PHARMACOGNOSTICAL AND PHYTOCHEMICAL INVESTIGATION OF EUPHORBIA PROSTRATA AIT.
HTML Full TextPHARMACOGNOSTICAL 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±5oC 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.
Macroscopic characters | Root | Stem | Leaf | |||
Fresh | Dried | Fresh | Dried | Fresh | Dried | |
Condition | Tap root system
slender |
Tap root system
slender |
Prostrate, slender, internode | Prostrate, slender, internode | Pubscent, opposite, broad, oblong, glabrous above | Opposite, broad, oblong, glabrous above |
Length | 8 to 12 cm | 5 to 10 cm | 17 to 20 cm | 15 to 18 cm | 2.5 to 5 mm | 2 to 3.5 mm |
Breadth | 0.2 to 0.4 cm | 0.1 to 0.25 cm | 0.3 to 0.5 cm | 0.2 to 0.4 cm | 2 to 4 mm broad
|
1.5 to 3 mm broad |
Colour | Pink at base,
pale yellowish downwards, creamish from inner side |
Light yellowish outside and
off from inner side |
Green or sometime
purple tint from outside, creamish inner side |
Purple tint, slightly green
patches, creamish from inner side |
Green, purplish red | Purplish red |
Branching | One primary long vertical root, some secondary root at stem base | One primary long vertical root, some secondary root at stem base | Branched | Branched | Opposite, petiole short, margin serrulate | Opposite, petiole short, margin serrulate |
Fracture | Tough | Tough | Fibrous | Fibrous | Fibrous | Fibrous |
Taste | Tasteless | Tasteless | Tasteless | Tasteless | Tasteless | Tasteless |
Odour | Not characterstic | Not characterstic | Not characterstic | Not characterstic | Not characterstic | Not characterstic |
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.
FEATURES | |||
Stem | Root | Leaf | Powder (Whole plant) |
Unicerate multicellular trichomes | Endodermis | Epidermis | Unicerate multicellular trichomes |
Cortex | Cork cells | Vascular bundles | Lignified sclerenchymatous Cells |
Pericyclic fibers | Cambium | Stomata (anomocytic) | Spiral lignified vessels |
Parenchymatous pith | Short medullary rays | Glandular hairs | Starch granules |
Cork Cells | |||
Lignified fibers | |||
Stomata (anomocytic) | |||
Parenchymatous cells |
TABLE 3: QUANTITATIVE MICROSCOPY OF LEAVES OF E. PROSTRATA AIT.
Determination | Range | Mean* |
Upper epidermal stomatal index | 11.7-18.7 | 15.2 |
Lower epidermal stomatal index | 17.6-26.3 | 21.95 |
Vein islet number | 2-5 | 3.5 |
Vein termination number | 5-13 | 9 |
*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.
Heavy metal | Concentration(ppm) | |
Lead | 0.0041 | |
Cadmium | 0.0026 | |
Nickel | 0.0093 | |
Mercury | 0.0005 | |
Iron | 0.4264 | |
Copper | 0.511 | |
Sodium | 1.52 | |
Magnesium | 0.690 | |
Arsenic | 0.0019 |
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.
Parameter | Determined values* |
Ethanol soluble extractives | 12±0.9 (% w/w) |
Water soluble extractive | 34±1.0 (% w/w) |
Petroleum Ether soluble | 7±0.4 (% w/w) |
Total ash | 10±0.3 (% w/w) |
Water soluble ash | 9.1±0.26 (% w/w) |
Acid insoluble ash | 0.9±0.1 (% w/w) |
Sulphated ash | 2±0.2 (% w/w) |
Moisture Content | 14±0.7 (% w/w) |
Swelling Index | 2.7 ml/mg |
Foaming Index | Less than 100 |
Crude Fiber content | 50.8% |
*Mean value of three counts
TABLE 6: QUALITATIVE CHEMICAL TESTS OF E. PROSTRATA AIT.
Phytoconstituents | P | C | A | Aq |
Alkaloids | – | – | – | – |
Flavonoids | – | – | + | + |
Carbohydrates | – | – | + | + |
Sterols | – | + | + | – |
Glycosides | – | + | + | – |
Saponins | – | – | + | + |
Phenolic compounds and Tannins | – | – | + | + |
Proteins and Amino acids | – | – | – | + |
Resin | – | – | – | – |
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
Treatment | Colour | ||
Visible | Short UV (254 nm) | Long UV (365 nm) | |
Powder as such | Yellow Green | Light Green | Brownish Black |
Powder + 1N Hydrochloric acid | Dark Green | Brownish Black | Black |
Powder + Picric acid | Yellowish Green | Yellowish Green | Black |
Powder + Acetic Acid | Dark Green | Dark Green | Reddish Black |
Powder + 50% Sulphuric Acid | Greenish Black | Dark Brown | Brownish Black |
Powder + 1N Sodium Hydroxide | Green | Purple Green | Purple Black |
Powder + Methanol | Yellowish Green | Dark Green | Purple Black |
Powder + 1N Nitric Acid | Green | Dark Green | Black |
TABLE 8: HISTOCHEMICAL COLOUR REACTIONS ON E.PROSTRATA POWDER
Reagents | Test for | Color change in histochemical reaction | Degree of change | Inference |
Potassium iodide & Iodine solution | Proteins | Light yellow | Black | Absent |
Iodine solution | Starch | Blue black | Brownish Black | Present |
Aqueous potassium hydroxide solution (10%)+ Sulphuric acid | Suberin | Brown | Green | Absent |
Ferric chloride solution | Tannins | Black | Greenish Black | Present |
Wagner’s reagent | Alkaloids | Yellow | No change | Absent |
Dragendroff’s reagent | Alkaloids | Orange | No change | Absent |
Dilute Sulphuric acid
|
Sterols
|
Dark red
|
Red changes to
Black |
Present |
Chloroform+ Dilute sulphuric acid | Sterols | Dark red | Reddish black | Present |
Libermann Burchard’s reagent | Terpenes | Pink | Dark pink | Present |
REFERENCES:
- Singla AK, Pathak K. Anti-inflammatory studies on Euphorbia prostrata. J Ethnopharmacol 1989;27(1-2):55-61.
