Impact factor: 0.3397/ICV: 4.10 248 PHARMACOGNOSTICAL

Impact factor: 0.3397/ICV: 4.10 248

Krunal et al. / Pharma Science Monitor 5(3) Supl-1, Jul-Sep 2014, 248-255

Pharma Science Monitor 5(3) Supl-1, Jul-Sep 2014

PHARMACOGNOSTICAL STANDARDIZATION OF SAPTAPARNA (ALSTONIA

SCHOLARIS R. Br.) STEM BARK

Kamlesh Bhogayata1, B. R. Patel2, Jignesh Kevalia3, Bhakti Chhaya4, Bharatkumar M. Gohel5,

Krunal Doshi6* 1Assistant Professor, Dept. of Dravyaguna, Sheth J. P. Govt. Ayurved College, Bhavnagar Gujarat, India. 2Assistant Professor, Dept. of Dravyaguna, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India. 3Head, Dept. of Pharmacognosy, Indian Institute of Ayurvedic Pharmaceutical Sciences, 4Assistant Professor, Dept. of Shalakya, Sheth J. P. Govt. Ayurved College, Bhavnagar Gujarat, India. 5Assistant Professor, Dept. of Preventive and Social Medicine, PDU Govt. Medical college, Jamnagar road, Rajkot, Gujarat, India. 6Lecturer, Dept. of Dravyaguna, Indian Institute of Ayurvedic Pharmaceutical Sciences, Gujarat Ayurved University, Jamnagar.

ABSTRACT The bark of Alstonia scholaris R. Br. (Apocynaceae) locally known as ‘Saptaparna’ is reported to have anticancer, anti-helminthic, anti-diarrhoeal, anti-asthamatic, anti-malarial etc., activities. The present work represents the study carried out for quality control of herbal drugs which comprises of macroscopy, microscopy, powder characters and histochemical study of stem bark of A. Scholaris R. Br. Result showed the stem bark was dark grey to brown in colour without any odour and bitter in taste. Microscopy revealed the presence of cork, cambium, phloem, medullary rays and stone cells. Powder microscopy of the sample drug exhibited the occurrence of cork cells, stone cells, prismatic crystals, starch grains, phloem, fibers etc. The presence of starch grains, prismatic crystals, stone cells and fibers were also showed by histochemical study. The present study on pharmacognostical investigation of A. scholaris R. Br. bark will be helpful in developing standards for quality, purity and sample identification of this plant. KEYWORDS: Alstonia scholaris R. Br., Saptaparna, Stem Bark, Macroscopy, Microscopy, Histochemical study. INTRODUCTION

Pharmacognosy has become one of the pillars in areas like pharmacy, medicine, natural product

chemistry and many others allowing scientists to recognize the importance of plants as sources of

medicines. This approach has initiated active research programs either to isolate new lead

compounds or to produce standardized extracts.[1] For this it is very necessary to evaluate various

qualitative and quantitative parameters, which may be helpful in setting standards for particular

medicinal plant/parts of the plant. With the help of these standards one can easily identify and

characterize an individual drug, which may play a major role in maintaining quality and purity of

PHARMA SCIENCE MONITOR

AN INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES

Journal home page: http://www.pharmasm.com



that particular drug and its formulation and prevent it from being adulterated by drug of same or

other genius having low potency.[2]

The present study deals with the standardization of one such medicinal plant Saptaparna-

