Legal Test for Digitalis

(i) Modified brontrager test: 1 g sample + 5 ml dilute HCl + 5 ml iron chloride →10 min. in a water bath → cool (at room temperature) →filter → filter the filter extract with carbon tetrachlor or benzene + equal amount of ammonia solution →pink to red appearance → presence of anthraquinone. Patients with digitalis resistance may require higher doses and higher serum levels than usual (e.g. patients with hyperthyroidism). 0.7 ml (Note: This volume does not allow for repeat tests.) Raymond`s test: Cardenolides give a purple color when they react with m-dinitrobenzene and methanolic potassium hydroxide. (1) Antimony trichloride test: Produce an alcoholic extract of the drug → evaporate → dry → extract with chloroform + a saturated solution of antimony trichloride in chloroform, which contains 20% acetic anhydride → pink color when heated → the presence of steroids and triterpinoids. 5. Specific test for the identification of cardiac glycoside: (a) Baljet test (b) Legal test (c) Keller-Killiani test (d) 3,5-dinitrobenzoic acid test Baljet test: A sodium picrate solution is added to a section of the digitalis sheet. It shows a yellow to orange color.

Keller: This test may be interfering if the sample is taken from a person who consumes a supplement containing a high dose of biotin (also called vitamin B7 or B8, vitamin H or coenzyme R). It is recommended to ask all patients indicated for this test for biotin supplementation. Patients should be advised to stop using biotin at least 72 hours before a sample is collected. Specific tests: For anthraquinone glycosides type C Killani test: Cook 1g of finely reduced digitalis powder with 10 ml of alcohol (70%) for 2-3 min. The extract is filtered. Add 5 ml of water and 0,5 ml of thick lead acetate solution to the filtrate. Shake well and separate the filtrate. The filtrate is treated with an equal volume of chloroform and evaporated to extract.

Then the extract is dissolved in glacial acetic acid and after cooling, 2 drops of iron chloride solution are added. Transfer this content to a test tube containing 2 ml of sulphuric acid. A reddish-brown layer, which acquires a blue-green color after standing, is observed due to the presence of digitoxosis. ANSWERS: 1. B and C 2. Steroidal glycoside 3. Cardiac glycoside 4. Flavonoid extract 5. Test for 3,5-dinitrobenzoic acid 6. NaOH 7.

Anthraquinone glycoside 8. Saponin glycoside 9. Cynophores glycoside 10. Borntrager test a and b: Boil the powdered drug with dilute sulfuric acid, filter it and add chloroform to the filtrate. Shake well and collect the organic layer. Add a few drops of strong ammonia solution, shake lightly and hold the test tube aside for a few minutes. The lower layer of ammonia takes on a pink or red color. This test is due to the presence of anthraquinones and is negative in reduced forms of anthraquinones, i.e. anthranols.

The use of foxglove to increase the contractile strength of a failing heart has been known since 1799, but the medicinal use of foxglove (Digitalis purpurea) and squid dates back to antiquity. The active ingredients of these plants, as well as Strophanthus species, are cardiac glycosides, whose aglycones (or genins) are steroids with a butyrolactone substituent in position 17. Examples are digoxygenin (41; R = OH) and digitoxygenin (41; R = H). The genins of squid and closely related compounds, toad venoms (found in the skin secretions of some toads), have α-pyrone rings instead of butyrolactone as in bufaline (42). The conformation of cardiotonic steroids differs from that of androgens and corticosteroids in that the cyclic compounds are c/d and (usually) a/b cis. The most common sugars are β-d-glucose, α-l-rhamnose, β-d-digitoxose, and β-d-cymarose, but the cardiotonic activity is due to aglycone. The presence of the heterocyclic ring is not essential, and synthetic compounds with an unsaturated ester or acrylonitrile group at position 17 are also active 〈74JPS1649〉. ii) Foam test: 1 g of drug sample + 10 to 20 ml of water → well stirred →foam → the presence of saponins.

