Beginning at a very young age, today’s students are schooled at home (and just about everywhere else) by television Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 1 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET ANTIOXIDANTS FROM AMAZONIAN VEGETABLES: A POTENTIAL SOURCE OF NATURAL MEDICINE Mariana A. Perez-Chavez, Juan C. Ramirez-Gonzalez and Luisa D. Sanchez-Flores National University of the Peruvian Amazon, Iquitos Peru Abstract: The Amazon rainforest is home to a vast diversity of plant species, many of which have been used for medicinal purposes by indigenous peoples for centuries. This study investigated the antioxidant activity of plant species collected from the Tamshiyacu-Tahuayo Communal Regional Conservation Area (ACR CTT) in Peru. The antioxidant activity of the plant extracts was determined using the DPPH assay. The results showed that the plant extracts had significant antioxidant activity, with some extracts exhibiting more activity than commercial antioxidants. The findings of this study suggest that the plant species from the ACR CTT have potential as sources of natural antioxidants. Keywords: Amazon rainforest, Medicinal plants, Antioxidant activity, DPPH assay, Natural antioxidants 1. Introduction Certain Amazonian plant species among its many utilities that ethnobotany mentions, is its use as medicinal plants that have been used by the inhabitants of this region since time immemorial and, thanks to the studies of various researchers, it has become known and valued for a proper use of them (Duke & Vasquez, 1994) The town of Tamshiyacu is the capital of the district of Sargento Lores in the Loreto Region and its primary forests belong for the most part to the Tamshiyacu - Tahuayo Communal Regional Conservation Area of Peru (ACR CTT). Located in zone 18 of the UTM projection system, between coordinates 680 075 E, 9 528 176 N and 768 162 E, 9 444 073 N. It has an area of four hundred twenty thousand and eighty hectares with two thousand five hundred square meters (420,080, 25 ha), (Gobierno Regional de Loreto, 2011); (Shoobridge, et al., 2004). The antioxidant activity is considered as the sequestration of free radicals, which are found in excess, in the human organism they are considered to cause various pathologies that include aging, cancer, arteriosclerosis and other diseases in humans, (Ibarra et al., 2011). The main organic molecules provided by the plant species that manage to sequester these free radicals are mainly phenolic compounds, some alkaloids and vitamins A, E and C, (Coronado, et al., 2015). The objective of the present study was to perform the evaluation of the antioxidant activity of the foliar samples of 31 plant species of the Amazonian forest of the Tamshiyacu locality and to select the three best with high activity, 2. Materials and Methods 2.1. Materials Collection of plant material: Leaves of 31 plant species were collected in the vicinity of the town of Tamshiyacu and are shown in Table 1. The exsicatas were deposited in the Herrerense herbarium of the Research Institute of the Peruvian Amazon (IIAP). Evaluation of antioxidant activity. with the samples of the dried and powdered leaves, the methanolics extracts were prepared with some concentrations of 0.25; 0.1; 0.05 and 0.01 mg/mL. To determine the antioxidant activity, we used the UV/Vis spectrophotometer equipment, Specturlamb brand 22 pc. (KERLAB). In a 1.5 mL polystyrene mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 2 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET cuvette, 25 μL of the methanolic extract and 975 μL of 0.1 mmol DPPH solution were added, then the absorbance was measured at a wavelength of 517 ηm, the readings were performed for 5 minutes with 30 second intervals, all reactions were performed