Male Carica Crude Solvent Extracts

manful genus genus Carica Crude Solvent ExtractsMale pawpaw tree have flowers that grow on short stalks (Chin et al., 2001). Carica pawpaw is a native of the tropics of America, perhaps from S break throughhern Mixico and neighouring central American (Morton, 1987). According to Eno et al. (2000), pawpaw is the fruits, of the plant. Carica papaw belong to the genus carica. Preliminary qualitative and denary phytochemical analysis of ethanol and aqueous draw offs of C. papaya tree showed the presence of many phytocompounds. These choices were found to inhibit these eight shield micro-organisms Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, genus Aspergillus niger, Penicillium notatum, Fusarium solani and Candida albican.(Okoye, 2011). proximate analysis of leaves of C. papaya withal showed appreciable quantity of ash content, sore protein, crude fat, crude fibre, carbohydrate and high calorific value (Okoye et al., 2011). In this usher in travel, the author intends to study the preliminary pharmaceutical constituents of crude solvent extracts of flower and stalk of virile person Carica papaya since little or no work had been done in this atomic number 18a. The aim and objective of this work is to determined the phytocompounds present in flower and stalk of male Carica papaya to extract the crude solvent extracts of the flower and stalk of male C. papaya and to find out whether the solvent extracts can inhibit the addition of ten pathogenic fungi. Carica papaya is composed of many biological active compounds, many of which are found concentrated in the latex, which is present in parts of the plant (Madrigal et al., 1980).Within Carica papaya plants, the do using of bio-actives will vary with position of plant, age of plant and cutivar. Also, concentration of bioactive differs between male, hermaphrodite and female plants. Female plants exude more latex than hermaphrodite and male plants. Carica papaya latex is rich in cystein proteinases which are proteolytic enzymes (caricain, chymopapain, papain and glucylendopeptidase) these constitute 80% of latex enxymes. Other enzymes present are glycosyl hydrolases (-1, 3-glucanases, chitiriases and lyzozynus) protease inhibitors (cystertin and ghtaminylcyclotrunsferces and lipases (Moussaoul et al., 2001). It was reported that intake of two table spoons of pulverized papaya seeds mixed with hot water system supply twice per day is used in the handed-down management of diabetes and obesity (Adeneye and Olagunju, 2009). Carica papaya (pawpaw) contains the enzyme papain, a protease used for tendering meat and other proteins (Morton, 1987). The fruits are popularly used and processed into juice and wine, and also cooked as vegetable (Gragson, 2001). The seeds are medically important in the treatment of sickle cell disease and poisoning related disorder. The lead tea or extract had a reputation as a tumor destroyer agent. The flesh green tea is anti septic while the brown dried leaves are best served as tonic and blood purifier (Ezugwu, 2008). Due to its antioxidant and fibre content, it is used in treatment of ailments such as chronic indigestion, overweighing, obesity, high blood force (Everetta, 2003).Sample Collection and PreparationFlower stalk of male Carica papaya was collected from Adazi-enu in Anaochia Local Government Area of Anambra State, Nigeria. It was dried at a lower place air and mild sun-shine, for about three weeks and ground into powders. The ground sample was thence kept in a clean polyethylene bottle until require for analysis. Phytochemical and the extr achievement of the active components are determined by the methods outlined by Harbon (Harbon, 1973). The antifungal activity of flower and stalk of male C. papaya was determined by agar well diffusion method (Okeke et al., 2001). The zone of inhibition was recorded to the nearest size in mm (Norrel, 1997). After extraction of the active components dev elopment three incompatible solvents separately (Ethanol, Water and N-hexane), the solvent extracts were evaporated to dryness at about 67, 98 and 66C respectively in a water bath separately. 