PHARMACOGNOSTICAL AND PHYTOCHEMICAL INVESTIGATION OF EUPHORBIA PROSTRATA AIT.

PHARMACOGNOSTICAL AND PHYTOCHEMICAL INVESTIGATION OF EUPHORBIA PROSTRATA AIT.

Surendra Kr. Sharma*, Joginder Singh and Sumitra Singh

Department of Pharmaceutical Science, Guru Jambheswar University of Science and Technology, Hisar-125001, Haryana, India

ABSTRACT

Euphorbia prostrata Ait.(Euphorbiaceae) is a small, prostrate, annual green herb sometime purple tint in colour. It is widely distributed globally and used as antihemorrhoidal, anti-inflammatory, analgesic, hypolipidemic, antidiabetic, antidirroheal, antiasthmatic and for various skin diseases. The microscopy of root shows the presence of obliterated cork cells, phelloderm, cortex, endodermis, phloem, medulary rays and xylem; the stem shows the presence of multicellular trichome, cuticle, epidermis, cortex, endodermis, pericycle, phloem, latex canal, xylem and pith; the leave reveal the presence of multicellular, multiseriate glandular hairs, epidermis, vascular bundles, stomata anomocytic and anisocytic. The vein islet number and vein termination number have also been determined. The powder study reveals the presence of epidermal cells, trichomes, parenchymatous cells, pollen grains, vessels, fibers and stomata. The water soluble, alcohol soluble and petroleum ether extractive values were determined. The total ash, water soluble ash, acid insoluble ash and sulphated ash were also observed. Preliminary phytochemical studies revealed the presence of flavonoids, tannins, glycosides and saponins in the alcoholic extract of the plant. This is the first report on pharmacognostical studies on this plant.

INTRODUCTION: Euphorbia prostrata Ait. (Euphor-biaceae) is small annual herb found all over India especially in foot hills of Himalayas. It is native to the West Indies and certain parts of South America and also widely naturalized in many other parts of the world. The two varities found are red and green ¹. These are branched prostrate with many stems spreading from the roots, slender upto 20 cm long, leaves green but occasionally purplish red ². It is a reputed drug in the Indian System of Medicine and used in treatment of many diseases of skin ^{3, 4}, digestive system ⁵, antiasthmatic ⁶, antidiabetic ⁷, haemorrhoids ⁸ etc. It is also used traditionally as snake bite remedy ⁹.

The various types of phytoconstituents reported in Euphorbia prostrata Ait. are like glucoside, galactoside, β-sitosterol, compesterol, stigmasterol, cholesterol, apigenin, luteolin, apigenin-7-glucoside, luteolin-7-glucoside, gallic acid, ellagic acid and tannins ^{2, 10}.

MATERIAL AND METHODS: The plant was collected from the campus of Guru Jambheshwar University of Science and Technology, Hisar, Haryana and identified by Dr. H.B Singh at NISCAIR, New Delhi vide reference no. NISCAIR /RHMD /Consult /-2009- 10/1282/86. The herb was air dried, packed and stored for study.

 Pharmacognostic Studies:

Macroscopic: Macroscopic studies were done using simple microscope. The taste, odour, shape of plant parts of fresh and dried plant was observed ¹¹.

Microscopic: Anatomical sections surface preparation of stem, root, leaves for the microscopy were carried out ^11-13. The microscopical features were observed under Zeius Trinocular Microscope (Germany). Quantitative microscopy of leaf was also carried out and upper and lower stomatal number, vein islet number and vein termination number were observed ^{14, 15}.

Powder Study: Plant was oven dried at 40±5^oC to make it moisture free and powdered form with the help of electric grinder and powder was passed through sieve no. 60. Standard methods were followed to study the powder characteristics ¹¹. Flourescent study and various histochemical reaction studies were carried out on plant powder ^{15, 16}.

Physical Evaluation: The physiochemical parameters such as water, alcohol, petroleum ether soluble extractive values, percentage of total ash, water soluble ash, acid insoluble ash and sulfated ash, loss on drying, swelling index, foaming index, bitterness value, crude fiber content and heavy metals concentration were performed and calculated as per WHO guidelines ¹⁷.

