Spectrophotometry Uv-Vis and TLC Data for Phytochemical of Ethanolic Extract Moringa oleifera

Description

The UV-Visible spectrophotometric analysis of Moringa oleifera extract revealed characteristic absorption peaks indicating the presence of phenol, flavonoid, alkaloid, tannin, and saponin compounds, while Thin Layer Chromatography (TLC) profiling steroids and terpenoids.

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Phytochemical quantification of Moringa oleifera ethanolic extract was performed using UV-Visible spectrophotometry to assess flavonoid, phenolic, tannin, alkaloid, and saponin contents. For flavonoid determination, 50 mg of dried extract was reacted with 5% sodium nitrite, 10% aluminum chloride, and 1 M sodium hydroxide, then diluted and measured at 510 nm. Total phenolic content was analyzed by mixing 50 mg of extract with Folin–Ciocalteu reagent and sodium carbonate, followed by measurement at 760 nm. Tannin quantification involved ether extraction, followed by reaction with Folin–Ciocalteu reagent and sodium carbonate, and absorbance reading at 760 nm. Alkaloid analysis involved acidification of the extract with 2 N HCl, followed by chloroform washing and neutralization with 0.1 M NaOH. The mixture was then treated with phosphate buffer, BCG reagent, and chloroform, stirred, and the organic phase was evaporated under nitrogen. The residue was reconstituted and analyzed at 470 nm. Saponin content was assessed by hydrolyzing the extract with 25% sulfuric acid under autoclaving, followed by ether extraction, reconstitution with water, and treatment with 4-anisaldehyde and 50% sulfuric acid. The final absorbance was measured at 435 nm. For qualitative phytochemical profiling, Thin Layer Chromatography (TLC) was performed to detect steroids and terpenoids. A 50 mg sample was dissolved in ethanol, sonicated, vortexed, centrifuged, and macerated. A 100 μL aliquot was applied to a silica gel 60 F254 TLC plate alongside β-sitosterol standard. The plate was developed using ethyl acetate and toluene (93:7 v/v) as the mobile phase. After development, Liebermann–Burchard reagent was sprayed onto the plate, which was then heated at 110 °C. Detection was carried out at 340 nm under UV light. For qualitative phytochemical profiling, Thin Layer Chromatography (TLC) was performed to detect steroids and terpenoids. A 50 mg sample was dissolved in ethanol, sonicated, vortexed, centrifuged, and macerated. A 100 μL aliquot was applied to a silica gel 60 F254 TLC plate alongside β-sitosterol standard. The plate was developed using ethyl acetate and toluene (93:7 v/v) as the mobile phase. After development, Liebermann–Burchard reagent was sprayed onto the plate, which was then heated at 110 °C. Detection was carried out at 340 nm under UV light.

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