Thick section of the digitalis sheet or part of the drug containing cardiac glycoside, when soaked in sodium picrate solution, it forms a yellow to orange color in the presence of aglycones or glycosides. In general, steroidal glycosides are used without exception in the therapeutic area mainly for two vital reasons, namely: (1) Libermann-Burchard test: alcoholic extract of the drug → evaporated → dry →extract with CHCl3 + a few drops of acetic anhydride + conc.salphuric acid (from the side wall of the test tube) → appearance of a purple ring → blue color → presence of sterol group in the drug. The alcoholic extract of the drug was evaporated to dryness and extracted with CHCl3, a few drops of acetic anhydride followed by conc. H2 SO4 of the side wall of the test tube with CHCl3 extract. The formation of a ring of purple to blue color at the junction of two liquids indicates the presence of steroid units. To the alcoholic extract of the drug, the same volume of water and 0.5 ml of strong lead acetate solution was added, shaken and filtered. The filtrate was extracted with chloroform of equal volume. The chloroform extract was evaporated to dryness and the residue was dissolved in 3 ml of glacial acetic acid, followed by the addition of a few drops of FeCl3 solution. The resulting solution was placed in a 2 ml conc test tube.

H2SO4. A reddish-brown layer forms, which becomes bluish-green after standing due to digitoxosis. 4. The vanillin HCl test is used to identify: (a) cardiac glycoside (b) anthraquinone glycoside (c) flavonoid glycoside (d) cynophore glycoside 2. Salkowaski test used to identify which glycoside unit used in a drug sample? a) Stelagglycoside b) Anthraquinone glycoside c) Cynophorglycoside d) Flavonoid glycoside i) Test for 3,5-dinitrobenzoic acid: Alcoholic solution from the drug sample + a few drops of NaOH + 2% solution of 3,5-dinitrobenzoic acid →pink color→ indicates the presence of cardiac glycosides. The drug is sprayed and extracted with 50% ethanol at low temperature, followed by the addition of a lead acetate solution to remove impurities, precipitates are removed by centrifugation, the cardiac glycoside present in the supernatant is extracted with chloroform, the chloroform extract is evaporated under vacuum, and the left residue (cardiac glycoside) is purified by chromatography. i) Sodium picrate test: small amount of drug sample → humidification with water in a conical flask + a few drops of conc. H2SO4 → appearance of brick color → presence of cynophoric glycoside. (i) Hemolysis test: One drop of blood on slides + a few drops of Aq.

Saponin solution → appearance of ruptured red blood cells → presence of saponin glycoside. Trichloroacetic acid test: drug + saturated trichloroacetic acid → precipitate colored → the presence of triterpenes. A glycoside solution is treated with a small amount of Kedde reagent (mixing equal volumes of a 2% solution of 3,5-dinitrobenzoic acid in menthol and a 7.5% aqueous solution of KOH). The development of a blue or purple color, fading in 1-2 hours, indicates the presence of cardinodes. To 1 g of the drug was added 5 ml of diluted HCl followed by 5 ml of ferric chloride (5% w / v). Boil for 10 min in a water bath, cool and filter, the filtrate is extracted with carbon tetrachloride or benzene and add the same volume of ammonia solution, formation of pink to red color due to the presence of anthraquinone. This is a type C of anthraqui-non-glycosides used. b. Based on aglycone: Based on the presence of the nature of aglycan groups, the classes are as follows: • sterol and triterpene glycosides • anthraquinone glycosides • cyanogenic and cyanophoric glycosides • saponin glycosides • flavone glycosides • coumarin glycosides • steroidal or cardiac glycoside (iii) vanilin HCl test: alcoholic solution of the drug sample + HCl vanillin → pink appearance→ presence of flavonoid To a solution glycoside, add a solution of antimony trichloride and trichloroacetic acid, then heat the mixture. The appearance of blue or purple color indicates the presence of cardenolides and bufanolides 1. General tests to identify sterol and triterpenoid glycosides in a drug sample (a) Borntrager test (b) Antimony trichloride test (c) Tetranitromethane test (d) b and c tests (i) Cuprocyanate test: saturate the filter paper in a freshly prepared solution of guaic resin + dissolved in ethanol → dry. → this filter paper be brought into contact with the dilute CuSO4 solution → place it in contact with the drug sample → generation of HCN gas with the appearance of spots → the presence of cynogenetic glycoside.