The Ruthenium Red Test Biology Essay

This test is subjected to identification of carbohydrates. 1 w/v dispersion of test substance was prepared in distilled water. Test mixture was combined with a small amount of molisch’s reagent in a test tube. After mixing, small amount of concentrated sulphuric acid was slowly added down the sides of the sloping test tube to obtain violet ring at junction.

5.2.1.4. Ninhydrin test

This test is subjected to identification of proteins. 2 ml aqueous solution of test substance and 1 ml alcoholic ninhydrin solution was mixed in test tube and heated at 80° C for 5 min to obtain red color.

5.2.1.5. Test with iodine

This test is subjected to identification of non reducing polysachharides (starch). 1 % w/v dispersion of test substance was prepared in distilled water. 3 ml test solution and few drops of dilute Iodine solution were mixed to obtain blue color after cooling and no color after heating.

5.2.1.6. Biuret test

This test is subjected to identification of proteins. 1 % w/v dispersion of test substance was prepared in distilled water. 3 ml test solution, 1 ml 4% sodium hydroxide and 2 drops of copper sulphate solution were mixed to obtain violet to pink.

5.2.1.7. Salkowski test

This test is subjected to identification of steroids. 1 % w/v dispersion of test substance was prepared in distilled water. 2 ml test solution, 2 ml chloroform and 2 ml conc. sulphuric acid were mixed and shaken well to obtain red colored chloroform layer and greenish yellow fluorescence in acid layer.

5.2.1.8. Baljet test

This test is subjected to identification of glycosides. 1 % w/v dispersion of test substance was prepared in distilled water. 2ml test solution and 2 ml sodium picrate was mixed in test tube to obtain yellow to orange color.

5.2.1.9. Shinoda test

This test is subjected to identification of flavonoids. 1 % w/v dispersion of test substance was prepared in distilled water. 3 ml test substance, 5 ml 95% ethanol, 2 drops of Conc. HCl and 0.5 g magnesium turnings were mixed to obtain pink color.

5.2.1.10. Wagner’s test

This test is subjected to identification of alkaloids. 1 % w/v dispersion of test substance was prepared in distilled water and filtered. 3 ml test solution filtrate was mixed with 2 drops of Wagner’s reagent to obtain reddish brown precipitate.

5.2.1.11. Test with acetic acid solution

This test is subjected to identification of alkaloids. 1 % w/v dispersion of test substance was prepared in distilled water. 2 ml test solution and acetic acid solution were mixed to obtain red color.

5.2.2. Physicochemical studies123

5.2.2.1. Viscosity

1% w/v solution of test substance was prepared in distilled water and 0.1 N HCl separately. Solutions were kept at 37 ± 2 °C for 6 hours. Viscosity of test solutions determined using Brookfield viscometer using spindle no. 3 at 50 rpm (Model No. DV-E).

5.2.2.2. pH determination124

1 % w/v dispersion of test substance in distilled water was prepared by shaking for 30 min and the pH determined using a pH meter (Elico).

5.2.2.3. Swelling Index88

Swelling index is the volume in ml occupied by 1 g of a drug, including any adhering mucilage, after it has swollen in an aqueous liquid for 4 hours.

1 g test substance (Fine powder) was placed in 25 ml ground glass stoppered measuring cylinder with graduations in 0.5 ml divisions. The test substance was moistened with 1.0 ml of 96% ethanol and 25 ml distilled water was added. Cylinder was closed using stopper. Cylinder was shaken every 10 min time interval for 1 hour and then was allowed to stand for 3 hours. Volume occupied by the test sample was determined in ml after decanting the supernatant clear liquid. Swelling index was calculated from average of three tests. Same experiment was performed using 25 ml 0.1 N hydrochloric acid instead of distilled water. Swelling index was calculated using following equation.

Vt = Volume occupied by test substance after hydration, at time 3 hrs.

V0 = Volume occupied by test substance before hydration, at time zero.

5.2.2.4. Bulk density123

Bulk Density was determined using Borosil 50 ml measuring cylinder. The volume of 5 gm mucilage powder was noted. Value of Bulk density was calculated by,

5.2.2.5. Tapped density123

Tapped Density was determined using Borosil 50 ml measuring cylinder. The volume of known weight 5gm was noted. The cylinder was given 250 taps on using tapped density apparatus. Value of Tapped density was calculated by

5.2.2.6. Carr’s compressibility index123

Carr’s compressibility index of AER mucilage powder was calculated from respective tapped density and bulk density by,

5.2.2.7. Loss on drying88

In flat bottom dish, 50 mm in diameter and 30 mm in height, 1 g fine powdered test substance dried in oven at 100-105 C for 3 hours. The test substance was allowed to cool in desiccator over diphosphorus pentoxide for 24 hours and weighed. Percentage loss on drying was calculated by following equation.

Weight of empty dish = A g

Weight of test substance taken = Y g

Weight of dish containing test substance after drying and placing in desiccator = B g

Weight of test substance after drying = (B – A) g