in triplicate. The inhibition of DPPH radical sequestration by increasing solutions of the extracts was determined by the following expression, (Sotero et al., 2011). % Inhibition DPPH = [(Ac – Am)/Ac] 100 Where: Ac, is the absorbance of the control (0.1 mmol of DPPH), and Am, is the absorbance of the sample (increasing solutions of the extracts) in a time n. 2.2. Methods Phenolic compounds: For the extraction of phenolic compounds, the technique of (Valls et al., 2000) is as followed: 0.5 g of the sample is weighed and extracted successively with 3 volumes of 25 ml of ethanol acidulated with 1% formic acid. The extract is concentrated in a rotary evaporator at 40 °C. The dry residue is redissolved in a 50% methanol solution acidified with a formic acid solution, and brought to a volume of 10 mL. This is stored for subsequent analyzes. Table 1. List of 31 plant species collected in the vicinity of the town of Tamshiyacu, Loreto-Peru. mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 3 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET Registry Collector Order/families Species Georeference UTM 1 PAA Malpighiales/ Euphorbiaceae Sapium sp. 18M 0714538 9555251 2 PAA Magnoliales/ Myristicaceae Virola. sebifera 18M 0714538 9555249 3 PAA Magnoliales/ Annonaceae Oxandra sp. 18M 0714501 9555241 4 PAA Magnoliales/ Myristicaceae Virola. sebifera 18M 0714495 9557062 5 PAA Magnoliales/ Myristicaceae Iryanthera cf. laevis 18M 0714497 9555242 6 PAA Magnoliales/ Annonaceae Cymbopetalum cf.longipes 18M 0714451 9555237 7 PAA Rosales/ Moraceae Ficus cf. americana 18M 0704508 9560836 8 PAA Fabales/ Fabaceae Parkia cf. multijuga 18M 0704522 9560848 9 PAA Rosales/ Moraceae Brosimum parinaroides. 18M 0704985 9561001 10 PAA Gentianales/ Apocynaceae Couma macrocarpa. 18M 0705215 9561180 11 PAA Magnoliales/ Annonaceae Xylopia cf. benthamii 18M 0705289 9561177 12 PAA Malpighiales/ Caryocaraceae Caryocar glabrum 18M 0705951 9558757 13 PAA Magnoliales/ Annonaceae Guatteria cf. hyposericea 18M 0705964 9558755 14 PAA Sapindales/ Anacardiaceae Tapirira guianensis 18M 0705988 9558712 15 PAA Magnoliales/ Myristicaceae Virola cf. surinamensis 18M 0706268 9559258 16 PAA Gentianales/ Apocynaceae Llacmelleasp. 18M 0711608 9557071 17 PAA Magnoliales/ Annonaceae Guatteria cf. flabellata 18M 0711617 9557063 18 PAA Fabales/ Fabaceae Dialium cf. guianense 18M 0711612 9557072 19 PAA Rosales/ Moraceae Helicostylis cf. tomentosa 18M 0711610 9557062 20 PAA Rosales/ Moraceae Helicostylis cf. turbinata 18M 0711623 9557049 21 PAA Santalales/ Olacaceae Mnquartia guianensis 18M 0711855 9556913 22 PAA Laurales / Siparunaceae Siparuna cf. sessiliflora 18M 0711854 9556909 23 PAA Rosales/ Moraceae Ficus sp. 18M 0711842 9556901 24 PAA Alismatales/ Araceae Dracontium cf. amazonense 18M 0711830 9556904 25 PAA Malpighiales/ Clusiaceae Vismia cf. macrophylla 18M 0711852 9556899 26 PAA Caryophyllales/ Nyctaginaceae Neea cf. divaricata 18M 0711864 9556887 27 PAA Fabales/ Fabaceae Zygia cf. macribridei 18M 0711868 9556888 28 PAA Fabales/ Fabaceae Diaium sp. 18M 0706244 9561454 mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 4 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET 29 PAA Gentianales/ Apocynaceae Malouetia cf. naias 18M 0706538 9561642 30 PAA Magnoliales/ Annonaceae Unonopsis cf. sitipitata 18M 0706764 9561927 31 PAA Ranunculales/ Menispermaceae Curarea cf. toxicofera 18M 0706669 9561534 Anthocyanins and total flavonoids: The determination of anthocyanins and total flavonoids is performed by UV/Vis spectrophotometry in 1mL of the extract prepared for the phenolic compounds by reading the absorbance at 535 ηm and 374 ηm respectively, after dilution of the samples. To perform the calculations, the molar extinction coefficient of malvidin-3-glucoside is used: 29500 L/mol