Alstonia scholaris R. Br. is an important medicinal plant in folklore medicine. The plant belongs

to family Apocynaceae and is native to India. It grows throughout India, in deciduous and ever-

green forests and in plains. Juice of leaves and tincture of the bark acts as a powerful

galactogogue and also used in cases of snake bite. Milky juice of the plant is applied on wounds

and ulcers. The bark is bitter, acrid, astringent, digestive, laxative, thermo genic, antipyretic,

galactogogue, cardiotonic and tonic. It is useful in abdominal disorders, fevers, leprosy, skin

diseases, chronic and foul ulcers, asthma, bronchitis and helminthiasis.[3] The bark extract

induces the cellular immune response at low doses and inhibited the delayed type of

hypersensitivity reaction at high doses.[4] The methanolic extract of this plant exhibits

pronounced anti plasmodial activity.[5] The alkaloid fraction of A. scholaris was found to have

potential anticancer agent.[6] It’s bark extract showed chemo preventive potential against skin

tumor genesis in swiss albino mice.[7] Ethanolic extracts of A. scholaris possess powerful in-vitro

antioxidant activity.[8]

Barks from trees and shrubs constitute an important source of drugs used in Indian System of

Medicine. Just after collection and on drying, morphologically or organoleptically several barks

resemble each other in many respects and may exhibit few minor variations in their size, shape,

outer and inner surface, colour, mode of fracture, odour, tastes, etc. However, microscopical

characters performs the major parts and assists us greatly for the correct identity. Hence, an

attempt has been made to ensure properties of Saptaparna (Alstonia scholaris R. Br.) stem-bark

through pharmacognostical study.

MATERIALS & METHODS

Collection of Sample

Bark of Alstonia scholaris R. Br. was collected from the periphery Gujarat state by Pharmacy

department of IPGT & RA, GAU, Jamnagar. Further the authenticity of the sample was

confirmed by the experts of Gujarat Ayurved University and by comparing their characters

mentioned in various floras.

Preservation

The drug was thoroughly washed with running water and cut into small pieces and preserved in

the solution containing [Formalin: Glacial Acetic Acid: Ethanol 70 % in 5: 5: 90]. While some

pieces were dried on sun and powdered for studying its powder microscopy.



Preparation of illustrations

The section were taken by free hand and their photo graphs were taken by using Canon digital

camera attached to Zeiss microscope and powder characters were drawn with the help of Camera

Lucida lying at the Pharmacognosy department of I.P.G.T. & R.A., Gujarat Ayurved University,

Jamnagar.

Macroscopic & organoleptic evaluation

The characters were studied systematically as per the methods described in the text books of

pharmacogosy. The specimen were observed as such with naked eyes or with the help of

dissecting microscope.[11] Various parameters of the plant material, such as size, shape, colour,

odour and taste of the stem bark were also recorded.[9, 10, 11]

Microscopic evaluation

Thin free-hand sections of the stem bark were made and washed with chloral hydrate solution.

The stain was made with phloroglucinol and conc. HCl solution. Diagnostic characters in TS and

powder of stem bark of A. scholaris R. Br. was studied with and without staining.

Microphotographs were taken using Carl Zeiss binocular microscope.[11]

Histochemical Test

Histochemical tests were performed to detect the kinds of cell wall like lignin, primary

metabolites like starch grains, ergastic substances like crystals of calcium oxalate and calcium

carbonate, etc. present in the sample.[12]

RESULT & DISCUSSIONS

Macroscopic & Organoleptic characters

Pieces are curved or flat or occasionally quilled, measuring 8 – 12cm in length, 5 - 8cm broad

and 5 – 15mm in thickness, externally uneven, transversely and longitudinally fissured, marked

with numerous patches of whitish to silver coloured lenticels; surface internally smooth,

longitudinally striated; fracture outer hard and short, inner granular; fractured surface shows

narrow cork and wide phloem zone traversed by latex canals; colour externally dark grey –

brown, internally brownish buff – dark grey, young bark is somewhat paler; odour not

characteristic; taste is bitter and slightly astringent. (Fig. 1)



FIGURE 1 – MACROSCOPIC CHARACTERS OF INTERNAL AND EXTERNAL

SURFACE OF A. SCHOLARIS R. Br. STEM BARK

Microscopic evaluation:

Diagrammatic section: Diagrammatic TS of bark shows outer wide multi layered cork, lying

underneath the cortex and a very large region of inner phloem traversed with laticeferous canals.