(iii) Test HCl vanilin: alcoholic solution of the drug sample + HCl vanillin → appearance of pink color→ presence of flavonoid 10. Which color ring appears in Salkowaski`s test on connecting two mixed liquids? (a) Yellow (b) Red (c) Pink (d) A and b Libermann-Burchard test: To a solution of the drug in glacial acetic acid, a drop of concentrated sulfuric acid is added. A color change occurs from pink to red, purple and blue to green. The reaction is due to the steroid content. Make sure the patient does not take digitoxin instead of digoxin. Digitoxin is also an active component of the digitalis sheet. The 2013 ACCF/HCA guidelines for the management of heart failure suggest a therapeutic range of 0.5 to 0.9 ng/ml for digoxin. Ninety percent of nontoxic patients have values ≤2.0 ng/ml, 87% of toxic patients have values >2.0 ng/ml. Values >3.0 ng / ml in adults strongly indicate overdose; However, digitalis values should always be interpreted in light of clinical and chemical data. Older and smaller patients need less digoxin. In the elderly, proportionately lower doses are recommended.1 The leading cause of digoxin toxicity in the elderly is reduced kidney function. Maintenance doses should be adjusted according to glomerular filtration rate.1 Renal failure, hypercalcaemia, alkalosis, myxedema, hypomagnesemia, recent MI and other acute heart diseases, hypokalaemia and hypoxia may increase susceptibility to the toxic effects of digoxin.

6. Coumarin glycoside fluorescence test reagent: (a) KOH (b) NaOH (c) FeCl3 (d) CuSO4 (i) Legal test: Alcoholic extract of the drug + equal amount of water + 0.5 ml of strong lead acetate solution → shake well →filter → filtrate, extract with equal volume CHCl3→CHCl3 extract evaporate → dryness → rest dissolve with 2 ml of pyridine and sodium nitroprusside + NaOH solution to make it alkaline → color Rose → indicates the presence of aglycone or glycoside. 4. The HCl vanillin test is used in the identification of: (a) cardiac glycoside (b) anthraquinone glycoside (c) flavonoid glycoside (d) cynophores glycoside (i) Shinoda test: alcoholic extract of 1 g of drug + rotation of magnesium + diluted HCl → appearance of red color → presence of flavonoids. • Alcoholic extract of 1 g of drug sample + zinc twist + Dil. HCl → appearance of dark red color → turns into magenta color → the presence of dihydroflavonoids( another type of flavonoid glycoside). 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. Keller kiliani test: This test is specific to the digitoxosis content. Processing time is defined as the usual number of days from the date a sample is taken for analysis until the result is communicated to the order provider.

In some cases, additional time is required for additional confirmation or reflex testing. Test schedules may vary. (2) Tetranitromethane test: alcoholic extract of medicinal products + tetranitromethane solution →formation of yellow colour → the presence of .sterol and triterpenoid. Cardeinolines give a positive answer to all of the above tests. But bufadeinodes only give a positive test for the Libermann-Burchard test. Modified Borntrager test: It uses ferric chloride with dilute hydrochloric acid to induce oxidative hydrolysis. The released anthraquinones are extracted with carbon tetrachloride and give a pink-pink to cherry red color when their solution is stirred with dilute ammonia. 2.

Salkowaski test to identify which glycoside unit in a drug sample? (a) Stelaglycside (b) Anthraquinone glycoside (c) Cynophore glycoside (d) Baljet Test flavonoid glycoside: This includes sodium picrate solution. A yellow color is observed, which changes to orange. 3. Legal test to identify which glycoside: (a) anthraquinone glycoside (b) coumarin glycoside (c) cardiac glycoside (d) all (i) hemolysis test: One drop of blood on slides + a few drops of Aq. Saponin solution → appearance of ruptured red blood cells → presence of saponine lycoside. (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 the sterol group in the drug. (ii) Feeriferrocyanide test: transfer 1 g of sample + 5 ml of alcoholic → KOH to an aqueous solution of FeSO4 and FeCl3 → store at room temperature for 10 minutes. → the entire 20% HCl solution → appearance of Prussian blue color → the presence of cynogenetic glycosides. 7.

Modified Borntrager test to identify which glycosides? (a) Cardiac glycoside (b) Flavonoid glycoside (c) Saponin glycoside (d) Anthraquinone glycoside (ii) Ammonia test: For the alcoholic solution of 1 g of drug sample, when the filter paper is soaked and then exposed to ammonia vapor, the appearance of a yellow spot on the filter paper indicates the presence of flavonoids. ii) Baljet test: section of the drug, cardiac glycoside (thick cut of the digitalis sheet) →soaked in sodium picrate solution → appearance of yellow to orange color → indicate the presence of aglycan units. The bis(hydroxycoumarin) derivative dicoumarol (45), which was first identified as the causative agent of sweet clover disease in cattle, is used as an anticoagulant in the treatment of thrombosis. Similar compounds with other groups of compounds have been used, and warfarin (46), which was first introduced as a rodenticide, is also useful.