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<b>: </b>The present study was aimed to validate the phenolic, flavonoids, alkaloids and tannins profile of <i>Aspleniuma ethiopicum </i>(Burm. f.) Becherer methanolic extracts using HPTLC (High-performance thin-layer chromatography). The chromatographic separation was performed using the standard method. The separation methodology was optimized and phytoconstituents of <i>A. aethiopicum </i>methanolic extracts were separated. The methanolic extract of <i>A. aethiopicum</i> showed several bands identified as known phenolic compounds. The obtained flavonoids profiles were identified. HPTLC alkaloids profile of <i>A. aethiopicum</i> revealed 11 types of alkaloids. The tannin profile of methanolic extracts of <i>A. aethiopicum</i> demonstrated 11 different types of tannins. The observed profiles will be used as phytomarker to identify the chemical constituents of <i>A. aethiopicum </i>methanolic extracts. These profiles will act as a fingerprint of <i>A. aethiopicum</i> and help us to distinguish from its adulterants. The observed profile will help us to identify the crude drugs and improve the therapeutic potentials of <i>A. aethiopicum</i>.