MACROSCOPICAL, ANATOMICAL AND PHYSICO- CHEMICAL STUDIES OF EUPHORBIA HIRTA LINN. GROWING WIDELY ON EASTERN UTTAR PRADESH REGION OF INDIA

Traditional knowledge and ethno-botanical use of plants have been widely acknowledged all over the world. Certain people of India, using whole plant of Euphorbia hirta L. as an effective remedy for varities of diseasese. The documentation of traditional knowledge from eastern Uttar pardesh area reveals that it is highly widely used in Ayurveda. Pharmacological and physicochemical details of E.hirta not available in authentic litrature including API (Ayurvedic Pharmacopea of India) So the validation and standardisation of whole plant of Euphorbia hirta. was carried out to establish its macroand microscopical standards, physicochemical parameters, preliminary phytochemical investigation and TLC profiles to evaluate the characters of the plant. Macroscopic characters of E. hirta leaves shows composition of leaf is simple with dark green color about 2-6cm. long in size, T.S. of the leaf revealed the presence of stomata, upper and lower epidermis, vascular bundle. Powder characteristics revealed the presence of starch gran-ules, covering trichomes. The various diagnostic characteristic of leaf powder was coarse, dark green, which revealed the presence of lignified xylem vessel,anomocytic type of sto-mata, Physicochemical parameters were also evaluated. Preliminary phytochemical analysis indicated a high percentage of quercetine and flavonoids and this may be one of the reasons behind the anti diabatic and hepatoprotective activity of this plant. The above parameters, which are being reported for the first time in this plant, are significant towards establishing the pharmacognostic standards for future identification and authentication of genuine plant material used Ayurveda for various diaeases in eastern Uttar Pradesh.


Introduction
As a result of the side effects associated with synthetic drugs, people started looking back at the ancient healing systems like Ayurveda, Siddha and Unani. Herbal drugs play an important role in health care programs especially in developing countries. However, obstacle behind the acceptance of alternative medicines in developed countries is the lack of documentation and stringent quality control. So the documentation and standardization of the raw materials used in herbal medicine is very essential for the world wide acceptance of this system of medicine. 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. Pharmacognostic standardisation of plant material include its morphological (organoleptic), anatomical and biochemical characteristics. 1 The Eastern Uttar Pradesh is using many plants by by Ayurvedic practicner for their health care and day to day ailments but many of them are not yet scientifically validated. This plant called dudhi as an effective remedy for various ailments. It is identified as Euphorbia hirta Linn. of the family Euphorbiaceae. , they prefer methanolic extract of whole plant of E. hirta Linn.as a remedy for various disease 2-3 As far as the available literature is concerned, this plant has not yet been scientifically validated. E. hirta Linn. asthma herb plant in English, is a slenderstemmed, annual hairy plant with many branches from the base to top, spreading upto 40 cm in height, reddish or purplish in color. Leaves are opposite, elliptic -oblong to oblong-lanceolate, acute or subacute, dark green above; pale beneath, 1-2.5 cm long, blotched with purple in the middle, and toothed at the edge. The fruits are yellow, three-celled, hairy, keeled capsules, 1-2 mm in diameter, containing three brown, four-sided, angular, wrinkled seeds. Its distributed throughout the hotter parts of India and Australia, frequently found in waste places along the roadsides. 4  Microscopic studies were carried out by preparing thin hand section of leaf and stem with chloral hydrate solution, stained with Phloroglucinol-hydrochloric acid (1:1) and mounted in glycerine. 5 Histochemical studies and powder microscopy were carried out to know about the inclusions and detailed anatomical characters of the material. 6

Physico-chemical Evaluations:
Moisture content, total ash, acid-insoluble ash, alcohol and water-soluble extractive values were carried out as described in Indian Pharmacopoeia. 7-8

Preliminary Phytochemical Screening:
The methanolic, petroleum ether, chloroform and aqueous extract of E. hirta Linn. was subjected to tests for the presence or absence of the major class of compounds by standard methods. 9 2.5 TLC profile: Powdered E. hirta Linn., 10 g was extracted by refluxing with aqueous and methanol (50 ml × 3) sequentially for a period of 30 minutes each and the combined extract of each were filtered and concentrated to 10 ml. Apply 10 ml each extract as bands at a height of 10 mm from the base of a 5 × 10 cm coated silica gel plate 60 F254 using CAMAG Automatic sampler (ATS4) and developed up to 80 mm from the base of the plate in an Automatic Developing Chamber (CAMAG ADC2) using the mobile phase toluene :ethyl acetate: acetic acid (3:7:1) v/v for aqoues, Toluene :Ethyl acetate: Formic acid (3:1:1) v/v for methanol extract. Dry the plate in air and profile pictures were taken after derivatization with anisaldehyde sulphuric acid reagent (ANS)

