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EVALUATION OF PHYTOCHEMICAL CONSTITUENTS AND ANTIBACTERIALACTIVITY OF CARICA PAPAYA AND HIBISCUS ROSA SINENSIS AGAINST

PSEUDOMONAS AERUGINOSA AND AEROMONAS HYDROPHILA

Vandana Singh, S. A. John and K. P. Rao

ABSTRACTA study was conducted to investigate the phytochemical constituents and antibacterial activity of Carica papaya and Hibiscus rosa sinensisleaf extracts, using three different solvents (Distilled water, ethanol and butanol) through soxhlet extraction. Phytochemical screening ofthe dried leaves of papaya (Carica papaya) and China rose (Hibiscus rosa sinensis) revealed the presence of bioactive tannins, flavonoids, andsaponins, which have been linked to their antibacterial properties. Antibacterial activity of the aqueous and ethanolic extracts of plantmaterials were tested by disk diffusion method against Pseudomonas aeruginosa and Aeromonas hydrophilla. In both plants alkaloids,flavonoids, phenols and saponins were present in enormous amount. Total alkaloids (70.0-59.7 mg/g leaves), total phenols (36.2-40.2 µgg-1fw), total saponins (53.8-65.7 µg g-1fw), among all solvents used, Ethanolic extract of Carica papaya and Hibiscus rosa sinensis containedmore alkaloids, phenols and saponins and showed comparatively more antibacterial activity with zone of inhibition (6.0-9.0 mm), thanother solvents.

Keywords: Phytochemical, Antibacterial, Phenols, Alkaloids, Saponin

M.Sc. Student, Associate Professor, Assistant ProfessorDepartment of Biological Sciences,

Sam Higginbottom University of Agriculture, Technology And Sciences, Allahabad - 211007

INTRODUCTIONThis enables herbal medicines to be as effective as

conventional medicines, but also gives them the same potentialto cause harmful side effects. The range of pathogenic bacteriais wide and so is the variety of diseases caused by them. Despitethe existence of potent antimicrobial agents, resistant or multi-resistant strains are continuously emerging, imposing the needfor a continuous search and development of new drugs (Machadoet al., 2003).

Anti implant's activity of hibiscus flowers root ethanol extract.The anti hypertensive properties of petroleum ether, hydroalcohol, and chloroform of hibiscus flower extract (Siddiqui et al.,2006).

It has been used to treat digestive problems and intestinalworms as well as warts, sinusitis, eczema, coetaneous tuberculesand hardness of the skin. Green fruits are used to treat high bloodpressure, roundworm infection, dyspepsia, constipation,amenorrhea, skin disease, general debility and genito-urinarydisorders. Mucilage prepared from the Hibicus rosa-sinensis roothas been used in the treatment of coughs (Duke and Ayensu1985; Chopra et al., 1986).

Carica papaya is an evergreen shrub or small tree that growsbest in full sun to light shade. The plant likes lots of water andfertilizer in warm weather. It has been used locally in the treatmentof urinary tract infections (Aliyu, 2006).

Pseudomonas aeruginosa is a Gram-negative, aerobic, rod-shaped bacterium with unipolar motility. An opportunistic humanpathogen, P. aeruginosa is also an opportunistic pathogen ofplants. P. aeruginosa is the type species of the genusPseudomonas.

Pseudomonas aeruginosa is increasingly recognized as anemerging opportunistic pathogen of clinical relevance. One of the

most worrying characteristics of P. aeruginosa is its low antibioticsusceptibility. Aeromonas hydrophila Members of the genusAeromonas are facultatively anaerobic, oxidase positive, gram-negative bacteria whose natural habitat is in the aquaticenvironment. Some species are pathogenic for animals andhumans (Martin-Carnahan and Joseph 2005). The mesophilicaeromonads have been commonly isolated from patients withgastroenteritis although their role in disease causation remainsunclear. They are also associated with sepsis and wounds, andwith eye, respiratory tract, and other systemic infections. Thestudy was conducted with the aim to analyse the phytochemicalconstituents and antibacterial activity of leaf extract of Caricapapaya and Hibiscus Rosa sinensis against Pseudomonasaeruginosa and Aeromonas hydrophila.

MATERIALS AND METHODSCollection of test organisms : The pathogenic bacterial strains

Pseudomonas aeruginosa (MCCB0034) and Aeromonashydrophila (MCCBR0006)) were obtained from the "MicrobialCulture Collection Bank", Department of Microbiology andFermentation Technology, Sam Higginbottom University ofAgriculture, Technology And Sciences, Allahabad.

Phytochemical screening : of alkaloids (Mayer's and Wagner'sreagent) flavonoids (10% lead acetate solution) saponins(Distilled water) tannins (Lead acetate) steroids (Chloroform,Conc. H2SO4 ) amino acids (Ninhydrin reagent) was carried.

Quantitative analysis of extracts : for total alkaloids (Dayanandet al., 1998) total phenols (Malick and Singh., 1980), totalsaponins (Brunner et al., 1984) was carried out. This wasfollowed by Screening of Antibacterial activity of extracts (Agarryet al., 2005)

The Allahabad Farmer, Vol. LXXIII, No. 4, October - December, 2017

Received 22-09-2014Accepted 07-09-2017

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RESULT AND DISCUSSIONQualitative analysis of extracts : The screening of Carica

papaya leaf extract showed the presence of Flavonoids, saponins,Alkaloids, Steroids in all three extracts while Tannins, Glycosides,shows negative result in distilled water and positive result inFlavonoids, Saponins, Alkaloids, and Steroids.

