Quantification of Scopoletin from Roots of Argyreia Speciosa (Linn. F) Sweet Using HPLC through the Concept of Design of Experiment.

BACKGROUND Roots of Argyreia speciosa (Linn. F) Sweet (family: Convolvulaceae) are used in Ayurveda to treat male reproductive and nervous system disorders. OBJECTIVE Isolation of scopoletin from the roots of Argyreia speciosa, development and validation of an analytical method using HPLC for the quantification of scopoletin from the root powder of Argyreia speciosa. METHOD Scopoletin was isolated from chloroform fraction prepared from hydrolyzed methanolic extract and identified using spectral studies. A reverse-phase HPLC based analytical method was developed and optimized using the DoE approach to estimate scopoletin from the roots of Argyreia speciosa. Scopoletin was separated and quantified using HPLC containing C18 column and a PDA detector. The optimized mobile phase was methanol: water (pH∼3.2) [25: 75, %v/v]. RESULTS The Box-Behnken design was used to optimize chromatographic parameters and the extraction procedure. The validation studies showed a linear relationship (r2=0.998) in the range of 1-40 µg/ml. The limit of Detection and Limit of Quantification were found to be 0.28 µg/ml and 0.84 µg/ml, respectively and the recovery values were found between 91.94 to 97.86%. The developed analytical method was found robust too. The amount of scopoletin was estimated to be 0.024 ± 0.0016%w/w from dried root powder. CONCLUSION The recorded chromatogram and amount of scopoletin determined would serve as one of the standardization parameters to access the quality of raw material containing Argyreia speciosa. HIGHLIGHTS Developed analytical method may be adopted as a part of the standardization procedure for Argyreia speciosa in the quality control laboratory.