International Journal of Advanced Research in Chemical Science (IJARCS) Volume 5, Issue 10, 2018, PP 21-29 ISSN No. (Online) 2349-0403 DOI: http://dx.doi.org/10.20431/2349-0403.0510005 www.arcjournals.org

Chemical Constituents of Methanol Leaf Extract of africana C.D. Adams by GC MS

Ahamefula Anselm Ahuchaogu1*, Bright Chinenyeze Onyekwere3, Emmanuel Udeala Godwin.I. Ogbuehi1, Godwin Uzoma Ihenetu2, John Bull Onyekachi Echeme2 1Department of pure and Industrial Chemistry, Abia State University, Uturu, Nigeria 2Department of Chemistry, Michael Okpara University of Agriculture, Umudike, Umuahia, Abia State, Nigeria 3Federal Polytechnic Nekede Owerri, Imo State

*Corresponding Author: Ahamefula Anselm Ahuchaogu, Department of pure and Industrial Chemistry, Abia State University, Uturu, Nigeria

Abstract: The therapeutic value of (Asteraceace), commonly known as ‘‘hemorrhage ’’ has been recognized as a component of traditional medication for the treatment of various human ailments. In Nigeria, a large number of medicinal have been used for many centuries for treating various diseases. Medicinal plants have been as remedies for human diseases because of its chemical contents of therapeutic value. Most traditional medicines are developed from nature. Thus plants remain a major source of medicinal compounds A. africana, which is widespread in , is used in traditional medicine to stop bleeding from wounds, clean the surfaces of sores. The current study was undertaken to evaluate the phytochemical constituents of Methanol leaf extract of Aspilia Africana C.D. Adams by Gas Chromatography –Mass Spectrometry. The GC-MS analysis revealed the presence of 34 bioactive compounds such as flavonoids, alcohols, aldehydes, aromatic compounds, fatty acid methyl esters, terpenoids, phenolics, and steroids that can be postulated for antibacterial activity. The prevailing compounds are: Betulin (24.912%), Acetic acid, 10,11-dihydroxy-3,7,11-trimethyl-dodeca-2,6-dienylester (8.087%), 2,5,5,8a-Tetramethyl-4-methylene-6,7,8, 8a-tetrahydro-4H,5H-chromen-4a-yl hydroperoxide (7.074 %), 2,6,8-Trimethylbicyclo[4.2.0]oct-2-ene-1,8- diol (7.848%), Cholestan-3-ol, 2-methylene-, (3,5)- (4.446%), 4-(2,6,6-Trimethylcyclohexa-1,3-dienyl)but-3- en-2-one (3.579 %), 3-Hydroxy-α-ionene (3.361 %), n-Hexadecanoic acid (3.253 %), 3,4,4-Trimethyl-3-(3- oxo-but-1-enyl)-bicyclo [4.1.0] heptan-2-one (2.219 %),. Keywords: GC-MS, photochemical constituent, methanol extract, anti-microbial, Aspilia africana

1. INTRODUCTION In developing countries, communities rely heavily on traditional herbal medicines in order to meet their primary health care needs. In many industrialized countries herbal medicines are gaining popularity as alternative and complimentary therapies. Some of the plants are used as food or medicine. These plants exhibit a wide range of biological and pharmacological activities such as anti- cancer, anti inflammatory, diuretic, oxytocic, laxative, antispasmodic, antihypertensive, anti-diabetic, and anti-microbial functions.[1]

Their role is twofold in the development of medicines and served as a natural blue print for the development of new drugs [2-3] Plants used for traditional medicine contain a wide range of substances that can be used to treat chronic as well as communicable diseases [4]. Medicinal plants represent a rich source of antimicrobial agents [2] [5].According to World Health Organization