Cortex: Detailed TS shows 80 – 100 rows of narrow, rectangular, thin to thick walled, yellow to

brown coloured cork cells at places thick walled with different lumen stone cells are seen, within

the cork a well-defined, thin walled, broadly rectangular, narrow cells or phellogen or cork

cambium; phelloderm or secondary cortex is thin walled, nearly cubical to rectangular, many

layered, traversed with cubical to oblong, groups of stone cells varies in size and with different

kind of lumen, sometimes with pits; phloem is very wide, parenchymatous, the cells being

discontinuous because of parallel arrangement of uni to biserriate, rectangular, thin walled

medullary rays, radially elongated in the portion nearer the wood but becomes broad towards the

distal ends, laticiferous canals traversed throughout the phloem region; prismatic crystals of

calcium oxalate and simple, spherical starch grains scattered throughout the section. (Fig. 2)

Partly cut TS of the bark of Alstonia scholaris R. Br. shows cork, cortex, and prismatic crystals along with starch grains. (Blue colour shows presence of starch grains).

Partly cut TS of the bark of Alstonia scholaris R. Br. shows cortex region with the arrangement of stone cells.

Partly cut TS of the bark of Alstonia scholaris R. Br. showing lignification of stone cells in cortex

region.



FIGURE 2 – MICROSCOPIC CHARACTERS OF A. SCHOLARIS R. Br. STEM BARK

Powder microscopy

A light brown powder with fine texture and without any characteristic

odour having bitter taste has following distinguished microscopical

characters:

1. The abundant groups of stone cells of various sizes, shapes and thickness

with distinct radiating pits and striations. 2. Plenty of sclereids with highly

thickened and striated walls with various sizes and shapes. 3. The abundant prismatic crystals of

calcium oxalate scattered as such and embedded in parenchymatous cells of phelloderm, phloem

and stone cells. 4. The fragments of parenchymatous tissue and tangentially- longitudinally cut

medullary rays of phloem, embedded with lactiferous canals which at places show bulging

enlargements and are filled with granular contents. 5. The fragments of cork in surface view and

in sectional view; in surface view the cells are thick walled, hexagonal to pentagonal in shape

while in sectional view, the cells are thick walled, rectangular to squarish in shape, often with

lignified and pitted walls. 6. The fragments of beaded hexagonal to elongated, lignified

parenchymatous cells of the phloem. 7. Simple, spherical to oval, starch grains scattered as such

and embedded in the parenchymatous cells. 8. Thick walled, striated, non lignified fibres

occasionally present tin the young bark only. (Fig. 3)

Partly cut TS of the bark of Alstonia scholaris R. Br. showing lignification

of stone cells in cortex region (enlarge view)

Partly cut TS of the bark of Alstonia scholaris R. Br. shows cortex region with broad parenchyma containing abundant starch grains and groups of 5-7 rows of

stone cells.

Partly cut TS of the bark of Alstonia scholaris R. Br. shows broad zone of

phloem consisting phloem parenchyma, phloem fibres, medullary

rays and latex cells.

Longitudinally and tangentially cut arrangement of medullary rays in

TLS view.

Phloem region in large view shows phloem parenchyma traversed with prismatic

crystals of calcium oxalate and starch grains.



FIGURE 3 – POWDER CHARACTERSTICS OF A. SCHOLARIS R. Br. STEM BARK

TABLE NO. 1 - COMPARATIVE STATEMENT OF STEM BARK AND ITS POWDER

FORM

Sr. Characters Stem bark of

A. scholaris R. Br.

Powder of

A. scholaris R. Br.

1. Taste Astringent & Bitter Bitter

2. Color Grayish brown (Ext.)

Light brown Creamiest white (Int.)

3. Odor Not characteristic Not characteristic

4. Surface Rough & Uneven (Ext.

Texture – fine Smooth (Int.)

