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Revista de Ciencias J. D. Yakobi-Hancock, L. A. Ladino and J. P. D. Abbatt [84] Weingartner, E., Burtscher, H. and Baltensperger, U. (1997). Hygroscopic properites of carbon and diesel soot particles, Atmospheric Environment Atmospheric Environment, 31(15), 2311–2327.
Facultad de Ciencias Naturales y Exactas Universidad del Valle [85] Yakobi-Hancock, J. D., L. Ladino, and J. Abbatt (2013). Feldspar minerals as efficient deposition ice nuclei, Atmospheric Chemistry and Physics, 13, 11,175– TOTAL PHENOLICS ANTIOXIDANT ACTIVITY AND
[86] Zhang, Q., Jimenez, J. L., Canagaratna, M. R., Allan, J. D., Coe, H., Ulbrich, I., PHYTOCHEMICAL PROFILE OF SOME PLANTS
Alfarra, M. R., Takami, a., Middlebrook, a. M., Sun, Y. L., Dzepina, K., Dunlea, FROM THE YOTOCO NATIONAL PROTECTED FOREST.
E., Docherty, K., DeCarlo, P. F., Salcedo, D., Onasch, T., Jayne, J. T., Miyoshi, T., VALLE DEL CAUCA, COLOMBIA
Shimono, a., Hatakeyama, S., Takegawa, N., Kondo, Y., Schneider, J., Drewnick, F., Borrmann, S., Weimer, S., Demerjian, K., Williams, P., Bower, K., Bahreini, Harlen Torres Castañeda
R., Cottrell, L., Griffin, R. J., Rautiainen, J., Sun, J. Y., Zhang, Y. M. and Worsnop, Universidad Nacional de Colombia D. R. (2007). Ubiquity and dominance of oxygenated species in organic aerosols in anthropogenically-influenced Northern Hemisphere midlatitudes, Geophysical Ana Julia Colmenares Dulcey José Hipólito Isaza Martínez
Research Letters, 34(13), L13801, doi:10.1029/2007GL029979.
Universidad del Valle [87] Zobrist, B., Marcolli, C., Koop, T., Luo, B. P., Murphy, D. M., Lohmann, U., Received: November 15, 2013 Accepted: December 17, 2013 Zardini, A., Krieger, U. K., Corti, T., Cziczo, D. J., Fueglistaler, S., Hudson, P. K., Thomson, D. S. and Peter, T. (2006). Heterogeneous ice nucleation in upper tropospheric aerosols, Atmospheric Chemistry and Physics, 6, 3115–3129.
Determining the antioxidant activity and the total phenolic content are routine procedures in most [88] Zobrist, B., Marcolli, C., Pedernera, D. A. and Koop, T. (2008). Do atmospheric natural product laboratories; however, when dealing with large number of samples, it is necessary aerosols form glasses?, Atmospheric Chemistry and Physics, 8, 5221–5244. to employ methods that allow a quick, easy and economical screening. The aim of this study is to determine the antioxidant activity and the total phenolic content of plant species as criteria for the selection of promising species. To reach this aim, we used twenty species from Yotoco National Protected Forest in Valle del Cauca, Colombia. The species Clidemia tococoidea and Miconia aeruginosa, showed the highest total phenolic content together with the best antioxidant activity in Jacqueline D. Yakobi-Hancock terms of DPPH radical scavenging activity. The excellent correlation (R2=0.9610) shown between Department of Chemistry, University of Toronto, Toronto, ON, Canada these parameters, demonstrated the utility of the process used as a method for primary screening and selection of promising species for phytochemical analysis at a preparative scale for this two assays.
Luis Antonio Ladino Keywords: antioxidant activity, phenols, DPPH, Folin-Ciocalteau.
Department of Chemistry, University of Toronto, Toronto, ON, Canada Jonathan P. D. Abbatt Department of Chemistry, University of Toronto, Toronto, ON, Canada Antioxidants may inhibit or retard biomolecules oxidation, through inhibition of the initiation and propagation steps of reactive oxygen species (ROS) mediated chain reactions [1], whose overproduction can lead to immuno-pathological phenomena related to oxidative stress. ROS are also involved in the generation of a wide variety of health disorders or conditions including inflammation [2, 3], cancer [3], atherosclerosis [4] and degenerative diseases [5, 6].