cm. Catechin and Proanthocyanidins: It is done by the vanillin test. 0.5 ml of the extract is mixed with 1.25 ml of vanillin in 1% methanol (w/v) and with 1.25 ml of 25% sulfuric acid (v/v) in methanol. The white is prepared simultaneously in the same way, but replacing the vanillin solution with methanol. It is left to rest for 15 minutes and then the absorbance reading is made at 510 ηm. Total phenolic compounds: Measurement of the Folin index is carried out, for which 40 μl of the extract prepared for phenolics are treated, with 0.5 ml of Folin-Ciocalteau reagent and 2mL of 20% sodium carbonate (w/v), and they are taken to 10 ml. After half an hour, the absorbance reading is carried out at 765 ηm. To establish the calibration, catechin standards of concentrations between 0 - 100 mg/L are used. Alkaloids: The method indicated by Shamsa et al., (2008), is used, 5 g dry pulverized sample is weighed and extracted with methanol in soxhlet equipment for 12 continuous hours. The extract was filtered and the methanol was separated in a rotavapor at 45 ºC, redissolved with 2N HCl and then filtered, 1 mL of this solution was transferred to a decanting pear and washed three times with 10 mL of chloroform. The pH of this solution is neutralized with 0.1 N NaOH, 5mL of BCG and 5mL of phosphate buffer are added, the mixture is shaken and the complex formed is extracted with 1, 2, 3, and 4mL of chloroform with stirring. The extracts are collected in a 10mL vial and then filled to volume with chloroform, in this solution is proceeded to perform the absorbance reading at 470 ηm. The data are quantified with a standard curve of atropine. Chromatographic fractionation and identification of polar molecules by gas chromatography coupled to mass spectrophotometry (CG-Ms). The chromatographic fractionation of methanolic extracts, in open column with silica gel No. 100, of the three species with high antioxidant activity was performed, and identification of the molecules by thin-layer chromatography, grouping the fractions with similar molecules, then they were subjected to Gas chromatography equipment with mass spectrometry, using Agilent Technologies 7890ª ® with an Agilent 122-5532 DB 5MS ® capillary column of 30 m, with internal diameter of 0.25 mm. The initial temperature of the oven was 100 °C/03 min, followed by a ramp of 20 °C/3 min up to 300 °C/19 min; the maximum temperature of the oven was 325 °C. A Split injection was used and the helium gas flow was 2 mL/min. The fragments for the analyses were recorded with the parameters for a scan of 50 to 500 m/z, Sotero et al., (2016). 3. Results Table 2 shows the antioxidant activity of the leaves of the 31 plant samples studied, ordered by families, where all the evaluations of the antioxidant activity of this species are summarized, it is observed that the three species that exceed 50 % inhibition of antioxidant activity at a concentration lower than 5.0 mg/ml are, Virola sebifera, Caryocar glabrum and Tapirira guianensis. Tables 5, 6 and 7 show the molecules identified by the GC-Ms of the extracts of the species Virola sebifera, Caryocar glabrum and Tapirira guianensis, respectively. mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 5 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET Table 2. Percentages of Inhibition of the methanolics extracts of the leaves of the 31 plant species under study at different concentrations, using the DPPH method. Concentration, mg/mL Family Species 5,0 0,5 0,25 0,1 0,05 0,01 mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml Annonaceae Oxandra sp. 32,59 10,01 12,11 18,35 18,37 17,67 Annonaceae Guatteria hyposericea 44,42 10,37 5,55 3,51 3,17 2,80 Annonaceae Cymbopetalum.longipes 26,19 0,05 0 0 0 0 Annonaceae Xylopia benthamii 43,70 9,46 0,51 0,43 0 0 Annonaceae Guatteria flabellata 45,56 26,74 7,10 3,68 2,40 1,51 Annonaceae Unonopsis sitipitata 3,15 0 0 0 0 0 Apocynaceae Couma macrocarpa. 