RP-HPTLC fingerprinting of secondary metabolites from Nephrolepis exaltata and Cycas revoluta

This study aims to develop a quantitative fingerprinting between two evolutionary close plant species Nephrolepis exaltata (L.) Schott and Cycas revoluta Thunb. The crude plant extracts were prepared in 1:1:1, v/v/v: Ethyl Acetate: n-Hexane: Methanol (AR) solvent system through cold extraction for 24 Hrs. The plant extracts were diluted (10 mg/mL) with a similar solvent system spotted (10 μL) on silica gel 60 F²⁵⁴ thin-layer chromatography plates. Five mobile phases (i) tetrahydrofuran (THF): toluene: formic acid: water (16:8:2:1) (ii) toluene: ethyl acetate: diethylamine (7:2:1) (iii) toluene: ethyl acetate: formic acid (7:3:0.1) (iv) n-hexane: ethyl acetate (7.2:2.9) (v) toluene: methanol (9:1) were used. The results were scanned at 254 nm, 366 nm, and in visible white light. This study gave minimum compact spots between Rf 0.06 to 0.051 corresponding to terpenoids, simultaneously for flavonoids spots were found between Rf 0.004 to 0.910. Total eight phenolic bands were found in Cycas revoluta among which five was found in leaf extract with Max RF of 0.033, 0.093, 0.273, 0.901 and 0.964 whereas three bands were found in stem extract with Max Rf of 0.007, 0.897 and 0.954. The presence of fifteen different types of flavonoids was determined by the flavonoids profile at Rf 0.004, 0.027, 0.047, 0.073, 0.134, 0.230, 0.303, 0.369, 0.369, 0.427, 0.514, 0.631, 0.703, 0.757, 0.846 and 0.910 for Nephrolepis exaltata. HPTLC fingerprint has shown several peaks with different Rf for different mobile phases. The fingerprinting would be helpful in the identification, and authentication of these species ecologically and therapeutically.