INTERNATIONAL JOURNAL OF PHARMACEUTICAL … 744.pdf · Research Article CODEN: IJPRNK IMPACT...

Research Article CODEN: IJPRNK IMPACT FACTOR: 1.862 ISSN: 2277-8713 Kumar S, IJPRBS, 2014; Volume 3(3): 509-518 IJPRBS

Available Online at www.ijprbs.com 509

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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Kumar S, Singh D, Dwivedi KN; IJPRBS, 2014; Volume 3(3): 509-518



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 .



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



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)



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



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



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



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



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



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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3. Kirtikara. K.R., Basu. B.D, Indian medicinal plants, Vol. E. Blatter and J.F. Caius editor. 2nd ed.

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