Botanical Features for Taxonomic Evidence of Erythroxylum monogynum Dried Leaf

Introduction: Erythroxylum monogynum belonging to family Erythroxylaceae is commonly found in southern parts of India. Traditionally, leaves of the plant possess high medicinal value and known for the treatment of diabetes, jaundice and to kill intestinal worms, which are scientifically reported. However, no elaborative pharmacognostic study of leaves has been performed yet. Objective: To pharmacognostically standardize the leaves by studying anato mical characters of transversal section and powdered leaves. Material and methods: Leaf samples were studied by macroscopy, microscopy, physicochemical and preliminary phyto chemical analysis as recommended by WHO. Results: Externally the leaves are elliptic lanceolate, alternate, base attenuate, apex obtuse and entire margin. Internally it shows the presence of paracytic stomata confined to lower epidermis, conjoint, collateral arc shaped vascular bundles encircled by sclerenchyma, lignified fibers and xylem vessels with spiral thickening. Powder microscopy revealed the presence of epidermis with papillae, phytochemical and physicochemical of powdered drug proved useful to differentiate the powdered drug material. Conclusion: The establishment of botanical features of leaf material will be useful in identification and standardization of Erythroxylum monogyum in obtaining quality formulations.


INTRODUCTION
Herbs represent major part in medicine for thousands of years and now herbal drugs play a significant role in health care management principally in developing countries.However, its acceptance is limited due to lack of standardization and stringent quality control.World Health Organization emphasized on standard ization and quality control of herbal drugs, according to which standardization complies the physicochemical evaluation of crude drug such as ash values, moisture content, extractive values, crude fiber.The quality aspects of the drug majorly include macro and micro scopic examination, qualitative and quantitative chemical evaluation and chromatographic fingerprinting. 1 Correct identification and quality assurance of plant material is indispensable to ensure reproducible quality of herbal medicine, which will contribute to its safety and efficacy. 2In this study, pharmacognostic standardi zation of Erythroxylum monogynum (E.monogynum) include its morphological, anatomical and biochemical characteristics.E.monogynum Roxb belongs to family Erythrox ylaceae, commonly known as Bastard sandal or Red cedar.It is commonly found in southern parts of India.In folklore medicine, parts of the plant are claimed to have medicinal benefits.The infusion of bark and wood is used as stomachic, diaphoretic, stimulant, diuretic and also in mild cases of dyspepsia and continued fever. 3he leaves extract are mixed with yogurt to kill intes tinal worms, where as leaf juice used for the treat ment of jaundice. 4,5E.monogynum scientifically reported for its antibacterial activity, 6 hepatoprotective 7 and antidiabetic activity. 8he objective of the present study is to reveal the pharmacognostical, physicochemical and phyto chemical aspects of leaves of E.monogynum which may help to develop quality, purity, authenticity of the drug and to distinguish from spurious and adul terated species

Plant Material
The plant, E.monogynum was collected from Khammam District (Telangana) during flowering season.The plant was authenticated by Botanical Survey of India, Hyderabad.The leaves of the plant were dried, powdered and passed through 40 mesh sieve and stored in an airtight container for future use.

Macroscopic analysis
Fresh leaves of the plant were noted for the following macroscopical character: Size and shape, color, odor, taste, surfaces, base, apex, margin, venation, lamina and presence or absence of petiole.

Microscopical analysis
For microscopical studies, the leaves were cut and removed from the plant and thin hand sections were prepared.The sec tions were cleared with chloral hydrate solu tion, stained with phloroglucinolhydrochloric acid (1:1) and toludiene blue.Some hand sections were also examined in glycerine. 9,10The following characteristics were observed for their presence or absence: epidermal cells, type of stomata and trichomes.The trans verse sections of the fresh leaves through the lamina and the midrib are also cleared, mounted and observed.The powdered drug of the leaves was separately treated with solution of phloroglucinolhydrochloric acid (1:1), acetic acid and iodine solution to identify the presence of lignified fibers, calcium oxalate crystals and starch grains 9 .