Nowadays, there is great interest in natural antioxidants, especially from plant sources, as they are seen as potentially therapeutic agents for diseases caused by ROS and useful as nutraceuticals, due to its positive impact on human health [7, 8]. These properties have been attributed to different types of phenolic compounds which have a growing interest in Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco the food industry. In agriculture, species with high content of phenolic compounds have been shown to possess antimicrobial [9], cytotoxic [10] and the insecticide properties Dried and powdered aerial parts (5 g) of each species were successively extracted three times with chloroform and acetone-water (70:30) in a Branson Scientific ultrasonic In this research, we use the advantages of techniques based on absorbance reading bath at room temperature for 15 minutes. The extracts were decanted and filtered through in microplates, for quick, simple and inexpensive primary screening of twenty plant Whatman filter paper No. 1. The solvent was evaporated at low pressure in an IKA species from Reserva Nacional Forestal Bosque de Yotoco, requiring only an incubator RV10 Control V rotary evaporator at 40 °C and the water extracts were dried in a lab and conventional microplate reader. The use of a microplate-based method requires small with a freeze-dryer. The extraction yield was measured and the result was expressed as a amounts of sample and reagents and it allows simultaneous measurement of multiple percentage (%).
replicates to calculate the effective concentration of samples, which is required to scavenge 50% of DPPH free radicals (FRS ).
2.3 Phytochemical screening
Screening of chemical constituents was carried out by using test tube chemical 2 Materials and methods
methods. Extracts were analyzed for the presence of alkaloids, saponins steroids, tannins, flavonoids and phenolics according to standard methods [13].
2.1 Plant materials
2.4 Antioxidant activity assays
The species were collected in August 2010 at "Reserva Forestal Nacional de Yotoco" (Table 1), located at the municipality of Yotoco, Valle del Cauca, Colombia. Those 2.4.1 Qualitative DPPH radical scavenging assay using thing layer chromatography species were identified at the collection site by Professor Eugenio Escobar, Botanist of Qualitative screening for antioxidant activity was done using the 1,1-diphenyl-2- "Universidad Nacional de Colombia", Palmira, Colombia [12]. Voucher specimens picryl hydrazyl (DPPH) (Sigma-Aldrich) radical assay. A thin-layer chromatoplate of were prepared and deposited at the Luis Sigifredo Espinal Tascón Herbarium CUVC at each extract on silica gel 60 F (10 x 20 cm, Merck) was developed with hexane-acetone Universidad del Valle, and their identity was confirmed by Dr. Philip A. Silverstone-Sopkin. (8:2) and n-butanol-acetic acid-water (7:2:1) as mobile phase for the chloroform and aqueous extracts respectively. DPPH radical test was performed directly on thin-layer Table 1. List of plant species from Yotoco National Protected Forest.
chromatography (TLC) plates by spraying with DPPH methanol solution (0.2% w/v).
2.4.2 Quantitative DPPH radical-scavenging assay DPPH free radical scavenging activity was determined in triplicate by the method Banara glauca (Kunth) Benth Piper aequale Vahl of Sdiri, et al. [14], with slight modifications. Extracts (100 µL) at two-fold serial Casearia megacarpa Sw.
Piper imperials (Mic.) DC.
dilutions (0.5-512 mgL-1) were mixed with 100 µL of a 132 mgL-1 DPPH solution Piper augustum Rudge prepared in methanol. After 1 hour of reaction, the absorbency of the mixtures was read Besleria solanoides Kunth Piper setosum Trel.
at 520 nm (Metertech, AccuReader M965+ microplate reader). Different concentrations (0.5-512 mgL-1) of ascorbic acid and quercetin (Sigma-Aldrich) were used as positive Clidemia tococoidea Palicourea thyrsiflora controls and run in parallel. The results were expressed as a percentage of radical (DC.) Gleason (Ruiz-Pav) DC.
scavenging activity (%FRS) according to the equation: Miconia acuminifera Triana Psychotria compta Standl.
