Ethiopian Health and Nutrition Research Institute (EHNRI)

Addis Ababa, Ethiopia

TITLE: Chemical screening method for the detection Origin Date: March,2010
of Free Anthraquinones in medicinal plants
Document Number: TMMRD-TP#9 Revision status: Current
Section: Phytochemistry Page: 1 of 5

Purpose This procedure provides instructions how to perform Chemical screening method for the
detection of free Anthraquinones in medicinal plants.

Abbreviations NA Not Applicable

C Concentration
V Volume

Materials Reagents
Benzene
Ammonia (10%)

Reagents preparation: Refer manual for phytochemical screening of medicinal

plants pages # 72-79

Reagents stability and storage: keep in a refrigerator for not more than 3
months
Supplies
Filter paper
cotton
test tubes
funnels
volumetric flasks 100ml
beakers 250ml
droppers
washing bottles
amber bottles
spatula
glass rod
Labeler
Pencil
Pipettes

Equipment
Analytical Balance
Vortex shaker
Refrigerator

Prepared by: Kissi Mudie Authority: EHNRI

Healthy, productive and prosperous Ethiopians

 Ethiopian Health and Nutrition Research Institute (EHNRI)
Addis Ababa, Ethiopia

TITLE: Chemical screening method for the detection Origin Date: March,2010
of Free Anthraquinones in medicinal plants
Document Number: TMMRD-TP#9 Revision status: Current
Section: Phytochemistry Page: 5 of 5

Sample Sample type Amount Transport and Stability

required Storage
Hydroalcoholic 0.1 gram At temperature Depends on the
extract less than 00C type and part of
plant

Limitations: Decomposition

Sample retention: 3-12 hours

Special Safety - Refer to National Safety Guidelines

Precautions

Quality Control Level Stability Frequency Preparation

Control (y/n)

NA Control preparation:

Procedure Step Action

1 Weigh 0.1gram of extracted sample in a test tube.
2 Add 10ml of Benzene.
3 Shake vigorously.
4 Filter the suspension through cotton into a test tube.
5 Add 5ml of 10% ammonia solution to the filtrate solution and shake.
6 Observe the color of the solution in the ammonia phase

Calculation C1V1=C2V2 (Dilution Law)

Result The red, pink or violet color appears in the ammonia phase indicates the presence of
Interpretation free anthraquinones.

Limitations none

Prepared by: Kissi Mudie Authority: EHNRI

.

Healthy, productive and prosperous Ethiopians

 Ethiopian Health and Nutrition Research Institute (EHNRI)
Addis Ababa, Ethiopia

TITLE: Chemical screening method for the detection Origin Date: March,2010
of Free Anthraquinones in medicinal plants
Document Number: TMMRD-TP#9 Revision status: Current
Section: Phytochemistry Page: 5 of 5

Procedural
Notes

Anthraquinone is the oxidized form of anthracene nucleus in its different oxidation

Principle state. The glycosides upon hydrolysis yield aglycones that are di, tri, or tetra-
hydroxyl anthraquinones or modifications of these compounds. Anthraquinones with
carboxylic group are isolated as salts of sodium or potassium since they are soluble
in aqueous basic solutions (Na2CO3, NaHCO3, NaOH, etc…). Anthraquinones with
hydroxyl groups in β-position form phenolites with aqueous solutions of sodium
carbonate and sodium hydroxide while anthraquinones with hydroxyl groups in α-
position form phenolites only with sodium hydroxide.

Clinical Utility To detect the presence of free anthraquinones in medicinal plants

Related
Procedures Manual for Phytochemical Screening of Medicinal Plants
and Manual for Qualitative Chemical Analysis Of Medicinal Plants
Documents

1. Harborne , J.B.(1973).Phytochemical methods, A guide to modern

Reference
techniques of plant analysis, Chapman and hall, London,
2. Yilma Desta (1988). Manual for qualitative analysis of medicinal plants.
Department of Traditional Medicine, Ministry of Health, Addis Ababa
3. Bu’ lock, J.D (1965). The Biosynthesis of Natural Products: An
introduction to secondary metabolites .MC Graw-Hill publishing
Company Ltd, England.
4. Trease, G.E and Evans , W.C (1966). Pharmacognosy. 14th edin., WB
sounders company Ltd. London.

Prepared by: Kissi Mudie Authority: EHNRI

.

Healthy, productive and prosperous Ethiopians

 Ethiopian Health and Nutrition Research Institute (EHNRI)
Addis Ababa, Ethiopia

TITLE: Chemical screening method for the detection Origin Date: March,2010
of Free Anthraquinones in medicinal plants
Document Number: TMMRD-TP#9 Revision status: Current
Section: Phytochemistry Page: 5 of 5

Name Signature and Date

Prepared by: ------------------------ -----------------------------

Reviewed by: --------------------------- -----------------------------

Approved by: -------------------------- ------------------------------

(Director of Traditional and Modern Drug Research Directorate)

Prepared by: Kissi Mudie Authority: EHNRI

.

Healthy, productive and prosperous Ethiopians

 Ethiopian Health and Nutrition Research Institute (EHNRI)
Addis Ababa, Ethiopia

TITLE: Chemical screening method for the detection Origin Date: March,2010
of Free Anthraquinones in medicinal plants
Document Number: TMMRD-TP#9 Revision status: Current
Section: Phytochemistry Page: 5 of 5

Declaration
I, the undersigned laboratory personnel, certify that I am conducting every steps of
the procedures incorporated in this SOP after a prior reading.

Name Signature and Date

1. _______________________ ______________________
2. _______________________ ______________________
3. _______________________ ______________________
4. _______________________ ______________________
5. _______________________ ______________________

Prepared by: Kissi Mudie Authority: EHNRI

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Healthy, productive and prosperous Ethiopians