Biochemical Technology Topic: Identification of Saponins

1. Preparation of test solution:

1) Take 1 g of saponin pieces in a large test tube (or small beaker), add 15 ml of distilled water, immerse in a 30-90 ° water bath for 15 minutes, filter, and use the filtrate for identification.

2) The yam aqueous extract was prepared by adding 0.5 g of potato chips.

3) Take 0.5g of potato chips in a large test tube or a small conical flask and add 10ml of 95% ethanol to the water bath for 15 minutes, and the filtrate is for identification.

2. Identification test:

(1) Hemolysis test: Take a small piece of filter paper, add a drop of saponin infusion at the place of care, and then add a drop to the same place after the operation. If it is repeated until a few drops are added, no spray blood test after drying. Liquid (take bovine blood, sheep blood or rabbit blood, stir with a glass rod or cotton swab, remove the agglutinated blood protein, add a dilution of pH 7.4 phosphate buffer), and observe the red back after a few minutes. Whether there is no red yellow (or transparent) spot in the bottom (the origin of the soap solution in the center). (This reaction can also be carried out in a test tube. After the saponin in the herbal immersion liquid of the blood cell test solution is dissolved, the blood cell liquid is changed from turbidity to clarification. Further, it can be carried out on a glass slide, and the state before and after the blood cell rupture and dissolution is observed under a microscope. ).

(2) Foam test: 2 ml of yam immersion liquid and saponin immersion liquid were placed in test tubes. Shake it vigorously for one minute, and observe whether the two tubes have persistent foam in 10 minutes.

(3) Concentrated sulfuric acid test of acetic anhydride (Liebermann-Burchard reaction), 5ml of saponin immersion liquid, evaporated to dryness in a water bath in an evaporating dish, dissolved in 1 ml of acetic anhydride, dripped in a dry colorimetric plate, and slowed from the edge Slowly add 1 drop of concentrated sulfuric acid and observe the color change.

Another 5 ml of dioscorea immersion liquid was placed in an evaporating dish, evaporated to dryness on a water bath, 1 ml of acetic anhydride solution was added, and poured into a colorimetric plate. (Test tube) A few drops of concentrated sulfuric acid were added along the tube wall to observe whether there was any interface between the interfaces. What is the purple-red ring?

(4) Chloroform-concentrated sulfuric acid test (Salkowski reaction): Take 2 ml of diosgenol infusion solution, evaporate to dryness on a water bath, dissolve in 1 ml of chloroform, transfer to a dry small test tube, carefully add 1 ml of concentrated sulfuric acid along the wall, and the chloroform layer becomes red. Or blue, the sulfuric acid layer has green fluorescence, indicating steroidal saponin.