Extraction and Phytochemical Analysis of Secondary Metabolites From Achyranthes Aspera Linn

Abstract: Background: Apamarga (Achyranthes aspera Linn. (Family Amaranthaceae) is a common medicinal herbwith ancient roots in Ayurvedic, Unani and Siddha systems of healing. It is used in tropical and subtropical regions forthe treatment of inflammation, respiratory tract disease, gastrointestinal disorders, dermatologic conditions, kidneydisease and as an antidote to snake envenomation. Despite this high ethnopharmacological recognition, a potentialwidecomparative phytochemical survey of five different plant organs — roots, stems, leaves flowers and seeds usingcontemporary analytical methodology has not been carried out previously. Objective: To sequentially extract, isolateand characterize the secondary metabolite profiles of each major organ/part of this plant such as roots, bark, fruits andseed using polarity-gradient solvent extraction followed by qualitative/quantitative phytochemical screening(phytochemicals including alkaloids–the most dominant), chromatographic isolation in crude form and spectroscopicidentification. Materials and methods: Root, stem, leaf, flower, and seed shade-dried plant materials were successivelyextracted using petroleum ether, chloroform, ethyl acetate, methanol and water. By using conventional methods such asmaceration and Soxhlet extraction alongside green solvents, including ultrasound-assisted extraction (UAE) andmicrowave-assisted extraction (MAE). Standard qualitative phytochemical screening. Major constituents were isolatedby silica gel column chromatography and their structures elucidated using UV, FTIR, 1H/13C NMR and massspectrometry. Results: MAE gave the highest extract percentages for all studied plant parts (24.3–30.1% w/w formethanol), which were significantly higher than those obtained by maceration (17.1–23.0%). The columnchromatography afforded fourteen characterized compounds which were identified as β-sitosterol, chlorogenic acid,betaine, achyranthine, ecdysterone, ecdysone, lupeol, oleanolic acid, rutin and quercetin and caffeic acid andstigmasterol. Conclusions: A. aspera has chemically diverse and part-specific secondary metabolite profile that furthervalidates its traditional uses inpharmacotherapy. Plant Parts: Leaves are best for phenolic and flavonoid extraction; rootsand seeds are optimal for alkaloids, ecdysteroids, and saponins. However, MAE is advised to be the extraction techniqueof choice for high yield low solvent consumption. The isolated compounds and quantitative result sets lay the scientificgroundwork for quality control of herbal products while providing insight for future bioassay-directed pharmacologicalstudies.Keywords: Achyranthes aspera; Apamarga; phytochemical screening; sequential extraction; column chromatography;GC-MS; alkaloids; ecdysteroids; saponin s’; flavonoids terpenoids medicinal plants natural products.