The HPTLC Validated Method Development for the Quantification of Naringin from the Partially Purified Labisia Pumila Dichloromethane Extract

Abstract

Partially purified fraction E, dichloromethane extract of leaves from Labisia Pumila has been shown to possess anti-ulcer, anti-inflammatory, anti-asthmatic activities among others. Naringin has been identified by LC-MS and HPLC as the major compound present in fraction E. The present study reports the development and validated TLC densitometric method for the quantification of Naringin present in Fraction E. ICH guidelines were followed to develop this method. CAMAG-HPLTC system comprising of a TLC Visualizer and Linomat 5 sample applicator was used in this study. The separation was performed using TLC aluminum plates pre-coated with silica gel 60 F254. Optimized mobile phase consisted of Methanol: Ethyl Acetate (60:40 v/v). Win-CATS-V 1.2.3 software and Video Scan were used to identify and quantify Naringin at 366nm in fluorescence mode. The peak heights at Rf 0.648 ± 0.01 gave a linear range from 200-1000µg/ml correlation coefficient R2± SD = 0.973 ± 0.024. The LOD and LOQ were found to be Naringin 0.74 ± 0.29 µg and 7.73 ± 0.26 µg, respectively. Repeatability gave CV % < 2.0. We recovered 92.56% of Naringin and 642.4 mg/g (64.24%) was found to be present in Fraction E. The HPTLC method was found to be reproducible, accurate and convenient for rapid screening of bioactive constituents present in Fraction E. This developed method will be used for analysis and quality control of drug formulations containing Labisia Pumila.