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

Abstract

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.

Simultaneous Estimation of Eugenol and Scopoletin by UV-Spectroscopic Method Using in-house Avipattikar churna

“…11 Furthermore, the other literature review revealed that there are various methods available for the detection and quantification of eugenol alone or along with other constituents using UV-spectroscopy, 12 High-performance liquid chromatography 13 and LC-MS. 14 Likewise, scopoletin was estimated by UV-spectroscopy, 15 HPTLC 16 and High-performance liquid chromatography. 17 Hence, from the literature review it is clear that there is no method reported in pharmacopoeias for simultaneous analysis of eugenol and scopoletin by UV spectroscopy in Avipattikar churna.…”

Simultaneous Estimation of Eugenol and Scopoletin from the in House <i>Avipattikar Churna</i> by RP-HPLC Method and Estimation of Scopoletin from Different Extracts of <i>Jalap</i>

“…Analysis was conducted using a RP-ODS C8 (250*4.5 mm). From the literature, UV detection and injection volume (20 µl) were optimized by conducting various trials [13][14][15] .…”

Estimation of scopoletin from a polyherbal formulation composition using HPLC coupled with fluorescence detector through the concept of Design of Experiment

“…An analytical method for the estimation of scopoletin from Argyreia nervosa using HPLC coupled with UV detector was also reported. [ 11 ] Scopoletin was estimated from commercial herbal drug product containing asiaticoside along with scopoletin using HPLC coupled with UV/FD (fluorescence detector). [ 12 ] Scopoletin was estimated from rat plasma using HPLC.…”

“…This can be attributed to the development of analytical methods to estimate scopoletin from different plant extracts using HPLC coupled with a fluorometric detector, but comprising gradient mobile phase. [11,16,31,32] The fluorescence signal is obtained as a combination of excitation and emission wavelengths, which helps in the selective detection of the compound of interest. Moreover, the fluorescence detector possesses a better signal-to-noise ratio (S/N) as compared to UV detector which makes them more sensitive.…”