Mayer s reagent

The alkaloids test was conducted by implementing the method of Mayer, Wagner, and Dragendorff. Three milliliters of Meniran extract were placed in a porcelain cup, added with 5 mL 2 M HCl, then stirred and cooled at 25°C. Then, the cold sample was added with 0.5 g NaCl, then stirred and filtered. The filtrate obtained was mixed with three drops of 2 M HCl, then separated into four parts; A, B, C, and D. Filtrate A was used as a void, filtrate B was mixed with Mayer’s reagent (Merck, Germany), filtrate C was added with Wagner’s reagent (Merck), and filtrate D was used for the confirmation test. If a precipitate was formed on the Mayer and Wagner reagents added filtrates, it indicated the presence of alkaloids. In addition, a confirmation test was conducted by adding 25% ammonia into filtrate D to reach pH 8-9. Then, it was added with chloroform and evaporated over the water bath. The next step was adding 2 M HCl; it was then stirred and filtered. Then, the generated filtrate was divided into three parts; filtrate A was used as a void, filtrate B was tested with Mayer’s reagent, while filtrate C was tested with Dragendorff’s reagent (Merck). Any precipitate formation indicated the presence of alkaloids [19 ].

The filtrate obtained from maceration was evaporated using the rotary evaporator Heidolph Basis Hei-VAP HL (Germany), while antioxidant activities were determined using the spectrometer UV/VIS Shimadzu Pharma Spec UV-1700 Series. The medium used for brine shrimp growth was prepared using a small tank consisting of two compartments filled with sea water. Hexane, ethyl acetate, and methanol solvents for extraction were obtained via distillation. Iron (III) chloride, hydrogen chloride, sulfuric acid, sodium hydroxide, ammonia, magnesium powder, acetic anhydride, chloroform, Mayer's reagent, sodium carbonate, and methanol p.a. were purchased from Merck KGaA (Darmstadt, Germany). Gallic acid, Folin–Ciocalteu, and 2,2-diphenyl-1-picrylhydrazyl were purchased from Sigma Chemical Company (St. Louis MO, USA).

Methanol, ethanol, double distilled water, iron (III) chloride, gelatin, ammonia, chloroform, hydrochloric acid, Mayer’s reagent, Dragendorff’s reagent, magnesium, amyl alcohol, ether, vanillin-sulfuric acid reagent, sodium hydroxide, and Liebermann–Burchard reagent were purchased from Merck (Kenilworth, NJ, USA), without further purifications.