Estimation of total saponin (µg g-1fw) leaves of Carica papayaand Hibiscus rosa sinensis The saponin were quantified andtabulated in table 2. Showed significant increase in phenoliccontent was recorded in T3 Butanol (33.9µg g-1fw) followed byT1 distilled water (9.6µg g-1fw). All the treatments were foundstatistically significant. T2 is a best result (53.8µg g-1fw). Thesaponin were quantified and tabulated in table 2. Showedsignificant increase in phenolic content was recorded in T3Butanol (31.7 µg g-1fw) followed by T1 distilled water (21.3 µg g-

1fw). All the treatments were found statistically significant.T2 isa best result (65.7 µg g-1fw).

Table. 1: Qualitative Analysis of phytochemical constituentsof the Papaya (Carica papaya) and China rose(Hibiscus rosa sinensis) leaves

Papaya China rose(Carica papaya) leaves (Hibiscus rosa sinensis)

Phytochemicals Solvents SolventsD.W. Ethanol Butanol D.W. Ethanol Butanol

Tannins - + + + + +Flavonoids + + + + + +Saponins + + + + + +Alkaloids + + + + + +Glycosides - + - + + -Steroids + + + + + +

Table - 2: Estimation of total Alkaloids (mg/gm) leaves ofCarica papaya

Alkaloids Phenols SaponinSolvents (mg/gm leaves) (µg g-1fw) (µg g-1fw)

Carica Hibiscus Carica Hibiscus Carica Hibiscuspapaya rosa papaya rosa papaya rosa

sinensis sinensis sinensis D.W. Extract 15.8 25.7 16.5 15.0 9.6 21.3Ethanol Extract 70.0 59.9 36.2 40.1 53.8 65.7Butanol Extract 24.6 41.7 12.1 36.8 33.9 31.7

Table - 3 : Effect of Carica papaya and Hibiscus rosasinensis Extract on Zones of inhibition ofHibiscus rosa sinensis against the P. aeruginosaand A. hydrophilla (mm)

Solvents Zone of inhibition (mm)Carica papaya Hibiscus rosa sinensis

Control 0.0 0.0D.W. Extracts 4.0 6.5Ethanol Extracts 6.0 9.0Butanol Extracts 5.0 7.5

Estimation of total Alkaloids (mg/gm) leaves of Hibiscusrosasinensis

The alkaloids were quantified and tabulated in table 2 showedsignificant increase in alkaloids content was recorded intreatments T1 distilled water (25.7 mg/gm) followed by T3butanol (41.7 mg/gm) T2 ethanol (25.7 mg/gm). All the treatmentswere found statistically significant. However, ethanolic extract wasfound superior over all the treatment.

Estimation of total Phenols (µg g-1fw) leaves of Carica papayaand Hibiscus rosa sinensis The phenols were quantified andtabulated in table 2 showed significant increase in phenoliccontent was recorded in T3 Butanol (12.1µg g-1fw) followed byT1 distilled water (16.5µg g-1fw). All the treatments were foundstatistically significant treatments. The phenols were quantifiedand tabulated in table 2 showed significant increase in phenoliccontent was recorded in T3 Butanol (36.8µg g-1fw) followed byT1 distilled water (15.0µg g-1fw). All the treatments were foundstatistically significant.T2 is a best result (40.1µg g-1fw). Thepresence of phenolics compounds in the leaves of Hibiscus rosasinensis and Carica papaya indicate that these plants are anti-bacterial agents. Phenolic compound as electron donors arereadily oxidized to form phenol to ion or quinine, an electronaccepter. This give rise to practical uses prorogated phenol isused as cleaning agent (Salah et al, 1995).

Antibacterial activity of extracts:The antibacterial activity of all the plant extracts of Carica

papaya against the P. aeruginosa and A.hydrophilla shown intable 3. It was found that maximum antibacterial activity wasshown by ethanol with (6.000 mm) of inhibition zone followed bybutanol with (5.000 mm) and distilled water with (4.000 mm). ThusEthanolic extract was best among all the treatments.

Zones of inhibition of Hibiscus rosa sinensis against the P.aeruginosa and A. hydrophilla (mm): (Igbinosa et al., 2009; Stray,1998).

CONCLUSIONIt is concluded from this study that the leaves of Carica

papaya and Hibiscus rosa sinensis have antibacterial propertiesagainst Pseudomonas aeruginosa and Aeromonas hydrophila.The present study suggests that among all solvents used, theEthanolic extract of Carica papaya and Hibiscus rosa sinensiscontained more alkaloids, phenols and saponins and showedcomparatively more antibacterial activity than other solvents. Theantibacterial activity analysis of Carica papaya and Hibiscus rosasinensis extracts against the choosen organisms the inhibitionzone diameter produced, ethanolic extract shows best result. Forboth microorganism Hibiscus rosa sinensis have more inhibitionzone of Carica papaya. The result depicted the use of theseplants in treating antibacterial infection and shows that Caricapapaya and Hibiscus rosa sinensis could be exploited for newpotent antibacterial agents.

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The Allahabad Farmer, Vol. LXXIII, No. 4, October - December, 2017