Pharmacognostical and Preliminary Phytochemical Studies on the Stem Bark of Soymida febrifuga (Roxb)

Malarkodi Velraj¹*, Armstrong¹, Ravichandran¹, Jeyakumari¹, Hemalatha¹, Vijayalakshmi¹ and Srikanth.J²

¹Department of Pharmacognosy, VELS College of Pharmacy, VELS University, Pallavaram, Chennai-600 043, Tamil Nadu, India.

²Department of Pharmacology, Sri Ramachandra College of Pharmacy, Sri Ramachandra University, Porur, Chennai-600 116, Tamil Nadu, India.

ABSTRACT

Soymida febrifuga is commonly known as Shem in Tamil and Indian red wood in English. In Ayurveda the bark is used as a good cure for ulcer, leprosy and dysentery. In unani the bark is used as an astringent to the bowel and useful in fever. The bark is also used in vaginal infection, rheumatic swellings, oedema, wounds, dental problems, uterine bleeding and malarial fever. There was no report on the pharmacognostic study of the plant and hence the present investigation deals with the anatomical, microscopical, powder microscopy, physio-chemical, fluorescence analysis, Thin layer chromatography and high performance thin layer chromatography. The study revealed the presence of drugs of calcium oxalate, tannins dilated phloem rays, sieve elements and schlerids. Phytochemical studies showed the presence of steroids, terpenoids, flavanoids and tannins.

KEYWORDS: Soymida febrifuga, TLS, RLS

INTRODUCTION:

Soymida febrifuga (Roxb) Meliaceae syn Swietemia febrifuga is a very large tree of 70-80 feet high. Leaves are paripinnate, leaf lets are 6 pairs, opposite, ovate-oblong, entire, obtuse or retuse. Flowers are greenish-white in auxiliary or terminal panicles. Sepals-5 imbrigate. Petals-5 clawed. Stamens-10 in a staminal tube, ovary 5 – locular, ovules numerous per locule, pendulous, stigma 5 – angled. The bark is refrigerant, anthelmintic, aphrodisiac, laxative good for sore throat and used in advanced stages of dysentery and in diarrhea. Bark contains a resinous bitter principle and is a popular drug in indigenous medicine ^{1, 2}.

MATERIALS AND METHODS:

The bark was collected from Tirupati and authenticated by Dr.K.Madhavachetty Department of botany; Sri Venkateswara university and given the accession number 1312 and a voucher specimen is deposited in the department of Pharmacognosy, Vel’s college of pharmacy. Fresh plants were used for macroscopic and microscopic studies. The plant parts were fixed in FAA and dehydrated with graded series of tertiary butyl alcohol. Microtome stem were stained with toludine blue as per standard method ³. Powder microscopy was done according to standard procedure ⁴. Physiochemical standard were done according to WHO guidelines ⁵. Fluorescence analysis was done by standard procedure ⁶. Histochemical reaction was followed from crom well ⁷. TLC was followed using standard procedure from Wagner⁸. Quantitative microcopies were followed from Trease and Evans ⁹.

RESULTS AND DISCUSSION:

Macroscopic Features: (fig 1and2)

The older bark is compact, thick fibrous pieces which are rusty, grey or brown colour externally red on the inner surface. Bark obtained from young tree is straight or somewhat curved; half tubular quills an inch or more in diameter and about ½ an inch in thickness. It exhibits furrows or crack but numerous small corky warts. These form little elliptic scars or rings brown in centre. The inner side and edges of quills are of a bright reddish brown. The bark has a fibrous fracture, without any distress odour but with a bitter astringent taste.

Fig 1: Pm=Periderm, PeF=periderm flake

Fig 2: Ph=Phloem, Pe=Periderm

Microscopic Characters:

The bark is divided into outer bark and inner bark. Outer bark is the periderm part of the bark. It is 250μm thick and consists of dark staining outer part of phellem and a wide light stained Phelloderm (fig.3). The phellem cells are suberised. Phelloderm is wider, thin walled and living tissue. It is 400μm thick and consists of narrow and wide tubular cells which alternated in radial files.

Fig 3: Pm=Periderm, Pd=Phellem, CPh=Collapsed Phloem

The inner bark consists of two regions i.e. outer zone of crushed and collapsed phloem tissue and inner zone of intact phloem.

The outer collapsed phloem (fig.4) much dilated with wavy phloem rays. The rays become further distortered in the outer region towards the periderm. The sieve elements are crushed into dark patches. Tannin content and calcium oxalate crystals are heavily deposited into collapsed phloem. Phloem sclerenchyma are scattered into small masses in this region.

Fig 4: CPh=Collapsed Phloem, NCPh= Non Collapsed Phloem, PhR=Phloem ray

The inner zone of intact –phloem lies outer to the secondary xylem (fig 5). It is narrow and comprise of intact sieve elements. Phloem rays are narrow and straight. Small blocks of phloem sclerenchyma or sclerids are seen in regular tangential bands (fig 6).

Fig 5:CPh=Collapsed Phloem, PhR=Phloem ray

Fig 6: PhR=Phloem ray, PhS=Phloem sclereid, SC=Sieve cells

TLS View of the Bark:

In TLS the ray serration, ray dimension, their arrangement, structure and dimension of the sieve tube and phloem parenchyma cells were studied. Medullary rays are not studied. They are wide semi seriate occasionally biseriate comprising one type of cells which are squarish or rectangular in shape. These rays are 220-700μm high and 20 - 100μm wide.

The sieve tube numbers are narrow and short. They are 300-320μm in height and 30-40 μm in width. The sieve plate is simple and oblique. The companion cells are narrow and occur in vertical row of many cells (fig 7).

