Development and Validation of RP-UHPLC and HPTLC Methods for a Polyherbal Active Ingredient Boswellia serrata

Abstract

The aim of present research work was to create a quick, accurate, and reliable RP-UHPLC and HPTLC method for estimating Boswellic acid from Boswellia Serrata extracts and formulations. RP-UHPLC method has been developed and validated for the determination of Boswellic acidin extract and in formulation. The RP-UHPLC analysis was performed on the Cosmosil C18, (250mm x 4.6mm i.d., 5 µm) in isocratic mode, at ambient temperature using Methanol : Water (0.1 % OPA) (80:20, v/v) as mobile phase; flow rate was set at 1.0mL/min. The detection was carried out at 250nm. The retention time for Boswellic acid was found to be 12.5 min. Linearity of Boswellic acid followed in the concentration range of 10-60µg/mLand correlation coefficient was found 0.9997. The HPTLC method was performed on 10cm x 10cm aluminium plate coated with 0.2 mm layer of silica gel 60 F254 (E. Merck, Germany). Samples were applied to the plate as bands width 6 mm by using Camag (Muttenz, Switzerland) Linomat-5 applicator fitted with 100µl syringe (Hamilton, Switzerland). The rate of application was constant at 150nl sec-1 and space between two bands was 14mm. The mobile phase consisted of toluene: ethyl acetate: methanol (4:1:0.5). Densitometric scanning was performed at 254nm using Camag TLC 3-Scanner. The average Rf is 0.54. The calibration curve was linear in concentration range of 500-3000ng/spot and correlation coefficient is 0.9996. The Limit of detection was calculated and it was found to be 5.42ng and Limit of quantification was found to be 16.45ng. The developed RP-UHPCL and HPTLC method was validated as per ICH Q2 (R1) analytical method validation guidelines. All developed methods could be used for routine analysis of Boswellic Acidin the dosage form.