Bioassay- guided fractionation and anti-fungal activity studies on pisonia grandis r
International Journal of Pharmacognosy and Phytochemical Research 2013; 5(1); 1-3
Bioassay- Guided Fractionation and Anti-Fungal Activity Studies on
*Shubashini K. Sripathi, Poongothai G.
Department of Chemistry, Avinashilingam University for Women, Coimbatore Tamilnadu, India. ABSTRACT Bioassay- guided fractionation of ethanol extract of leaves of Pisonia grandis was studied for its anti-fungal activity for the microorganism Candida albicans, Aspergillus niger, Pencillium citrinum and Monascus purpureus by disc diffusion method. The ethanol extract showed good anti-fungal activity for Monascus purpureus compared to standard clotrimazole. Keywords: Pisonia grandis, Nyctaginaceae, Monascus purpureus, Clotrimazole. INTRODUCTION
combined. A portion of these two layers are concentrated
separately and subjected to anti-fungal study. The rest of
distributed throughout India and is a widespread
the CHCl3 layer was distilled completely; the residue was
evergreen commonly grown lettuce tree1. Leaves, stem
dissolved in 10% aqueous ethanol for further extraction
and root of this species are extensively used by the tribals
with pet-ether. The LLE with pet-ether is continued until
in the preparation of several folk medicines. It has been
the organic layer was colorless. Then the entire organic
extensively used in Indian traditional medicine as an
and aqueous layer was combined and distilled by vacuum
antidiabetic, anti-inflammatory agent, and used in the
and the residue was stored for anti-fungal study.
treatment of an algesia, ulcer, dysentery and snake bite2-7.
A column of silica gel (400 g) built in CHCl3 and was
The plant has been studied by different workers with
eluted with CHCl3, Chloroform- methanol mixtures of
special reference to its pharmacological activity but no
increasing polarity. The homogeneity of the fractions was
isolation of phytochemicals has been reported8. Also no
examined by TLC and similar fractions were combined
report on the antifungal effects of Pisonia grandis exists.
and tested for anti-fungal activity (Table 2). Three
This paper reports the anti-fungal effects of its extracts
compounds isolated from the column were also tested for
and with clotrimazole as reference drug.
their anti-fungal activity. Anti-Fungal assay : The anti-fungal activity was assayed
MATERIALS AND METHODS
by Disk diffusion method. Sabouarud’s dextrose agar is
Collection of plant material: The plant material (leaves)
the most suitable medium because fungal growth is
was collected during January- March 2009 in the local
favored by a high sugar concentration and is relatively
areas of Coimbatore, Tamilnadu, India. The identity of
tolerant to acidity (pH 5.4) 9. Nutrient agar plates were
plant material was confirmed by the taxonomist Dr.
prepared when the agar medium was amended with
C.Kunhikannan, Scientist D, Biodiversity Division,
complexes separately. Then the medium was warm and
poured into petri plates. After solidification of the
Coimbatore. The leaves were dried in shade and cut into
medium, mycelia discs (6 mm dia) of the test fungi were
small pieces and then used for phytochemical study.
inoculated at the centre of the plates. Diameter of the
Preparation of leaf extract: Air dried pieces of leaves of
inhibition zone for each fungus was measured after 48 hrs
Pisonia grandis were extracted with 100% ethanol for 6
hour at reflux temperature. The extract was filtered; thefiltrate was evaporated to one tenth volume under reduced
RESULTS AND DISCUSSION
pressure to get a greenish black pasty solid (sample A).
Table 1 shows the results of anti-fungal activity. The leaf
Fractionation and column chromatography Procedure: A
ethanol extract and the various fractionated portion of the
small portion of the sample A was set aside for testing its
leaf extract showed varying degrees of inhibition against
anti-fungal activity and the rest was macerated with equal
all the fungal stains but only the ethanol soluble extract of
volume of water and extracted with equal volumes of
Pisonia grandis possesses maximum anti-fungal activity
chloroform (CHCl3). The Liquid liquid extraction (LLE)
with CHCl3 was continued until the CHCl3 layer was
clotrimazole. Hence the ethanol extract of the leaves was
colorless. Then the entire CHCl3 and aqueous layer were
selected for column chromatographic analysis. *Author for correspondence: Email: [email protected]Shubashini K. Sripathi, Poongothai G./ Bioassay- Guided Fractionation…
Table 1 Anti-fungal screening result (Zone of inhibition in mm)Sample
Table 2 Anti-fungal screening result for column fractions and for isolated compounds against Monascus purpureus
Table 2 shows anti-fungal screening result for column
Alloxan-induced diabetic rats. International journal
fractions against Monascus purpureus and column
of Applied Research in natural products. 2009;
fractions 4, 5,6,7,8 and 11 showed a higher zone of
inhibition than the standard Clotrimazole. The Minimum
Radha R, Arokiyaraj S, Agastian P, Balaraju K,
Inhibitory Concentration (MIC) values of these column
Mohan Kumar R, Bula P. Phytochemical analysis
fractions were also shown in Table 2. This study also
and anti-inflammatory activity of Pisonia grandis
revealed the potentially active fractions from which the
R.Br. Journal of Biomedical and Pharmacology.
active principles would be isolated in the same
Prabu D, Nappinnai M, Ponnudurai K, Prabhu K. Evaluation of Wound-Healing Potential of PisoniaACKNOWLEDGEMENTS grandis R.Br: A Preclinical Study in Wistar Rats.
The authors thank the authorities of Avinashilingam
The International Journal of Lower Extremity
University for women, Coimbatore, India for providing
necessary facilities and technical assistance. Authors are
Anbalagan N, Rajinikanth KN, Kishore Gnanasam S,
also grateful to The SAIF, Indian Institute of Technology
Thomas Leonard J, Balakrishna K, Ramachandran S,
Madras, India for the spectral data.
Sridhar SK. Analgesic, Anti-inflammatory andDiuretic Activities of Pisonia grandis. Journal ofREFERENCE Natural product sciences. 2002; 8(3):97-99.
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Buenz EJ, Johnson HE, Beekman EM, Motley TJ,
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IJPPR, Vol-5, Issue 1, March- May 2013, 1-3
Shubashini K. Sripathi, Poongothai G./ Bioassay- Guided Fractionation…Herbal: Volume I. Journal of Ethnopharmacology.
IJPPR, Vol-5, Issue 1, March- May 2013, 1-3
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