phytochemical of pterospermum

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This article can be downloaded from www.ijpbs.netP - 356

ISSN 0975-6299 Vol 3/Issue 1/Jan Mar 2012

RESEARCH ARTICLEInternational Journal of Pharma and Bio Sciences

PHARMACOGNOSTIC AND PRELIMINARY PHYTOCHEMICAL

INVESTIGATIONS ONPTEROSPERMUM ACERIFOLIUMWOOD

RASIKA D. BHALKE *1

AND SUBODH C. PAL2

1Department of Pharmacognosy, Sanjivani College of Pharmaceutical Education and Research,

Kopargaon, Maharashtra, India-423603.2N. D. M. V. P. Ss College of Pharmacy, Nashik, (MS), India-422002

PHARMACOGNOSY

RASIKA D. BHALKE

Department of Pharmacognosy, Sanjivani College of Pharmaceutical Education and

Research, Kopargaon, Maharashtra, India-423603.

ABSTRACT

Pterospermum acerifolium plant is considered to be laxative, anthelmintic, stomachic andused in inflammation, blood disorders, ulcers and leprosy. The wood was studied forpharmaconostic evaluations, including examination of morphological and microscopiccharacters, determination of ash values and extractive values. The morphological studiesrevealed that the wood is yellowish in color with smooth texture and characteristic odour.Qualitative studies indicated the presence of uni to biseriate medullary rays, pittedxylem fibers, big vessels in groups, xylem parenchyma, pith, calcium oxalate crystals,starch grains, lignified fibers. The total ash, acid insoluble ash and water-soluble ashvalues were observed to be 3.3%, 1.9 % and 1.1 % respectively. The wood wassuccessively extracted with petroleum ether, chloroform, ethyl acetate and methanol inincreasing order of polarity. Preliminary phytochemical investigation showed the presence

of alkaloids, glycosides, tannins, triterpenoids, carbohydrates and flavonoids. Totalphenolic content was determined using a spectrophotometric technique, based on theFolin-Ciocalteau reagent, and calculated as gallic acid equivalents GAE/g. These findingswill be useful towards establishing pharmacognostic standards on identification, purity,quality and classification of the plant, which is gaining relevance in plant drug research.


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KEYWORDS

Pterospermum acerifolium, Pharmacognosy, Drug standardization, Sterculiaceae

INTRODUCTION

Pterospermum acerifolium (L) Willd (Family:Sterculiaceae) commonly known as Dinner platetree (English) and Muchukunda (Hindi), is alarge deciduous tree of about 24 m height and2.5m girth. Flowers are large 12-15 cm indiameter, axillary, solitary or in pairs. It is widelydistributed in North Canada and in many partsIndia i.e. river banks of sub-Himalayan tracts,

Dehradun, West Bengal, Assam and Manipur1-2

.In traditional system of medicine, the flowers areused as a general tonic, anti tumor agent,analgesic and for the treatment of diabetes,gastrointestinal disorders, leprosy, bloodtroubles, bronchitis, cough, cephalic pain,migraine and inflammation. The wood is used ashaemostatic and antimicrobial agent 3-6. Themain aim of the present investigation is to studythe macro, microscopic and some otherpharmacognostic characters and

physicochemical standards of wood ofPterospermum acerifolium which could be usedto prepare a monograph for the properidentification of the plant.

MATERIALS AND METHODS

Samples of wood of Pterospermum acerifoliumwas collected from Nasik district of Maharashtraand authenticated at Botanical Survey of India,Pune, where a sample (voucher number-

RASPTA1) has been deposited.

Qualitative InvestigationThe macroscopic features of the fresh wood ofPterospermum acerifolium were determinedusing the method of Evans7. Anatomical sectionsand powdered samples for the microscopy andchemo-microscopy were carried out according tomethods outlined by Brain and Turner8.

Quantitative InvestigationThe moisture content, ash and extractive valuesof the powdered wood samples and thequantitative microscopy on the anatomicasection were carried out as described in theIndian Pharmacopoeia 8 and Khandelwal 9.

Preliminary Phytochemical Investigation

The preliminary phytochemical investigation wasdone by the standard chemical tests of Evansand Brain and Turner10.

