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PHARMACOGNOSTICAL AND PHYTOCHEMICAL EVALUATION OF BALANITES AEGYPTIACA LINN. DELILE. SEED.
KUMAR S1, SINGH D2, DWIVEDI KN1
1. Department of Dravyaguna, I.M.S, B.H.U, Varanasi, India.
2. Department of Kayachikitsa, I.M.S, B.H.U, Varanasi, India
Accepted Date: 19/06/2014; Published Date: 27/06/2014
Abstract: The present work deals with the pharmacognostical and preliminary phytochemical
studies on the seeds of Balanites aegyptiaca Linn. Delile1. Pharmacognostical parameters for
the seeds of Balanites aegyptiaca Linn. Delile were studied with the aim of drawing the
pharmacopoeial standards for this species. Macroscopical and Microscopical Characters,
physico-chemical constants, quantitative microscopy parameters, extractive values with
different solvents, fluorescence analysis of dry powder, its reaction after treatment with
chemical reagents under visible light and UV light at 254 nm and 366 nm.
Preliminary phytochemical screening on the seeds of Balanites aegyptiaca Linn. Delile were
studied. The determination of these characters will help future researchers in
their Phytochemical as well as Pharmacological analyses of this species.TLC profile was
established for successive extracts of the seed using TLC system (Figure 8).
Keywords: Balanites aegyptiaca, Pharmacognosy, Phytochemical, Macroscopic, Microscopic.
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Corresponding Author: DR. SANJEEV KUMAR
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Kumar S, Singh D, Dwivedi KN; IJPRBS, 2014; Volume 3(3): 509-518
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INTRODUCTION
Since ages, plants have remained important sources of medicines in our country, which is
evidenced through their uses in traditional system of medicine i.e. Ayurveda, Siddha, Unani,
Homeopathy and Chinese (Mukherjee, 2010). The earliest documentation about the usage of
plant remedies comes from India as evident from Rigaveda (2000 BC)2 where Aushadhi sukta
includes a good number of plants for various ailments. Realizing the importance of medicinal
plants as a natural source of newer medicines, now the world is moving towards the plant
based medicine or phytomedicines that repair and strengthens bodily systems (especially the
immune system, which can then properly fight foreign invaders) and help to destroy offending
pathogens without toxic side effects. The major limitation of modern medicine, being its
adverse effects has opened new doors to Ayurveda, where physicians of the traditional systems
of medicine are using plants from many years. In developed countries too, people are seeking
alternative to modern therapies. In order to make sure the safe use of these medicines, a
necessary first step is the establishment of standards of quality, safety and efficacy. Keeping
this fact in to consideration, the attempts were made to establish physicochemical standards of
the plant Balanites aegyptiaca (Linn) Delile. Balanites aegyptiaca (Linn) Delile (Hindi- Hingen,
Hingan, Hingol, Hingot)3 belonging to family Simaroubaceae.
MATERIAL & METHOD
Drug was collected by rural area of district Itawah, U.P and identified by the teacher of
Dravyaguna department in Faculty of Ayurveda B.H.U Varanasi. Macroscopic and microscopic
evaluation was carried out with seed. Seed was pulverized in the mechanical grinder to a
moderate fine powder to carry out microscopic studies and was stored in a well closed
airtight vessel for further analysis. All reagent and chemicals used for the study were of
analytical grade4,5.
PHYSICO-CHEMICAL EVALUATION
MACROSCOPIC AND MICROSCOPIC EVALUATION
MACROSCOPIC CHARACTERSTIC OF SEED POWDER OF BALANITES AEGYPTIACA LINN. DELILE.
In present study the Powder of seeds of Ingudi (Balanites aegyptiaca.Linn.Delile) were
investigated for its macroscopic and microscopic characterstics.
MATERIALS:
1. Powdered seeds of Ingudi (Balanites aegyptiaca.Linn.Delile).
2. Petri dish .
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METHOD:
5 gm powder was taken in a petri dish and examined with naked eye.
MICROSCOPIC CHARACTERSTIC OF SEED POWDER OF BALANITES AEGYPTIACA LINN. DELILE.:
The seeds of Ingudi (Balanites aegyptiaca.Linn.Delile) were pulverized in to fine powder. The
powder was investigated for their microscopic characterstics.
MATERIALS:
1. Powder of Ingudi (Balanites aegyptiaca.Linn.Delile).
2. Phloroglucinol
3. Conc. HCl
4. Microscope
5. Glycerine
6. Slide & Coverslip
7. Watch glass
METHOD:
5 gm powder of seeds was boiled separately with chloral hydrate solution in small quantity.
