4.3.3. Phenolic Compounds (Vegetable Tannins) (FeCl<sup>3</sup> Test)

An amount of 2 mg of the extracts was dissolved in 2 mL of water or ethanol, and then three drops of 2.5% FeCl<sup>3</sup> in water were added. The appearance of a red, blue-violet, or green precipitate was considered positive.

### 4.3.4. Steroids and Terpenes (Salkowski Test)

An amount of 2 mg of each extract was dissolved in 2 mL of chloroform, and subsequently, 2 mL of H2SO<sup>4</sup> was added. A positive result was considered for sterols and methyl sterols when a red-brown ring was formed at the interface.

### 4.3.5. Carbohydrates

### Molish Test

Molish's reagent (1% alpha-naphthol in ethanol) was added dropwise to 2 mg of the extracts and then 2 mL of H2SO<sup>4</sup> was added through the walls of the tube. The test was considered positive when a purple-colored ring formed at the interface.

### Coumarin Test

An amount of 2 mg of the extracts was dissolved in 2 mL of ethanol and 10% NaOH was added dropwise. The test was considered positive if a yellow coloration was present and if it disappeared when the solution is acidified.

### Lactone Test

An amount of 2 mg of the extracts was dissolved and 2 mL of an alcoholic solution of 10% NaOH was added. The test was considered positive when there was a turn to yellow or orange color that was lost or disappeared when a few drops of HCl were added, indicating the presence of a lactonic ring.

### 4.3.6. Sesquiterpene Lactones (Baljet Test)

A amount of 2 mg of the extracts was dissolved in 2 mL of ethanol, and three drops of the mixed solution were added, with a positive result being indicated if it turned from orange to dark red. The 1:1 mixed solution consisted of Solution A, which contained 1% C6H3N3O<sup>7</sup> in ethanol and Solution B, which contained 10% NaOH.
