Life Science Archives (LSA)

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1 Available online at Life Science Archives (LSA) ISSN: Volume 2; Issue - 2; Year 2016; Page: Research Article ROLE OF Piper nigrum L. IN THE HEALTH CARE SYSTEM OF HUMAN BEINGS S. Manjusha* and J. Lohidas, Department of Botany and Research Centre, Scott Christian College (Autonomous,) Nagercoil , Kanyakumari district, Tamil Nadu, India. Abstract The Piper nigrum L. plant materials such as (leaf and seed) were collected and allowed to shade dry to remove moisture content. The dried samples were used for further studies. The powdered plant materials were filled separately in the thimble and extracted successively using a soxhlet extractor with distilled water, acetone, chloroform, DMSO, ethanol and aqueous. All the extracts were subjected to systematic phytochemical screening for the presence of phytochemical constituents. Carbohydrates, Amino Acids, Proteins, Chloride, Alkaloids, Tannins, Phlobatannins, Steroids, Phenolic compounds, saponins are traced. Antimicrobial activity of the plant extracts were tested by agar well diffusion method against four bacterial pathogens (two Gram positive and two Gram negative strains) such as E. coli, K. pneumoniae, B. cereus and Staphylococcus aureus. In this assay, the leaf and seed of Piper nigrum extracts showed inhibition activities on Gram negative bacterium such as E. coli and K. pneumoniae. The aqueous extract showed inhibitory activity only on Gram negative bacterium B. cereus and S. aureus. Article History Received : Revised : Accepted : Introduction Medicinal plants are being used since ancient time without knowing about its active ingredients. The healing power of certain plant was understood and accepted before mankind discovered the existence of microorganisms (WHO, 2005). The wide spread use of herbal remedies and health care preparations obtained from commonly used traditional herbs and medicinal plants have been raised due to the occurrence of natural products with medicinal properties. Even though pharmacological industries have produced a number of new antibiotics in the last three decades, resistance to * Corresponding author: S. Manjusha Department of Botany and Research Centre, Scott Christian College (Autonomous,) Nagercoil. Key words: Soxhlet extractor, Piper nigrum L. and Antibacterial activity these drugs by microorganisms has also increased (Gislene et al., 2000). Diseases are the major causes of death in the developing countries and accounts to 50 % of it. The extensive use of the antibiotics to control these diseases has led to the emergence of multidrug resistance (Westh et al., 2004). Nowadays, the use of antibiotics to control diseases is producing adverse toxicity to the host organs, tissues and cells. The toxicity produced by the antimicrobial agents can be cured or prevented or antagonized using herbs. Herbal medicines are in great demand in both developed and developing countries as a source of wide biological and medicinal activities, high safety margins and lesser costs. Herbal molecules are safe and will overcome the resistance produced by the pathogens as they exist in a combined form or in a

2 S. Manjusha /Life Science Archives (LSA), Volume 2, Issue 2, Page 514 to 518, pooled form of more than one molecule in the protoplasm of the plant cell. Some herbs were known to prevent cancer. Some herbs have antibacterial and antifungal properties that are useful for clinical use. Some of the in vitro studies have been conducted, in which herbal extracts were given to clinical drug resistant strains and different serotype strains of infection (Amin and Kapadnis, 2005). Phytochemicals are compounds that occur naturally in plants. They contribute to the color, flavor and smell of plants. In addition, they form part of a plant s natural defense mechanism against diseases. Their therapeutic values to human health and disease prevention have been reported (Okwu, 2004). The genus Piper (L.) contains more than 1000 species they grow in tropical and subtropical rain forest, but the most well known species are P. nigrum. 51 cultivars of P. nigrum have been reported from the tropical and subtropical regions of India (Nisar Ahmad et al., 2012). Piper nigrum is famous as the spices king due to its pungent quality, member of Piperaceae family. They are sometimes called Indian Long Pepper, is a flowering vine in the family Piperaceae, cultivated for its fruit, which is usually dried and used as a spice and seasoning. It is a close relative of the black pepper plant, and has a similar, though generally hotter, taste. The root and fruit of Piper nigrum are used in palsy, gout and lumbago. The fruits have a bitter, hot, sharp taste, tonic to the liver, stomachic, emmenagogue, abortifacient, aphrodisiac and digestive (Sashidhar, 2002). 2. Materials and methods Selection of Plant material In this present study, three plants such as Piper nigrum (leaf and seed) was selected. It was shade dried, powered and passed through sieve no. 40 and stored in air tight container and used for further extraction. Preparation of plant extracts All the samples were subjected to soxhlet extraction using five solvents such as Acetone, Chloroform, Dimethyl sulfoxide, Ethanol and Distilled water. Each 5 grams of plant material was filled separately in the thimble and extracted successively with 60 ml of solvents using a soxhlet extractor for three hours. After solvent evaporation, each of these solvent extract was weighed and preserved in room temperature until further use. Qualitative analysis phytochemical constituents All the plant extracts were subjected to systematic phytochemical screening for the presence of chemical constituents like Carbohydrates, Amino Acids, Proteins, Chloride, Alkaloids, Tannins, Phlobatannins, Steroids, Phenolic compounds and Saponins (Harborne, 1999). Anti-microbial activity assay Antimicrobial activities were determined by Agar well diffusion method. Four bacterial pathogenic strains such as Two Gram Positive strains (Bacillus cereus and Staphylococcus aureus) and Two Gram negative strains (Escherichia coli and Klebsiella pneumoniae) were used in this investigation. 3. Result and Discussion Many plants are used as the source of medicine and they have been playing an important role in the health services around the globe (Thomson, 2010). About 75 % of the world s population relies on plant and their extracts for health care and plants used in traditional medicine contain a wide range of ingredients that can be used to treat many infectious diseases (Kunwar, 2008). This present study was aimed to screen the antimicrobial properties and pharmaceutically important bioactive substances from Piper nigrum plants. The plant parts such as dried leaves and seeds of Piper nigrum were subjected to soxhlet extraction using five solvents viz., acetone, chloroform, dimethyl sulfoxide, ethanol and distilled water. Preliminarily the plant extracts were subjected to systematic phytochemical screening for the presence or absence of chemical constituents. In phytochemical screening, the leaf of Piper nigrum showed positive result for amino acid, chloride, alkaloids, flavanoids, tannin and

