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

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RESEARCH ARTICLE

Evaluation of Ephedrine Content and Identification of Elite Chemotype(s) of Ephedra gerardiana (Wall.) from Kashmir Himalayas Mridul Kant Chaudhary1 • Ankita Misra1 • Sharad Srivastava1

Received: 12 July 2019 / Revised: 3 October 2019 / Accepted: 20 November 2019  The National Academy of Sciences, India 2019

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 lg/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

Significance’ Statement The present study is to evaluate microscopic characters, pharmacognostical standards and quantification ephedrine in Ephedra gerardiana through HPTLC and to identify the elite chemotype in Ephedra gerardiana collected from Kashmir Himalayas. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s40011-019-01153-2) contains supplementary material, which is available to authorized users. & Sharad Srivastava [email protected] 1

Pharmacognosy Division, CSIR-National Botanical Research Institute, Lucknow, U.P. 226001, India

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. Keywords Ephedra gerardiana  HPTLC  Chemotype  Ephedrine

Introduction Ephedra gerardiana Wall. (Ephedraceae) is an evergreen shrub, profusely branched and mainly found in the xerophytic and salty habitat. The genus Ephedra has worldwide occurrence up to altitudes of 2400–5000 m and is found in South America, Europe and temperate parts of Asia. In India, Ephedra gerardiana is found in the Alpine Himalayas from Kashmir to Sikkim, Chamba, Spiti and Ladakh [1, 2]. The species has been used