Physico-chemical and Florescence analysis
According to WHO guidelines parameters determined for the powdered leaves are Loss on drying, total ash, acid insoluble ash, water soluble ash.Petroleum Ether soluble extractive, Chloroform extractive value, alco hol soluble extractive (90% ethanol) and watersoluble extractive values were performed to find out the amount of ether, Chloroform, alcohol and water soluble constituents respectively. 11Powdered leaf material was analyzed by treating with various organic/inorganic reagents like NaOH, HCl, HNO 3 and H 2 SO 4 under visible light, short ultraviolet light (254nm) and long ultraviolet light (366nm). 12

Preliminary phytochemical analysis
The airdried leaves of E.monogynum were made into coarse powder.The powdered material was extracted by maceration technique using methanol as solvent for overnight.The obtained extract was filtered, evaporated to dryness and subjected to qualitative chemical tests according to standard procedures. 13

Macroscopic characteristics
Macroscopically, E.monogynum has pinnately compound leaves, alternate in arrangement.The leaflets were 78 pairs and shaped obovate or oblan ceolate, base attenuate or cuneate, apex obtuse, margin entire, secondary veins are pinnately arranged.The leaf size is 24 cm long, 0.8 to 1cm wide and petiolate where petiole is 0.30.6 cm in length.Fresh leaves are green, odorless and have bitter taste (Figure 1).

Microscopical characteristics
The transverse section of a leaf of E.monogynum shows dorsi ventral characters.The important characteristics in the midrib regions are as follows: The midrib slightly protrudes convexly at the dorsal side and strongly projects at the ventral side.The epidermis is composed of single layer of barrel shaped cells.Adaxial epidermis is without stomata but abaxial side consists of paracytic stomata and rounded papillae.Collenchyma consists of 23 layers of cells which are present below both upper and lower epidermis.Intercellular spaces are present between collenchyma cells.It is followed by cortex composed of 3 -4 layered oval -circular parenchymatous cells.Conjoint, collateral, arc shaped vascular bundles are present which are encircled by lignified sclerenchyma fibres.Xylem vessels are lignified, composed of spiral thickening (Figure 2).T. S. of laminar region shows both upper and lower epidermis.Palisade cells are compact, elongated and confined to lower portion of upper epi dermis which is followed by loosely arranged spongy parenchymatous cells.Papillaeelongated cells are present below lower epidermis (Figure 3).

Powder Characteristics
Powder is dull greenish in color, with bitter taste and faint odor.The leaf powder showed fragments of epidermis with paracytic stomata, epidermal    cells with papillae in surface view.Xylem consisting of vessels with spiral thickening was observed.Fragments of sclereids and scattered palisade cells of laminar portion are the characteristic features of powder micros copy for the leaf of E.monogynum (Figure 4).

Physicochemical constants
Physicochemical analysis of leaf viz.loss on drying, ash value and extractive value are presented in Table 1.The fluorescence analysis of leaf powder observed under day light, short ultraviolet light (254nm) and long ultraviolet light (366nm) mentioned in Table 2.

Preliminary phytochemical analysis
Qualitative analysis of leaves of E.monogynum petroleum extract, chloroform extract, methanolic extract and aqueous extract showed the presence of alkaloids, flavonoids, saponins, phenols, terpenoids and alkaloids in respective tests which are mentioned in Table 3

DISCUSSION
Ethnomedically, the leaves of E.monogynum were used by local people in the treatment of some ailments without standardization.Pharma cognostic standardization plays an important role in identification, authentification and determining the level of adulteration, which help in establishing quality control of crude drugs. 14In this study, pharma cognostic standards for leaves of E.monogynum are carried out for the first time.
The microscopical characters of the leaf which serves as diagnostic parameters, revealed the presence of conjoint, collateral, arc shaped vascular bundles containing spiral shaped xylem vessels encircled by lignified sclerenchyma fibers.The palisade parenchyma confined to lower portion of upper epidermis.Physicochemical analysis of the drug could be useful for identification of adulterants and also in compilation of a suitable monograph.Ash values are validity parameters to judge the identity or purity of the crude drug.Extractive value mainly used to determine the exhausted or adulterated drugs along with estimation of specific components soluble in particular solvents. 15In this study higher extractive value of methanol than water indicates more methanolic soluble components.The fluorescence analysis of drug extract helps to identify the drug with specific fluorescent colors and to resolve doubtful specimens.
Phytochemical evaluation is to determine the class of compounds present in the plant parts, which also suggests possibility of any potent pharmacological active constituents in determination of biological assays.ss 16

CONCLUSION
The present study established pharmacognostic standar dization of leaves of E.monogynum, which can serve as an important source to determine the quality and purity of the drug.This work also could be useful in com pilation of a suitable monograph and further establishing standard data in pharmaceutical preparation from leaves of E.monogynum.

Figure 1 :
Figure 1: Macroscopic characters of leaves of E. monogynum

Figure 4 :
Figure 4: Powder microscopy of E.monogynum leaf A) Stomata confined to lower epidermis B) Papillae C) Xylem vessels D) Scattered palisade parenchyma