R. Hari Babu et al. /JPR:BioMedRx: An International Journal 2013,1(10),947-951 Research Article ISSN: 2321-4988 Available online through http://jprsolutions.info Phytochemical screening of underutilized species of

R. Hari Babu* and N. Savithramma Dept of Botany, S.V. University, Tirupati – 517 502, Andhra Pradesh, India

Received on:17-07-2013; Revised on:16-08-2013; Accepted on:21-09-2013

ABSTRACT Phytochemicals are important constituents of to improve economic values of species. These studies were neglected in the members of poaceae. Hence the aim of the study to screen the South Indian grass plants of poaceae for phytochemical constituents. The results obtained in the present study suggest that preliminary phytochemical analysis detuned the presence of Alkaloids, Coumarins, Flavonoids, Glycosides, Lignin’s, Phenols, Quinones, Saponins, Steroids, Tannins, and Terpenoids. Results of quantitative phytochemical analysis revealed that high content of Alkaloids, followed by Flavonoids, Phenols, Saponins and Tannins receptively. The results suggest that the species posses the properties for curing various ailments and may be the source for potential antioxidant, anti-inflammatory and antimicrobial activities. Which may leads to isolation of new and novel compounds.

Key words: Grasses, Secondary metabolites, Poaceae and Phytochemicals

INTRODUCTION The grasses that yield food and fodder are well known from times traditional medicine system but yet have to be explored. Hence the immemorial and considerable amount of research work has been car- aim of this study is to examine the phytochemical composition of ried on them. But the grasses that yield therapeutically important aqueous extracts of twenty poaceae grass species. Preliminary phy- products are among the least studied in the poaceae family. tochemical screening of the plants is primarily an important aspect in Phytochemicals are naturally occurring, biologically active chemical finding the chemical constituents in plants. compounds in plants. They act as a natural defense system for host plants and provide colour, aroma and flavour. These are non-nutritive Alkaloids which are one of the largest groups of phytochemicals in plant chemicals that have protective or disease preventive proper- plants have amazing effects on humen based on their toxicity against ties1. Many antioxidant compounds naturally occurring in plant source cells of foreign organisms. Therefore, the compounds detected may have been identified as free radicals or active oxygen scavengers2. be responsible for the antibacterial activity of the herb extracts. Fla- Phytochemicals like carotenoids, flavonoids and polyphenols, they vonoids exhibited a wide range of biological activities such as anti- posses antioxidant activity and protect our cells against oxidative oxidant, anti-inflammatory, antimicrobial, anti-angionic, anticancer and damage and reduce the risk of developing certain types of cancer. anti-alergic 6 7 3. The flavonoids have aroused considerable interest The majority of the active compounds are phenolics, vitamin C, vita- recently because of their potential beneficial effects on human health. min E, tannins and carotenes3. Sources of natural antioxidants are They have been reported to have antiviral, anti allergic, anti platelet, primarily plant phenolics such as flavonoids that exhibit antioxidant, anti inflammatory, antitumor, anti carcinogenic and anti ageing prop- antimicrobial, anticarcinogenicity and other biological activities 4. erties 8 9. Flavonoids may help provide protection against these dis- eases by contributing along with antioxidant vitamins and enzymes. World plant biodiversity is the largest source of herbal medicine and Flavonoids are free radical scavenger, super antioxidant and potent still about 60 – 80% world population rely on plant based medicines water soluble which prevent oxidative cell damage and have strong which are being used since the ancient ages as traditional health care anti cancer activity 10. Flavonoids apart from their antioxidant protec- system. It is now clear that, the medicinal value of these plants lies in tion effects, inhibit the initiation, promotion and progression of tu- the bioactive phytochemical constituents that produce definite physi- mour 10 11. Saponins which are one of the active constituents involved ological effects on human body. These natural compounds formed in plant disease resistance because of their antimicrobial activity 12. the base of modern drugs as we use today 5. Plants belonging to the Traditionally, saponins are subdivided into triterpenoid and steroid family Poaceae are mainly grasses. These grasses are included in glycoside. Tannins are phenolic compound which act as primary an- tioxidants or free radical scavengers13. According to 14, several phe- *Corresponding author. nolic compounds like tannins present in cells of plant are potent in- R. Hari Babu hibitors of many hydrolytic enzymes such as proteolytic macerating Research Scholar enzymes used by plant pathogens. In addition, herbs that have tannins as their main components are astringent in nature. Department of Botany S.V. University, Tirupati-517 502 Chittoor (Dt), Andhra Pradesh, India.