(WHO) more than 80 % of the world's population relies on traditional medicine for their primary healthcare needs. Medical plants contain large varieties of chemical substances, which possess important therapeutic properties that could be utilized in the treatment of human diseases. The potential of higher plants as source of new drugs is still largely unexplored. Among the estimated 250,000 –500,000 plant species, only a small percentage has been investigated phytochemically and the fraction submitted to biological or pharmacological screening is even smaller [6]. Aspilia africana is one of such medicinal plants, which are fast gaining recognition. It is a semi woody herb occurring throughout the regions of the savannah and tropical Africa on wastelands [7]. It

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 21 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS is known by various names among the Nigerian populace (Orangila in Igbo, Tozalin in Hausa, Yunyun in Yoruba and "Edemedong" in Efik). The plant has been reported in literature to possess antimicrobial [8], haemostatic [9], anti-inflammatory [10] and anti-fertility [11] activity. The wound healing and anti-ulcer activity of Aspilia africana n-hexane and methanolic extracts have also been reported [12-13][10]. In South-eastern Nigeria, all the parts of Aspilia Africana are widely used in traditional medicine, the leaves are claimed to be effective in the treatment of a number of health conditions. The infusion from the crushed leaves is applied on the wound, throughout many African communities, to stop bleeding and for cleaning the surfaces of sores. Historically, Aspilia africana was used in Mbaise and most Igbo speaking parts of South eastern Nigeria to prevent conception, suggesting potential contraceptive and anti-fertility properties[14] Taxonomical classification of Aspilia Africana L Kingdom : Plantae Phylum : Spermatophyta Subphylum : Angiospermae Order : Family : : Aspilia Species : Aspilia africana

Fig1. Aspilia africana leaves and flowers 2. MATERIALS AND METHODS 2.1. Collection and Authentication of Plant Material Wild growing Aspilia africana plants fresh leaves were collected from Abia state university, Uturu Nigeria. farmland on 26th July, 2018 and identified by Dr E.O Emmanuel, a plant taxonomist. Voucher specimens [1769] were deposited at the herbarium section of Department of Plant Science and Biotechnology, Abia State University. 2.2. Preparation of Leaf Extracts The Fresh harvested plant leaves were washed with tap water and rinsed with sterile distilled water. The fresh plant leaves sample was air dried on the laboratory bench for ten days at temperature below 30 0C to avoid decomposition of thermo labile compounds. The sample was milled using and electric blender to coarse powder and powdered sample was kept in a clean closed container pending