1, 2, 3, 4 and 5mg of dry ethanolic, n-hexane and water extracts were weighed into five different labeled evidence tubes differently. Then 10ml of the corresponding solvents used for extraction was added to the dried extracts to make 0.1,0.2,0.3,0.4 and 0.5mg/ml concentrations of the extracts.The MIC of flower and stalk of male Carica papaya were found out by using 0.1, 0.2, 0.3, 0.4, and 0.5mg/ml of each extract which were added to test tubes containing 1ml of sterile medium. The tubes were then inoculated with a drop of microbial suspension and incubated for 48 hours at 25C. Then 0.1, 0.2, 0.3, 0.4, and 0.5mg/ml of amphotericin B (for A. flavus, F. verticilloides, A. parasiticus, F. oxysporum and Fluconazole (for all candida was used for positive control and water for negative control respe ctively. The MIC value was determined, macroscopically after incubation in comparison with the growth and sterility control. MFC the plates (petri-dishes) were divided into six different sections and labeled with the different concentration on the base of the plates, these were used to plate out the contents of each tube with the respective sections of the plate. The plates were incubated for 18 24 hours at 37C after which the MFC were recorded. Three replicates were done for each extract concentration and control against the fungi.Results Tables 1 Qualitative Phytochemical Analysis of Flower and Stalk of Male Carica papaya pattern of phytocompoundInference AlkaloidFlavonoidSaponinTanninsTerpenoidsSteroidsPhenolCardiac glycosides+++++++++Key + = present, = absentTable 2 Quantitative Estimates of Phytochemical Constituents of Flower and Stalk of Male Carica papayaClass of phytocompoundInference AlkaloidFlavonoidSaponinTanninsTerpenoidsSteroidsPhenolCardiac glycosides0.530.010.860.0 20.370.022.600.010.210.010.080.01Nil1.870.02Table 3 Antifungal activities of crude solvent extracts of flowers and stalk of male Carica papayaText organisms(fungi)Conc. of extract(mg/ml)Average Diameter (mm) of InhibitionZone+ve control for all Candida Fluconazoleothers Anaphotericin-ve Control distilWaterDistill H2OEtOHN-HexaneAspergillusniger0.11.9 0.20NANA16.26 0.25NA0.22.8 0.10NANA19.00 0.10NA0.33.1 0.022.33 0.02NA21. 6 0.21NA0.44.04 0.103.21 0.01NA23.2 0.28NA0.55.00 0.023.98 0.102.41 0.0224.80 0.01NAMicrosporumgypseum0.1NANANA8.00 0.02NA0.2NANANA8.60 0.10NA0.3NA2-33 0.02.NA8.60 0.06NA0.42.61 0.013.21 0.01NA9.98 0.22NA0.53.22 0.013.89 0.10NA10.40 0.01NACandidaalbican0.12.40 0.017.5 0.022.00 0.0130.08 0.02NA0.22.63 0.018.2 0.012.88 0.0233.20 0.02NA0.33.91 0.029.00 0.033.12 0.0135.80 0.10NA0.44.62 0.029.97 0.013. 92 0.0137.00 0.03NA0.54.88 0.1011. 00 0.024.17 0.0230.28 0.17Aspergillusflavus0.11.21 0.012.80 0.20NA17.80 0.10NA0.21.90 0.103.65 0.10NA21. 00 0.20NA0.32.40 0.204.00 0.02NA23.23 0.10NA0.42.86 0.014.86 0.01NA25.00 0.05NA0.53.16 0.305.37 0.20NA28.4 0.10NAFusariumVerticilloides0.1NANANA7.00 0.30NA0.2NANANA7.90 0.02NA0.3NANANA9.00 0.30NA0.4NANANA9.58 0.01NA0.5NANANA10.22 0.07NAAspergillusparasiticus0.11.38 0.02NA3.73 0.0123. 80 0.2NA0.22.06 0.01NA4.02 0.3024.00 0.10NA0.32.97 0.02NA4.08 0.2025.61 0.50NA0.43.30 0.02NA4.60 0.0126. 00 0.20NA0.53.88 0.01NA4.88 0.0227.20 0.10NAFusariumoxysporum0.1NANANA18.37 0.40NA0.2NANANA20.00 0.20NA0.3NANANA21.56 0.10NA0.4NANANA22.26 0.10NA0.5NANANA24.00 0.01NACandidaglabrata0.11. 