Aqueous extract
Methanolic extract 2.6 Fluorescence analysis: Powder drug was treated with different reagent and was observed for Fluorescence under UV light. 2.7 Powdered drug reaction with different reagent: Powder drug was treated with different reagent and was observed from naked eye. 3. RESULTS 3.1 Morphological characters: is a slenderstemmed, annual hairy plant with many branches from the base to top, spreading upto 40 cm in height, reddish or purplish in color. Leaves are opposite, elliptic -oblong to oblonglanceolate, acute or subacute, dark green above; pale beneath, 1-2.5 cm long, blotched with purple in the middle, and toothed at the edge. The fruits are yellow, three-celled, hairy, keeled capsules, 1-2 mm in diameter, containing three brown, four-sided, angular, wrinkled seeds. 3.2 Anatomical characters: the detail and systemic pharmacognostical evaluation would give valuable information for the future studies. Macroscopic characters of E. hirta leaves shows composition of leaf is simple with dark green color having no odour about 2-6cm. long in size, shape is ovate, texture is hairy, apex is acute and midrib is distincts on both the side. T.S. of the figure-1) leaf revealed the presence of stomata,upper and lower epidermis, trichomes vascular bundle and collenchyma. Powder characteristics revealed the presence of starch gran-ules, sclariform vessels, covering trichomes, lignified fibres, pericyclic fibres, epidermial cells with trichomes.The various diagnostic characteristic of leaf powder was coarse, dark green, odourless, with bitter test which revealed the presence of lignified xylem vessel,anomocytic type of sto-mata, epidermal cells with stomata, pericyclic fibers, scaliform vessels and trichomes.The quantitative determination of some pharmacognostic parameters is useful for setting standard for crude drugs the vein islet, vein termination and other parameters determined in the quantitative microscopy are relatively constant for the plants and can be used to differentiated closely IJBAR (2012) 03(07) www.ssjournals.com related spe-cies. The physical constant evaluation of drugs is an impor-tant parameter in detecting adulteration or improper handling of drugs.The moisture content of the drug is not too high, thus it could discourage bacteria, fungi or yeast growth. The total ash is particularly important in evaluation of purity of drugs.

Physicochemical parameters:
The value of loss on drying at 1100 C showed the presence of moisture content in the sample, which is 8.5 %. The total ash, acid insoluble ash and water soluble ash were found to be 5.5 %, 2.20 and 4% respectively. The ash contents showed the amount of inorganic matter present in the sample and the acid insoluble ash almost within 2.2 %, which expresses low siliceous matter present in the sample. Extractive values are tabulated in Table -1

Preliminary phytochemical investigation:
The result of phytochemical analysis tabulated in Table-2  Table- -+  +  +  2  Carbohydrates  ----3 Glycosides --+ + 4 Phenolic compounds and Tannins   --+  +   5  Flavonoids  -+  +  +  6 Terpenoids --+ + 7 Saponins --+ + 8 Sterols Proteins ----10 Resins -- 0il / Fats ---3.6 TLC profile: Well resolved TLC profiles were recorded: For the future reference and identity of the plant material.Aqueous extract TLC profile showed four spots under UV 254 nm, after derivatization in visible light). Methanolic extract the profile showed three spots under UV 254 nm, All the above parameters, which are being reported for the first time in this plant, are significant towards establishing the pharmacognostic standards for future identification and authentication of genuine plant material.The result obtained from TLC are depicted in Table-3

Discussion and Conclusion
WHO has emphasised the need to ensure quality control of the raw materials used for Ayurvedic medicines by using modern techniques and by applying suitable parameters and standards. In the present study various standardization parameters such as macroscopy, microscopy (histochemical and powder), physicochemical standards, preliminary phytochemical investigation and TLC profiles in aqueous and methanol extracts were studied, which are being reported for the first time in this plant and could be helpful in authentication and preparation of a suitable monograph for the proper identification of of E.hirta Linn. for the future.