JPR:BioMedRx: An International Journal Vol.1 Issue 10.October 2013 947-951 R. Hari Babu et al. /JPR:BioMedRx: An International Journal 2013,1(10),947-951 MATERIAL AND METHODS can play the role in pharmacological activities as anti-inflammatory, analgesic, anti-oxidant, antifungal and immunostimulant providing a Collection and identification of plant material key role of flavonoids to their biological actions8. Glycoside sub- Fresh leaves of Alloteropsis cimicina, Aristida hystrix, Aristida stances are absent in seven grasses and found in thirteen grasses setacea, Brachiaria racemosa, Chloris barbata, Cymbopogon among selected twenty species. They are important in medicine be- coloratus, dactylon, Dactyloctenium aegyptium, Digitaria cause of their aciton on heart and are used in cardic insufficiency31. saguinalis, Echinochola colonum, Eleusine indica, Lignins are present in eleven species of selected plants. Lignins are amabilis, Eragrostiella bifaria, Heteropogon contortus, Panicum significant components in the global carbon cycle; the resistance of repens, Perotis indica, Pycreus flavidus, Setaria pumila, Sporobolus lignin to microbial degradation enhances its persistence in soils 32. coromandelianus and Sporobolus wallichii were collected from the different locations of South India, during the month of December, Twelve species among twenty selected grasses posses phenol com- 2011. The leaves were washed thoroughly 2-3 times with running tap pounds. Heteropogon contortus is the rich source of phenol com- water, leaf material was then air dried under shade after complete pounds followed by Chloris barbata.21 28 26 studied the phenols in shade drying the plant material was used for phytochemical analysis. Cymbopogon citrates and Imperata cylindrical. The higher amount of phenols are important in the regulation of plant growth, develop- Preparation of extract ment and disease resistance. It can be used as fungicide, pesticides, The grinded leaf material of 5g was crushed in 25 ml of sterile water, antiseptic and disinfectant and in the manufacture of resins. Plant boiled at 50-60oC for 30 minutes on water bath and it was filtered phenols may interfere with stages of Cancer process. Potentially re- through Whatman No. 1filter paper. Then filtrate was centrifuged at sulting in a reduction of cancer risks 33. Over the last few years, 2500 rpm for 15 minutes and filtrate was stored in sterile bottle at 50C 15 several experimental studies have revealed biological and pharmaco- for further use . logical properties of phenolics compounds, especially their anti-in- flammatory activity, antiviral and cytotoxic activity34. Phenolics are Preliminary screening and quantification of phytochemicals active in curing kidney and stomach problems as well as helpful as The condensed extracts were used for preliminary screening of anti-inflammatory in action35. Phenols are reported antitumour agents phytochemicals by following the methods for alkaloids, lignin, phenols 36 16 17 and to exhibit antiviral and antimicrobial activities , hypotensive ef- and steroids ; Coumarins and flavonoids ; glycosides, terpenoids 37 13 18 fects and antioxidant properties . Phenols are said to offer resistance and saponins and tannins . Quantification of alkaloids and sa- to diseases and pest in plants38. ponins 15; flavonoids 19 and phenols and tannins 20. Quinones are present in seven plants of selectd species. Quinones RESULTS AND DISCUSSION are known to act as mobile electron carriers within the lipid phase of Qualitative phytochemical studies revealed