Phytochemical Investigation on Stem Bark of Holoptelea integrifolia (Roxb.)

Jyoti Sharma, S.C. Sharma, Y.S. Sarangdevot and Bhupendra Vyas

B.N. College of Pharmacy, Udaipur (Rajasthan)

ABSTRACT:

The present study deals with the pharmacognostical investigation on stem bark of Holoptelea integrifolia (Roxb.) belongs to Ulmaceae family. This species is used traditionally for the treatment of edema, diabetes, leprosy, skin diseases, intestinal disorders and piles. Pharmacognostical evaluation such as macroscopical and microscopical characters, ash values, extractive values and phytochemical investigation of hydroalcohoic extract were carried out. Phytochemical studies shows the presence of carbohydrates, mucilages, alkaloids, flavonoids, terpenoids, steroids, tannins and glycosides.

KEYWORDS: Holoptelea interifolia, macroscopical, microscopical, phytochemical investigation.

INTRODUCTION:

Holoptelea integrifolia (Roxb.) belonging to the family Ulmaceae. It is commonly known as Indian Elm tree. Holoptelea integrifolia is a large deciduous tree distributed throughout the greater part of India up to an altitude of 2,000 feet¹. Ethnomedically used for the treatment of inflammation, acid gastritis, dyspepsia, flatulence, colic, intestinal worms, vomiting, wounds, vitiligo, leprosy, filariasis, diabetes, hemorrhoids, dysmenorrhoea and rheumatism². It is an important pollen allergen of India³ and sensitizes almost 10% of the atopic population⁴.

The bark and leaves are bitter, astringent, thermogenic, anti-inflammatory, digestive, carminative, laxative, depurative, anthelmintic and repulsive^5-7.

Recently, the activities such as anti-inflammatory^8,9, adaptogenic¹⁰ antidiarrhoeal¹¹, anthelmintic^12,13, wound healing¹⁴, antioxidant^14,15, antimicrobial ^15-19, antitumor²⁰and antidiabetic²¹ activity have been reported in this plant.

MATERIALS AND METHODS:

Collection and identification of plant:

Stem bark of Holoptelea integrifolia was collected from campus of B.N. College of pharmacy, in the month of june 2011. The tree was identified by Dr. J.S. Rathore, professor in botany; B.N.P.G. college, Udaipur (Rajasthan). The stem bark was dried under shade and then powdered and stored in air tight container. Fresh stem bark was used for macroscopic and microscopic investigations.

Extraction:

Extraction of stem bark powder of Holoptelea integrifolia was performed by cold maceration procedure using hydroalcoholic (50% alcohol) solvent. The crude drug powder was intimated with whole of the menstruum in a closed round bottom flask with occasional shaking for 7 days. After 7 days the marc was filtered and the solvent was evaporated on water bath. The drug: menstruum ratio was taken as 1:10²².

Physico-chemical analysis:

The total ash, water soluble ash, acid insoluble ash, water soluble extractive value,alcohol soluble extractive were obtained by employing standard methods of analysis as described in Pharmacopoeial literature .

Phytochemical analysis:

The qualitative analysis of stem bark extract of Holoptelea integrifolia was carried out for the identification of various chemical constituents i.e. carbohydrates, mucilages, proteins, alkaloids, flavonoids, terpenoids, steroids, tannins, glycosides and saponins.

RESULTS AND DISCUSSION:

Morphology:

Fresh stem bark was subjected to macroscopical identification based on colour, odour, taste, form, size and fracture of the stem bark.

Organoleptic evalution:

The macroscopic characteristics of the stem bark of Holoptelea integrifolia were seen it was found that the colour of stem bark is silver green. Thickness of fresh bark is about 8 to 30 mm and dried bark 6 to 18 mm. Longitudinal fissures and irregular rectangular rhytidomes are present on external surface of stem bark. Inner surface of bark was found to be white when fresh and dark brownish when dried. Fracture is hard, irregular and fibrous. Stem of Holoptelea integrifolia having astringent taste. The shape of stem bark is channelled.

Microscopy:

Transverse section of stem bark:

T.S. of stem bark (Fig 1and 2) of Holoptelea integrifolia shows cork in the outer layer. Two types of cork cells were seen, the outer cork was found 3-5 layered, thin walled tangentially elongated rectangular. Cells of inner cork 3-5 layered, thick walled, tangentially elongated and rectangular in shape. Cork cambium is 2-3 layered cells. Outer cortex was found very thick, composed of rectangular cells tangentially elongated, thin walled and compactly arranged having crystals. Crystals were rhomboidal or squarish shaped. Secondary phloem containing lignified fibers, the fibers were found to be polygonal, thick walled with small lumen. Cells of phloem contain prismatic crystals.