Cleaved powder was removed in three separate watch glasses respectively and stained with
one drop each of phloroglucinol and conc. HCL. A little of the treated powder was mounted in
dilute glycerine and the slide was observed under microscope at low power.
The morphological studies and the analysis of the seed powder revealed that, it is creamish
white in colour with aromatic odour.
STANDARDIZATION OF SEED OF BALANITES AEGYPTIACA LINN. DELILE.:
In the present study, the seeds of Ingudi (Balanites aegyptiaca.Linn.Delile) were collected from
Itawah region in UP. After authentication, seeds were dried at room temperature until they
were free from the moisture and subjected to physical evaluation with different parameters.
The parameters used for evaluation are nature, odour, colour, taste and texture, Determination
of petroleum ether soluble extractive value, ethanol soluble extractive value, water soluble
extractive value, loss on drying, determination of total ash, acid insoluble ash, water insoluble
ash, fluorescence analysis of the drug, determination of foreign matter by different procedure6.
PHYTOCHEMICAL SCREENING
Qualitative examination of inorganic matters & determination of heavy metals was done as per
reported methods. The dried powdered bark was subjected to preliminary phytochemical
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screening for qualitative detection of phytoconstituents. For further studies extraction was
done by soxhlet apparatus. The shade dried seeds of Ingudi (Balanites aegyptiaca .Linn.Delile)
family Simarubaceae7 ) were reduced to fine powder (#40 size mesh) and around 45 gm of
powder was subjected to successive hot continuous extraction (soxhlet) with petroleum ether
( 60- 800C), ethanol and distilled water. Each time before extracting with next solvent the
powdered material was air dried in hot air oven below 500C. After the effective extraction , the
solvent was distilled off, the extract was then concentrated on water bath and the extract
obtained with each solvent was weighed. Its percentage was calculated in terms of air dried
weight of plant material, Colour and consistency of the extract was noted.
QUALITATIVE CHEMICAL INVESTIGATION OF EXTRACT
Qualitative tests were conducted for all the extracts of seeds of Balanites aegyptiaca
.Linn.Delile to identify the various phytoconstituents.
RESULTS
MACROSCOPIC EVALUATION
The Macroscopic characters found are discussed in Table 1.
EXTERNAL MORPHOLOGY OF SEED OF BALANITES AEGYPTIACA LINN. DELILE.
External morphology and seed of Balanites aegyptiaca .Linn.Delile. are shown in (Figure 1,2)
TRANSVERSE SECTION OF SEED OF BALANITES AEGYPTIACA LINN. DELILE. SHOWS:(Figure 7)
1. Epidermis: large, thick walled lignified polygonal tabular cells with slit like pits.
2. Epidermal trichomes: thick walled, bend, twisted, lignifed with about 10 longitudinal ribs on
the limb.
3. Inner layer of epidermis with collapsed cells.
4. Endosperm: thick walled cellulosic parenchyma of isodiametric cells.
5. Embryo and endosperm both contain aleurone grains and fixed oil globules.
MICROSCOPIC CHARACTERSTICS OF POWDERD DRUG:
TRICHOMES: Entire or fragments of trichomes, lignified. Base is pitted and broad. Apex is
rounded. Ridges are present on the surface.
ENDOSPERM: Polygonal cellulosic cells, with oil globules and aleurone grains. Calcium oxalate
crystals and starch grains are absent. (Figure 4,6)
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PHYSICO-CHEMICAL EVALUATION
The physicochemical studies and Successive extractive values of Seeds of Balanites aegyptiaca
Linn. Delile. Summarized in Table 2 and Table 3.
PHYTOCHEMICAL SCREENING
The results demonstrated presence of Saponin glycosides , Volatile oils, Proteins, Amino Acid,
Fat & Oils, Steroids flavanoids, Tannins and Phenolic Compounds, mainly in the seeds of
Balanites aegyptiaca. The presences of various phytoconstitutes in various extracts are
summarized in Table 4.
Table1: Macroscopic Characteristic of seed powder of Ingudi (Balanites aegyptiaca. Linn.
Delile).
