Evidence-Based Complementary and Alternative Medicine

Research Article

Antibiofilm Activity of Azadirachta indica and Catharanthus roseus and Their Synergistic Effects in Combination with Antimicrobial Agents against Fluconazole-Resistant Candida albicans Strains and MRSA

Table 1

Phytochemical screening methods as described by Visweswari et al. [23] and Neglo et al. [24].
PhytochemicalTestObservation
AlkaloidWagner’s reagent (I2/KI) was used. Minute quantity of extracts was dissolved in dilute HCl and filtered. Few drops of Wagner’s reagent (I2/KI) were added to about 2 mL of the filtrate.Formation of brownish/red precipitate was used to determine the presence of alkaloid.
FlavonoidsSulphuric acid (H2SO4) test was done by treating a fraction of the extract with concentrated H2SO4.Formation of orange color was used to detect the presence or absence of flavonoids.
SteroidsLiebermann–Burchard test was used. Four milligrams of the extracts was treated with 0.5 ml of acetic anhydride and 0.5 mL of acetic acid. Concentrated H2SO4 was slowly added.The development of a reddish/brown color indicated the presence of steroids.
TerpenoidsLiebermann–Burchard test was used. Four milligrams of the extracts was treated with 0.5 ml of acetic anhydride and 0.5 mL of acetic acid. Concentrated H2SO4 was slowly added.The development of a blue-green color indicated the presence of terpenoids.
SaponinsThis was tested for using foam test. Exactly 0.5 g of the plant extract was dissolved in 2.5 mL of distilled water. The mixture was shaken vigourously.The presence of foam indicated the presence of saponins.
TanninsFerric chloride test was used to test for the presence of tannins. An exact amount of 0.5 g of the extract was boiled in 20 mL of distilled water and filtered afterwards. Few drops of 0.1% of FeCl3 were added.The presence of brownish-green, brownish-black, or blue-black color was used to detect the presence of tannins.
GlycosidesBenedict’s test was used for the detection of glycosides. Precisely, 0.5 g of plant extract was dissolved in 5 ml of distilled water. Exactly 2 mL of Benedict’s solution was heated and 8 drops of the dissolved sample were added and allowed to boil for 5 minutes.Formation of brick-red precipitate indicated the presence of glycosides.