DISCUSSION

DISCUSSION

Hemidesmus indicus (L.) R.Br. is an important drug of Indian System of

Medicine and used in medicines since time immemorial. A number of phytochemical

studies have been carried out and many of the compounds reported are mainly
19-27
triterpenoids, glycosides, organic acids, aldehydes etc . Various pharmacological

activities have been reported such as anticancer, hepatoprotective, anti snake-venom

activity, anti-ulcerogenic, antiatherogenic, anti-inflammatory, analgesic, antileprotic and

antimicrobial etc 28-64. Although many works have been carried out on this plant but there

are scanty reports on specific markers for this plant, HPLC profile of the compounds and

standardization. Hence in the present study an attempt had been made to isolate and

standardize the constituents which are unique to this plant and characterize them as

markers which are chemically defined constituents and can serve as a powerful tool for

standardization.

Isolation of the phytoconstituents

The methanolic root extract of Hemidesmus indicus was refluxed with acetone.

The acetone extract was then fractionated using silica gel column chromatography (60-

200 mesh). Review of literature indicates that column chromatography using silica gel is

the most popular method for isolation 19-27. Totally four different column chromatography

has been done with different solvent systems which resulted in the isolation of four

different compounds. These were given codes as HI-01, HI-02, HI-03 and HI-004.

Department of Pharmacognosy - PESCP 72

DISCUSSION

Analysis of isolated compounds

Determination of Melting point and solubility profile were done for the isolated

compounds. New HPLC methods were developed to analyze the purity of the

compounds. The HPLC method indicated that the compound HI-01 was 99.41 % pure,

HI-02 was 32.88 % pure, HI-03 was 46.82% pure and HI-004 was 99.84 % pure. Hence

depending upon the purity only HI-01 and HI-004 were taken for the further

characterization.

Characterization of the isolated compounds

The integral values of NMR spectroscopy, the values where the bands obtained

in the IR spectroscopy, the CHN analysis and the Mass spectroscopic analysis clearly

suggested that the compound HI-01 was found to be a pentacyclic triterpene acetate. The

chemical name suggested for this was 20(30)-Ursen-3-acetate. The other compound that

is HI-004 was found to be an aromatic aldehyde and the chemical name suggested for this

compound was 2-Hydroxy-4-Methoxy benzaldehyde 72, 73.

Estimation of isolated compounds

In order to check the suitability of the isolated compounds as markers, HPLC

analytical methods were developed. The distinct peaks at the retention time of 1.694 min.

and 26.021 min. (corresponding to the same value of HI-01 and HI-004 respectively) in

HPLC chromatograms of acetone fraction was observed indicating that the method was

suitable and sensitive. The acetone fraction of the MeOH extract was found to contain

14.34 % of HI-01 and 12.94 % of HI-004. Hence these compounds can be termed as

markers and the extract is said to be standardized.

Department of Pharmacognosy - PESCP 73