19,48 7,18 4,10 3,26 0,55 1,78 Apocynaceae Llacmelleasp. 42,11 17,09 10,75 10,90 10,90 10,50 Apocynaceae Malouetia. naias 27,84 9,52 13,27 21,55 0 0 Anacardiaceae Tapirira guianensis 79.41 23.53 20.59 11.76 11.76 8.82 Araceae Dracontium amazonense 25,82 12,69 7,72 7,11 7,08 8,50 Caryocaraceae Caryocar glabrum 74,25 29.41 23.53 14.71 11.76 5.88 Clusiaceae Vismia macrophylla 23,87 10,43 7,27 6,23 5,83 6,32 Euphorbiaceae Sapium sp. 40,56 7,03 11,98 10,78 11,88 16,04 Fabaceae Parkia multijuga 6,82 1,40 2,81 1,50 0,85 0,10 Fabaceae Dialium guianense 11,41 3,17 2,08 0,71 0,26 0 Fabaceae Zygia macribridei 11,49 0 0,27 0 0 0 Fabaceae Diaium sp. 30,48 5,55 4,09 2,11 2,18 2,42 Menispermaceae Curarea toxicofera 22,85 2,91 1,37 0 0 0 Moraceae Brosimum parinaroides. 17,89 3,92 0,84 1,33 1,46 0,36 Moraceae Helicostylis turbinata 22,21 12,94 13,78 2,36 2,85 4,39 Moraceae Ficus americana 9,85 0 0 0 0 0 Moraceae Helicostylis tomentosa 13,10 1,99 4,71 4,56 4,67 4,22 Moraceae Ficus sp. 34,48 23,94 24,97 24,72 18,53 19,23 Myristicaceae Virola. sebifera 81.08 40 32.43 24.32 8.11 5.41 Myristicaceae Virola sebifera 43,16 12,25 9,96 6,52 0,94 0,12 Myristicaceae Iryanthera laevis 35,22 6,42 5,87 -2,54 -5,89 -6,06 Myristicaceae Virola surinamensis 46,19 16,80 13,06 7,44 8,52 6,57 Nyctaginaceae Neea divaricata 10,33 0 0,98 0 0 0 Olacaceae Mnquartia guianensis 42,25 8,38 4,20 1,65 1,41 2,00 Siparunaceae Siparuna sessiliflora 21,32 4,64 2,01 2,03 2,52 2,78 Table 3. Phenolic compounds present in the leaves of three plant species with a high percentage of antioxidant activity. mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 6 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET Species Anthocyanins Flavonoids Phenolics Catechins and proanthocyanidins mg /100g mg /100g mg/100g mg/100g Virola sebifera 71,38 143,90 18580,87 0,16 Caryocar glabrum 93,95 144,33 15180,71 0,18 Tapirira guianensis 41,94 144,12 11568,78 0,10 Table 4. Total Alkaloids present in the leaves of three plant species with a high percentage of antioxidant activity. Species Total, alkaloids mg/kg Total, alkaloids mg/100g Virola sebifera 36,03 3,60 Caryocar glabrum n.d n.d Tapirira guianensis 74,00 7,40 Table 5. Molecules found by CG-MS in the methanol fractions of the leaves of the 2PAA specie (Virola sebifera) F1-2PAA Nº Retention time, min Molecules Prob. % Area 1 9.99 (5a) preganane-3,20a-diol, 14a- [4-methyl- 3-oxo- (1-oxa-4azabuten-1,4- (diyl)] diacetate 63.4 11.5 2 11.67 4-piperidine acetate, 1-acetyl-5-ethyl-2- [3- (2-hydroxyethyl- 1H-indol-2-yl] -o-methyl-methyl 66.8 23.2 3 11.35 octadecane, 3-ethyl-5- (2-ethylbutyl) 64.0 23.2 4 14.61 1H-inden-1-one, 2,3-dihydro-5,8- dimethoxy-3-methyl 64.1 12.1 5 16.62 Folic acid 61.8 19.9 F2- 2PAA 1 40.16 2'-methylene, bis 6- (1-dimethylethyl) -4- methyl-phenol 85.0 0.01 mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 7 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET 2 42.63 3 ', 8,8', trimethoxy-3-piperidine-2,2'- binaphthalene-1,1 ', 4,4'tetrone 70.0 0.01 3 47.70 3,5-di-tert-butyl-4-hydroxyanisole 69.5 98.42 F4-2PAA 1 10.91 Copaene 88.50 2.73 2 12.12 Cariopilene 93.20 12.09 3 14.65 1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1- (1- methylethyl) (1Scis) -naphthalene 91.20 3.08 4 14.77 1,3,3,4-tetrahydro-1,6-dimethyl-4-81- methletiy) - (1Scis) naphthalene 86.60 1.02 5 14.94 3-methoxymethyl.2,5,58a-tetramethyl- 6,7,8,8a-tetrahydro-5chromene) 76.00 1.65 6 16.33 (-) espatulenol 88.60 7.32 7 16.44 cariopilene oxide 87.90 2.71 8 16.56 Globulol 53.00 0.91 9 16.83 8S, 14-cedran-diol, 77.60 1.35 10 17.86 Cubenol 85.50 0.59 11 18.11 espatulenol 79.90 1.43 12 18.90 a-cadinol 81.70 0.70 13 19.60 