties. The quality control of the in-house Balarista and marketed formulations was assessed by microbial count, aflatoxin content, high-performance thin-layer chromatography, and Fourier-transform infrared fingerprint analysis. The in vitro antioxidant properties of the formulation were studied by 2, 2-diphenyl-1-picrylhydrazyl, 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid), hydrogen peroxide, nitric oxide scavenging and reducing power assay. Total bacterial count in in-house Balarista and M1 was found within the standard limit. Aflatoxin content was found less than 5.0 ppb in all formulations. Fourier-transform infrared fingerprint spectra of different fraction of M1 formulation revealed an excellent common peak ratio with in-house Balarista, suggesting the use of prescribed raw materials in the formulations. However, the poor common variation peak ratio with other marketed formulations indicates the use of improper raw materials. The high-performance thin-layer chromatography profile of chloroform fraction of in-house and marketed formulations revealed five spots each while ethyl acetate fraction of in-house Balarista and M3 showed six spots each. The highest 2, 2-diphenyl-1-picrylhydrazyl and nitric oxide radical scavenging activity was achieved by in-house Balarista, 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) by M4, hydrogen peroxide by M1 and reducing power by M2 formulation. The antioxidant activity of the formulation could be due to the presence of considerable amount of phenolic and flavonoid content. The formulation's comprehensive chemical analysis by Fouriertransform infrared and high-performance thin-layer chromatography profile can be used as standards for future reference.

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