Isolation, Simultaneous Quantification of Taxifolin and Taxifolin-3-O-rhamnoside and Validation by RPHPLC

Background: Taxifolin (TA) is a flavonoid that has antioxidant, hepatoprotective, antialzheimer, anti-hyperglycemic, cardiovascular, anti-inflammatory, anti-psoriatic and antialzheimer properties. Taxifolin 3-O-rhamnoside (TAR), a glycoside of taxifolin, has antioxidant, anticonvulsant, anticancer, anti-inflammatory, and immunosuppressive effects. Objectives: The goal of this study is to separate TA and TAR from Smilax china Linn. rhizomes, as well as to develop and validate a technique for simultaneous measurement of TA and TAR. Materials and Methods: The hydro alcoholic extract of the rhizome of S. china yielded TA and TAR. HPLC system was fitted with a C₁₈ column (shim-pack) of size 150 mm x 4.6 mm; 5μ, with suitable eluting mixture of methanol: water (90:10 v/v) at a flowing rate of 1 ml/min and at 254 nm wavelength for identification of peaks. Lab solution software was used to establish the quantification method for above-mentioned chemical compounds. Results: TAR and TA were separated using the proposed technique at Rt 2.917 and 3.924 min respectively. Over the range of 0.1-0.8 μg/ml, calibration curves were produced with a linear relationship R2 > 0.9941 and 0.9963, respectively. The relative standard deviation was less than 2%. The percentage recoveries were shown to be between 97 and 102.1. TA and TAR had detection limits of 0.156 and 0.077 μg/ml; quantification limits were 0.473 and 0.234 μg/ml respectively. The technique that was devised was simple, sensitive and specific. Conclusion: The newly designed RPHPLC technique has improved specificity, precision, and accuracy. The quality of S. china and other dietary supplements containing these two flavonoids may be successfully assessed by quantifying these two flavonoids and their glycosides.