Abstract

DEVELOPMENT OF VALIDATED HPTLC METHOD FOR QUANTIFICATION OF LUTEOLIN IN ECLIPTA ALBA AND ITS FORMULATION

Kavita Munjal, Rashmi Uniyal, Ashok Kumar, Anchal Arora, Apeksha Gupta, Abdul Haye and Vinod Kumar Gauttam*

ABSTRACT

Objectives: A simple, sensitive, precise, robust and rapid high performance thin layer chromatography (HPTLC) method for analysis of luteolin in Eclipta alba extract as well as in the formulation was developed and validated. Method: The mobile phase system consisted of toluene- ethyl acetate- formic acid (5: 3.5: 0.1, v/v/v). Densitometry analysis was carried out in the absorbance/reflectance mode at the wavelength of 350 nm at room temperature (25 ± 2°C). Result: The system was found to give compact bands for luteolin (Rf value of 0.34±0.02). The linear regression analysis data for the calibration plots showed good linearity (r2 = 0.99601 ± 0.002) in the concentration range 200–1200 ng spot-1. The method was validated according to the International Conference on Harmonization (ICH) guidelines. The limits of detection and quantitation were 20 and 60 ng spot-1, respectively. Conclusion: The method has been successfully applied in the analysis of herbal extract and market formulations containing luteolin.

Keywords: Densitometry, Flavonoids, Herbal formulation, HPTLC, Luteolin, Validation.