Histochemical characters

1. Test for Starch grains:

Mount a section of sample in water, add a drop of Iodine, blue colour shows the presence of

starch grains.

2. Test for Crystals of Calcium oxalate:

Cork in sectional view Cork in surface view Part of phloem in

tangential - section

Prismatic crystals, Starch grains,

Parenchyma filled with prismatic crystals.

Beaded phloem parenchyma,

Longitudinal cut fragments of laticiferous

canal

Fibers Sclereides

Stone cells



They are insoluble in glycerin and phenolic reagents but soluble in most acids commonly used

hydrochloric acid. By adding2 - 3 drops of hydrochloric acid crystals of calcium oxalate gets

dissolved.

3. Test for fibres and stone cells:

Fibres of Sclerenchyma and stone cells gives pink to red colour when stain with Phloroglucinol

and HCl.

CONCLUSION

Standardization of herbal drugs is a topic of great concern because of the great variability

derived from heterogeneous sources. Alstonia scholaris R. Br. is a plant with large medicinal

advantages and it’s stem bark plays a major role in these medicinal properties. Thus the present

study will provide referential information for correct identification and standardization of this

plant material through pharmacognostical study as basic tool for identification.

REFERENCES

1. Ameenah and Gurib-Fakim: Review medicinal plants traditions of yesterday and drugs of

tomorrow. Molecular Aspects of Medicine. 2006; 27:1–93.

2. Doshi KA, Acharya RN, Harisha CR and Preeti P: Pharmacognostical evaluation of the

wild and cultivated variety of Eranda (Ricinus communis Linn.) root. Phcog J 2012; 4:

54-58.

3. Kirtikar KR and Basu BD: Indian Medicinal Plants. Lalit Mohan Basu Publishers,

Allahabad, India, Vol. I, 2002: 111-114.

4. Iwo MI, Soemardji AA, Retnoningrum DS and Sukrasno UUM: Immunostimulating

effect of pule (Alstonia scholaris L. R. Br., Apocynaceae) bark extracts. Clinical

emorheology and Microcirculation 2000; 23: 177-183.

5. Keawpradub N, Kirby GC, Steele JCP and Houghton PJ: Antiplasmodial activity of

extracts and alkaloids of three Alstonia species from Thailand. Planta Medica 1999; 65:

690 - 694.

6. Jagetia GC and Baliga MS: Evaluation of anticancer activity of the alkaloid fraction of

Alstonia scholaris (Sapthaparna) in vitro and in vivo. Phytotherapy Research 2006; 20

(2): 103-109.

7. Baliga MS: Alstonia scholaris Linn R Br in the Treatment and Prevention of Cancer:

Past, Present, and Future. Integrative Cancer Therapies 2010; 9 (3): 261-269.



8. Arulmozhi S, Mazumder PM, Ashok P and Narayanan LS: In vitro antioxidant and free

radical scavenging activity of Alstonia scholaris Linn. R. Br. Iranian Journal of

Pharmacology and Therapeutics 2007; 6: 191-196.

9. Kokate CK, Purohit AP and Gokhale SB. Pharmacognosy. Nirali Prakashan, Pune,

Edition 2, 2008: 63.

10. Anonymous: Ayurvedic Pharmacopoeia of India. Govt. of India, Ministry of Health of

Family Welfare, New Delhi, Edition 1, Vol-2, Part-2, 2008: 15-17.

11. Khandelwal KR: Practical Pharmacognosy - Techniques and Experiments. Nirali

Prakashan, Pune, Edition 9, 2002: 24-29,149-156.

12. Krishnamurthy KV: Methods in the Plant histochemistry. Vishwanandhan Pvt Limited,

Madras, 1988: 1-77.

For Correspondence Krunal Doshi Email: [email protected]

Documents

Impact factor: 0.3397/ICV: 4.10 248 PHARMACOGNOSTICAL