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 22 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS extraction. 50 g of pulverized leaf material was mixed with 250 ml of solvent (95 % methanol ) and kept in rotary shaker at 100 rpm overnight and filtered with Whatman No.1 filter paper The extract was concentrated under reduced pressure using Digital Heidolph Rotary evaporator (4000 series) and the supernatant plant extract was decanted after complete removal of the solvent 2.3. GC –MS Analysis 2.3.1. Preparation of Extract Two micro liters of the methanol leaf extract was employed in GC–MS analysis, for the identification of compounds. 2.3.2. GC – MS Analysis Conditions The GC-MS analysis of the methanol leaf extrac of Aspilia africana was carried out using a HP 7890 GC instrument integrated with an Agilent 5975C MSD mass spectrometer (Aligent, Santa Clara, CA, USA). The capillary column was an Agilent HP-5MS (30.m x 0.25mm i.d. x 0.25 NM film thickness), helium (Purity > 99.999 %) was used as the carrier gas, and the flow rate was 1 ML/min. The injector temperature was 2500c, and the injection mode was splitless. The G.C oven temperature was held at 50 0C for 5min, which was increased to 210 0at a rate of 30C/min, maintained at 210 0Cfor 3 min, and finally increased to 230 0C at 150C/min. The mass spectrometer conditions were as follow: [12:13:14] ionization energy, 70 Ev; ion Source temperature, 230 0C; quadrupole temperature, 150 0C; quadrupole mass spectrometer scan range 30 – 500 atomic mass units (amu); solvent delay time 2.8 min. 2.4. Components Identification The components of the chloroform extract of Ageratum conyzoides was identified by matching the peaks with computer Wiley Ms. libraries and confirmed by comparing mass spectra of the peaks and those from literature (Ahuchaogu et al., 2018.) 3. RESULTS AND DISCUSSION Gas chromatography – mass spectrometry (GC-MS) is a method that combines the features of gas liquid chromatography and mass spectrometry to separate different substances within a test sample based on their retention time [16]. In recent times, GC-MS has become well recognized as a key technological platform for secondary metabolite profiling in both plant and non-plant materials [17] The bioactive components present in the leaf methanol extract of the plant were identified by GC–MS analysis. The gas chromatogram shows the relative concentrations of various compounds eluted as a function of their retention time (Fig. 2). Identification of the compounds was done by comparing their mass spectra and retention indices with those given in the literature and with the authentic samples. The methanol extract of the leaves of Aspilia africana on GC-MS analysis showed thirty four. The 34 compounds were identified namely; 1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl)-(1.173%), Indolizine (1.093%), Falcarinol (0.955%), Caryophylla-4(12),8(13)-dien-5α-ol (0.861 %), 3-Hydroxy-α-ionene (2.194%), 4-(2,6,6-Trimethylcyclohexa-1,3-dienyl)but-3-en-2-one (3.579 %), 1b,5,5,6a-Tetramethyl- octahydro-1-oxa-cyclopropa[a]inden-6-one (0.816 %), 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro- 4,4,7a-trimethyl-,(R)-(1.574%),2,5-Octadecadiynoic acid, methyl ester (1.320%), Panaxydol (1.391 %), Cyclododecanone, 2-methylene-(1.614 %), 5,8,11,14-Eicosatetraenoic acid, methyl ester, (all-Z)- (2.101%), 3-Hydroxy-α-ionene (3.361%), Cholestan-3-ol, 2-methylene-,(3,5)-(4.446%), 3,4,4-Trimet hyl-3-(3-oxo-but-1-enyl)-bicyclo[4.1.0]heptan-2-one(2.219%),2-Methyl-4-(2,6,6-trime thylcyclohex- 1-enyl)but-2-en-1-ol (1.332%), [1,1'-Bicyclopropyl]-2-octanoic acid, 2'-hexyl-, methyl ester (0.79%), Cholestan-3-ol, 2-methylene-, (3,5)- (2.400 %),7-Hydroxyfarnesen (2.518%},2,5,5,8a-Tetramethyl-4- methylene-6,7,8,8a-tetrahydro-4H,5H-chromen-4a-ylhydroperoxide (7.074 %),1-Oxaspiro[2.5]octa ne,5,5-dimethyl-4-(3-methyl-1,3-butadienyl)-,(2.564%), 2,6,8-Trimethylbicyclo [4.2.0]oct-2-ene-1,8- diol (7.848 %), 17-Octadecynoic acid (1.367 %), 2,5-Octadecadiynoic acid, methylester (1.355%), Acetic acid, 10,11-dihydroxy-3,7,11-trimethyl-dodeca-2,6-dienyl ester (8.087 %),9,12-Octadecad ienoyl chloride, (Z,Z)-(1.111), Cyclopropanebutanoic acid,2-[[2-[[2-[(2-pentylcyclopropyl)methyl] cyclopropyl] methyl]cyclopropyl]methyl]-, methyl ester(1.648%), n-Hexadecanoic acid (3.253%), 8, 11,14-Eicosatrienoic acid, (Z,Z,Z)- (0.816 %), 3,7,11,15-Tetramethyl-2-hexadecen-1-ol(1.488%), 8,11,14-Eicosatrienoic acid, (Z,Z,Z)-(0.879%), Tributyl acetylcitrate (0.888%),1H-Cycloprop [e]azule n-7-ol,decahydro-1,1,7-trimethyl-4-methylene-,[1ar-(1aα,4aα,7β, 7aβ,7bα)]-(0.973%), Betulin (24.912%).

