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Dr. Mohammed N. Sabir, BSc. Pharm, MSc., Ph.D. Pharmacog.

Lecturer, Dept. of Pharmacognosy and Pharmaceutical Chemistry
College of Pharmacy, University of Sulaimani
mohammed.sabir@univsul.edu.iq

Experimental pharmacognosy I
First semester, third-year pharmacy, 2021 – 2022

Experiment No. 1

Extraction of anthraquinone glycosides and caffein from senna leaves and coffee beans
by maceration and percolation respectively

The maceration process is widely applied for the extraction of natural products from their crude materials.
This process showed advantages compared to other procedures, these include, simplicity, cost-efficient,
applicability, and being devoid of heat involvement. However, some drawbacks were also parallel to the use
of this method like poor productivity. To overcome this problem, high quantities of crude materials are used
for the extraction. Other disadvantages include a longer extraction duration. In some cases, yield improvement
is performed by applying some physical enhancements like agitation (shaking), and medium temperature
elevation.
Other extraction procedures include percolation, a fast, economic, and simple technique applied for the
extraction of a variety of natural products. Despite its low yield, this method is still widely applied.
Several medicinal plants are showing therapeutic benefits, some of which were used as a source for
medications, among these plants is Senna. The dried leaves of Cassia alexandrina, Cassia acutifolia, C.
angustifolia were used for centuries for the treatment of constipation. Nowadays, the dried plant leaves are the
source of important anthraquinone glycosides, which are believed to be responsible for their laxative action.
On the other hand, coffee beans are widely used as beverage, and as a source of caffeine (an alkaloid).
Caffeine is used in some pharmaceutical preparations and demonstrates energetic and antioxidant activity.
Both, the anthraquinone glycosides and caffeine are dissolved in polar solvents. Generally, water and ethanol
are widely used for this purpose. Decreasing the medium pH, results in the possibility of breaking the glycosidic
linkage and release the aglycon part.
This experiment aims to extract bioactive glycosides and alkaloids from the dried Senna leaves, and
powdered coffee beans using maceration and percolation methods respectively.

Extraction procedures

1. Using mortar and pestle, grind the dried senna leaves to a coarse powder and transfer to a dry watch glass,
and weigh (10) grams.
2. Weigh (5) grams of powdered coffee beans.
3. Prepare 150 mL [ethanol: water (1: 1 v/v)] to be used as the extraction solvent.
4. Soak the crude senna powder in two clean glass beakers with 50 mL of the extraction solvent (5 gm of
each).
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5. Place the first glass vessel (marked as A) on the lab bench and leave for two hours.
6. Put the other glass beaker on an orbital shaker, and adjust the rotation to 100 rpm, at room temperature
for two hours (mark as B).
7. Prepare a clean glass conical, glass funnel, and a Whatman No.1 filter paper.
8. Place the filter paper in the glass funnel, and place them over the glass funnel (Marked as C). Place the
weighted coffee powder in the funnel and compact by applying low pressure with a glass rod.
9. Start the extraction by pouring 100 mL hot distilled water over the powdered coffee at a slow rate.
10. Collect the filtrate, and filter again using another filter paper, and preserve the solution in refrigeration.
11. Filter the A and B crude senna extracts under vacuum (repeat the filtration if necessary) until a clear
solution is obtained.
12. Add 3N hydrochloric acid (HCl) in a dropwise manner to both extracts until no more precipitate appears.
13. Filter both crude extracts, and add distilled water to enhance the filtration. Neutralize the medium by
adding ammonia, and adjust the pH of the solution to 7.0.
14. Collect the mark, and expose it to dry at room temperature in the hood.
15. Weight the dried marks and calculate the percentage yields of both A and B as shown below,

% Yield = [obtained dry mark / crude plant weight] X 100

16. Write your report, compare the results obtained from both methods, explain the difference in the yield.
Use previously reported yields to support your results.
17. Preserve the dried marks for further analysis.

Materials
1. Glass beaker (200 mL).
2. Glass conical flasks (250 mL).
3. Glass funnel (100 mL).
4. Glass rod.
5. Watch glass.
6. Measuring cylinder.
7. Whatman No. 1 filter paper.
8. Coffee bean powder.
9. Dried senna leaves.
10. Ethanol 99.9%.
11. Distilled water.
12. Hydrochloric acid.
13. Ammonium hydroxide solution.
14. Reagent bottle.
15. Marker and pen.
16. Orbital shaker.
17. Hot plate.
18. Clamp.
19. Vacuum motor.
20. Notebook.
21. Nitrile gloves.
22. Mask.
23. Sensitive electric balance.