Miconia aeruginosa Naudin Psychotria hazeni Standl.
%FRS = [(A – A )/A ]x100 Miconia prasina (Swartz) DC.
Psychotria longirostris where A is the absorbency of DPPH radicals without sample or positive control (Rusby) Standl.
and A is the absorbency of DPPH radicals with sample or positive control. Efficient Psychotria macrophylla concentration of samples and positive controls that inhibits 50% of the DPPH radicals (FRS ) was calculated and expressed as mgL-1.
Erythroxylum citrifolium A. St-Hill 2.5 Estimation of total phenolics (TP)
Siparuna aspera (Ruiz-Pav.) Total phenolic content (TP) of aqueous extracts were determined according to Lacistema aggregatum Siparuna gigantotepala S.S. method of Sdiri, et al. [14] with some modifications. An appropriately diluted sample (P.J. Bergius) Rusby (100 µL) was mixed with 50 µL of 20% v/v Folin-Ciocalteau reagent (Sigma-Aldrich). Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco the food industry. In agriculture, species with high content of phenolic compounds have been shown to possess antimicrobial [9], cytotoxic [10] and the insecticide properties Dried and powdered aerial parts (5 g) of each species were successively extracted three times with chloroform and acetone-water (70:30) in a Branson Scientific ultrasonic In this research, we use the advantages of techniques based on absorbance reading bath at room temperature for 15 minutes. The extracts were decanted and filtered through in microplates, for quick, simple and inexpensive primary screening of twenty plant Whatman filter paper No. 1. The solvent was evaporated at low pressure in an IKA species from Reserva Nacional Forestal Bosque de Yotoco, requiring only an incubator RV10 Control V rotary evaporator at 40 °C and the water extracts were dried in a lab and conventional microplate reader. The use of a microplate-based method requires small with a freeze-dryer. The extraction yield was measured and the result was expressed as a amounts of sample and reagents and it allows simultaneous measurement of multiple percentage (%).
replicates to calculate the effective concentration of samples, which is required to scavenge 50% of DPPH free radicals (FRS ).
2.3 Phytochemical screening
Screening of chemical constituents was carried out by using test tube chemical 2 Materials and methods
methods. Extracts were analyzed for the presence of alkaloids, saponins steroids, tannins, flavonoids and phenolics according to standard methods [13].
2.1 Plant materials
2.4 Antioxidant activity assays
The species were collected in August 2010 at "Reserva Forestal Nacional de Yotoco" (Table 1), located at the municipality of Yotoco, Valle del Cauca, Colombia. Those 2.4.1 Qualitative DPPH radical scavenging assay using thing layer chromatography species were identified at the collection site by Professor Eugenio Escobar, Botanist of Qualitative screening for antioxidant activity was done using the 1,1-diphenyl-2- "Universidad Nacional de Colombia", Palmira, Colombia [12]. Voucher specimens picryl hydrazyl (DPPH) (Sigma-Aldrich) radical assay. A thin-layer chromatoplate of were prepared and deposited at the Luis Sigifredo Espinal Tascón Herbarium CUVC at each extract on silica gel 60 F (10 x 20 cm, Merck) was developed with hexane-acetone Universidad del Valle, and their identity was confirmed by Dr. Philip A. Silverstone-Sopkin. (8:2) and n-butanol-acetic acid-water (7:2:1) as mobile phase for the chloroform and aqueous extracts respectively. DPPH radical test was performed directly on thin-layer Table 1. List of plant species from Yotoco National Protected Forest.
chromatography (TLC) plates by spraying with DPPH methanol solution (0.2% w/v).
2.4.2 Quantitative DPPH radical-scavenging assay DPPH free radical scavenging activity was determined in triplicate by the method Banara glauca (Kunth) Benth Piper aequale Vahl of Sdiri, et al. [14], with slight modifications. Extracts (100 µL) at two-fold serial Casearia megacarpa Sw.
