Pharmacognostic quantification of flavonoids by high performance thin layer chromatography and in vitro cell line study on developed herbal formulation from Cestrum nocturnum plant extract

Abstract

Background: Herbal therapy is eventually based on the knowledge and practitioner skills. In India, local plants, as well as shrubs, can be procured easily due to influence of ancient Ayurveda. Taking into due considerations, we investigated Cestrum nocturnum (CN) Linn. commonly known as Raat rani. The shrub has been reported to have a wide arena of ethnopharmacological relevance. Objective: In the current research studies, our aim was to develop controlled release herbal tablets from the CN extract followed by in vitro screening on colon cancer cell line and correlate the quantified phytoconstituents such as rutin and quercetin by high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC). Materials and Methods: The leaves were macerated in methanol. The extract after preliminary phytochemical screening was subjected to sophisticated methods such as HPTLC and HPLC to elucidate rutin and quercetin which were found to positive by thin layer chromatography. To develop herbal tablets, the extract was subjected to excipient profile compatibility which was confirmed by Fourier transmission infrared (FTIR) spectrometry. Wet granulation method was used to develop the formulation. The developed herbal tablets were screened for colon cancer targeted cell line (HCT 15) using sulforhodamine B assay method with four dose levels of different concentrations. The results were analyzed as a percentage of growth inhibition 50%. Results: HPTLC confirmed the presence of spots of rutin (4.15% w/w) and quercetin (10.61% w/w) in comparison to the standard at Rf value 0.47 and 0.29, respectively. HPLC chromatogram showed the presence of rutin and quercetin with retention time 5.391 min and 6.698 min, respectively, in comparison to standard at 360 nm. The FTIR revealed no interaction of drug and excipients since the peaks of both are observed to be identical. Unsatisfactory results were obtained for the formulation on the selected cell line. Conclusion: In future, the world needs to explore, screen and develop herbal formulations which can be found to be compatible for cancer chemotherapy.