Pharm Methods, 2016; 7(2): 121-126 A multifaceted peer reviewed journal in the field of Pharm Analysis and Pharmaceutics www.phmethods.net | www.journalonweb.com/phm Original Article Pharmaceutical Methods, Vol 7, Issue 2, Jul-Dec, 2016 121 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 fame ionization as a detector. Nitrogen at a fow 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 quantifcation 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. Correspondence: B. M. Gurupadayya, Department of Pharmaceutical Chemistry, JSS College of Pharmacy, JSS University, Sri Shivarathreeswara Nagar, Mysuru-570 015, Karnataka, INDIA. Telephone: +91-821-2548353 Fax Number: +91-821-2548359 Mobile Number: +91-7411006136 E-mail: bmgurupadayya@jssuni.edu.in DOI : 10.5530/phm.2016.7.18 INTRODUCTION Acorus calamus is an aromatic medicinal plant which is commonly known as ‘Sweet fag’. It has a very good medicinal values. 1 It is been used fromolden times of use in both traditional Indian and Chinese systems of medicine. 2 In earlier days it was used as a brain tonic for rejuvunation of the brain and nervous system. It showed wide variety of pharmaco- logical actions in ayurvedic system of medicine like anti-hemorrhoidal, used for reliving cold sensation on skin, used as an adjunct to decoction enemas, it is also used as natural substances that remove fat from the body and also used in cleansing agent in nasal therapy. 3 Acorus calamus was even used in traditional folk medicine of America and Indonesia for diseases like diabetes and diarrhea. 4 It is also used in the treatment of various ailments like appetite loss, bronchitis, cough, chest pain, cramps, diarrhea, digestive disorder, depression, fever, hemorrhoids, infammation, numbness, rheumatisim, skin diseases, tumors and as antidotes for several poisoning. 5-7 Various parts of Acorus calamus have its own pharmacological impor- tance in treatment of diferent diseases. Of all the parts most of the experimental studies have been carried on leaves, roots and stem of the plant. 8 Te rhizomes of the plant has a long history of usage in many countries, it been used almost from 2000 years in India and China. Chinese have used this rhizome for the treatment of swelling and con- stipation. Native Americans have also used this plant rhizome as an anesthetic for toothache and headache. Even in Ayurveda system of medicine rhizomes of Acorus calamus are considered to possess aphrodisiac, aromatic, antispasmodic, anthelmintic, bitter tonic, carminative, diuretic, emetic, laxative, expectorant, and stimulant properties. Acorus calamus are even used in the management of various ailments like asthma, abdominal tumors, bronchial catarrh, chronic diarrhea, cough and dysentery. 9 Acorus calamus rhizomes are even used in traditional system of medicine like in case of new born children it is given in form of vasambu(rhizome paste) along with honey for proper brain development, better visual power, increased seminal power and proper speech ability. Afer the birth of newborns it is applied on the tongue in the form of paste along with the ghee, gold and water to improve the grasping power. It also stimulates the nervous system, giving relief from depression. It is useful in improving speech in stammering. 10 Vacha taken along with milk improves intellectual, grasping and memory power. 11 A detailed literature review indicated that there are few analytical methods reported like HPTLC, HPLC, GC-MS and LC-MS 12-15 for quantifcation of various constituents of Acorus calamus. But till date there are no reported methods for quantifcation of β-asarone (Figure 1) in ethanolic, ethyl acetate, aqueous extracts from the Rhizomes of Acorus calamus. So in this concern,we have frst extracted β-asarone from its rhizomes and its antimicrobial evaluation by well difusion method. Followed by its quantifcation through GC-FID method. Te GC-FID method employed was very simple, highly sensitive, does not require any derivitization of the sample and also short runtime. Chemical Constituents Many number of chemical constituents have been reported from the leaves, rhizomes along with essential oil of Acorus calamus. 16 Te oil of Acorus calamus rhizomes were found to contain diferent concentration of alcohol, aldehyde, ester, furan, hydrocarbon, ketone, N-containing miscellaneous like α-asarone, β-asarone, α- pinene, β- pinene, calamene, Kavyashree Banda Vijayakumar, Gurupadayya Bannimath*, Venkata Sairam Koganti, Vishwanathan Balasubramanya Iyer Department of Pharmaceutical Chemistry, JSS College of Pharmacy, JSS University, Mysuru-570015, INDIA.