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2018; 8(5-s):332-337
ABSTRACT
Boerhavia diffusa Linn. (Nyctaginaceae), commonly known as ‘Punarnava’ is a perennial creeping herb widely studied and has a
long history of uses by the tribal people and in Ayurvedic and Unani medicines. Our previous study showed that in different extracts
of Boerhavia diffusa (ethanol, methanol, petroleum ether, aqueous and hexane), methanolic extract had significant anti-
oxidant activity, but the active components present in that extracts are still unclear. In this study, Boerhavia diffusa Linn was
investigated for the bioactive compounds present in its methanolic extract. Separation and purification of the compounds in the most
active methanol extract was done using a combination of column chromatography and thin layer chromatography. The compound
was identified using gas chromatography and mass spectrophotometry. The resulted compound anthraquinone which is also called as
9, 10 anthracenedione. In addition to their known use as natural dyes, it have several biological activities like antitumor,
antiinflammatory, diuretic , antiarthritic, antifungal, antibacterial, antimalarial and antioxidant activities . This justifies the use of
this plant in traditional medicine and indicates a promising potential for the development of medicinal agents from Boerhavia
diffusa.
Keywords: Antioxidant, Boerhavia diffusa Linn, Anthraquinone, Column chromatography, Thin layer Chromatograhy, GC-MS.
Article Info: Received 17 Sep, 2018; Review Completed 08 Oct 2018; Accepted 09 Oct 2018; Available online 15 Oct 2018
Cite this article as:
Kanagavalli U, Sadiq AM, Isolation and characterization of bioactive compound anthraquinone from methanolic
extract of Boerhavia diffusa linn., Journal of Drug Delivery and Therapeutics. 2018; 8(5-s):232-237
DOI: http://dx.doi.org/10.22270/jddt.v8i5-s.1987
cytotoxicity, combined with simplicity, specificity, and procedures or protocols to isolate and characterize
speed9. certain bioactive molecules. This could be due to
different parts (tissues) in a plant, many of which will
In vitro methods are usually more desirable than in vivo
produce quite different compounds, in addition to the
assays because animal experiments are expensive, take
diverse chemical structures and physicochemical
more time, and are prone to ethical controversies. There
properties of the bioactive phytochemicals10.
are some factors that make it impossible to find final
Purification of the Bioactive Molecule. complex phytochemicals, which make a good separation
difficult. Therefore, increasing polarity using multiple
Many bioactive molecules have been isolated and
mobile phases is useful for highly valued separations.
purified by using paper thin-layer and column
Thin-layer chromatography has always been used to
chromatographic methods. Column chromatography and
analyze the fractions of compounds by column
thin-layer chromatography (TLC) are still mostly used
chromatography. Silica gel column chromatography and
due to their convenience, economy, and availability in
thin-layer chromatography (TLC) have been used for
various stationary phases. Silica, alumina, cellulose, and
separation of bioactive molecules with some analytical
polyamide exhibit the most value for separating the
tools11.
phytochemicals. Plant materials include high amounts of
Figure 2: Thin layer chromatogram identified spot at different fractions using chloroform:methanol(9:1) solvent system.
The visualized spots of the components in the Determination of the structure of certain molecules uses
chromatoplate were marked and the Rf value of each data from a wide range of spectroscopic techniques such
spot was calculated by the formula: Rf = distance as UV-visible, Infrared ( IR), Nuclear Magnetic
travelled by the sample (cm)/distance travelled by the Resonance (NMR), a n d mass spectroscopy. The basic
solvent (cm).The several fractions gave a many spot at principle of spectroscopy is passing electromagnetic
different Rf on silica gel 60 F254 pre-coated aluminum radiation through an organic molecule that absorbs some
plate, of 0.2 mm thickness using Chloroform: Methanol of the radiation, but not all. By measuring the amount of
(9:1) as the developing solvent system. Visualization absorption of electromagnetic radiation, a spectrum can
was carried out by dipping the plate in vanillin- be produced. The spectra are specific to certain bonds in
sulphuric acid reagent and heating at 105°C till the color a molecule. Depending on these spectra, the structure of
of the spot appeared (Figure:2). Thin layer the organic molecule can be identified. Scientists mainly
chromatogram fractions in long and shot UV is shown in use spectra produced from either three or four
figure 3 and 4. regions—Ultraviolet (UV), Visible, Infrared (IR), radio
frequency, and electron beam for structural
Structural Clarification of the Bioactive Molecules
clarification12.
Figure 5: GCMS Chromatogram fractions isolated from methanolic extract of Boerhavia diffusa containing
anthraquinone.
Table 1: GCMS Data of bioactive compounds detected with their percentage area and retention time.
The results pertaining to GC-MS analysis of the isolated identify the nature and structure of the compounds. The
fraction from methanolic extract of Boerhavia diffusa large compound fragments into small compounds giving
lead to the identification of a bioactive compound rise to appearance of peaks at different m/z ratios. These
anthraquinone. This compound is identified through mass spectra are fingerprint of that compound which can
mass spectrometry attached with GC at Retention time be identified from the data library.
(34.834). The detected compound present in the entire
These results reveals that the plant has the antioxidant
herb of Boerhavia diffusa by the GC-MS was shown in
compound anthracenedione (Also called as
[Table 1 ].
Anthraquinone) and further investigation required to
The GC-MS spectrum confirmed the presence of identify the cardioprotective activity of this compound
bioactive components with different retention times as through biochemical parameters, gene expression study
illustrated in [Figure 5, 6]. The mass spectrometer and histopathological study.
analyzes the compounds eluted at different times to
ISSN: 2250-1177 [336] CODEN (USA): JDDTAO
Kanagavalli et al Journal of Drug Delivery & Therapeutics. 2018; 8(5-s):332-337
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