that the aqueous extract the membrane in photosynthetic and respiratory electron transport of plant species contained phytoconstituents like Alkaloids, chains39. The function of quinones is essential for proton-based en- Coumarins, Flavonoids, Glycosides, Lignins, Phenols, Quinones, ergy coupling and specific binding sites, allowing only two- electron Saponins, Steroids, Tannins and Terpenoids (Table 1). Seven spe- transfer steps, prevent damage of the cell by free-radical formation40. cies of selected plants posses Alkaloid compounds in aqueous ex- tracts. The highest levels of alkaloids were found in Cymbopogon Saponin content are present in maximum number of selected species coloratus 1.317 mg/g dw followed by Cynodon dactylon. Alkaloids showed. Maximum levels of saponins were present in Heteropogon are found in Cymbopogon and Cynodon dactylon 21 22. Most of the contortus 2.81 and lower level in Sporobolus coromandelianus 0.005 plants have alkaloids in different organs with different chemical mg/g dw. The results are agrement with21 22 in Cymbopogon coloratus, configurations 23 . Alkaloids are beneficial chemicals to plants with Cynodon dactylon. Traditionally saponins have been extensively predator and parasite repelling and physical state. Number of alka- used as detergents as pesticides and mollusicides, in addition to their loids were isolated from dicots and using as efficient drugs. Coumarins industrial applications as foaming and surface active agents and also are present in eleven plants among selected twenty grasses, which have beneficial health effects11. Saponins has relationship with sex reduce the impact of grazing animals. Though the compound has a hormones like oxyticin, which is a sexharmone involved in controlling pleasant sweet odor, it has a bitter taste, and animals tend to avoid it. the onset of labour pains in female and the subsequent release of Reports on coumarin activity includes anti-HIV, anti-tumor, anti-hy- milk41. Saponins enhance nutrient absorption and aid in animal diges- pertension, anti-arrhythmia, anti-inflammatory, anti-osteoporosis, an- tion. They are bitter to taste, and so, can reduce plant palatability or tiseptic and analgesic. It is also used in the treatment of asthma and even imbue them with life threatening animal toxicity42. There is tre- lymphedema 24. mendous, commercially driven promotion of saponins in traditional medicine preparation43, where oral administration might be expected Among twenty selectd plants fourteen species possessing Fla- to lead the hydrolysis of glycosides from terpenoid and obviation of vonoids. Alloteropsis cimicina containg maximum levels of fla- many toxicity associated with the intact molecule. Saponin protects vonoids followed by Heteropogon contortus. Flavonoids are re- the plant against microbes fungi and viral diseases44. 45reported that, corded in the grass of Cynodon dactylon 22, Sacchrum spontaneum25, saponins had expectorant action which is very useful in the manage- Imperata cylindrical26, Pennucetum purpureum27 and Cymbopogon ment of upper respiratory tract inflammation and Saponins have been citrates28. They also inhibit microbes which are resistance to antibi- found to possess hypocholesterolemic property for the control of otics29. Flavonoids are free radical scavengers and have strong anti- high blood lipids46. cancer activity30 evidence of their inherent ability to modify the body’s reaction to allergies, virus and carcinogens. flavonoid rich species Tannins are present in ten plants among selected species. Maximum