Fluorescence analysesThese analyses were carried out as per thestandard procedures 11. In the present study, thepowdered wood were treated with variouschemical reagents like aqueous 1N Sodiumhydroxide, alcoholic 1N sodium hydroxide, 1Nhydrochloric acid, 50% sulphuric acid and 50%

nitric acid and their extracts were subjected tofluorescence analysis in day light and UV light(254 nm and 366 nm).

Estimation of Total Phenol12

Ten milligram of standard gallic acid wasdissolved in 100 ml distilled water in a volumetricflask (100g/ml of stalk solution). From theabove stalk solution 0.5 to 2.5 ml of aliquotswere pipetted out into 25 ml volumetric flasksTen ml of distilled water and 1.5 ml of folin

ciocalteus reagent (diluted according to the labespecification) were added to each of the abovevolumetric flasks. After 5 min, 4 ml of 20%sodium carbonate solution was added and thevolume was made up to 25 ml with distilled waterand incubated at room temperature for 30 minand the absorbance of the solution was recordedat 765 nm and a standard curve of absorbanceverses concentration of gallic acid (50-250 g)


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was plotted. One gram of the powdered drugwas extracted with 70% methanol (15x3 times),filtered, pooled and the volume was adjusted to50 ml with 70% methanol in a volumetric flask.

From the stock solution, suitable quantity of theextract was taken into a 25 ml volumetric flaskand 10 ml of water and 1.5 ml of Folin Ciocalteureagent were added to it. The mixture was keptfor 5 min, and then 4 ml of 20% sodiumcarbonate solution was added and made up to25 ml with double distilled water. The mixturewas incubated at room temperature for 30 minand the absorbance was recorded at 765 nm in aspectrophotometer. Percentage of totalphenolics was calculated from calibration curve

of gallic acid (50-250 g) plotted using the aboveprocedure and total phenolics were expressedas milligrams of gallic acid equivalent per gramof dry weight (mg GAE/g dw).

RESULTS AND DISSCUSSION

The wood is yellowish when freshly cutand turns into brown-colored when exposed.Microscopically transverse section showedpresence of uniseriate to biseriate medullaryrays of about more than 50 cells high and oneto two cells wide, thick walled pitted, arrangedin radial rows. Xylem fibers were observed.Vessels are big, found either single or in groupsof few. Small polygonal, less thickened xylemparenchymatous cells were observed. Pith wasobserved at the center. Powder charactersshowed presence of lignified fibers, calciumoxalate crystals, starch grains and vessel.

The drug extract in solvents likepetroleum ether, chloroform, ethyl acetate andmethanol exhibits specific colour reactions whentreated with different reagents, thereby indicatingthe presence or absence of certainphytochemicals in the drug. Preliminaryphytochemical investigation showed thepresence of alkaloids, glycosides, tannins,

triterpenoids, carbohydrates and flavonoids asshown in Table 1. From the phytochemicascreening it can be concluded that plant extractshave potential secondary metabolites which can

be used to discover bioactive natural productsthat may serve as a leads for the development ofnew pharmaceuticals for therapeutic needsSuch screening of various natural organiccompounds and identifying active agents is theneed of the hour, because successful predictionof lead molecule and drug like properties at theoutset of drug discovery will pay off later in drugdevelopment.

For the gallic acid, the curve absorbanceversus concentration is described by the

equation y = 0.464x + 0.0181 (R2 = 0.999)Here, y = absorbance and x = concentration. Theamount of total phenolics was found to be 32.25mg GAE/g of dry material. The estimation ofphenolic content of P. acerifolium was doneusing FolinCiocalteu reagent that producedblue colour. It was observed that more thenumber of hydrogen donating groups in thephenolic compounds; more the intensity of bluecoloured complex that indicated the higher totaphenol content. The quantitative determinations

of some pharmacognostic parameters are usefufor setting standards for crude drugs.The physical constant evaluation is animportant parameter in detecting adulteration orimproper handling of the drug. Various ashvalues are important to determine the purity ofthe drug i.e the presence or absence of foreigninorganic matter. Since the plantP.accerifolium is useful in the traditionamedicine for the treatment of some ailmentit is important to standardize it for use as a

drug. The pharmacognostic constants for thewood of this plant, the diagnostic microscopicfeatures and the numerical standards reportedin this work could be useful for the compilationof a suitable monograph for its properidentification.


