III.

Anthraquinone glycosides are natural products found in many medicinal plants, such as rhubarb and
senna, and are known for their laxative and anti-inflammatory effects. One of the commonly used
methods for screening anthraquinone glycosides is the Borntrager’s test. Here’s how it works:

A. Borntrager's test:

Procedure:

Take the powdered plant material in a test tube and add 10 ml of dilute hydrochloric acid (HCl).

Heat the mixture in a water bath for about 30 minutes.

Cool the mixture and add an equal volume of chloroform.

Shake the mixture vigorously, and allow the layers to separate.

Transfer the lower chloroform layer to a clean test tube.

Add a few drops of ammonia solution to the chloroform layer and shake.

Observe for the appearance of a pink or red color in the ammoniacal layer.

Result:

If the test is positive, i.e., a pink or red color appears in the ammoniacal layer, it indicates the presence
of anthraquinone glycosides in the plant material.

B. Modified Borntrager’s test:

The modified Borntrager’s test is a more sensitive and reliable method for screening anthraquinone
glycosides. The procedure is similar to the original Borntrager’s test, but with some modifications. Here’s
how it works:

Procedure:

Take the powdered plant material in a test tube and add 10 ml of dilute hydrochloric acid (HCl).

Heat the mixture in a water bath for about 30 minutes.

Cool the mixture and add an equal volume of chloroform.

Shake the mixture vigorously, and allow the layers to separate.

Transfer the lower chloroform layer to a clean test tube.

Add a few drops of 10% potassium hydroxide (KOH) solution to the chloroform layer and shake.

Observe for the appearance of a pink or red color in the chloroform layer.

Result:

If the test is positive, i.e., a pink or red color appears in the chloroform layer, it indicates the presence of
anthraquinone glycosides in the plant material.

Both Borntrager’s test and modified Borntrager’s test are simple and effective methods for screening
the presence of anthraquinone glycosides in plant materials. However, it is important to note that a
positive result from these tests does not necessarily confirm the presence of anthraquinone glycosides,
as other compounds may also produce a similar reaction. Further tests, such as thin-layer
chromatography (TLC) or high-performance liquid chromatography (HPLC), may be required for
confirmation

X.

Thee strain test, also known as the”spot test, is a simple and quick screening test that can be used to
determine the presence of fixed or volatile oils in a substance. It is commonly used in the field of
essential oil analysis, as well as in the testing of other natural products like herbal extracts.

Procedure:

Take a clean piece of filter paper or chromatography paper.

Place a small amount of the sample to be tested in the center of the paper.

Allow the sample to evaporate for a few minutes.

Observe the stain left behind on the paper.

Results:

Fixed oils will leave a permanent grease stain on the paper, which will not evaporate.

Volatile oils will evaporate quickly and leave behind a transient stain or ring on the paper.

The strain test is a useful screening tool for identifying the presence of fixed or volatile oils in a sample.
However, it does not provide information about the identity or purity of the oil, nor does it distinguish
between different types of oils. Further testing, such as gas chromatography or mass spectrometry, may
be necessary to fully characterize the oil.

References:
CWang, Z., Ma, P., Xu, L., He, C., Peng, Y., & Xiao, P. (2013, October 26). Evaluation of the content
variation of anthraquinone glycosides in rhubarb by UPLC-PDA. Chemistry Central Journal; BioMed
Central. https://doi.org/10.1186/1752-153x-7-170

Tewari, A., Singh, V. P., Yadav, P., Gupta, G., Singh, A., Goel, R. K., Shinde, P. S., & Mohan, C. V. (2014,
October 1). Synthesis, biological evaluation and molecular modeling study of pyrazole derivatives as
selective COX-2 inhibitors and anti-inflammatory agents. Bioorganic Chemistry; Elsevier BV.
https://doi.org/10.1016/j.bioorg.2014.05.004