a-N-normetadol 64.20 0.51 14 21.65 isoaromadendreno epoxide 84.10 1.11 15 42.63 Diisooctyl 1,2-benzenedicarbilate 92.30 1.81 16 47.22 13-docosenamide (Z) 88.30 29.13 17 47.61 3,5-di-tert-butyl-4-hydroxyanisole 69.30 25.07 18 49.91 7-Acetoxy-3-methoxy-2- (3,4-dimethoxy penyl) -4-chromen4-one) 68.40 0.80 F5-2PAA 1 19.62 1,9,5-cycloheptatriene, 6-methyl-1- (6- methyl) -1,3,5cycloeptatriene-il) 82.00 0.61 2 42.63 Diisooctyl 1,2-benzenedicarboxylate 94.10 1.75 3 47.68 3,5-di-tert-butyl-4-hydroxyanisole 69.60 97.65 F6-2PAA 1 12.12 Cariofillene 87.40 52.31 mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 8 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET 2 14.65 2,3,5,6,8a-hexahydro-4,7-dimethyl-1- (1- methyl ethyl) - (1Scis) or cadine-3,9-diene- naphthalene 82.40 10.54 3 16.34 Espatulene 88.00 37.15 Table 6. Molecules found by CG-'MS in the methanol fractions of the leaves of 12PAA specie (Caryocar glabrum) F2-12 PAA Nº Retention time, min Molecules Prob. % Area 1 11.17 7,8-epoxylanstan-ol, 3-acetoxy 63.1 0.95 2 11.172 4,3-ethyl-5-octadecane 66.9 1.79 3 14.624 72.1 0.62 4 18.872 1,2,8-trimethyl-4-propenyl- (E) - naphthalene 80.9 0.70 5 19.64 1,1''bisphenyl, 2,2 ', 5,5'-tetramethyl 81.3 1.78 6 20.939 1,1'-dodecylidene bis (methyl) - benzene 65.4 1.78 7 42.638 Mono (2-ethyl hexyl) 1,1'- benzenedicarboxylate 98.1 93.19 F3 - 12PAA 1 14.6 2,4-bis (1,1-dimethyl-ethyl-phenol) 73.3 1.37 2 42.6 diisooctyl 1,2-benzene dicarboxylate 96.6 98.6 F4-12PAA 1 14.61 3,5-bis (1,1-dimetyl ethyl)-phenol 78.9 2.10 2 43.63 diisooctyl 1,2-benzene dicarboxylate 96.5 97.0 F5-12PAA 1 14.61 7,8-epoxylanostan-11-ol, 3 acetoxy 60.3 3.71 2 16.8 chlorotetracycline 62.2 3.62 3 42.6 Diisoctyl 1,2-benzenedicarboxylate 95.7 92.59 F6-12PAA 1 23.7 9 H-fluorene, 9-methylene 95.9 11.23 2 30.7 fluoranthene 95.7 33.14 3 32.12 pyrene 93.2 24.65 4 41.5 benzantrene 90.3 6.48 5 42.63 Diisoctyl 1,2-benzenedicarboxylate 94.2 24.47 mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 9 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET Table 7. Molecules found by CG-'MS in the methanolic fractions of the leaves of the 14PAA specie (Tapirira guianensis) F1 -14PAA Nº Retention time, min Molecules Prob. % Area 1 14.02 decahydro-4a-methyl-1-methylene-7- (1- methylethylidene) -4 a-trans-naphthalene 80.7 6.7 2 14.81 2,4-bis (1,1-dimethylethyl) -phenol 76.2 11.19 3 19.65 naphthalene, 1,2,3-trimethyl-4-propanil 70.5 5.84 4 47.27 Vitamin E 80 76.17 F2-14PAA 1 10.93 copaene 81.2 12.41 2 13.83 2-isopropyl-4a, 8-dimethyl-1,2,3,4,4a, 5,6,7octahydronaphthalene 86.2 8.62 3 13.94 Eudesna-4 (14), 11dieno 67.8 2.89 4 14.03 1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7- (1- methylethyl enyl) - [1R - (1a, 7a, 8a, a)] - naphthalene (synonimous 2valenceno) 96.8 33.67 5 14.73 a-panasinsene 89.3 42.42 F3-14 PAA 1 11.58 2,4-bis (1,1-dimethylethyl) -phenol 69.2 2.944 2 14.83 α -patchulene 83.4 9.13 3 13.93 valenceno 88.6 27.87 4 14.03 a-panasinseno 88.6 39.596 5 14,72 diisooctylphthalate 95.0 20.407 F4 - 14PAA 1 13.859 α -patchulene 74.2 2.011 2 14.058 (+) valencene 91.4 4.086 3 42.659 diisooctylphthalate 96 93.903 mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 10 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET 4. Discussion Test for antioxidant activity. Based on the results presented in Table 2, it can be stated that of the 31 species studied, 9.7% exceeded 60% inhibition, while 16.1% exceeded 45%. The families with better results, besides those mentioned, of inhibition were Annonaceae, Apocynaceae and a species of Euforbiaceae, with inhibition greater than 40%. These results are within the expected, since researchers who worked with 25 plants from Colombia, found similar or higher percentages of inhibition, and those who also consider that the Euforbiáceaes have good