Phytochemical Analysis Lab Report

Phytochemical Screening

A brief summary of the experimental conditions for various methods of phytochemical analysis for plants material

Secondary metabolites Name of test Methodology Result Reference

1) Alkaloid Dragendorff’s test Spot a drop of extract on a small piece of precoated TLC plate. Spray the plate with Dragendorff’s reagent Orange spot (Kumar et al., 2007); Wagner test Add 2ml filtrate with 1% HCl + steam. Then add 1ml of the solution with 6 drops of Wagner’s reagent. Brownish-red precipitate (Chanda et al., 2006). TLC method 1 Solvent system: Chloroform: methanol: 25% ammonia (8:2:0.5).

Spots can be detected after spraying with Dragendorff reagent Orange spot (Mallikharjuna et al.,

2007).

2) Anthraquinone Borntrager's test Heat

A yellow solution with

NaOH, turns colorless with dilute HCl (Onwukaeme et al., 2007).

5)Phenol Phenol test Spot the extract on a filter paper. Add a drop of phoshomolybdic acid reagent and expose to ammonia vapors. Blue coloration of the spot (Kumar et al., 2007);

6)Saponin Frothing test /

Foam test Add 0.5ml of filtrate with 5ml of distilled water and shake well. Persistence of frothing (Parekh and Chanda,

2007).

7)Steroid Liebermann-

Burchardt test To 1ml of methanolic extract, add 1ml of chloroform, 2-3ml of acetic anhydride, 1 to 2 drops of concentrated sulphuric acid. Dark green coloration (Kumar et al., 2007). - To 1 ml of extract, add 2 ml acetic anhydride and 2 ml concentrated sulphuric acid H2SO4. Color change to blue or green (Edeoga et al., 2005).

8)Tannin Braemer’s test 10% alcoholic ferric chloride will be added to 2-3ml of methanolic extract

(1:1) Dark blue or greenish grey coloration of the solution (Kumar et al., 2007);

(Parekh and Chanda,

2007).

9)Terpenoid Liebermann-

Burchardt test To 1ml of methanolic extract, add 1ml of chloroform, 2-3ml of acetic anhydride, 1 to 2 drops of concentrated sulphuric acid. Pink or red coloration (Kumar et al.,

Water: Water is universal solvent, used to extract plant products with antimicrobial activity. Though traditional healers use primarily water but plant extracts from organic solvents have been found to give more consistent antimicrobial activity compared to water extract. Also water soluble flavonoids (mostly anthocyanins) have no antimicrobial significance and water soluble phenolics only important as antioxidant compound (Das K et al.,2010).

2. Acetone: Acetone dissolves many hydrophilic and lipophilic components, is miscible with water, is volatile and has a low toxicity, it is a very useful extractant, especially for antimicrobial studies where more phenolic compounds are required to be extracted. A study reported that extraction of tannins and other phenolics was better in aqueous acetone than in aqueous methanol (Das K et al.,2010, Eloff JN.,1998) . Both acetone and methanol were found to extract saponins which have antimicrobial activity (Ncube