Piper imperials (Mic.) DC.
dilutions (0.5-512 mgL-1) were mixed with 100 µL of a 132 mgL-1 DPPH solution Piper augustum Rudge prepared in methanol. After 1 hour of reaction, the absorbency of the mixtures was read Besleria solanoides Kunth Piper setosum Trel.
at 520 nm (Metertech, AccuReader M965+ microplate reader). Different concentrations (0.5-512 mgL-1) of ascorbic acid and quercetin (Sigma-Aldrich) were used as positive Clidemia tococoidea Palicourea thyrsiflora controls and run in parallel. The results were expressed as a percentage of radical (DC.) Gleason (Ruiz-Pav) DC.
scavenging activity (%FRS) according to the equation: Miconia acuminifera Triana Psychotria compta Standl.
Miconia aeruginosa Naudin Psychotria hazeni Standl.
%FRS = [(A – A )/A ]x100 Miconia prasina (Swartz) DC.
Psychotria longirostris where A is the absorbency of DPPH radicals without sample or positive control (Rusby) Standl.
and A is the absorbency of DPPH radicals with sample or positive control. Efficient Psychotria macrophylla concentration of samples and positive controls that inhibits 50% of the DPPH radicals (FRS ) was calculated and expressed as mgL-1.
Erythroxylum citrifolium A. St-Hill 2.5 Estimation of total phenolics (TP)
Siparuna aspera (Ruiz-Pav.) Total phenolic content (TP) of aqueous extracts were determined according to Lacistema aggregatum Siparuna gigantotepala S.S. method of Sdiri, et al. [14] with some modifications. An appropriately diluted sample (P.J. Bergius) Rusby (100 µL) was mixed with 50 µL of 20% v/v Folin-Ciocalteau reagent (Sigma-Aldrich). Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco Then, 50 µL of sodium carbonate solution (1,6% p/v) was added to the mixture. The mixture The results of the phytochemical screening indicated the presence of terpenes, was incubated for 1 hour at 60 °C and then was allowed to stand at room temperature for steroids and phenols in all samples; however, the specific assays for alkaloids, flavonoids, 15 min. The absorbance was read at 650 nm. A standard calibration curve of gallic acid soaps, hidrolyzable and condensed tannins showed more restricted distribution at specific (1-32 mgL-1) was plotted. Results were expressed as gallic acid equivalents (GAE)/g dried level (Table 2). Table 2. Phytochemical screening of plant species from "Reserva Nacional Forestal Bosque de Yotoco".
2.6 Statistical analysis
Terpenoids
Condensed Hydrolysable Alcaloids
and steroids
Results were given as the mean±SD for at least three replicates for each sample. Statistical analysis was performed using GraphPad Prism 5® (GraphPad Software Inc., San Diego, California, USA). FRS were calculated using nonlinear regression with one phase exponential decay calculation.
3 Results and discussion
3.1 Extraction yields and phytochemical profile
In order to determine the antioxidant potential and phenolic content of twenty plant species from Yotoco National Protected Forest, for which extractions were performed with chloroform and acetone-water (70:30). S. gigantotepala showed the highest overall yield, followed by C. tococoidea, E. citrifolium, P. setosum, P. augustum, P. compta and other species (Fig. 1). - No appreciable evidence; + weak evidence; ++ strong evidence 3.2 Estimation of the antioxidant potential and total phenolics
TLC analysis showed that most of aqueous extracts possessed potent antioxidant activity, in contrast with low activity exhibited by the chloroform extracts. The ability of the extracts to scavenge DPPH radicals was also investigated at various concentrations to determine the FRS values. As shown in Figure 2, the extracts possessed substantial dose-dependent antioxidant activity. Some species, such as C. tococoidea and M. aeruginosa exhibited similar behavior to quercetin (Figure 2). In others species as B. glauca, almost complete DPPH radical scavenging was observed at concentrations higher than 50 mgL-1. This activity was comparable to that of ascorbic acid and quercetin, which were used as control antioxidants [15]. Interestingly, P. longirostris has a higher activity than S. gigantotepala at concentrations less than 50 mgL-1. However, at concentrations higher than 50 mgL-1 the activity of S. gigantotepala is greater.
Figure 1. Extraction process yields (for species complete names, see table 1).
Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco Then, 50 µL of sodium carbonate solution (1,6% p/v) was added to the mixture. The mixture The results of the phytochemical screening indicated the presence of terpenes, was incubated for 1 hour at 60 °C and then was allowed to stand at room temperature for steroids and phenols in all samples; however, the specific assays for alkaloids, flavonoids, 15 min. The absorbance was read at 650 nm. A standard calibration curve of gallic acid soaps, hidrolyzable and condensed tannins showed more restricted distribution at specific (1-32 mgL-1) was plotted. Results were expressed as gallic acid equivalents (GAE)/g dried level (Table 2). Table 2. Phytochemical screening of plant species from "Reserva Nacional Forestal Bosque de Yotoco".
2.6 Statistical analysis
Terpenoids
Condensed Hydrolysable Alcaloids
and steroids
Results were given as the mean±SD for at least three replicates for each sample. Statistical analysis was performed using GraphPad Prism 5® (GraphPad Software Inc., San Diego, California, USA). FRS were calculated using nonlinear regression with one phase exponential decay calculation.
3 Results and discussion
3.1 Extraction yields and phytochemical profile
In order to determine the antioxidant potential and phenolic content of twenty plant species from Yotoco National Protected Forest, for which extractions were performed with chloroform and acetone-water (70:30). S. gigantotepala showed the highest overall yield, followed by C. tococoidea, E. citrifolium, P. setosum, P. augustum, P. compta and other species (Fig. 1). - No appreciable evidence; + weak evidence; ++ strong evidence 3.2 Estimation of the antioxidant potential and total phenolics
TLC analysis showed that most of aqueous extracts possessed potent antioxidant activity, in contrast with low activity exhibited by the chloroform extracts. The ability of the extracts to scavenge DPPH radicals was also investigated at various concentrations to determine the FRS values. As shown in Figure 2, the extracts possessed substantial dose-dependent antioxidant activity. Some species, such as C. tococoidea and M. aeruginosa exhibited similar behavior to quercetin (Figure 2). In others species as B. glauca, almost complete DPPH radical scavenging was observed at concentrations higher than 50 mgL-1. This activity was comparable to that of ascorbic acid and quercetin, which were used as control antioxidants [15]. Interestingly, P. longirostris has a higher activity than S. gigantotepala at concentrations less than 50 mgL-1. However, at concentrations higher than 50 mgL-1 the activity of S. gigantotepala is greater.
Figure 1. Extraction process yields (for species complete names, see table 1).
Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco Folin-Ciocalteau total phenolics ranged from 34 to 480 mg GAEg-1 DE. In average, the species showed a high phenolic content, particularly M. aeruginosa, P. setosum and C. tococoidea (Figure 4). The comparison between FRS ) and TP showed that there was a statistically significant strong correlation (R2=0.9610; Y=(891.9+6.957)e(0,01668X)+6.957; 95% confidence intervals; 15 degrees of freedom) (Figure 5), suggesting, that the phenolic compounds present in the extracts could be responsible for the observed DPPH radical scavenging activity, since they can readily donate hydrogen atom to the radical, and even more, if they bear catechol moieties [16]. Figure 2. DPPH free radical scavenging activity (FRS).
In DPPH assay, the FRS values range from 4.4 mgL-1 (M. aeruginosa aqueous extract) to 486 mgL-1 (B. solanoides aqueous extract) (Figure 3). As it is known, the lower the FRS value the higher the antioxidant activity of plant extract. Also, the FRS of aqueous extracts from C. tococoidea (6.0 mgL-1), P. longirostris (9.0 mgL-1), B. glauca -1), E. citrifollium (20.9 mgL-1), P. setosum (24.0 mgL-1) and S. gigantotepala Pi Ps Pc Ph Pl Pm Sa Sg
(31.5 mgL-1) are considered to have a strong free radical scavenging activity.
Figura 4. Folin-Ciocalteau estimated total fenolics (TP) of the 20 species studied.