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 23 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

Among the identified compounds n-, hexadecanoic acid is reported to have anti-inflammatory , Antioxidant, hypocholesterolemic nematicide, pesticide, anti-androgenic flavor, hemolytic,5-Alpha reductase inhibitor , potent mosquito larvicide [18], Betulin:antibacterial, antioxidant, antitumor, cancer-preventive, immunostimulant, chemopreventive, lipoxygenase-inhibitor, pesticide [19], Indolizine; the biological activities include antimicrobial activity, antioxidant activity, anti- inflammatory activity , anticonvulsant activity, enzymes inhibition, activity and activity as calcium entry blocker [20]. Panaxynol, antitumor [21]. Falcarinol: antimicrobial properties [22-23] The presence of various bioactive compounds justifies the use of the whole flower for various ailments by traditional practitioners

Fig2. GC-MS Chromatogram of Methanol extract of the leaves of Aspilia Africana Table1. Components detected in the plant of methanol extract of Aspilia africana MW: Molecular Weight, RT: Retention Time FORMU SN RT COMPONENT MW % LA 1 16.931 1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl C7H7NO2 137 1.173 2 18.928 Indolizine C8H7N 117 1.093 3 22.560 Falcarinol C17H24O 244 0.955 4 23.986 Caryophylla-4(12),8(13)-dien-5α-ol C15H24O 220 0.861 5 24.045 3-Hydroxy-α-ionene C13H20O2 208 2.194 6 24.456 4-(2,6,6-Trimethylcyclohexa-1,3-dienyl)but-3-en-2-one C13H20O 192 3.579 7 25.018 1b,5,5,6a-Tetramethyl-octahydro-1-oxa-cyclopropa[a]inden-6-one C13H20O2 208 0.816 8 25.748 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)- C11H16O2 180 1.574 9 25.983 2,5-Octadecadiynoic acid, methyl ester C19H30O2 290 1.320 10 27.098 Panaxydol C17H24O2 260 1.391 11 27.551 Cyclododecanone, 2-methylene- C13H22O 194 1.614 12 28.223 5,8,11,14-Eicosatetraenoic acid, methyl ester, (all-Z)- C21H34O2 318 2.101 13 28.457 3-Hydroxy-α-ionene C13H20O2 208 3.361 14 29.254 Cholestan-3-ol, 2-methylene-, (3,5)- C28H48O 400 4.446 15 29.691 3,4,4-Trimethyl-3-(3-oxo-but-1-enyl)-bicyclo[4.1.0]heptan-2-one C14H20O2 220 2.219 16 29.774 2-Methyl-4-(2,6,6-trimethylcyclohex-1-enyl)but-2-en-1-ol C14H24O 208 1.332 17 30.672 [1,1'-Bicyclopropyl]-2-octanoic acid, 2'-hexyl-, methyl ester C21H38O2 322 0.79% 18 30.857 Cholestan-3-ol, 2-methylene-, (3,5)- C28H48O 400 2.400 19 30.991 7-Hydroxyfarnesen C15H24O 220 2.518 2,5,5,8a-Tetramethyl-4-methylene-6,7,8,8a-tetrahydro-4H,5H- 20 238 31.695 chromen-4a-yl hydroperoxide C14H22O3 7.074

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 24 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