Longitudinal section of stem bark of Holoptelea integrifolia:

Longitudinal section of stem bark (Fig 3 and 4) of plant Holoptelea integrifolia shows secondary phloem. Lignified fibers having polygonal cell. Sieve plate and sieve tube were shown in phloem fibers.

Physico-chemical analysis:

The ash value is a criterion to judge the identity or purity of crude drug. The ash is the residue remaining after incineration, the ash content of the drug represents inorganic salts naturally occurring in drug. Different ash values i.e total ash, water soluble ash and acid insoluble ash of the stem bark of Holoptelea integrifolia were determined as per pharmacopoeial literature, the values are shown in Table 1.

Extractive values represent the amount of active constituents extracted with solvents from a known amount of plant material. The water soluble extractive and alcohol soluble extractive values were determined as per pharmacopoeial literature. Extractive values of stem bark of Holoptelea integrifolia are shown in Table 1.

Percentage Extractive yield:

Extraction of stem bark powder of Holoptelea integrifolia was performed by maceration process using hydroalcoholic solvent alcohol: water (50:50). Percent extractive yield shown in Table 2.

Phytochemical analysis:

The qualitative analysis of stem bark extract of Holoptelea integrifolia was carried out for the identification of various chemical constituents. The hydroalcoholic extract was found to contain carbohydrates, mucilages, alkaloids, flavonoids, terpenoids, steroids, tannins and glycosides. The results obtained are shown in Table 3.

Fig 5: Stomata

TABLE 1: Physico-chemical parameters of stem bark of Holoptelea integrifolia

Total ash

(%W/W)

Acid insoluble ash

(%W/W)

Water soluble ash

(%W/W)

Water soluble extractive

(%W/W)

Alcohol soluble extractive

(%W/W)

6.25

1.973

0.6979

11.265

5.56

TABLE 2: Percent extractive yield

S.No

Solvent

Weight of Powder drug

Weight of Dried Extract

% Yield

Colour

Odour

Consistency

Hydroalcoholic

(Alcohol: water) (50:50)

100 gm

9.0277 gm

9.027%

Blackish brown

Characteristic

Sticky

TABLE 3: Phytochemical analysis of the stem bark extract of Holoptelea integrifolia.

S.NO	Chemical Constituents	Chemical Tests	Result
1.	Carbohydrates	Molisch’s Test	Positive
		Fehling Test	Positive
		Benedict’s Test	Positive
2.	Mucilages	Ruthenium red Test	Positive
2.	Mucilages	With aqueous KOH	Positive
3.	Proteins	Xanthoprotic Test	Negative
3.	Proteins	Ninhydrin’s Test	Negative
4.	Alkaloids	Mayer’s Reagent	Positive
		Wagner’s Reagent	Positive
		Dragondoff’s Reagent	Positive
		Hager’s Reagent	Positive
5.	Flavonoids	Shinoda Test	Positive
		Alkaline reagent Test	Positive
		Lead acetate Test	Positive
6.	Terpenoids and Steroids	Liberman Burchard Test	Positive
		Salkowski Test	Positive
		Copper acetate test	Positive
7.	Saponins	Foam Test	Negative
7.	Saponins	Froth Test	Negative
8.	Tannins	Potassium dichromate	Positive
		ferric chloride Test	Positive
		Salt of lead,copper,tin.	Positive
		Gelatine Test	Positive
9.	Glycosides	Modified Borntrager’s Test	Positive
9.	Glycosides	Legal’s Test	Positive

CONCLUSION:

The stem bark of Holoptelea integrifolia has been studied to compare and give report on pharmacognostical, preliminary phytochemical studied on it. This will help correct identification of this plant for future references.

REFERENCES:

1. The Wealth of India. Council of Scientific and Industrial Research, New Delhi. 1959; Vol-5:109-10.

2. Sharma PC, Yelene MB, Dennis TJ. Database on Medicinal Plants used in Ayuarveda. Central Council for Research in Ayurved and Siddha, Gov. of India, New Delhi 2005; Vol-2: 171-3.

3. Singh AB and Kumar P. Aeroallergens in Clinical Practice of Allergy in India: An Overview. Annals of Agricultural and Environmental Medicine 2003; 10: 131-6.