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Table 1Physico-chemical characters of the powdered leaves of P. acerifolium

Evaluation parameters Value (% w/w)*

Moisture content 5.1

Total ash value 3.3

Water-soluble ash value 1.1

Acid-insoluble ash value 1.9

Extractive values

a) Petroleum ether (40 600C) 1.29

b) Chloroform 2.06

c) Ethyl acetate 4.94

d) Methanol 9.12*Mean value of six counts

Table 2Preliminary phytochemical investigation of various extracts of leaves of P. acerifolium

Test for activeConstituents

Petroleumether extract

Chloroformextract

Ethylacetateextract

Methanolextract

Steroids + + - -

Triterpenes + - - -

Saponine ++ + - -

Alkaloids - - - +

Tannins - - - +

Flavonoids - - + +

Glycosides - - + +

Table 3Fluorescence analysis of powdered drugs under ultra-violet light

Powder and reagent Color in ordinary light Color in UV light.

Powder Brown Dark brown

Powder+ Nitrocellulose Faint brown Faint greenish.


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Powder+ NaOH in Methanol Brownish Orange brown

Powder+ NaOH in Methanol +Nitrocellulose

Blackish brown Dark green

Powder+ 1N NaOH in water Dark reddish brown Brown

Powder+ 1N NaOH in water+Nitrocellulose

Dark reddish brown Yellowish

Powder + HCl Yellowish Green

Powder + HCl + Nitrocellulose Faint yellow Faint brown

Powder+ HNO3 Faint brown Brown

Powder+ H2SO4 Faint yellowish Dark yellow

Figure. 1T.S. ofPterospermum acerifolium wood

Figure. 2T.S. of Pterospermum acerifolium wood showing lignified xylem region


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Figure . 3T.S. of Pterospermum acerifolium wood showing central pith

Figure. 4

T.S. of Pterospermum acerifolium wood showing xylem region

Figure 5

prismatic calcium oxalate crystal

REFERENCES

1. The wealth of India, A dictionary of Indianraw materials and Industrial product.

Publications and Information DirectorateCSIR, New Delhi. 8, pp: 308-311. (2005).


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2. Kritkar KR and Basu BD. Indian medicinalplants. 2nd ed. Bishen Singh and MahendraPal Singh publishers, Dehradun, India. pp:373-376. (1998).

3. Agarwal VS. Drug Plants of India, Vol.II,Kalyani Publishers, New Delhi, pp: 590-591. (1964).

4. Chopra IC and Chopra RN. Glossary ofIndian Medicinal Plants, Drug ResearchLab, CDRI, Lucknow, pp: 206-207. (1956).

5. Sengupta KN. Headache: The Ayurvedicsystem of Medicine, 1st ed. Vol-I, NeerajPublishing House, New Delhi, pp: 428-443.(1984).

6. Singh VK, Govil JN, Sharma H and Singh

G. Recent progress in medicinal plant andEthnomedicine and Pharmacognosy, vol-VII, Studium Press Lic, New Delhi, pp: 247.(2003).

7. Evans WC. Trease and EvansPharmaconosy, 14th ed., W.B. SoundersCompany Ltd., London, pp: 545-546.(1996).

8. Brain KR and Turner TD. The PracticaEvaluation of Phytopharmaceuticals, WrightScience Technica, Bristol, pp: 81-82(1975).

9. Anonymous. The Indian Pharmacopoeia4th ed., Vol. II, The Controller ofPublications, Government of India, NewDelhi, pp. A-53, A-54; A-089. (1996).

10. Khandelwal KR. Practical Pharmacognosy10th ed., Nirali Prakashan, Pune, pp. 162-65. (2003).

11. The Ayurvedic Pharmacopoeia of India,Part I, Vol III, Government of India, Ministryof Health and Family Welfare., New Delhi.,235. (1999).

12. Singelton, V., R., Orthifer, R., Lamuela-Raventos, R., M., '' Analysis of total phenolsand other oxidation substrates andantioxidants by means of Folin-Ciocalteureagent'' Methods in Enzymology. 299, pp152-178. (1999

phytochemical of pterospermum

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