21 31.897 1-Oxaspiro[2.5]octane, 5,5-dimethyl-4-(3-methyl-1,3-butadienyl)- C14H22O 206 2.564 22 32.190 2,6,8-Trimethylbicyclo[4.2.0]oct-2-ene-1,8-diol C11H18O2 182 7.848 23 32.929 17-Octadecynoic acid C18H32O2 280 1.367 24 33.088 2,5-Octadecadiynoic acid, methyl ester C19H30O2 290 1.355 Acetic acid, 10,11-dihydroxy-3,7,11-trimethyl-dodeca-2,6-dienyl 25 298 33.465 ester C17H30O4 8.087 C H Cl 26 18 31 298 33.851 9,12-Octadecadienoyl chloride, (Z,Z)- O 1.111 Cyclopropanebutanoic acid, 2-[[2-[[2-[(2- 27 pentylcyclopropyl)methyl]cyclopropyl]methyl]cyclopropyl]methyl] 374 34.782 -, methyl ester C25H42O2 1.648 28 35.655 n-Hexadecanoic acid C16H32O2 256 3.253 29 38.264 8,11,14-Eicosatrienoic acid, (Z,Z,Z)- C20H34O2 306 0.816 30 38.507 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C20H40O 296 1.488 31 39.103 8,11,14-Eicosatrienoic acid, (Z,Z,Z)- C20H34O2 306 0.879 32 41.317 Tributyl acetylcitrate C20H34O8 402 0.888 1H-Cycloprop[e]azulen-7-ol, decahydro-1,1,7-trimethyl-4- 33 220 41.653 methylene-, [1ar-(1aα,4aα,7β,7aβ,7bα)]- C15H24O 0.973 34 57.558 Betulin C30H50O2 442 24.912

137 100 66 117 100

50 O N NH 50 90 O 94 109 44 80 119 63 0 39 51 40 50 60 70 80 90 100 110 120 130 140 150 27 43 55 58 69 74 78 84 0 (mainlib) 1H-Pyrrole-2,5-dione, 3-ethenyl-4-methyl- 20 30 40 50 60 70 80 90 100 110 120 130 (mainlib) Indolizine 55 84 100 100 91 OH 115 134 43 41 50 129 50 79 109 39 55 91 119 77 141 159 77 29 103 43 69 HO 67 27 53 123 137 95 155 173 31 45 58 72 89 97 152 187 201 217 229 243 0 0 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 (mainlib) (S,Z)-Heptadeca-1,9-dien-4,6-diyn-3-ol (mainlib) 2-Cyclohexen-1-ol, 2-methyl-5-(1-methylethenyl)-, cis- 177 100 109 100

O 50 O 43 50 91 105 HO 91 135 81 125 55 77 105 121 149 55 69 147 27 65 159 192 93 137 175 15 87 57 71 83 157 0 165 190 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 0 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 (mainlib) 3-Buten-2-one, 4-(2,6,6-trimethyl-1-cyclohexen-1-yl)- (mainlib) 3-Hydroxy-α-ionene 111 100 41 100 O H O 50 43 55 O 137 50 109 180 27 67 67 95 H 81 41 95 124 152 O 55 59 65 81 85 103 165 125 151 0 137 165 179 193 208 0 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 (mainlib) 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)- (mainlib) 1b,5,5,6a-Tetramethyl-octahydro-1-oxa-cyclopropa[a]inden-6-one

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 25 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

121 91 100 55 100 OH 105 91 117 41 O 41 50 65 77 O 50 55 79 105 133 67 131 O 29 145 29 51 147 51 159 161 175 177 191 184 197 213 231 242 259 0 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 (mainlib) 2,5-Octadecadiynoic acid, methyl ester (mainlib) Panaxydol 79 41 100 100 91 O O 67 55 67 50 105 50 41 81 55 119 29 95 150 43 69 109 133 98 123 O 71 137 194 29 161 175 203 151 87 187 220 31 37 45 51 165 176 15 232 247 259 275 287 318 0 0 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 (mainlib) Cyclododecanone, 2-methylene- (mainlib) 5,8,11,14-Eicosatetraenoic acid, methyl ester, (all-Z)- 105 69 100 100 41 119 81 81 95 55 91 OH 50 69 50 133 175 105 H 29 190 H H 208 121 147 HO 135 149 H 175 161 189 203 227 245 260 315 329 400 0 0 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 (mainlib) Cholestan-3-ol, 2-methylene-, (3β,5α)- (mainlib) 2-Methyl-4-(2,6,6-trimethylcyclohex-1-enyl)but-2-en-1-ol