Fig 7: PhP=Phloem parenchyma, PhR=phloem ray, ST=Sieve tube

RLS View of the Bark:

In RLS view the phloem rays are seen in horizontal wide bands consisting of rectangular cells. Phloem sclerenchyma occurs in vertical parallel pillars (fig 6). The periderm consists of wide bands of radially arranged rectangular narrow and wide Phelloderm cells (fig 8).

Fig 8: Cr=Crystals, PhS=Phloem sclereid, PhR=Phloem ray

Crystal Distribution:

Calcium oxalate crystals are abundant in the collapsed phloem tissue. There are two types of crystals seen in the phloem (fig 9). Druses and prismatic crystals are equally abundant. The crystals occur mostly in the phloem parenchyma cells. The crystals are large and each crystal occupies the entire lumen of the cell. When seen in LS view the crystals are vertical continuous strands (fig 10). Prismatic crystals are mostly associated with the sclerid masses. The prismatic crystals are up to 50μ thick and druses are 40μm thick.

Fig 9: Scl=Sclerids, Dr=Druses, PC=Prismatic crystals

Fig 10: Dr=Druses, PC=Prismatic crystals

Powder Microscopy:

The powder of the bark is reddish in colour with aromatic odour. The taste is bitter and the texture is coarse.

These fibre sclerids cells are fibre like and elongated with tapering ends. But their walls and pits are like sclerids. The walls are thick with wide lumens and pits are canal like and simple. The cells are narrow or wide and are about 950μm long and 30-40 μm thick. When the sclerids are seen under the polarized light microscope they appear bright indicating that the walls are lignified (fig 11and12). Crystals are abundant in powder. They are calcium oxalate druses and prismatic type. The druses are scattered in the powder and the prismatic crystals occur in strands.

Fig 11: FSc= Fibre sclereids

Fig 11: FSc= Fibre sclereids

Transverse section of the bark was treated with routine chemicals and results are tabulated. It gave positive test for steroids, tannins, terpenoids and flavonoids and results are tabulated in Table no: 1

Table No 1:Chemical Tests of the transverse section of the bark
S.No	Reaction	Colour Observation	Inference
1 2 3 4 5 6	Section placed in weak iodine solution Section placed in 10% FeCl3 and a little of Na2CO3 Section placed in Dragendroffs reagent Section placed in a drop of 5% aqueous KOH solution Section placed in caustic alkali and HCl Section placed in a drop of H2SO4	Absence of bluish patches Bluish colour appeared Presence of orange colour A strong yellow colour appeared. Yellowish patches appeared. No yellow or red colour appeared.	Absence of starch Presence of tannins Presence of alkaloids Presence of flavonoids Presence of calcium oxalate Absence of saponins or lipids

The fluorescence analysis was observed in day light and UV light at 254nm using various solvents and tabulated below for the crude powder in Table no: 2.

Table No 2: Fluorescence analysis of powder stems bark of Soymida febrifuga.
Material or treatment	Colour in day light	Observation under UV light
Drug powder + 1N HCl Drug powder + 50% HCl Drug powder + 50% H2SO4 Drug powder + 50% HNO3 Drug powder + 1N NaOH Drug powder + 1N NaOH Drug powder + Al.NaOH Drug powder + water	Brown Brown Brown Brown Red Red Reddish brown Brown	Green Green Green Green Black Black Black Green

Physiochemical Standards:

The physiochemical standards studies like ash value, extractive value were carried out and observations are recorded in Table no: 3.

Table No 3: physiochemical standards
S.No	Parameters	Result
1. 2. 3. 4. 5. 6.	Total ash Acid insoluble ash Water soluble ash Extractive value Alcohol soluble extract Water soluble extract	9%w/w 3%w/w 0.7%w/w 3.5%w/v 5.8%w/v 4.2%w/v

Preliminary Phytochemical Analysis:

The Preliminary phytochemical analysis of the stem bark of Soymida febrifuga is tabulated in Table No: 4.

Table No 4: Preliminary Phytochemical Analysis
Test	Extracts
	Aqueous	Ethanol
Alkaloids	+	+
Carbohydrates	+	+
Glycosides	-	-
Sugar	-	-
Steroids	-	+
Saponins	+	-
Tannins	+	+
Phenolic compounds	-	-
Proteins	-	-
Terpenoids	+	+
Flavanoids	+	+
Anthocyanin	-	-
Quinones	-	-

Thin Layer Chromatography:

TLC of the ethanolic extract was developed in n butanol: acetic acid: water (4:1:5) which showed 1 spot at an Rf value of 0.8928 and with chloroform: methanol (10:1) showed one spot at an Rf value 0.434. HPTLC finger print profile of ethanol extract showed 8 spots with an Rf value of 0.16, 0.25, 0.48, 0.51, 0.60, 0.77, 0.84 and 0.90 at 5μl. At 10 ml it showed 7 spots with an Rf value of 0.16, 0.26, 0.46, 0.59, 0.76, 0.84, 0.90.

CONCLUSION:

The present data generated would help in the authentication of the drug both in dry and powder form. The macroscopic, microscopic characters and the quantitative standards would be useful for ranging down pharmacopoeial standards. The different spots observed in TLC and HPTLC finger print profile will be definitely useful for deciding the purity, quality of the drug to lay down standard.

REFERENCE:

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3. O'Brien TP, Feder N and McCully ME. Polychromatic staining of plant cell walls by toludine blue O. Protoplasma.1964; 59:367-373.

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Received on 25.09.2009

Accepted on 15.10.2009

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 1(3): Nov.-Dec.. 2009, 213-216