35 0.012. 60 0.014. 30 0.0122.00 0.01N a0.22.29 0.034.31 0.032. 10 0.0224.1 8 0.20NA0.32 .96 0.025.65 0.012.80 0.0326.00 0.02NA0.43. 12 0.037. 04 0.013. 20 0.0227.86 0.30NA0.53. 84 0.018. 52 0.023. 90 0.0129.00 0.10NACandida parasilosis ticum0.13.80 0.024.20 0.022.02 0.0123.00 0.10NA0.24.22 0.024.86 0.012.23 0.0223.86 0.20NA0.34.92 0.015.12 0.022 .77 0.0124.12 0.02NA0.45.28 0.025.66 0.013. 12 0.0426.00 0.10NA0.55. 88 0.015.91 0.013.93 0.0127.88 0.30NACandidatropicalis0.13.48 0.20NA0.10 0.022 1.00 0.20NA0.24.01 0.01NA2.1 5 0.022 1.95 0.10NA0.34.69 0.023.66 0.022.54 0.0123.00 0.10NA0.45.20 0.015.00 0.033.23 0.2023.86 0.02NA0.55.87 0.016.91 0.013. 87 0.0425.00 0.03NATables 4 Minimum Inhibitory Concentration (MIC)Minimum inhibitory concentration (mg/ml)Test organisms (Fungi)Water extract (MIC)Ethanol extract (MIC)Hexane extract (MIC)Aspergillus niger0.05 0.010.25 0.010.45 0.01Microsporum gypseum0.35 0.020.25 0.03NICandida albican0.05 0.020.05 0.010.05 0.01Aspergillus flavus0.05 0.020.05 0.01NIFusarium verticilloidesNININIAspergillus parasiticus 0.01NI0.05 0.01Fusarium oxysporumNININICandida glabrata0.05 0.01 0.010.05 0.01Candida parasilosis 0.05 0.010.05 0.010.05 0.01Candida tropicalis0.05 0.010.25 0.010.05 0.01NI No InhibitionTables 5 Minimum fungicidal Concentration (MFC) of Ten FungiMinimum fungicidal concentration (mg/ml)Test organisms (Fungi)Water extract(MFC)Ethanol extract(MFC)N-Hexane extract (MFC)Aspergillus niger0.040.010.150.030.230.02Microsporum gypseum0.300.010.250.01NICandida albican0.020.010.020.010.020.02Aspergilus flavus0.050.010.050.02NIFusarium verticilloidesNININIAsperigillus parasiticus0.050.01NI0.030.01Fusarium oxysporumNININICandida glabrata0.050.030.030.010.020.01Candida parasilosis 0.050.010.030.010.050.02Candida tropicalis0.030.010.050.010.050.01 Key NI No InhibitionDiscussion Table 1 and 2 showed both qualitative and quantitative hazard of phytocompounds present in the flower and stalk of male Carica papaya. The phytochemical constituents of the flower and stalk are as follows alkaloids 0.53 0.01%, floaonoid 0.08 0.02% saponins 0.37 0.02% tannins 2.06 0.01%, terpenoid 0.02 0.01%, steroids 0.08 0.01, phenol nil, cardiac glycoside 1.87 0.02%. The flower and stalk of male Carica papaya contain an appreciable quantity of alkaloids. It coul d be used to remedy some diseases, depending on the type of alkaloids it contains. The presence of alkaloids signified the possession of medicative values within the flower and stalk. Pure isolated alkaloids are used as basic medicinal agents and fungicidal properties. The presence of steroids in the sample indicates that it could be used to decrease the presence of cholesterol in the bloodstream. Saponins have been used in the treatment of cardiovascular disorders, they facilitate and ease the process of digestion and encourage the growth of beneficial bacteria within the intestine. Saponins also contribute to health and efficiency of the immune system by binding to germs and other pathogens as they enter the body. In this sense they function as natural anti-bodies. They are found tobe antimicrobial and are particularly effective when used to treat yeast a infections. The presence of cardiac glycosides in the flower and stalk of male Carica papaya indicates that it could be used, in the treatment of congestive heart failure and cardiac arrhythmia.Table 3 portrayed the result of antifungal activities of three solvent extracts of the flower and stalk of male Carica papaya on ten micro-organisms (fungi) investigated in this work. They are A. niger, M. gyspseum, C. albican, A. flavus, F. verticilloides, A. paraciticus, F. oxysporum, C. glabrata, C parasilosis and C. tropicalis. Five different concentrations of aqueous, ethanolic, and