S.No Parameters Observation of seeds
1 Nature Coarse powder
2 Colour Creamish yellow
3 Odour Aromatic
4 Taste Bitter
5 Texture Rough & fibrous
6 Size Uneven sized coarse particles
Table No 2: Phisico-Chemical Properties of Seeds of Balanites aegyptiaca.Linn.Delile
S.no Parameters Observation
Seeds
I Physical tests
Nature Coarse powder
Colour Creemish white
Odour Aromatic
Taste Bitter
II Extractive value (% w/w)
Pet. ether 2
Hydro-alcohol 1.1
Aqueous 2.5
III Loss on drying (%w/w) 9
IV Foreign matter Nil
V Fluorescence No fluorescence
VI Ash value (% w/w)
Total ash 4.5
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Acid insoluble ash 6.5
Water insoluble ash 2
Table No 3: Succesive Extractive Values of Seeds of Balanites aegyptiaca.Linn.Delile.
S.No Extracts Nature of Extract Weight (gm) % Yield w/w
I Petroleum Ether Oily 40 40
II Hydro-alcohol Viscous 22.7 22.7
III Aqueous Powder 19 19
Table No 4: Phytochemical Investigation of Extracts of Seeds of Balanites aegyptiaca. Linn.
Delile
S.No Chemical Tests Powder drug
Tests for Carbohydrates
I Molish’s test -ve
Test for Volatile Oils
I Odour +ve
II Filter paper Stain test +ve
III Solubility test +ve
Test for Proteins
I Biuret test +ve
II Xanthoproteic test +ve
III Precipitation test with: +ve
(i) Lead acetate solution 5% +ve
(ii) CuSO4 solution 5% +ve
Test for Amino acids
I Ninhydrin test +ve
Test for Fat & Oils
I Solubility test +ve
II Filter paper stain test +ve
Test for Steroids
I Salkowaski test +ve
II Liebermann-burchard test +ve
III Liebermann’s reaction +ve
Test for Saponin Glycosides
I Foam test +ve
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II Haemolytic test +ve
Test for Tannins & Phenolic Compounds
I 5% Fecl3 solution +ve
II Lead acetate solution +ve
III Dilute iodine solution +ve
IV Dilute HNO3 +ve
V Dilute KMnO4 solution +ve
VI Potassium dichromate solution +ve
VII Bromine solution +ve
VIII Acetic acid solution +ve
IX Gelatin solution +ve
X NH4OH + AgNO3 +ve
XI NH4OH + potassium ferricyanide +ve
Test for Vitamin C -ve
Figure1 Balanites aegyptiaca.Linn.Delile Figure2 Seeds of Balanites aegyptiaca.Linn.Delile
POWDER MICROSCOPY OF SEED OF BALANITES AEGYPTIACA LINN DELILE.:
Lignifed Tissues:
Xylem (Vascular
Bundle)
Figure 3: Lignified tissue: xylem (Vascular Bundle) Phloroglucinol + Conc. HCL
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Figure 4: Oil globules Present in the Cells of Endosperm. Sudan Red III, RED
Figure 5: Hemicellulose- Endospermic Wall (Plasmodesma) Dil.Iodine + Conc. Sulpuric
Acid, Blue Colour.
Figure 6: Aleurone Grain Present in the cells of Endosperm Alc. Picric Acid, Yellow Colour.
Oil globules Present in the
Cells of Endosperm
Hemicellulose- Endospermic
Wall (Plasmodesma)
Aleurone Grain Present in
the cells of Endosperm
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Figure 7: Transverse section of seeds of Balanites aegyptiaca.Linn.Delile.
Pet. Ether Extract Hydroalcoholic Extract
Figure 8 Thin Layer Chromatography of Balanites aegyptiaca.Linn.Delile.
Oil Globules
Endospermic Wall
Trichomes
Aleurone grains
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DISCUSSION AND CONCLUSION
‘Pharmacognosy’ is meant by identification of drugs by its every aspect, habit, cultivation,
procurement, micro and macroscopic characters, physical and chemical properties etc. In
present study pharmacognostical standards have been established with regards to seed of
Ingudi (Balanites aegyptiaca Linn.Delile, Local name Hingot). Powder microscopy of seeds
showed the presence of trichomes and endosperms containing polygonal cellulosic cells (Figure
5) and xylem containing Lignified cells (Figure 3) with oil globules, and aleurones grains, Calcium
oxalate crystals. The physical evaluation furnished different ash values, extractive values in
different solvents. Total ash, acid insoluble ash and water soluble ash values were also
determined. The phytochemical investigation shows the presence of Saponin glycosides,
Volatile oils, Proteins, Amino Acid, Fat & Oils, Steroids, Flavanoids, Tannins and Phenolic
Compounds in the seeds of Balanites aegyptiaca. Study was carried out in order to assess the
quality of seeds of Balanites aegyptiaca Linn. Delile and also to detect the adulteration and
substitution etc., which may be helpful to researchers in future.
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