12 Document heading A HPTLC method for the identification of ferulic acid from Lycopodium clavatum Sharad Srivastava, Adarsh Pratap Singh, Ajay Kumar Singh Rawat * Pharmacognosy and Ethnopharmacology Division, National Botanical Research Institute, Lucknow, India Asian Pacific Journal of Tropical Biomedicine (2012)S12-S14 Asian Pacific Journal of Tropical Biomedicine journal homepage:www.elsevier.com/locate/apjtb * Corresponding author: Ajay Kumar S ingh Rawat, Pharmacognosy and Ethnopharmacology Division, National Botanical Research Institute, Lucknow, India. Tel: +91 9415764994; 9415082210 Fax: +91 522 2207219 E-mail: pharmacognosy1@rediffmail.com 1. Introduction Ferulic acid, a phenolic acid has wide distribution in the plant kingdom and is more bioavailable than other dietary flavonoid and monophenolics studied [1] . It has been proved to be a potent antioxidant, anti-inflammatory, and is reported to terminate free radical chain reaction and reduce the risk for coronary heart diseases [2-6] . Chemical investigations of the lycopods have centered on the alkaloids first discovered by Bodeker [7] and further by several workers [8-16] . Although these botanically interesting plants, of which there are possibly 483 species, are common elements of the flora of many parts of the world, including Europe and North America, they have not been examined extensively from either a biochemical or a chemical standpoint. It is therefore of interest and has prompted us to study, in more detail, the ferulic acid content of Lycopodium clavatum. In view of its great importance worldwide, a simple and high-precision method has been developed to estimate ferulic acid using high performance thin layer chromatography (HPTLC) in this plant, which is not yet isolated by any researcher and will be utilized for chemotaxonomic significance of lycopods. 2. Materials and methods 2.1. Plant Material The plant sample of Lycopodium clavatum ( Family: Lycopodiaceae) was collected from the Kotayagiri of Nilgiri Hills, Ootacamund ( Tamilnadu) , India in the month of January, 2006. Authenticated specimens were deposited in the Institute’s herbarium (LWG 221424, 2006). 2.2. Chemicals and reference marker compound Reagents used were from Merck (Germany) and standards viz. ferulic acid was from Sigma-Aldrich ( Steinheim, Germany) supplied through their Indian authorized agents at Lucknow, India. 2.3. Extraction of plant material for analysis Air dried ( 45-55 曟 ) powdered whole plant sample of Lycopodium calavtum (1.0 g) in triplicate were extracted separately with 3 暳10 mL methanol. E xtracts were concentrated under vacuum, redissolved in methanol, filtered and finally made up to 100 mL volume with ARTICLE INFO ABSTRACT Article history: Received 13 January 2012 Received in revised form 21 January 2012 Accepted 22 March 2012 Available online 28 April 2012 Keywords: Lycopodium clavatum Ferulic acid HPTLC Objective: To develop a simple precise and novel method for the identification and quantification of bioactive molecules ferulic acid in Lycopodium clavatum. Methods: Bioactive molecules ferulic acid in Lycopodium clavatum was indentified and estimated by the assay combined separation and quantitative estimation of the analyte on silica gel 60F 254 HPTLC plates with visualization under UV and scanning at 320 nm. Results: HPTLC studies showed the separation and determination of ferulic acid (0.443%) in Lycopodium clavatum. The result of recovery ranged in 95%-97% for ferulic acid. Conclusions: This method is simple, sensitive, economic, novel and first time reported from this species. Therefore, ferulic acid might be a useful chemotaxonomic marker for the genus Lycopodium and other lycopods. Contents lists available at ScienceDirect