73 100 45 O 55 67 50 81 O 95 109 123 137 151 165 178 192 207 224 237 291 322 0 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 (mainlib) [1,1'-Bicyclopropyl]-2-octanoic acid, 2'-hexyl-, methyl ester 43 100 43 100 69 OH OH 109 O 50 55 50 55 93 81 95 123 29 O 69 162 135 149 163 79 105 119 137 205 147 159 177 191 59 177 187 202 220 220 238 0 0 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 (mainlib) 7-Hydroxyfarnesen (mainlib) 2,5,5,8a-Tetramethyl-4-methylene-6,7,8,8a-tetrahydro-4H,5H-chromen-4a-yl hydroperoxide 124 100 41 100 109 81 93 55 69 109 119 50 43 50 OH O 206 OH 29 135 82 163 191 41 150 55 91 164 173 27 69 149 136 179 0 0 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 (mainlib) 1-Oxaspiro[2.5]octane, 5,5-dimethyl-4-(3-methyl-1,3-butadienyl)- (mainlib) 2,6,8-Trimethylbicyclo[4.2.0]oct-2-ene-1,8-diol

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 26 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

91 100 55 100 81 105 O 117 O 41 67 41 79 95 50 55 50 OH 67 131 O 29 60 109 145 51 29 123 159 177 15 135 149 160 170 181 192 202 213 223 234 247 261 277 191 0 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 (mainlib) 17-Octadecynoic acid (mainlib) 2,5-Octadecadiynoic acid, methyl ester 43 67 100 100 81 59 OH 41 55 81 71 93 O

50 50 95 55 O O 134 OH 29 105 121 Cl 143 161 110 121 29 179 135 149 262 202 220 238 280 14 163 177 191 205 216 234 245 280 298 0 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 (mainlib) Acetic acid, 10,11-dihydroxy-3,7,11-trimethyl-dodeca-2,6-dienyl ester (mainlib) 9,12-Octadecadienoyl chloride, (Z,Z)- 43 55 74 100 73 100 41 60 67 O 81 O 57 95 50 O 50 29 69 OH 107 121 135 83 129 149 270 97 256 168 227 115 157 171 213 185 199 264 277 298 334 143 185 194 227 239 0 0 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 (mainlib) Cyclopropanebutanoic acid, 2-[[2-[[2-[(2-pentylcyclopropyl)methyl]cyclopropyl]methyl]cyclopropyl]methyl]-, methyl ester (mainlib) n-Hexadecanoic acid

43 81 95 79 100 123 100 55 67 71 O 55 93 41 50 OH 50 OH 109 107 29 121 150 29 135 137 278 208 179 15 163 177 189 219 235 246 263 277 289 306 151 193 0 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 (mainlib) 8,11,14-Eicosatrienoic acid, (Z,Z,Z)- (mainlib) 3,7,11,15-Tetramethyl-2-hexadecen-1-ol 185 100

O OO 43 129 259 O O 50 O 57 O 29 O 157 112 139 213 273 329 84 168 231 0 30 60 90 120 150 180 210 240 270 300 330 360 390 (mainlib) Tributyl acetylcitrate 41 43 100 100 O 29 55 70 205 OH 119 91 50 39 57 50 105 159 83 69 79 147 55 187 71 131 67 81 177 220 98 86 110 45 57 85 173 31 45 65 93 121 136 154 0 0 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 (mainlib) 1H-Cycloprop[e]azulen-7-ol, decahydro-1,1,7-trimethyl-4-methylene-, [1ar-(1aα,4aα,7β,7aβ,7bα)]- (mainlib) 2-Decenal, (Z)-

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 27 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

