Identification, quantification and comparative assessment of costunolide in wild, cultivated and in vitro callus culture of Saussurea lappa by HPLC technique

Saussurea lappa is an endangered medicinal plant. Serious efforts are needed to conserve it by rapid in vitro technique. Plant tissue culture is one of the best way for its conservation. Its root contains a bioactive compound, costunolide which possess anti-carcinogenic, anti-ulcer, antimicrobial and anti-inflammatory etc properties. A simple high performance liquid chromatographic (HPLC) method for identification and quantification of costunulide was established. Shoot and root derived calli were harvested after seven and fifteen days. The ethanolic extracts of calli, cultivated and wild plant roots, petiole and lamina were used. Standard compound, Costunolide and plant materials were extracted into 70% ethanol and separated on a Perkin Elmer, 200 series using reverse phase column C18 (25 cm x 4.6 mm, 5 pm) with UV visible detector at 225 nm. The mobile phase was HPLC grade methanol and water (70: 30 v/v) at flow rate of 1ml minute-1. The total analysis time per sample was set 20 minutes. The establish method showed a good linerity (R2 = 0.988). Costunolide was identified only in roots and root derived calli extracts. Wild root (root c) produced high quantity 1.257 µgml-1 of Costunolide followed by fifteen days old (callus b) callus 1.119 µgml-1 and seven days old callus 1.118 µgml-1 (callus c) while the quantity from cultivated root (root b, root a) was 1.105 µgml-1, 1.102 µgml-1 respectively.