Evaluation of Ephedrine Content and Identification of Elite Chemotype(s) of Ephedra gerardiana (Wall.) from Kashmir Himalayas

Abstract

Ephedra gerardiana Wall. (Ephedraceae) is an evergreen xerophytic shrub and is used in Indian and Chinese folk medicine since ancient times. It is used in asthma, hay fever, rheumatism, syphilis and as CNS stimulant due to its bioactive metabolite ephedrine. In this study, a validated HPTLC method was developed for the quantification and identification of the elite chemotype of E. gerardiana from Kashmir Himalayas. Ten samples were collected from different regions and their botanical and pharmacognostical standards were established. HPTLC method was developed and validated on pre-coated silica gel 60 GF254 plates in a tertiary solvent system of methylene chloride–ammonia–propan-2-ol (0.5:1.5:8 v/v), and quantification was done after derivatization with ninhydrin reagent. The method developed for quantification of ephedrine was linearly calibrated and found to be specific, repeatable and robust. LOD and LOQ were 36.33 and 110.10 µg/spot. Precision studies (inter-day and intra-day) are within the standard limit of ICH guidelines, and RSD (%) is not more than 5%. Mean recovery of ephedrine at three spiking levels of 25, 50 and 75% was 103.17%. The quantification of ephedrine within the population varies from 0.31 ± 0.05 to 0.81 ± 0.02% dry wt. basis, and in cluster analysis, two elite chemotypes viz. EPH-1 and EPH-5 were identified. These findings will help in setting the standard parameter for this plant in herbal Pharmacopoeia. The study will aid in the identification of potential sources of high ephedrine containing germplasm which will be useful to industry for quality raw material in herbal drug development.