Development of a Novel, Rapid and Validated HPTLC Protocol for the Quantitative Estimation of Marrubiin from the Extract of Marrubium vulgare Linn

Abstract

Aim: To develop a novel, simple, precise and rapid HPTLC protocol for the analysis of marrubiin (a furan labdane diterpene) in herbal extracts and formulations. Methodology: The marrubiin can be quantified by performing the HPTLC on silica gel F254 plates using toluene: ethyl acetate: acetic acid (5:4:1) as mobile phase.  The developed method was validated as per ICH guidelines. Results: The protocol was found to be linear in the concentration range of 40-400 ng/spot and simultaneous comparison of Rf (0.47 ± 0.05) and overlapping UV spectra of samples confirm the specificity of the method. The limit of detection (LOD) and limit of quantification (LOQ) of biomarker were found to be 15 and 40 ng by the selected method. The low value of % relative standard deviation (less than 2) in peak area on analyzing the sample on same and different days ensures the precision of the developed method. Further the recovery of more than 95% of the marrubiin affirms the accuracy of developed analytical method. Conclusion: It can be concluded that the developed protocol could be beneficial for the qualitative and quantitative analysis of the marrubiin in herbal extracts and formulations.