135 60 100 100 73 O

41 OH 50 57 50 71 129 150 29 HO 69 39 87 91 45 115 172 77 107 117 15 31 89 101 135 143 153 15 27 39 43 51 55 59 65 73 81 86 127 0 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 (mainlib) n-Decanoic acid (mainlib) Phenol, 2-methyl-5-(1-methylethyl)- Fig3. GC-MS Chromatogram of the extract 4. CONCLUSION In the present study thirty four chemical constituents have been identified from Methanolic extract of the plant leaves of Aspilia africana by Gas Chromatogram-Mass spectrometry (GC-MS) analysis. The presence of various bioactive compounds on the methanol leaf extract can be attributed to its widely use in folklore medicines, especially in arresting wound bleeding, inhibiting the growth of microbial wound contaminants and accelerating wound healing which suggest good potentials for use in wound care. However isolation of individual phytochemical constituents and subjecting it to biological activity will definitely give fruitful results. It could be concluded that Aspilia africana contains various bioactive compounds. So it is recommended as a plant of phytopharmaceutical importance

REFERENCES [1] R. Meenatchi Ammal and G. Viji Stella Bail (2013) GC-MS Determination of Bioactive Constituents of Heliotropium indicum Leaf Journal of Medicinal Plants StudiesVol. 1 Issue. 6 ;30-33 [2] Mohamed E. Zain, Amani S. Awaad, Monerah R. Al Othman, and Sahar K. Al Dosary.(2014) Phytochemical analysis and antimicrobial activity of medicinal plants against pathogenic microorganisms. Life Science Journal. 11(7) 350-354 [3] Asres, K.; Taddese, S.; Gebremariam T. (2033). In vitro antimicrobial activities of some selected topically applied medicinal plants of Ethiopia. Ethio Pharm J. 2003; 21: 34-45 [4] Saleh AI, Mohammad Al-Dosari S, Abdul M, Alsheikh M, Maged Abdel-Kader S.( 2009) Evaluation of the Hepatoprotective Effect of Fumaria parviflora and Momordica balsamina from Saudi Folk Medicine Against Experimentally Induced Liver Injury in Rats. Res J Med Plants; 3(1):9-15 [5] Awaad, Amani S.; Soliman, G. A.; El-Sayed, Dalia F.; El-Gindi, Omimah D., and Alqasoumi, S. I. (2012).Hepatoprotective activity of Cyperus alternifoliuson carbon tetrachloride–induced hepatotoxicity in rats. Pharmaceutical Biology, 50(2): 155–161 [6] Mohannad G. AL-Saghir, (2009). Antibacterial Assay of Cinnamomum cassia (Nees and the Nees), Nees ex Blume Bark and Thymus vulgaris L. Leaf Extracts against Five Pathogens. Journal of Biological Sciences, 9: 280-282. [7] Kazeem Olasunkanmi Ajeigbe,Seyi Samson Enitan., Dayo Rotimi Omotoso and Olayemi Olutobi Oladokun (2013) Acute Effects of Aqueous Leaf Extract of Aspilia Africana C.D. Adams on Some Haematological Parameters in Rats Afr J Tradit Complement Altern Med. 10(5): 236–243 [8] Macfoy C A, Cline E I.( 1990) In-vitro antibacterial activities of three plants used in traditional medicine in Sierra-leone. J Ethnopharmacol.;28(3):232–237. [9] Achonye E L (1976). A pharmacological investigation of the haemostatic action of pressed leaf extract of Aspilia latifolia (Compositae) Pharmacology & Toxicology Department, University of Nigeria. pp. 23–31. B. Pharm. thesis. [10] Okoli C O, Akah P A, Okoli A S.( 2007) Potential of leaves of Aspilia africana (Composite) in wound care: an experimental evaluation. Biomedical Centre Complimentary and Alternative Medicine.7(24):101– 109 [11] A Eweka, A Eweka (2008) . Anti-Ulcer Effect Of Aspilia Africana (Asteraceae) Leaf Extract On Induced Duodenal Ulcer Of Adult Wistar Rats (Rattus Norvegicus) – A Histological Study. The Internet Journal of Alternative Medicine.( 8 ) 1: 102- 111 [12] Ajeigbe K.O, Onifade A.A,Omotoso .R, Enitan S.S and Olaleye S.B (2014)Anti-ulcerogenic Activityof Aspilia AfricanaLeaf Extract: Roles of Gastric Acid, Oxidative Stress and Neutrophil Infiltration Afr. J. Biomed. Res.Vol.17;193-201