3 Acetone Chloroform DMSO Ethanol Aqueous Acetone Chloroform DMSO Ethanol Aqueous S. Manjusha /Life Science Archives (LSA), Volume 2, Issue 2, Page 514 to 518, steroids; seed of Piper nigrum showed positive result for chloride, alkaloid, flavonoid, tannin, phlobatannin, steroids and saponin. The phytochemical constituents are mainly responsible for the medicinal properties of the plant. In this study, the Piper nigrum contained amino acid, chloride, alkaloids, flavanoids, tannin, phlobatannin, steroids and saponin. Phytochemical analysis of ethanol extract of Piper nigrum showed the presence of tannins and alkaloids (Ganesh et al., 2014). Table 1: Phytochemical constituents of Piper nigrum leaf (ML) S. No. Phytochemicals Antibacterial activity of Piper nigrumseed (MS) 1 Carbohydrate Amino acid Protein Chloride Alkaloids Flavonoids Tannins Phlobatannins Phenols Steroids Saponin Table 2: Phytochemical constituents of Piper nigrum seed (MS) S. No. Phytochemicals 1 Carbohydrate Amino acid Protein Chloride Alkaloids Flavonoids Tannins Phlobatannins Phenols Steroids Saponin Antibacterial activity of Piper nigrum leaf (ML) Antimicrobial activity of the plant extracts were tested by Agar well diffusion method against four bacterial pathogens viz., E. coli, K. pneumoniae, B. cereus and S. aureus. The leaf and seed of Piper nigrum extracts showed inhibition activities on Gram negative bacterium such as E. coli and K. pneumoniae. The aqueous extract showed inhibitory activity only on Gram negative bacterium B. cereus and S. aureus. The inhibitory activity of P. nigrum was previously reported