(mgL g/L)
R2 = 0.9610 Ps Pc Ph Pl
Figure 5. Correlation between DPPH fre radical scavenging (FRS ) and total phenolics (TP) stimated
Figure 3. FRS of 20 species studied after 1 hour of reaction.
by Folin Ciocalteau method. Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco Folin-Ciocalteau total phenolics ranged from 34 to 480 mg GAEg-1 DE. In average, the species showed a high phenolic content, particularly M. aeruginosa, P. setosum and C. tococoidea (Figure 4). The comparison between FRS ) and TP showed that there was a statistically significant strong correlation (R2=0.9610; Y=(891.9+6.957)e(0,01668X)+6.957; 95% confidence intervals; 15 degrees of freedom) (Figure 5), suggesting, that the phenolic compounds present in the extracts could be responsible for the observed DPPH radical scavenging activity, since they can readily donate hydrogen atom to the radical, and even more, if they bear catechol moieties [16]. Figure 2. DPPH free radical scavenging activity (FRS).
In DPPH assay, the FRS values range from 4.4 mgL-1 (M. aeruginosa aqueous extract) to 486 mgL-1 (B. solanoides aqueous extract) (Figure 3). As it is known, the lower the FRS value the higher the antioxidant activity of plant extract. Also, the FRS of aqueous extracts from C. tococoidea (6.0 mgL-1), P. longirostris (9.0 mgL-1), B. glauca -1), E. citrifollium (20.9 mgL-1), P. setosum (24.0 mgL-1) and S. gigantotepala Pi Ps Pc Ph Pl Pm Sa Sg
(31.5 mgL-1) are considered to have a strong free radical scavenging activity.
Figura 4. Folin-Ciocalteau estimated total fenolics (TP) of the 20 species studied.
(mgL g/L)
R2 = 0.9610 Ps Pc Ph Pl
Figure 5. Correlation between DPPH fre radical scavenging (FRS ) and total phenolics (TP) stimated
Figure 3. FRS of 20 species studied after 1 hour of reaction.
by Folin Ciocalteau method. Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco [3] Li, L., Ishdorj, G. and Gibson, G. B. (2012). Reactive oxygen species regulation of autophagy in cancer: Implications for cancer treatment. Free Radical Biology This study shows that the species B. glauca, C. tococoidea, E. citrifolium, and Medicine 53: 1399–1410. M. aeruginosa, P. setosum, P. longirostris, and S. gigantotepala, significantly scavenged DPPH free radicals in vitro (Table 3). These suggest that the extracts of these species [4] Rojas, A., Figueroa, H., Re, L. and Morales, M. (2006). Oxidative stress at the vascular wall. mechanistic and pharmacological aspects. Archives of Medical could be used as a natural antioxidants source to limit free radical damage. Therefore, it Research 37: 436–448.
is worth to isolate the active secondary metabolites from these extracts and to do further research on the potential effectiveness of the plant extracts in preventing oxidative [5] Halliwell, B. (2001). Role of free radicals in neurodegenerative diseases: stress-mediated diseases in humans, animals and plants. Therapeutic implications for antioxidant treatment. Drugs Aging 18: 685–716.
Table 3. Promising species selected according to preliminary screening base on DPPH free radical
[6] Masia, M., Padilla, S., Bernal, E., Almenar, M. V., Molina, J. and Hernandez, I. scavenging activity and total phenolics. (2007). Influence of antiretroviral therapy on oxidative stress and cardiovascular risk: A prospective cross-sectional study in HIV-infected patients. Clinical Therapeutics 29: 1448–1455.
Banara glauca [7] Kitts, D. D., Wijewickreme, A. N. and Hu, C. (2000). Antioxidant properties of a North American ginseng extract. Molecular and Cell Biochemistry 203: 1–10.
[8] Nogochi, C. and Nikki, E. (2000). Phenolic antioxidants: A rationale for design and evaluation of novel antioxidant drugs for atherosclerosis. Free Radicals in Biology and Medicine 28: 1538–1546.
[9] Jaberian, H., Piri, K. and Nazari, J. (2013). Phytochemical composition and in Piper setosum vitro antimicrobial and antioxidant activities of some medicinal plants. Food Chemistry 136: 237-244.
[10] Diaz, L., Muñoz, D., Prieto, R., Cuervo, S., Gonzalez, D., Guzman, J. and Bhakta, S. (2012). Antioxidant, antitubercular and cytotoxic activities of Piper imperiale. Molecules 17: 4142-4157.
