Gas Chromatographic Method for Analysis β-Asarone in Rhizome extracts of Acorus calamus and Their Microbiological Evaluation

Abstract

Introduction: The aim of study was to develop a simple, sensitive and precise gas chromatographic method for the analysis of β-asaronein ethanolic, ethyl acetate, aqueous extracts from the rhizomes of Acorus calamus and validate according to current ICH guidelines. Followed by evaluation of antimicrobial activity of prepared extracts in comparison with penicillin disc. Methods: The β-asarone was one chief active constituent from the rhizomes of Acorus calamus. The ethanolic, ethyl acetate, aqueous extract of rhizomes was prepared and further evaluated for its antimicrobial activity against Streptococcus Mutants by agar well diffusion method. The GC method was used for the analytical determination of β-asarone. The sample was estimated using gas chromatography with flame ionization as a detector. Nitrogen at a flow rate of 1.18 mL/min was used as a carrier gas and total run time was 10 minutes. The injection port and detector temperature were set to 225˚C and 270˚C, respectively. The retention time of β-asarone was found to be 6.9 minutes. Results: The linearity of the developed method was tested in the range of 100 ng/mL-500 ng/mL for β-asarone, limit of detection and limit of quantification was found to be 22.78 and 69.05 ng/mL respectively and the percentage recovery was from 99.63- 100.64%. Conclusion: A simple, precise and accurate GC-FID method has been developed for the determination of β-asarone in ethanolic, ethyl acetate and aqueous extracts of rhizomes. Key words: Acoruscalamus , β-Asarone, Alcoholic, Aqueous, Ethyl acetate, Extracts, GC- FID, Microbiological evaluaiton, Stability studies.