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 28 Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS

[13] Nguelefack TB, Watcho P, Wansi S, Mbonuh N, Ngamga D, Tane P, Kamanyi A.(2005) The antiulcer effects of the methanolic extract of the leaves of Aspilia africana (Asteraceae) in rats. Afr J Trad CAM;2(3):233–237 [14] Oluyemi Kayode A.; Okwuonu Uche C.; Baxter D. Grillo & Oyesola Tolulope, O.,( 2007)"Toxic Effects of Methanolic Extract of Aspilia africana Leaf on the Estrous Cycle and Uterine Tissues of Wistar Rats" Int. J. Morphol., 25(3):609-614, [15] Ahamefula Anselm Ahuchaogu, Godwin .I. Ogbuehi, A I. Obike, Chisom Sampson Egedeuzu1, Okoronkwo Joseph Chukwu, John Bull Onyekachi Echeme (2018). GC-MS Analysis of Bioactive Compounds from Whole Plant Chloroform Extract of Ageratum conyzoides. International Journal of Medicinal Plants and Natural Products Vol. 4, Issue 2, 2018, PP 13-24 [16] D.B. Kell, M. Brown, H.M. Davey, W.B. Dunn, I. Spasic, and S.G. Oliver, Metabolic foot printing and systems biology: The medium is the message. Nat Rev Microbiol, 3, 2005, 557-65. [17] A.R. Fernie, R.N. Trethewey, A.J. Krotzky, and L. Willmitzer, Metabolite profiling: From diagnostics to systems biology. Nat Rev Mol Cell Biol, 5, 2004, 763-9. [18] Mustapha N. Abubakar and Runner R. T. Majinda (2016) GC-MS Analysis and Preliminary AntimicrobialActivity of Albizia adianthifolia (Schumach) and Pterocarpus angolensis (DC) Medicines. 3, 3; 1-9 [19] Michael Evanjaline R, Mohan VR (2018) Determination of bioactive components of caralluma umbellata haw. (apocynaceae) by gas chromatography and mass spectroscopy analysis.Asin journal of pharmaceutical and clinical researchVol 11, Issue 5, 2018 [20] Girija S.Singh and Edward. E.Mmatli (2011,)Recent progress in synthesis and bioactivity studies of indolizines. European Journal of Medicinal Chemistry 46, (11) 5237-5257 [21] Saita, T.; Katano, M.; Matsunaga, H.; Yamamoto, H.; Fujito, H.; Mori, M.(1993 ) The first specific antibody against cytotoxic polyacetylenic alcohol, panaxynol. Chem. Pharm. Bull41, 549–552. [22] Dawid, C., Dunemann, F., Schwab, W., et al. (2015). Bioactive C₁₇-polyacetylenes in carrots (Daucus carota L.): Current knowledge and future perspectives. J. Agric. Food Chem. 63(42), 9211-9222

[23] Leonti, M., Casu, L., Raduner, S., et al. (2010). Falcarinol is a covalent cannabinoid CB1 receptor antagonist and induces pro-allergic effects in skin Biochem. Pharmacol. 79(12), 1815-1826

Citation: Ahamefula Anselm Ahuchaogu et al. "Chemical Constituents of Methanol Leaf Extract of Aspilia africana C.D. Adams by GC MS” International Journal of Advanced Research in Chemical Science (IJARCS), vol. 5, no. 10, pp. 21-29, 2018.

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