Wagner s reagent

The reagents Luria–Bertani broth (Miller, Appleton, WI, USA) and Mueller–Hinton agar were procured from SRL Pvt. Ltd. (Mumbai, India), Himedia, whereas the ampicillin sodium injection was obtained from Saralife (New Delhi, India). Silica-gel-coated TLC plates, Wagner’s reagent, and Molisch reagent were procured from Merck, Darmstadt, Germany. All other reagents used were of analytical grade.

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 ].