Simultaneous Determination of Six Isoflavonoids in Radix Puerariae Lobatae from Different Habitats by HPLC

Abstract:

Objective: A reversed-phase high performance liquid chromatographic method (RP-HPLC) was developed for the simultaneous analysis of six isoflavonoids of Gegen, the roots of Puerariae lobatae. Method: HPLC determination was performed on a Kromasil C₁₈ column (4.6 mm×250 mm, 5 μm) and detected at 250 nm. The mobile phase was consisted of methanol and 0.1% citric acid solution with gradient elution.The flow rate was 1.0 mL·min^-1, and column temperature was kept at 30 ℃. Result: The method was proved to be linear in the ranges of 76.2-1 016, 65.8-1 316, 85.6-1 712, 49.98-499.8, 21.94-219.4, 48.5-485 ng for the six isoflavonoid,3'-hydroxypuerarin, puerarin, daidzin, genistin, ononin, daidzein, respectively. The average recoveries were 99.73% (RSD 0.6%), 100.48%(RSD 1.11%), 100.37%(RSD 0.79%), 100.28%(RSD 1.07%), 99.96%(RSD 1.22%),97.36%(RSD 1.59%),respectively. Conclusion: Results obtained showed that the method is convenient, accurate and reliable, can be applied to the simultaneous determination of the six isoflavonoids in Radix Puerariae lobatae.