4. Sharma S, Panzani RC, Gaur SN, Ariano R, Singh AB. Evaluation of cross-reactivity between Holoptelea integrifolia and Parietaria judaica. International Archives of Allergy and Immunology 2005; 136:103–12.

5. Warrier PK, Nambiar VPK, Ramakutty. Indian Medicinal Plants a Compendium of 500 Species, Orient Longman Private Limited 1995; Vol-3:162.

6. Prajapati ND, Purohit SS, Sharma AK. A Handbook of Medicinal Plants a Complete Source Book. Agrobias India Jodhpur 2003; 273.

7. Kare MA, Dhabe AS, Bhuktar AS. Studies on Crude Bark Drug - Holoptelea integrifolia (Roxb.) Planch. Journal of Experimental Sciences 2011; Vol-2(3): 09-11.

8. Kalpana and Upadhyay A. Anti-inflammatory evaluation of ethanolic extract of leaves of Holoptelea integrifolia, Planch. Annals of Biological Research 2010; 1 (2):185-95

9. Sharma S, Lakshmi KS, Patidar ACA, Dhaker S. Studies on Anti-inflammatory effect of aqueous extract of leaves of Holoptelea integrifolia, Planch in rats. Indian Journal of Pharmacology 2009; Vol-41(2): 87-8.

10. Shakti P, Bimlesh K, Jiban D, Prashant T, Manoj S, Mohanjit k, Amit M, Comparative Pharmacological Evaluation of Adaptogenic activity of Holoptelea integrifolia and Withania somnifera. International Journal of Drug Development and Research. 2011; 3(1): 84-98.

11. Shrinivas S, Lakshmi KS, Rajesh T. Evaluation of antidiarrhoeal potentials of ethanolic extract of leaves of Holoptelea integrifolia in mice model. International Journal of Pharma Tech Research 2009; 1: 832-6.

12. Nadella D and Paarakh PM. Evalution of Anthelmitic activity of Holoptelea integrifolia (Roxb.) planch. International Journal of Research in Ayurveda and Pharmacy 2010; 1(2): 637-41.

13. Kaur S, Kumar B, Puri S, Tiwari P, Divakar K. Comparative Study of Anthelmintic activity of aqueous and ethanolic extract of bark of Holoptelea integrifolia. International Journal of Drug Development and Research 2010; Vol-2(4): 758-63.

14. Reddy BS, Reddy KKR, Naidu VGM, Madhusudhana K, Agwane SB, Ramakrishna S, Diwan PV. Evaluation of antimicrobial, antioxidant and wound-healing potentials of Holoptelea integrifolia. Journal of Ethnopharmacology 2008; 115: 249-56.

15. Saraswathy A, Nandini DS, Ramasamy D. Antioxidant, Heavy Metals and Elemental Analysis of Holoptelea integrifolia Planch. Indian Journal of Pharmaceutical Sciences 2008; 70(5): 683.

16. Vinod NV, Shijina R, Dileep KV, Sadasivan C. Inhibition of Beta-Lactamase by 1,4-Naphthalenedione from Holoptelea integrifolia. Appl Biochem Biotechnology 2010; 160:1752–9.

17. Nadella D and Paarakh PM. Antibacterial activity of different extracts of Holoptelea integrifolia (Roxb.) planch. International Research Journal of Pharmacy 2011; 2(1): 111-3.

18. Singh M, Khatoon S, Singh S, Kumar V, Rawat AKS, Mehrotra S. Antimicrobial screening of ethnobotanically important stem bark of medicinal plants. Pharmacognosy Research 2010; 2(4): 254-7.

19. Rajbhandari M, Wegner U, Julich M, Schopke T, Mentel R. Screening of Nepalese medicinal plants for antiviral activity. Journal of Ethnopharmacology 2001; 7: 251–5.

20. Lakshmi KS, Sharma SS, Rajesh T, Chitra V. Antitumour activity of Holoptelea integrifolia on Dalton’s ascetic lymphoma in Swiss albino mice. International Journal of Green Pharmacy 2010; 44-7.

21. Sharma S, Khatri P, Pandey A, Jakhetia V,Chaturvedi L, Dwivedi N. Anti-diabetic screening leaves extract of Holoptelea integrifolia . International Journal of Pharma Research and Development 2010; 2: 66-71.

22. Jain NK and Sharma SN. A Textbook of Professional Pharmacy. Vallabh Prakashan 2001: 111.

Received on 11.04.2012

Modified on 29.04.2012

Accepted on 04.05.2012

Research Journal of Pharmacognosy and Phytochemistry. 4(3): May-June 2012, 178-181