Tinosporaside

The collected plant materials were shade dried for a day and then dried completely in an oven at 40°C. The plant materials were coarsely powdered using a rotary grinder and stored at 25°C airtight containers and powdered to 60 mesh when required for quantification of phytochemicals, antioxidant, and HPTLC analysis. Powdered dried plant samples (10 g) were extracted with methanol (4 × 25 mL, three times each for 3 days). The extracts were combined (separately for each samples), filtered, evaporated to dryness using a rotary evaporator, and then lyophilized. Accurately weighed extract (10 mg) was dissolved in 1 mL of methanol and filtered through a 0.45 μm filter membrane; the filtrate was used as sample solution. Standard solution of tinosporaside (1.0 mg/mL) and berberine (0.1 mg/mL) as reference biomarkers was also prepared in methanol. All chemicals used were of analytical grade. Berberine standard was procured from Sigma-Aldrich and tinosporaside from Dr. Rakesh Maurya, Scientist, CSIR-Central Drug Research Institute, Lucknow. All chemicals used were of analytical grade.

Sodium acetate, guanidine hydrochloride, and glacial acetic acid were purchased from SRL, India. Lipopolysaccharide (LPS), Carboxy Methyl Cellulose (CMC), Tinosporaside, Ferrozine (3-(2-Pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p′-di-sulfonic acid monosodium salt hydrate)^® and, Complete Freund’s Adjuvant (CFA) were purchased from Sigma-Aldrich, India.