Preliminary Phytochemical Studies: The dried plant was pulverized and 500 gm of plant sample was extracted successively with 4 litres ethanol using soxhlet apparatus for about 72 hours. Thereafter the marc was subjected to three consecutive aqueous extractions for 24 hours each. Each extract was concentrated and dried over anhydrous calcium chloride and kept aside for phytochemical investigation. The qualitative tests were carried out ^{12, 18}.

RESULTS AND DISCUSSION: Macroscopic characters of  E. prostrata shows that it is branched, prostrate with many stems spreading from the root, slender upto 20 cm long; leaves green but occasionally purplish red (fig. 1). The plant consist hairs occasionally present on the surface, no characteristic odour and taste (Table 1).

FIG. 1A: EUPHORBIA PROSTRATA AIT. HERB

FIG. 1B: EUPHORBIA PROSTRATA AIT. ROOT

FIG. 1C: EUPHORBIA PROSTRATA AIT. AERIAL PART

T.S. of the root shows the presence of the presence of cork cells, phelloderm, cortex, endodermis, phloem, medulary rays and xylem (fig. 2 A); the stem shows the presence of multicellular trichome, cuticle, epidermis, cortex, endodermis, pericycle, phloem, latex canal, xylem and pith (fig. 2 B); the leave reveal the presence of multicellular, multiseriate glandular hairs, epidermis, vascular bundles, stomata anomocytic and anisocytic (fig. 2 C; Table 2), upper and lower stomatal index is 11.7-18.7 and 17.6-26.3 respectively, vein islet number is found to be 2-5 and vein termination number is found to be 5-13 (fig. 2 D; Table 3).

 TABLE 1: THE MACROSCOPIC CHARACTERS OF ROOT, STEM AND LEAF OF E. PROSTRATA AIT.

FIG. 2A: T.S. OF EUPHORBIA PROSTRATA AIT. ROOT

FIG. 2B: T.S. OF EUPHORBIA PROSTRATA AIT. STEM

FIG. 2B: T.S. OF LEAF OF EUPHORBIA PROSTRATA AIT. THROUGH MIDRIB

FIG. 2D: DECOLORIZED LEAF ANATOMY

TABLE 2: OBSERVATIONS FROM T.S OF STEM, ROOT AND LEAF OF E. PROSTRATA AIT.

TABLE 3: QUANTITATIVE MICROSCOPY OF LEAVES OF E. PROSTRATA AIT.

*Mean value of 10 counts

The various diagnostic characters of plant powder was yellowish-green, tasteless with oily odour which shows the presence of epidermal cells, trichomes, parenchymatous patches, pollen grains, vessels, fibers, stomata (fig. 3).

FIG. 3: POWDER CHARACTERISTICS OF EUPHORBIA PROSTRATA AIT.

The physical parameters are necessary in detecting adulteration or improper handling of the drugs. The moisture content is not too high, thus it could discourage bacteria, fungi or yeast growth. Heavy metals concentration is within the limit which proves its non-toxic effects (Table 4). The total ash determines the purity of the drugs.

TABLE 4: HEAVY METAL CONCENTRATION IN EUPHORBIA PROSTRATA AIT.

Water soluble, alcohol soluble and petroleum ether extractive values were found to be 34±1.0%, 12±0.9% and 7±0.4% respectively. The total ash, water soluble ash, acid insoluble ash and sulphated ash were observed to 11±0.3%, 12±0.26%, 1±0.2% and 2±0.2% respectively. The total moisture content was found to be 14±0.7.

Swelling index, foaming index and crude fibre content were found to be 2.7 ml/mg, Less than 100 and 50.8%, respectively (Table 5). Qualitative tests reveal the presence of flavonoids, tannins, glycosides and saponins (Table 6). The fluorescent behavior and histochemical colour reaction studies were noted (Table 7, 8).This Pharmacognostical evaluation will provide valuable information for future studies.

 TABLE 5: PHYSIOCHEMICAL PARAMETERS OF E. PROSTRATA AIT.

*Mean value of three counts

TABLE 6: QUALITATIVE CHEMICAL TESTS OF E. PROSTRATA AIT.

P: Petroleum ether extract, C: Chloroform extract, A: alcoholic extract, Aq: Aqueous   extract. “+” Present, “-” Absent.

TABLE 7: FLUORESCENT BEHAVIOR OF E. PROSTRATA POWDER UNDER VISIBLE AND UV LIGHT

 TABLE 8: HISTOCHEMICAL COLOUR REACTIONS ON E.PROSTRATA POWDER

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Phytochemical evaluation