normal hexane extracts were used. At 0.1-0.5mg/ml concentration, aqueous extract showed some inhibitory effect on eight out of the ten test fungi. The fungi are A niger, M. gypseum, C. albican, A flavus, C. parasilosis, A. parasiticus, C. glabrata and C. tropicalis. No action was recorded on the two remaining test fungi F. verticilloides and F. oxysporum. At 0.1-0.2mg/ml concentration, ethanol extract shows some inhibitory effect on seven out of the ten test fungi. These are A, niger, M. gypseum, C. albican, A. flavus, C. glabrata, C. parasilosis and C. tropicalis. At 0.1-0.2mg/ml, ethanolic extract shows no action against three fungi. These are F. verticilloides, A. parasiticus, and F. oxysporum. At 0.1-0.5mg/ml, n-hexane extract indicated some inhibitory effect on six out of the ten test fungi. These are A, niger, C. albican, A. parasiticus, C. glabrata, C. parasilosis and C. tropicalis. It had no action on four fungi. These are M. gypseum, A. flavus, F. verticilloides and F. oxysporum.. Table 3 also showed the commercial drugs used as positive and negative control. Positive control using two different antibiotics specifically showed remarkable inhibitory effects on the ten test fungi while the negative control showed no action against the ten test fungi.Table 4 showed the results of the minimum inhibitory concentration (MIC) of the aqueous, ethanolic and n-hexane extracts of flower and stalk of male Carica papaya on the ten test fungi. The least MIC of the aqueous extract 0.05 0.01mg/ml was shown on five t est fungi These are A. niger, A. parasiticus, C. glabrata, C. parasilosis and C. tropicalis. MIC of the ethanolic extract 0.05 0.0lmg/ml was shown on four test fungi. These are, C. albican, A. flavus, C. glabrata, and C. parasilosis. MIC of the n-hexane extract 0.05 0.0lmg/ml was shown on five test fungi. These are C. albican, A. parasiticus, C. glabrata, C. parasilosis and C. tropicalis.Table 5 showed the result of minimum fungicidal concentration (MFC) of the aqueous, ethanolic and n-hexane extract of flower and stalk of male Carica papaya on ten test fungi. For aqueous extract, the least MFC 0.2 0.01mg/ml was shown on one test fungi,Candida albican. For ethanolic extract, the MFC 0.02 0.0lmg/ml was shown on one test fungi Candida albican. For n-hexane extract the least MFC 0.02 0.01mg/rnl was shown on one test fungi Candida glabrata.ConclusionThe analytical investigation showed that, the crude solvent extracts of flower and stalk of male Carica papaya have antigfungal effect on these microorganisms Aspergillus niger, Microsporum gypseum Candida albican, Aspergillus flavus, Aspergillus parasiticus, Candida glabrata, Candida parasilosis and Candida tropicalis. This implies that crude solvent extracts of flower and stalk of male Carica papaya can be used to cure the diseases caused by the above mentioned microorganisms. Bioactive ingredients responsible for the antimicrobial properties of the flower and stalk of male C. papaya should be elucidated. Male C. papaya Plant Flower and Stalk of male Carica papayaReferences1.Okoye, E.I., Orakwue, F.C. and Ebeledike, A.O. (2011).Nutritive Evaluation of Seeds of Gongronema latifolium (utazi) and leaves of Carica papaya.Anachem daybook, Vol. 5(1) pp. 985 988. ISSN 2006-5221.2.Okoye, E.I. (2011). Preliminary Phytochemical Analysis and Antimicrobial Activity of Seeds of Carica papaya. Journal of Basic Physical Research, Vol. 2, No. 1, Pp. 66 69. ISSN 2141- 8411.3.Okeke M.I., Iroegbu C.U., Eze. E.N., Okoli A. S and Esimone C.O. (2001) Evaluation of extracts of the root of Landolphia owerrience for antimicrobial activity. J. Ethnopharmacol, 78119-127.4.Norrel S.A and Messley K.E. 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