JPR:BioMedRx: An International Journal Vol.1 Issue 10.October 2013 947-951 R. Hari Babu et al. /JPR:BioMedRx: An International Journal 2013,1(10),947-951 - - + - - + + ------+ - + - - + - - + - + - + - + + + - - + - - - + + - + - - - + - - - - + - + - - + + - - + - - - - - + ++ - + + + + - ++ + + - + + - + + + + 0.310± 0.017 T annins - 0.012 ± 0.001 0.520 ± 0.004 - 0.720 ± 0.010 0.430 ± 0.032 0.003 ± 0.001 0.024 ± 0.002 - - 0.038 ± 0.031 - - - 0.172 ± 0.046 0.412 ± 0.011 - - + - + - + - + ------+ - - + + - + + - - + + - + + + + - - + - + + - - + Saponins - 0.212 ± 0.010 0.180 ± 0.019 - 0.646 ± 0.026 0.920 ± 0.030 0.601 ±0.004 1.120 ± 0.047 - 0.039 ± 0.011 0.316 ± 0.007 0.422 ± 0.001 - 2.81 ± 0.012 0.306 ± 0.016 - 0.443 ±0.011 0.807 ± 0.013 0.005 ± 0.001 0.125 ± 0.016 + - + + - + ++ - - + + + + - - + - + - - Phenols 0.021 ± 0.002 0.040 ± 0.006 - - 0.800 ± 0.021 0.650 ± 0.030 - 0.246 ± 0.041 0.102 ± 0.014 0.004 ± 0.001 0.410 ± 0.020 - - 1.210 ± 0.019 - 0.037 ± 0.007 0.081 ± 0.002 - - 0.013 ± 0.002 + - - - + + + + - + + + + - + + - - + + - - - + - + - ++ ++ + - - + + + + ++ + - ++ - + + 0.021 ± 0.001 0.470 ± 0.019 0.210 ± 0.040 0.098 ± 0.013 0.004 ± 0.002 0.120 ± 0.041 0.007 ± 0.001 0.012 ± 0.003 0.910 ± 0.016 0.011 ± 0.001 Flavonoids 1.040 ± 0.033 - - 0.502 ± 0.021 - 0.820 ± 0.020 0.130 ± 0.014 indicates Absent. + + - - + + + - + - - - - + + - + - + + ‘ ------’ , 0.005 ± 0.001 S.E 0.008 ±0.002 0.540 ± 0.083 0.244 ± 0.030 Alkaloids - 0.208 ± 0.011 - 1.317 ±0.053 0.720 ± 0.027 - 0.018 ± 0.001 - ± - - - + - - + + ------+ + - + - + T able-1: Preliminary phytochemical screening aquesous extract of Twenty selected Poaceae Grass species T able-2: Quantification of secondary metabolites from selected Poaceae Grass species (mg/g dw). Name of the species Alkaloids Coumarins Flavonoids Glycosides Lignin Phenols Quinones Saponins Steroids Tannin Terpenoids Alloteropsis cimicina Aristida hystrix Aristida setacea Brachiaria racemosa Chloris barbata Cymbopogon coloratus Cynodon dactylon Dactyloctenium aegyptium Digitaria saguinalis Echinochola colonum Eleusine indica Eragrostis amabilis Eragrostiella bifaria Heteropogon contortus Panicum repens Perotis indica Pycreus flavidus Setaria pumila Sporobolus coromandelianus Sporobolus wallichii : ‘+’indicates presence and ‘-’indicates absence Name of the species Alloteropsis cimicina Aristida hystrix Aristida setacea Brachiaria racemosa Chloris barbata Cymbopogon coloratus Cynodon dactylon Dactyloctenium aegyptium Digitaria saguinalis Echinochola colonum Eleusine indica Eragrostis amabilis Eragrostiella bifaria Heteropogon contortus Panicum repens Perotis indica Pycreus flavidus Setaria pumila Sporobolus coromandelianus Sporobolus wallichii Note V alues are the average of Triplicates S.No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 S.No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

JPR:BioMedRx: An International Journal Vol.1 Issue 10.October 2013 947-951 R. Hari Babu et al. /JPR:BioMedRx: An International Journal 2013,1(10),947-951 content found in Cymbopogon coloratus followed by Brachairia REFERENCES racemosa which are also reported in Cymbopogon citrates by21 28; 1. Ahmed F, Urooj A, J young pharm, 2010, 2, 160- 164. Sacchrum spontaneum by25; Imperata cylindrical25 26 27. Tannins 2. Zheng W, Wang SY, J. Agric. Food Chem, 2001, 49, 5165– contribute property of astringency i.e fasten the healing of wound 5170. and inflamed mucous membrane and have receive conciderable at- 3. Thitilertdecha N, Teerawutgulrag A, Rakariyatham tention in the fields of nutrition, health and medicine, largely due to NAntioxidant and antibacterial activities of Nephelium their physiological activity, such as antioxidant, antimicrobial and anti-inflammatory properties. Herbs that have tannins as their main lappaceum L.extracts. 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