Calamansi Lab Report

2.1. Plant Sample

Calamansi leaves were gathered within the vicinity of San Miguel, Bulacan. It was botanically identified and authenticated by a taxonomist at the National Museum, Botany Division, Manila.

2.2. Preparation of the Plant Extract

The leaves were washed thoroughly with distilled water. Then, it was oven-dried and pulverized into fine powder. 100g of the powdered leaves were weighed and macerated in 600mL ethanol in a stoppered flask for 24h. The solution was filtered and concentrated by a rotary evaporator.

2.3. Test for bioactive compounds A qualitative analysis for bioactive chemical constituents of calamansi leaves were carried out in aqueous extracts by utilizing the standard procedures described by Sofowora (1993); Trease

1.25mL of acetic acid and ferric chloride mixture was added to 2.5mL of the pure extract. Occurrence of blue-green color indicates the presence of glycosides.

Test for Saponins. 10mL of distilled water was added to 5 mL of the pure extract the mixture was shaken in a graduated cylinder for 15minutes. Occurrence of layer of foam or bubbles indicates presence of saponins.

Test for Phenols. 2.5mL of water was added to 2.5ml of the pure extract, then, ferric chloride, was also added, formation of blue or light green color indicates presence of phenols.

Test for Tannins. 2.5mL of ferric chloride solution was added to 2.5mL of the pure extract. Occurrence of dark blue or greenish-black color indicates presence of tannins.

Test for Anthocyanin. 1.25mL of Sodium hydroxide solution was added to 2.5mL of the extract. Formation of blue or green precipitates indicates presence of anthocyanin.

Test for Flavonoids. 2.5mL of Sodium hydroxide solution was added to 2.5ml of the extract. Occurrence of orange or intense yellow color indicates the presence of flavonoids.

Test for Alkaloids. 1.25 mL of Mayer’s reagent was added to 2.5mL of the pure extract. Formation of yellow precipitates indicates the presence of

The eggs were divided into 5 groups, each treatment was done in triplicates. Calamansi crude extract was reconstituted to three concentrations: (Treatment 1, 99μ of distilled water and 1μL of Calamansi crude extract) 1%, (Treatment 2, 95μ of distilled water and 5μL of Calamansi crude extract) 5%, and (Treatment 3, 90μ of distilled water and 10μL of Calamansi crude extract) 10%. Retinoic acid was used as a positive control, and ethanol as negative control. Approximately, 100μL of each treatment was placed in the filter paper. After the introduction of test solutions, the eggs were sealed with a tape and incubated for