[11] Kubo, I., Kinst-Hori, I., Nihei, K-I., Soria, F., Takasaki, M., Calderón, J. and Céspedes, C. (2003). Tyrosinase inhibitors from galls of Rhus javanica leaves and their effects on insects. Z. Naturforsch 58c: 719-725.
Authors thanks to Universidad del Valle (CI7852) and Universidad Nacional de [12] Escobar, E. (2001). Presentación de Yotoco "Reserva Natural" Flora: Plantas Colombia in Palmira for logistical support. Also thanks the financial support of this Vasculares. Palmira: Universidad Nacional de Colombia.
research by COLCIENCIAS through of the grant No. CT-557-2011.
[13] Harbone, J. B. (1973). Phytochemical methods. A guide to modern techniques of plant analysis. London:Chapman and Hall Ltd. pp, 49-188.
[1] Carocho, M. and Ferreira, I. (2013). A review on antioxidants, prooxidants and [14] Sdiri, S., Bermejo, A., Aleza, P., Navarro, P. and Salvador, A. (2012). Phenolic related controversy: Natural and synthetic compounds, screening and analysis composition, organic acids, sugars, vitamin C and antioxidant activity in the juice of methodologies and future perspectives. Food and Chemical Toxicology 51: 15-25. two new triploid late-season mandarins. Food Research International 49: 462-468.
[2] El Assar, M., Angulo, J. and Rodriguez-Maña, L. (2013). Oxidative stress and [15] Padayatty, S., Katz, A., Wang, Y., Eck, P., Kwon, O. and Lee, J. (2003). Vitamin vascular inflammation in aging. Free Radical Biology and Medicine 65: 380–401. C as an antioxidant: Evaluation of its role in disease prevention. Journal of the American College of Nutrition 22: 18–35.
Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza Total phenolics antioxidant activity and phytochemical profile of some plants from Yotoco [3] Li, L., Ishdorj, G. and Gibson, G. B. (2012). Reactive oxygen species regulation of autophagy in cancer: Implications for cancer treatment. Free Radical Biology This study shows that the species B. glauca, C. tococoidea, E. citrifolium, and Medicine 53: 1399–1410. M. aeruginosa, P. setosum, P. longirostris, and S. gigantotepala, significantly scavenged DPPH free radicals in vitro (Table 3). These suggest that the extracts of these species [4] Rojas, A., Figueroa, H., Re, L. and Morales, M. (2006). Oxidative stress at the vascular wall. mechanistic and pharmacological aspects. Archives of Medical could be used as a natural antioxidants source to limit free radical damage. Therefore, it Research 37: 436–448.
is worth to isolate the active secondary metabolites from these extracts and to do further research on the potential effectiveness of the plant extracts in preventing oxidative [5] Halliwell, B. (2001). Role of free radicals in neurodegenerative diseases: stress-mediated diseases in humans, animals and plants. Therapeutic implications for antioxidant treatment. Drugs Aging 18: 685–716.
Table 3. Promising species selected according to preliminary screening base on DPPH free radical
[6] Masia, M., Padilla, S., Bernal, E., Almenar, M. V., Molina, J. and Hernandez, I. scavenging activity and total phenolics. (2007). Influence of antiretroviral therapy on oxidative stress and cardiovascular risk: A prospective cross-sectional study in HIV-infected patients. Clinical Therapeutics 29: 1448–1455.
Banara glauca [7] Kitts, D. D., Wijewickreme, A. N. and Hu, C. (2000). Antioxidant properties of a North American ginseng extract. Molecular and Cell Biochemistry 203: 1–10.
[8] Nogochi, C. and Nikki, E. (2000). Phenolic antioxidants: A rationale for design and evaluation of novel antioxidant drugs for atherosclerosis. Free Radicals in Biology and Medicine 28: 1538–1546.
[9] Jaberian, H., Piri, K. and Nazari, J. (2013). Phytochemical composition and in Piper setosum vitro antimicrobial and antioxidant activities of some medicinal plants. Food Chemistry 136: 237-244.