antioxidant qualities (Mosquera et al. 2007). According to these results, the three with the best activities were selected: Virola sebifera (Myristaceae). Caryocar glabrum (Caryocarceae) and Tapirira guianensis (Anacardiaceae), which present an inhibition percentage of 81.08%, 74.25%, and 79.41% respectively, at the concentration of 5.0 mg/ml and the IC50 of each of them is 1.5 mg/ml 2.2 mg/ml, and 2.8 mg/ml, respectively. Phenols and total alkaloids. Several authors agree that the high concentration of phenolic compounds in a plant species has high antioxidant activity, this correlation is given for wine, which at higher phenolic concentration improves antioxidant activity. This is the case for tea (Camellia sinensis) (Benzie & Szeto 1999), from lime (Tilia argéntea) (Yildrim et al., 2000), among others. According to Table 3, it can be observed that the concentration of phenolic compounds is high in the species Virola sebifera 18580.87 g/100g followed by Caryocar glabrum 15180.71 mg/100g and Tapirira guianensis 11568.78 mg/100g. It is worth mentioning the very similar concentration of flavonoids and the high presence of anthocyanins, proven compounds with a high antioxidant effect. In the case of C. glabrum, researchers found molecules in the bark of this species, such as coumarins, (Aladul et al., 2007), as well as (Rodríguez et al., 2017), who found several anthocyanins in T. guianensis, such as quercitin, and derivatives thereof. It is observed that the concentration of alkaloids is present only in the species Virola sebifera and Tapira guianensis, but not in the Caryocar glabrum; indicative that these substances do not play an important role in this species, as antioxidants. Reducing its activity with greater certainty to the phenolic compounds present. On the other hand, in the other species Virola sebifera and Tapira guianensis, an appreciable concentration of these substances is observed, and according to (Chávez et al., 1996), they indicate that many alkaloids of different types of structures have been shown to be powerful inhibitors of singlet oxygen. Many of these compounds proved to be better inhibitors than the tertiary amine 1,4-diaza [2.2.2] bicycloctane (DABCO). Likewise, (Ibarra et al., 2011) found a high antioxidant activity in the alkaloidal fractions of Erytrina americana, and when isolating the molecule erisodin, found that it had an IC50 of 150 μg/mL. Molecules identified by CG-MS The molecules of interest found in the methanolic fractions were a) V. sebifera: folic acid, 3,5-diterbutio- 4hydroxyanisole (phenolic), caryophyllene (bicyclic sesquiterpene) and spatulenol (alcoholic sesquiterpene); b) C. glabrum: 2,4-bis (1,1-dimethylethyl) -phenol, 1,1'-benzenedicarboxylate of mono (2-ethyl hexyl), 9 H- fluorene, 9methylene, fluoranthene and pyrene (these last two, aromatic compounds) and c) T. guianensis: vitamin E, copaene (tricyclic sesquiterpene), patchulene and valencene both (sesquiterpenes). Likewise, observing the compounds currently recognized as antioxidants, there is an anisole derivative in fractions F2, F4 and F5 of V. sebifera, such as 3,5-di-tert-butyl-4-hydroxyanisole in fraction F3, F4 C. glabrum, phenolic derivatives, such as 2,4-bis (1,1-dimethylethyl-phenol) and 3,5-bis (1,1-dimethyl-ethyl) -phenol and in the F1 of T. guianensis, the Vitamin E. mailto:topacademicjournals@gmail.com Academic Journal of Science, Engineering and Technology Vol.6, Issue 1; January - Febuary 2021; 1252 Columbia Rd NW, Washington DC, United States https://topjournals.org/index.php/TAJES/index; mail: topacademicjournals@gmail.com 11 | A c a d e m i c J o u r n a l o f S c i e n c e , E n g i n e e r i n g a n d T e c h n o l o g y | https://topjournals.org/index.php/AJSET 5. 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