- The Ayurvedic Pharmacopoeia of India, Government of India, Ministry of Health and Family Welfare, New Delhi, Vol. 5(1), 28.
- Lin L. J., Marshall G. T. and Kinghorn A. D. The dermatitis-producing constituents of Euphorbia hermentiana latex. J Nat Prod 1983;46(5):723-31.
- Eberle M. M., Erb C., Flammer J. and Meyer P. Dermatitis and conjunctivitis after contact with Euphorbia myrsinites (wolf's milk extract)-a case report. Klin Monatsbl Augenheilkd 1999;215(3):203-4.
- Kamgang Rene et al. Activity of aqueous ethanol extract of Euphorbia prostrata Ait on Shigella dysenteriae type 1-induced diarrhea in rats. Indian J Pharmacol 2007;39(5):240-44.
- Sharma G.D. and Tripathi S.N.; Experimental evaluation of dugdhika (Euphorbia prostrata W.Ait.) for the treatment of `tamaka svasa' (bronchial asthma). Ancient Sci Life;3(3):143-50.
- Akhtar M.S., Q.M. Khan and Khaliq T. Effects of Euphorbia prostrata and Fumaria parviflora in normoglycaemic and alloxan-treated hyperglycaemic rabbits. Planta Med 1984;50:138-42.
- Bakhshi G.D., Langade D.G., Desai V.S. Prospective, Open Label Study of Euphorbia Prostrata Extract 100 mg in the Treatment of Bleeding Haemorrhoids. Bombay Hospital J 2008; 50(4):577-83.
- N. P. Manandhar. Ethnobotanical Notes on Certain Medicinal Plants Used by Tharus of Dang-Deokhuri District, Nepal. Int J Crude Drug Res 1985; 23(4): 153-59.
- The Wealth of India, Raw Materials. Council of Scientific and Industrial Research, New Delhi;Vol. 1,346-47.
- Khandelwal KR, Pawar AP, Kokate CK and Gokhale SB. Practical Pharmacognosy. Nirali Prakashan, Pune; 2001,19-153.
- Trease GE, Evans W C. Pharmacognosy. W.B. Saunders; 1996, Edn.15,516-47.
- Kokate C, Purohit A, Gokhale S. Practical Pharmacognosy. Vallabh Prakashan, New Delhi; 1994, Edn. 10,112-20.
- Kandalkar A.M.K., Manjunath K.P., Sholapur H.P., Patel A.M., Darade S.S. Phytochemical and Pharmacognostic Evaluation of Euphorbia hirta Linn. Leaves. J Pharm Research 2009; 2(3):349-52.
- Anbazhahi T., Kadavul K., Suguna G., Petrus A.J.A. Studies on the pharmacognostical and in vitro antioxidant potential of Cleome gynandra Linn. Leaves. Nat Prod Radiance. 2009;v8 (2):151-7.
- Gupta P, Vasudeva N, and Sharma S.K. Pharmacognostical Study and Preliminary Phytochemical Screening of the Roots of Tagetes erecta Linn. Roots. Hamdard Medicus 2009, 52(1).
- Quality Control Methods for Medicinal Plants. World Health Organization Geneva, A.I.T.B.S. Publishers and Distributors (Regd.), Delhi (India).
- Harborne JB. Phytochemical Methods-A Guide to Modern Technique of Plant Analysis, Champan and Hall, UK, 1998, Edn. 3:1-5.
Keywords:
Euphorbia prostrata Ait., Euphorbiaceae, Pharmacognostic, |
Phytochemical evaluation
Article Information
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1043-1048
742KB
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English
IJPSR
Surendra Kr. Sharma*, Joginder Singh and Sumitra Singh
Department of Pharmaceutical Science, Guru Jambheswar University of Science and Technology, Hisar-125001, Haryana, India
21 November, 2011
25 January, 2012
28 March, 2012
http://dx.doi.org/10.13040/IJPSR.0975-8232.3(4).1043-48
01 April, 2012