[10] Diaz, L., Muñoz, D., Prieto, R., Cuervo, S., Gonzalez, D., Guzman, J. and Bhakta, S. (2012). Antioxidant, antitubercular and cytotoxic activities of Piper imperiale. Molecules 17: 4142-4157.
[11] Kubo, I., Kinst-Hori, I., Nihei, K-I., Soria, F., Takasaki, M., Calderón, J. and Céspedes, C. (2003). Tyrosinase inhibitors from galls of Rhus javanica leaves and their effects on insects. Z. Naturforsch 58c: 719-725.
Authors thanks to Universidad del Valle (CI7852) and Universidad Nacional de [12] Escobar, E. (2001). Presentación de Yotoco "Reserva Natural" Flora: Plantas Colombia in Palmira for logistical support. Also thanks the financial support of this Vasculares. Palmira: Universidad Nacional de Colombia.
research by COLCIENCIAS through of the grant No. CT-557-2011.
[13] Harbone, J. B. (1973). Phytochemical methods. A guide to modern techniques of plant analysis. London:Chapman and Hall Ltd. pp, 49-188.
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Volumen 17 - Especial Química, 2013 Revista de Ciencias H. Torres, A. J. Colmenares and J. H. Isaza [16] Huang, D., Ou, B. and Prior, R. L. (2005). The chemistry behind antioxidant capacity assays. Journal of Agricultural and Food Chemistry 53: 1841–1856.
Facultad de Ciencias Naturales y Exactas Universidad del Valle Harlen Torres Castañeda Departamento de Ciencias Básicas, Universidad Nacional de Colombia, Palmira - Colombia EXTRACTION, CHEMICAL COMPOSITION AND ANTIMICROBIAL
ACTIVITY OF THE ESSENTIAL OILS OF PIPILONGO
(Piper tuberculatum) USING SUPERCRITICAL CARBON DIOXIDE
Ana Julia Colmenares Dulcey Departamento de Química, Universidad del Valle, Cali - Colombia Jaime Restrepo Osorio
Ana Julia Colmenares Dulcey
Luis E. Mora
Rubén Sánchez Andica
Universidad del Valle José Hipólito Isaza Martínez Departamento de Química, Universidad del Valle, Cali - Colombia Received: November 15, 2013 Accepted: December 23, 2013 Essential oils from pipilongo seeds (Piper tuberculatum) was extracted using supercritical carbon dioxide. The extraction was performed as a function of particle size of the grinded seeds. The highest yield (2,812%) was obtained with the smallest particle size. The chemical composition analysis of the oil by GC-MS led to identify 15 compounds, some of which are β-elemene, caryophyllene, β-farnesene, neophytadiene and piperine among others. The microbicide activity of the essential oil was determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays, showing that the growth of the bacteria Staphylococcus aureus and Bacillus subtilis was inhibited, and hence with a possible microbicidal effect, whereas for pseudomonas aeruginosa and Salmonella typhimurium showed no effect on their growth.
Keywords: Piper tuberculatum, essential oils, supercritical fluids, microbicide.
1 Introduction
The Piper tuberculatum species, known as pipilongo, belongs to the genus Piper (piperaceae) widely distributed in the tropical and subtropical regions of the world. Many of these species are considered as medicinal plants in Latin America as well as in the West Indies Region (Bezerra, et al. 2006; in Ordaz et al. 2011). According to the literature, these plants have been used as insecticides, antivirals, germicides and, particularly, as anti-fungus (Trindade et al. 2012). These biological properties are thought to be due to the presence of lignans and/or amides, such as alkylic or olefi n isobutylamides, fl avonoids, gamma-lactanes, butenolids and cyclohexane epoxids among others (Parmar et al. 1997; in Trindade et al 2012).
Piper tuberculatum is a 2 - 3 m tall bush, although sometimes it could reach a height of approximately 6 m. It has a thin trunk measuring 18 to 20 cm in diameter that branches out from the bottom, giving to the plant the typical bush appearance, thanks to their long and thin greyish branches. They have oblong alternate leaves that measure 4 to 12 cm Volumen 17 - Especial Química, 2013

Source: http://revistaciencias.univalle.edu.co/volumenes/vol_17N3_EE/HTorres.pdf