4 S. Manjusha /Life Science Archives (LSA), Volume 2, Issue 2, Page 514 to 518, against some bacteria such as Staphylococcus aureus, Bacillus cereus, Streptococcus faecalis, Pseudomonas aeruginosa, Salmonella typhi and Escherichia coli (Pavithra and Bhagya, 2009). The antibacterial activity of ethanol extract of Piper nigrum showed against Escherichia coli, Salmonella typhi, Proteus sp. and Staphylococcus aureus (Ganesh et al., 2014). The acetone and ethyl acetate extract of P. nigrum showed inhibition against Haemophilus sp, E. coli, B. subtilis, Enterobacter faecalis, S. aureus and Yersinia sp (Shanmugapriya et al., 2012). The leaf of Piper nigrum showed positive result for tannin in acetone extract; chloroform extract showed none of positive; DMSO extract showed positive for amino acid, chloride, tannin; ethanol extract showed positive for amino acid, tannin, steroids; aqueous extract showed positive for amino acid, chloride, alkaloids, flavanoids, tannin, steroids. The seed of Piper nigrum showed none of positive result in acetone extract; chloroform extract showed positive for alkaloid, steroid; DMSO extract showed positive for chloride, alkaloid, flavonoid, steroid; ethanol extract showed positive steroids only; aqueous extract showed positive for chloride, alkaloids, flavanoids, tannin, phlobatannin, steroids and saponin. The leaf of Piper nigrum acetone extract showed inhibitory activity on E. coli (13 mm), K. pneumonia (10 mm); chloroform extract showed inhibitory activity on E. coli (11 mm), K. pneumoniae (10 mm); DMSO extract showed pneumoniae (9 mm); ethanol extract showed pneumoniae (10 mm) and aqueous extract showed inhibitory activity on B. cereus (10 mm). The seed of Piper nigrum acetone extract showed inhibitory activity on E. coli (11 mm), K. pneumoniae (14 mm); chloroform extract showed inhibitory activity on E. coli (8 mm), K. pneumoniae (8 mm) and S. aureus (8 mm); DMSO extract showed pneumoniae (10 mm); ethanol extract showed pneumoniae (12 mm) and aqueous extract showed inhibitory activity on B. cereus (10 mm) and S. aureus (8 mm). The results of the present investigation showed organic solvent have good inhibition activity against pathogenic bacterial strains because the active principles were either polar or non-polar and were extracted only through successive organic solvents (Mohanasundari, 2007; Britto, 2001). Parekh and Chanda (2007) and Alam et al. (2009) have reported that methanol is highly potent solvent for extracting the phytochemicals from the plant material. It seems that the polarity of the solvent played an important role in extracting the plant components as well as their activity. In the study of Das et al. (2010 & 2012), the chloroform and methanol extracts were found to be more active than its petroleum ether and water extracts. The differential activity might be due to either difference in plant constituents in each extract or tolerance of the pathogens to different plant constituents. 4. Conclusion The present study concluded that, the preliminary screening of phytochemical constituents results demonstrated the presence of various bioactive metabolites. The antibacterial activity results showed the inhibitory activity of Piper nigrum. The results of the study also supports the traditional application of the plant and suggests that the plant extracts possess compounds with antibacterial properties that can be used as antibacterial agents in novel drugs for the treatment of pain relief, rheumatism, chills, flu, colds, muscular aches and fever. Further pharmacological evaluations, possible isolation of the therapeutic antibacterial from this plant are the future challenges. 5. References 1) Alam M.T, Karim M.M, Shakila Khan N Antibacterial activity of different organic extracts of Achyranthes aspera and Cassia alata. J. Sci. Res., 1: ) Amin M, Kapadnis B.P Heat stable antimicrobial activity of Allium ascalonicum against bacteria and fungi. Ind. J. Exp. Biology, 43:

5 S. Manjusha /Life Science Archives (LSA), Volume 2, Issue 2, Page 514 to 518, ) Britto J.S Comparative antibacterial activity study of Solanum incanum L., J. Bot Club, 18: ) Ganesh P, Suresh Kumar R and Saranraj P Phytochemical analysis and antibacterial activity of Pepper (Piper nigrum L.) against some human pathogens. Central European Journal of Experimental Biology, 3 (2): ) Gislene GF, Nascimento, Juliana Locatelli, Paulo C Freitas and Giuliana L Silva Antibacterial activity of plant extracts and phytochemicals on antibiotic resistant Bacteria. Brazilian Journal of Microbiology, 31: ) Harborne J. B Phytochemical Methods, Chapman and Hall Publications, London, ) Kirtikar, K. R and Basu, B. D Indian Medicinal Plants, Periodical Expert Book Agency, New Delhi, India. 8) Kunwar R.M and Bussmann, R. W Ethnobotany in the Nepal Himalaya. Journal of Ethnobiology and Ethnomedicine, 4 (24): ) Mohanasundari C, Natarajan D, Srinivasan K, Umamaheswar SA, Ramachandran A Antibacterial properties of Passiflora foetida L., a common exotic medicinal plant. African J. Biotechnol, 6: ) Nisar Ahmad, Hina Fazal, Bilal Haider Abbasi, Shahid Farooq, Mohammad Ali, Mubarak Ali Khan Biological role of Piper nigruml. (Black pepper): A review. Asian Pacific Journal of Tropical Biomedicine, ) Okwu D. E Phytochemicals and vitamin content of indigenous species of South Eastern Nigeria. J. Sustain Agric. Environ, 6: ) Pavithra VK, Bhagya L Antibacterial activity of black pepper (Piper nigrum Linn.) with special reference to its mode of action bacteria. Indian J Comp Microbiol. Immunol Infect. Dis., 30 (1): ) Parekh J, Chanda SV In vitro antimicrobial activity and phytochemical analysis of some Indian medicinal plants. Turkish J. Biol., 31: ) Sashidhar N. S Osmania University, Hyderabad, India. 15) Shanmugapriya K, Saravana PS, Harsha Payal S Peer Mohammed and Binnie Williams Antioxidant potential of pepper (Piper nigrum Linn.) leaves and its antimicrobial potential against some pathogenic microbes. Indian Journal of Natural Products and Resources, 3 (4): ) Thomson G. E Further consideration of Asian Medicinal plants in treating common chronic disease in West. Journal of Medicinal Plants Research, 4 (2): ) Westh H, Zinn CS, Rosdahl VT, Sarisa Study Group: An International multicenter study of Antimicrobial consumption in Staphylococcus aureus isolates from 15 hospitals in 14 countries. Microbial Drug Resistance, 10: ) WHO Traditional Medicine Strategy. World Health Organization, Geneva.