In-vitro anti-urolithiatic activity and simultaneous HPTLC quantification of berberine and palmatine in standardized extract of Thalictrum foliolosum DC.

Abstract

Thalictrum foliolosum DC. (Ranunculaceae) is a perennial plant distributed worldwide. In India it is used as a diuretic, anti-periodic, febrifuge, jaundice and in rheumatism. The present study was aimed to evaluate in-vitro anti-urolithiatic, anti-oxidant, anti-inflammatory activity along with concurrent quantification of berberine and palmatine through high performance thin layer chromatography (HPTLC). Anti-urolithiatic potential was evaluated via a number of different steps viz. nucleation, aggregation, whole urine assays and through kinetic study. The IC 50 value of nucleation and aggregation was found to be 114.314 and 67.625 µg/mL. Anti-oxidant activity was evaluated by DPPH and total antioxidant capacity with IC 50 value 54.96 µg/mL and 11.35 mg/ g anti-oxidant capacity. Anti-inflammatory potential was evaluated through inhibition of protein denaturation and antiproteinase activity, the IC 50 values were 94.61 and 60.51 µg/mL. Separation of berberine and palmatine was done on an aluminum TLC plate precoated with silica gel 60 GF 254 (10 × 10 cm), with mobile phase containing n-propanol-water-formic acid (90–10–1 v/v). Berberine and palmatine were identified at R f 0.44 ± 0.05 and 0.29 ± 0.06. Densitometric analysis was carried out at 350 nm for both the biomarkers in absorbance-reflectance mode. Berberine and palmatine concentrations were found to be 1.155 ± 0.035 and 0.111 ± 0.008 in plant on% dry weight basis. The result indicates that T. foliolosum has significant antioxidant and anti-inflammatory activity, and it is a potential candidate for phytotherapy against urolithiasis because of its ability to inhibit CaOx crystallization. Quantification of biomarkers will aid in quality standardization of raw material. • In-vitro anti-urolithiatic potential of Thalictrum foliolosum DC. root was evaluated. • In-vitro antioxidant and anti-inflammatory activity was also done to support the anti-urolithiatic potential of plant. • Two biomarkers berberine and palmatine were quantified through HPTLC.